Molecular evolution of the RNA polymerase II CTD.
In higher eukaryotes, an unusual C-terminal domain (CTD) is crucial to the function of RNA polymerase II in transcription. The CTD consists of multiple heptapeptide repeats; differences in the number of repeats between organisms and their degree of conservation have intrigued researchers for two decades. Here, we review the evolution of the CTD at the molecular level. Several primitive motifs have been integrated into compound heptads that can be readily amplified. The selection of phosphorylatable residues in the heptad repeat provided the opportunity for advanced gene regulation in eukaryotes. Current findings suggest that the CTD should be considered as a collection of continuous overlapping motifs as opposed to a specific functional unit defined by a heptad.


Treating cancer with selective CDK4/6 inhibitors.
Uncontrolled cellular proliferation, mediated by dysregulation of the cell-cycle machinery and activation of cyclin-dependent kinases (CDKs) to promote cell-cycle progression, lies at the heart of cancer as a pathological process. Clinical implementation of first-generation, nonselective CDK inhibitors, designed to inhibit this proliferation, was originally hampered by the high risk of toxicity and lack of efficacy noted with these agents. The emergence of a new generation of selective CDK4/6 inhibitors, including ribociclib, abemaciclib and palbociclib, has enabled tumour types in which CDK4/6 has a pivotal role in the G1-to-S-phase cell-cycle transition to be targeted with improved effectiveness, and fewer adverse effects. Results of pivotal phase III trials investigating palbociclib in patients with advanced-stage oestrogen receptor (ER)-positive breast cancer have demonstrated a substantial improvement in progression-free survival, with a well-tolerated toxicity profile. Mechanisms of acquired resistance to CDK4/6 inhibitors are beginning to emerge that, although unwelcome, might enable rational post-CDK4/6 inhibitor therapeutic strategies to be identified. Extending the use of CDK4/6 inhibitors beyond ER-positive breast cancer is challenging, and will likely require biomarkers that are predictive of a response, and the use of combination therapies in order to optimize CDK4/6 targeting.


A survey of the rock record of reptilian ontogeny.
Given the large diversity and long stratigraphical range of fossil reptiles, their development is a fundamental aspect of the evolution of ontogeny in vertebrates. Eggs, juveniles, embryos and growth series document different aspects of fossilized ontogenies. About three-fifths of the more than 850 available publications on these topics concern dinosaurs. Non-invasive imaging techniques have facilitated the study of embryos in ovo. Examination of ontogenetic trajectories is used to establish criteria to identify fossil growth series and solve taxonomic issues. Many morphological innovations in reptilian skeletal structures are associated with growth heterochronic changes, whereas sequence heterochronic changes remain largely unstudied but are a potential avenue of research. Relative age assessments via not only palaeohistology but also comparative anatomy have been used to reconstruct life history patterns in fossil archosaurs. Several fossil marine reptiles evolved viviparity convergently. Extinct adult phenotypes can reveal information on development, as in the discovery of polydactyly in diapsids, the examination of vertebral number evolution, and its relation to somitgenesis and Hox-gene boundaries, and signs of tissue regeneration provided by anatomical peculiarities following caudal autotomy.


Mutational analysis of the tyrosine phosphatome in colorectal cancers.
Tyrosine phosphorylation, regulated by protein tyrosine phosphatases (PTPs) and kinases (PTKs), is important in signaling pathways underlying tumorigenesis. A mutational analysis of the tyrosine phosphatase gene superfamily in human cancers identified 83 somatic mutations in six PTPs (PTPRF, PTPRG, PTPRT, PTPN3, PTPN13, PTPN14), affecting 26% of colorectal cancers and a smaller fraction of lung, breast, and gastric cancers. Fifteen mutations were nonsense, frameshift, or splice-site alterations predicted to result in truncated proteins lacking phosphatase activity. Five missense mutations in the most commonly altered PTP (PTPRT) were biochemically examined and found to reduce phosphatase activity. Expression of wild-type but not a mutant PTPRT in human cancer cells inhibited cell growth. These observations suggest that the mutated tyrosine phosphatases are tumor suppressor genes, regulating cellular pathways that may be amenable to therapeutic intervention.


A four-parameter system for rationalising the electronic properties of transition metal-radical ligand complexes.
A system of four principal parameters is reported that provides a unified description of the electronic and chemical properties of radical-ligand coordination compounds. This type of parametrisation applies to compositionally different types of radical-ligands, and the principal parameters rank in the following order: (a) coordination mode (metal-ligand orbital alignment) > (b) metal linkage > (c) ligand charge > (d) geometric strain (on orbital overlap). A series of group-10 metal complexes of an open-shell thiolate-arene-thiolate ligand suits to differentiate between three of the four effects in a clear-cut fashion, which allowed sorting these into a semi-quantitative order for the first time. Combined experimental and TD-DFT data aided in distinguishing structural effects from metal specific contributions such as relativistic effects. The applicability of spectroscopic and structure properties to serve as characteristic markers for comparison is discussed with regard to the large body of planar radical-ligand structures.


Workplace flexibility: from research to action.
Ellen Galinsky, Kelly Sakai, and Tyler Wigton explore the "time famine" among American workers-the continuing sense among employees of not having enough time to manage the multiple responsibilities of work and personal and family life. Noting that large shares of U.S. employees report feeling the need for greater workplace flexibility to enable them to take better care of family responsibilities, the authors examine a large-scale community-engagement initiative to increase workplace flexibility voluntarily. Using the 2008 National Study of the Changing Workforce as a primary source of data, the authors begin with an overview of the prevalence of flexibility in today's American workplace. They track which categories of employees have access to various flexibility options, as well as the extent to which employees with access to various types of flexibility use those options. Findings from the study indicate that the majority of employees want flexibility but that access to it varies, with more advantaged employees--those who are well educated, have high salaries, and work full time, for example--being doubly advantaged in having greater access to flexibility. A number of employers, say the authors, tend to be skeptical of the value of workplace flexibility and to fear that employees will abuse it if it is offered. But the study data reveal that most employees use flexibility quite conservatively. When the authors use their nationally representative data set to investigate correlations between access to workplace flexibility and a range of workplace outcomes especially valued by employers--employee engagement, job satisfaction, retention, and health--they find that employers as well as employees can benefit from flexibility. Finally, the authors discuss When Work Works, a large, national community-based initiative under way since 2003 to increase voluntary adoption of workplace flexibility. The authors detail the conceptual basis of the project's design, noting its emphasis on flexibility as one component of effective workplaces that can benefit employers, employees, and communities alike. Galinsky, Sakai, and Wigton conclude by drawing lessons learned from the project and briefly discussing the implications of using research to bring about workplace change.


Protein biomarkers discriminate Leishmania major-infected and non-infected individuals in areas endemic for cutaneous leishmaniasis.
A successful host immune response to infection is dependent upon both innate and adaptive immune effector mechanisms. Cutaneous leishmaniasis results in an adaptive Th1 CD4(+) T cell response that efficiently clears the parasite, but may also result in scaring. However the role of innate mechanisms during parasite clearance remains less well defined. We examined a unique cohort of individuals, living in a Leishmania major endemic region, that were stratified among 3 distinct clinical groups in a cross-sectional study. Specifically, patients were classified either as healed (n = 17), asymptomatic (23), or naïve to infection (18) based upon the classical Leishmanin Skin Test (LST) and the presence or absence of scars. Utilizing a multiplexed immunoassay approach we characterized the induced cytokine and chemokine response to L. major. A subset of innate immune molecules was induced in all groups. By contrast, T cell-associated cytokines were largely induced in exposed groups as compared to L. major-infection naïve individuals. Two exceptions were IL-17A and IL-12p70, induced and not induced, respectively, in naïve individuals. In addition, GM-CSF was more strongly induced in healed patients as compared to the other two groups. Surprisingly an IL-13 response was the best cytokine for classifying previously infected donors. Exploratory data analysis, utilizing principle component analysis (PCA), revealed distinct patient clusters of the healed and naïve groups based on the most differentially induced proteins. Asymptomatic previously infected individuals were more difficult to assign to a particular cluster based on these induced proteins. Analysis of these proteins may enable the identification of biomarkers associated with disease, leading to a better understanding of the protective mechanisms of immune response against leishmaniasis.


Association study designs for complex diseases.
Assessing the association between DNA variants and disease has been used widely to identify regions of the genome and candidate genes that contribute to disease. However, there are numerous examples of associations that cannot be replicated, which has led to skepticism about the utility of the approach for common conditions. With the discovery of massive numbers of genetic markers and the development of better tools for genotyping, association studies will inevitably proliferate. Now is the time to consider critically the design of such studies, to avoid the mistakes of the past and to maximize their potential to identify new components of disease.


Polarized expression of CD74 by gastric epithelial cells.
CD74 is known as the major histocompatibility complex (MHC) class II-associated invariant chain (Ii) that regulates the cell biology and functions of MHC class II molecules. Class II MHC and Ii expression was believed to be restricted to classical antigen-presenting cells (APC); however, during inflammation, other cell types, including mucosal epithelial cells, have also been reported to express class II MHC molecules. Given the importance of Ii in the biology of class II MHC, we sought to examine the expression of Ii by gastric epithelial cells (GEC) to determine whether class II MHC molecules in these nonconventional APC cells were under the control of Ii and to further support the role that these cells may play in local immune and inflammatory responses during Helicobacter pylori infection. Thus we examined the expression of Ii on GEC from human biopsy samples and then confirmed this observation using independent methods on several GEC lines. The mRNA for Ii was detected by RT-PCR, and the various protein isoforms were also detected. Interestingly, these cells have a high level expression of surface Ii, which is polarized to the apical surface. These studies are the first to demonstrate the constitutive expression of Ii by human GEC.


Chemical disinfection to interrupt transfer of rhinovirus type 14 from environmental surfaces to hands.
Rhinoviruses can survive on environmental surfaces for several hours under ambient conditions. Hands can readily become contaminated after contact with such surfaces, and self-inoculation may lead to infection. Whereas hand washing is crucial in preventing the spread of rhinovirus colds, proper disinfection of environmental surfaces may further reduce rhinovirus transmission. In this study, the capacities of Lysol Disinfectant Spray (0.1% o-phenylphenol and 79% ethanol), a domestic bleach (6% sodium hypochlorite diluted to give 800 ppm of free chlorine), a quaternary ammonium-based product (7.05% quaternary ammonium diluted 1:128 in tap water), and a phenol-based product (14.7% phenol diluted 1:256 in tap water) were compared in interrupting the transfer of rhinovirus type 14 from stainless steel disks to fingerpads of human volunteers upon a 10-s contact at a pressure of 1 kg/cm2. Ten microliters of the virus, suspended in bovine mucin (5 mg/ml), was placed on each disk, and the inoculum was dried under ambient conditions; the input number on each disk ranged from 0.5 x 10(5) to 2.1 x 10(6) PFU. The dried virus was exposed to 20 microliters of the test disinfectant. The Lysol spray was able to reduce virus infectivity by > 99.99% after a contact of either 1 or 10 min, and no detectable virus was transferred to fingerpads from Lysol-treated disks. The bleach (800 ppm of free chlorine) reduced the virus titer by 99.7% after a contact time of 10 min, and again no virus was transferred from the disks treated with it.(ABSTRACT TRUNCATED AT 250 WORDS)


Risk Factors for the Development of Nerve Palsy Following Primary Total Hip Arthroplasty.
Nerve palsy following total hip arthroplasty (THA) is a complication that worsens the functional prognosis. The present study analyzed the risk factors of nerve palsy following THA. The subjects of this study included 6,123 cases in which primary THA was performed under spinal anesthesia with cementless implants used in the posterolateral approach. Fourteen cases (0.22%) developed nerve palsy following THA, all of which involved palsy of the entire peroneal nerve region. The diagnoses included osteoarthritis due to subluxation (n=6), complete hip dislocation (n=3), osteonecrosis of the femoral head (n=2), primary osteoarthritis (n=1), osteoarthritis due to trauma (n=1), and multiple osteochondromatosis (n=1). Recovery from nerve palsy was confirmed in 10 cases; the longest recovery period was three years. A univariate analysis revealed significant differences in the osteoarthritis due to subluxation, osteonecrosis of the femoral head, complete hip dislocation, body weight and body mass index. However, none of the factors remained significant in multivariate analysis. Peroneal (ischiadic) nerve palsy following THA occurred in patients with osteonecrosis of the femoral head, complete hip dislocation, low body weight and a low body mass index. However, there were no cases of nerve palsy after the introduction of THA combined with shortening osteotomy of the femur for complete hip dislocation. patients. It is necessary to pay attention to direct pressure in cases of lower body weight and lower BMI because compression of the sciatic nerve during surgery and compression of the fibular head are considered to be risk factors.


Microfluidic modeling of the biophysical microenvironment in tumor cell invasion.
Tumor cell invasion, whether penetrating through the extracellular matrix (ECM) or crossing a vascular endothelium, is a critical step in the cancer metastatic cascade. Along the way from a primary tumor to a distant metastatic site, tumor cells interact actively with the microenvironment either via biomechanical (e. g. ECM stiffness) or biochemical (e.g. secreted cytokines) signals. Increasingly, it is recognized that the tumor microenvironment (TME) is a critical player in tumor cell invasion. A main challenge for the mechanistic understanding of tumor cell-TME interactions comes from the complexity of the TME, which consists of extracellular matrices, fluid flows, cytokine gradients and other cell types. It is difficult to control TME parameters in conventional in vitro experimental designs such as Boyden chambers or in vivo such as in mouse models. Microfluidics has emerged as an enabling tool for exploring the TME parameter space because of its ease of use in recreating a complex and physiologically realistic three dimensional TME with well-defined spatial and temporal control. In this perspective, we will discuss designing principles for modeling the biophysical microenvironment (biological flows and ECM) for tumor cells using microfluidic devices and the potential microfluidic technology holds in recreating a physiologically realistic tumor microenvironment. The focus will be on applications of microfluidic models in tumor cell invasion.


High impact of implementation on school-based smoking prevention: the X:IT study-a cluster-randomized smoking prevention trial.
Implementation fidelity describes how well an intervention is implemented in the real-world setting. Assessing implementation fidelity is essential in the understanding of intervention results. In most studies, implementation fidelity is measured insufficiently, though, not taking into account the complexity of the concept nor the intervention. The objective of the present study was to develop an overall quantitative measure of implementation fidelity, to examine the degree of implementation fidelity and the association of implementation and effect of a randomized school-based smoking prevention trial-the X:IT study. A cluster-randomized trial testing is a multi-component intervention to prevent smoking among adolescents in 94 Danish elementary schools (51 intervention, 43 control schools). Participants were grade 7 pupils (mean age 12.5 years). Data was collected by electronic questionnaires among pupils at baseline (n = 4161), the first follow-up (n = 3764), and the second follow-up (n = 3269) and among school coordinators at intervention schools at the first and second follow-up (50 and 39 coordinators). The intervention included three components: (1) smoke-free school grounds, (2) smoke-free curriculum, and (3) parental involvement, contracts, and dialogues. Implementation fidelity was assessed by four domains: adherence, dose, quality of delivery, and participant responsiveness. These were combined into an overall school-wise implementation index. The association of implementation and smoking was examined by logistic regression analyses. One fourth of the schools was characterized as high implementers of the program (all three components) at both first (12 schools, 24.0 %) and second follow-up (11 schools, 28.2 %). Implementation fidelity was strongly associated with smoking at the first and second follow-up, e.g., the odds for smoking at schools with high implementation both years were OR = 0.44 (95 % CI 0.32 to 0.68). Using an overall measure based on several aspects of implementation fidelity, we showed a negative graded association between implementation and smoking. This study suggests that higher degrees of implementation will improve the effect of the X:IT intervention. Studying the association between implementation and effect is extremely important; only by doing so, we can distinguish the quality of the intervention from the success of the implementation. Current Controlled Trials ISRCTN77415416.


CD4+CD8- T cells are the effector cells in disease pathogenesis in the scurfy (sf) mouse.
Mice hemizygous for the X-linked mutation, scurfy (sf), exhibit a fatal lymphoreticular disease that is mediated by T lymphocytes. To evaluate the respective roles of CD4 or CD8 single positive T cells in scurfy disease, neonates were treated with mAbs directed against the CD4 or CD8 molecules. Whereas mice treated with an anti-CD8 Ab developed lesions and succumbed to disease at the same time (17 days) as their untreated scurfy littermates, mice treated with an anti-CD4 Ab lived up to 11 wk before developing scurfy disease. To insure a more complete elimination of the T cell subsets, the scurfy mutation was bred onto beta 2-microglobulin (beta 2m)-deficient (CD8-less) and CD4-deficient transgenic mouse lines. Whereas there was little moderation of disease in beta 2m-deficient scurfy mice, CD4-deficient scurfy mice had markedly decreased scurfy lesions and a prolonged life span, similar to that of anti-CD4-treated sf/Y mice. Additionally, scurfy disease was transplanted into H-2-compatible nude mice through the adoptive transfer of CD4+CD8- T cells, but not CD4-CD8+ T cells. Flow-cytometric analysis revealed that sf/Y mice have an increased percentage of activated CD4+ T cells in their lymph nodes. In addition, there is an increase in the in vitro production of cytokines in the cultured splenocytes of CD8-less, but not CD4-less, scurfy mice. These data suggest that CD4+ T cells are critical mediators of disease in the scurfy mouse.


Glial interactions with neurons during Drosophila embryogenesis.
A monoclonal antibody (Mab5B12) demonstrating specificity for glial cells within the central and peripheral nervous systems of Drosophila has been used in combination with neural-specific antibodies to study the early organization of the Drosophila embryo. The embryonic central nervous system of Drosophila contains cells within the ventral midline that are recognized by monoclonal antibody 5B12. These cells are not recognized by either a polyclonal antiserum to horse radish peroxidase, which recognizes several antigens on the surface of Drosophila neurons, or Mab22C10, which recognizes an antigen specific to the peripheral nervous system. Mab5B12-positive cells lie dorsal both to the developing anterior and posterior commissures in each thoracic and abdominal segment and to the supraoesophageal commissure. They ensheath these commissures in later stage embryos. Other Mab5B12-positive cells lie dorsolateral to the CNS and send processes laterally to the lateral sensilla during axonogenesis in the PNS. These cells surround the axons of the intersegmental and segmental nerves. Other cells that line the advancing ectoderm during dorsal closure and surround the anal pads also express the Mab5B12 antigen. Neuronal cell cultures derived from Drosophila gastrulae contain cells expressing the Mab5B12 antigen. These cells can be found separate or in close association with neuronal clusters and their axons.


Efficient reverse transcription using locked nucleic acid nucleotides towards the evolution of nuclease resistant RNA aptamers.
Modified nucleotides are increasingly being utilized in the de novo selection of aptamers for enhancing their drug-like character and abolishing the need for time consuming trial-and-error based post-selection modifications. Locked nucleic acid (LNA) is one of the most prominent and successful nucleic acid analogues because of its remarkable properties, and widely explored as building blocks in therapeutic oligonucleotides. Evolution of LNA-modified RNA aptamers requires an efficient reverse transcription method for PCR enrichment of the selected RNA aptamer candidates. Establishing this key step is a pre-requisite for performing LNA-modified RNA aptamer selection. In this study three different reverse transcriptases were investigated towards the enzymatic recognition of LNA nucleotides. Both incorporation as well as reading capabilities of the LNA nucleotides was investigated to fully understand the limitations of the enzymatic recognition. We found that SuperScript® III Reverse Transcriptase is an efficient enzyme for the recognition of LNA nucleotides, making it a prime candidate to be used in de novo selection of LNA containing RNA aptamers.


Video capture of the circumstances of falls in elderly people residing in long-term care: an observational study.
Falls in elderly people are a major health burden, especially in the long-term care environment. Yet little objective evidence is available for how and why falls occur in this population. We aimed to provide such evidence by analysing real-life falls in long-term care captured on video. We did this observational study between April 20, 2007, and June 23, 2010, in two long-term care facilities in British Columbia, Canada. Digital video cameras were installed in common areas (dining rooms, lounges, hallways). When a fall occurred, facility staff completed an incident report and contacted our teams so that we could collect video footage. A team reviewed each fall video with a validated questionnaire that probed the cause of imbalance and activity at the time of falling. We then tested whether differences existed in the proportion of participants falling due to the various causes, and while engaging in various activities, with generalised linear models, repeated measures logistic regression, and log-linear Poisson regression. We captured 227 falls from 130 individuals (mean age 78 years, SD 10). The most frequent cause of falling was incorrect weight shifting, which accounted for 41% (93 of 227) of falls, followed by trip or stumble (48, 21%), hit or bump (25, 11%), loss of support (25, 11%), and collapse (24, 11%). Slipping accounted for only 3% (six) of falls. The three activities associated with the highest proportion of falls were forward walking (54 of 227 falls, 24%), standing quietly (29 falls, 13%), and sitting down (28 falls, 12%). Compared with previous reports from the long-term care setting, we identified a higher occurrence of falls during standing and transferring, a lower occurrence during walking, and a larger proportion due to centre-of-mass perturbations than base-of-support perturbations. By providing insight into the sequences of events that most commonly lead to falls, our results should lead to more valid and effective approaches for balance assessment and fall prevention in long-term care. Canadian Institutes for Health Research.


Calmodulin activation of the Ca2+ pump revealed by fluorescent chelator dyes in human red blood cell ghosts.
Ca2+ transport in red blood cell ghosts was monitored with fura2 or quin2 incorporated as the free acid during resealing. This is the first report of active transport monitored by the fluorescent intensity of the chelator dyes fura2 (5-50 microM) or quin2 (250 microM) in hemoglobin-depleted ghosts. Since there are no intracellular compartments in ghosts and the intracellular concentrations of all assay chelator substances including calmodulin (CaM), the dyes, and ATP could be set, the intracellular concentrations of free and total Ca [( Cafree]i and [Catotal]i) could be calculated during the transport. Ghosts prepared with or without CaM rapidly extruded Ca2+ to a steady-state concentration of 60-100 nM. A 10(4)-fold gradient for Ca2+ was routinely produced in medium containing 1 mM Ca2+. During active Ca2+ extrusion, d[Cafree]i/dt was a second order function of [Cafree]i and was independent of the dye concentration, whereas d[Catotal]i/dt increased as a first order function of both the [Cafree]i and the concentration of the Ca:dye complex. CaM (5 microM) increased d[Catotal]i/dt by 400% at 1 microM [Cafree]i, while d[Cafree]i/dt increased by only 25%. From a series of experiments we conclude that chelated forms of Ca2+ serve as substrates for the pump under permissive control of the [Cafree]i, and this dual effect may explain cooperativity. Free Ca2+ is extruded, and probably also Ca2+ bound to CaM or other chelators, while CaM and the chelators are retained in the cell.


A critical comparison of topology-based pathway analysis methods.
One of the aims of high-throughput gene/protein profiling experiments is the identification of biological processes altered between two or more conditions. Pathway analysis is an umbrella term for a multitude of computational approaches used for this purpose. While in the beginning pathway analysis relied on enrichment-based approaches, a newer generation of methods is now available, exploiting pathway topologies in addition to gene/protein expression levels. However, little effort has been invested in their critical assessment with respect to their performance in different experimental setups. Here, we assessed the performance of seven representative methods identifying differentially expressed pathways between two groups of interest based on gene expression data with prior knowledge of pathway topologies: SPIA, PRS, CePa, TAPPA, TopologyGSA, Clipper and DEGraph. We performed a number of controlled experiments that investigated their sensitivity to sample and pathway size, threshold-based filtering of differentially expressed genes, ability to detect target pathways, ability to exploit the topological information and the sensitivity to different pre-processing strategies. We also verified type I error rates and described the influence of overexpression of single genes, gene sets and topological motifs of various sizes on the detection of a pathway as differentially expressed. The results of our experiments demonstrate a wide variability of the tested methods. We provide a set of recommendations for an informed selection of the proper method for a given data analysis task.


The gunslinger effect: why are movements made faster when responding to versus initiating an action?
The authors replicated and extended results from the gunfight paradigm (A. Welchman, J. Stanley, M. Schomers, R. Miall, & H. Bulthoff, 2010a) in which participants moved faster when reacting to the perceived initiation of an opponent compared to initiating an action themselves. In addition to replicating these movement time effects, the authors found that time to peak velocity, peak velocity, and movement-endpoint dispersions were similarly impacted. The findings are discussed in terms of a triggering mechanism involved in ballistic and internally generated movements.


Polymer nanoparticle sizes from dynamic light scattering and size exclusion chromatography: the case study of polysilanes.
Dynamic light scattering (DLS) and size exclusion chromatography (SEC) are among the most popular methods for determining polymer sizes in solution. Taking dendritic and network polysilanes as a group of least soluble polymer substances, we critically compare and discuss the difference between nanoparticle sizes, obtained by DLS and SEC. Polymer nanoparticles are typically in poor solution conditions below the theta point and are therefore in the globular conformation. The determination of particle sizes in the presence of attractive interactions is not a trivial task. The only possibility to measure, aggregation-free, the true molecular size of polymer nanoparticles in such a solution regime, is to perform the experiment with a dilute solution of globules (below the theta point and above the miscibility line). Based on the results of our polysilane measurements, we come to a conclusion that DLS provides more reliable results than SEC for dilute solutions of globules. General implications for the size measurements of polymer nanoparticles in solution are discussed.


Differential gene expression and physiological changes during acute or persistent plant virus interactions may contribute to viral symptom differences.
Viruses have different strategies for infecting their hosts. Fast and acute infections result in the development of severe symptoms and may cause the death of the plant. By contrast, in a persistent interaction, the virus can survive within its host for a long time, inducing only mild symptoms. In this study, we investigated the gene expression changes induced in CymRSV-, crTMV-, and TCV-infected Nicotiana benthamiana and in PVX- and TMV-U1-infected Solanum lycopersicum plants after the systemic spread of the virus by two different high-throughput methods: microarray hybridization or RNA sequencing. Using these techniques, we were able to clearly differentiate between acute and persistent infections. We validated the gene expression changes of selected genes by Northern blot hybridization or by qRT-PCR. We show that, in contrast to persistent infections, the drastic shut-off of housekeeping genes, downregulation of photosynthesis-related transcripts and induction of stress genes are specific outcomes with acute infections. We also show that these changes are not a consequence of host necrosis or the presence of a viral silencing suppressor. Thermal imaging data and chlorophyll fluorescence measurements correlated very well with the molecular changes. We believe that the molecular and physiological changes detected during acute infections mostly contribute to virus symptom development. The observed characteristic physiological changes associated with economically more dangerous acute infections could serve as a basis for the elaboration of remote monitoring systems suitable for detecting developing virus infections in crops. Moreover, as molecular and physiological changes are characteristics of different types of virus lifestyles, this knowledge can support risk assessments of recently described novel viruses.


What Physical Health Means to Me: Perspectives of People with Mental Illness.
There are significant inequalities in physical health and life expectancy between people with and without a mental illness. Understanding perspectives of people with mental illness on personal meanings of physical health is essential to ensuring health services are aligned with consumer understandings, needs, and values. A qualitative exploratory study was undertaken involving focus groups with 31 consumers in The Australian Capital Territory, Australia. Participants were asked: "What does physical health mean to you?" Thematic analysis was applied to interview transcripts. Five themes are discussed, representing different emphases in the meaning of physical health: (1) physical and mental are interconnected, (2) absence of disease, (3) moving the body, (4) struggling for healthy diet, and (5) functioning and participation. Physical pain was a difficulty that arose across these themes. Mental health consumers see physical health as always connected with well-being. Nurses would benefit from been informed by consumer understandings of physical health. In addition, there should be more attention to quality of life measures of people with mental illness as these are more congruent with consumer perspectives on physical health than biomedical measures.


Phylogenetic analyses suggest that diversification and body size evolution are independent in insects.
Skewed body size distributions and the high relative richness of small-bodied taxa are a fundamental property of a wide range of animal clades. The evolutionary processes responsible for generating these distributions are well described in vertebrate model systems but have yet to be explored in detail for other major terrestrial clades. In this study, we explore the macro-evolutionary patterns of body size variation across families of Hexapoda (insects and their close relatives), using recent advances in phylogenetic understanding, with an aim to investigate the link between size and diversity within this ancient and highly diverse lineage. The maximum, minimum and mean-log body lengths of hexapod families are all approximately log-normally distributed, consistent with previous studies at lower taxonomic levels, and contrasting with skewed distributions typical of vertebrate groups. After taking phylogeny and within-tip variation into account, we find no evidence for a negative relationship between diversification rate and body size, suggesting decoupling of the forces controlling these two traits. Likelihood-based modeling of the log-mean body size identifies distinct processes operating within Holometabola and Diptera compared with other hexapod groups, consistent with accelerating rates of size evolution within these clades, while as a whole, hexapod body size evolution is found to be dominated by neutral processes including significant phylogenetic conservatism. Based on our findings we suggest that the use of models derived from well-studied but atypical clades, such as vertebrates may lead to misleading conclusions when applied to other major terrestrial lineages. Our results indicate that within hexapods, and within the limits of current systematic and phylogenetic knowledge, insect diversification is generally unfettered by size-biased macro-evolutionary processes, and that these processes over large timescales tend to converge on apparently neutral evolutionary processes. We also identify limitations on available data within the clade and modeling approaches for the resolution of trees of higher taxa, the resolution of which may collectively enhance our understanding of this key component of terrestrial ecosystems.


Public Reporting of Hospital-Specific Breastfeeding Measures.
Establishing breastfeeding in the first days of an infant's life is important for longer term success in breastfeeding. In 2009, New York State (NYS) was the second state to require maternity care facilities to collect infant feeding information and to publicly disseminate hospital-specific infant feeding statistics. Public reporting of these statistics as performance measures is a strategy to prompt hospitals to improve breastfeeding support. This qualitative study sought to explore how maternity care administrators and clinical staff responded to the mandate for publicly reported performance measures and whether they used this information to improve maternity care practices. This study used a stratified random sample of NYS hospitals with maternity care units. Participants were recruited by email and telephone calls. A total of 25 hospitals participated in the study, and 37 hospital administrators and staff completed in-depth interviews by telephone. The interviews were analyzed using an explanatory framework in NVivo 8. Publicly reported hospital-specific breastfeeding measures increased attention to breastfeeding performance. Hospital administrators and staff reported comparing their relative rankings to other hospitals in the state. Some hospitals used publicly reported breastfeeding measures to monitor performance, whereas others were prompted to generate additional measures for more frequent monitoring. Hospitals with relatively low breastfeeding statistics took certain actions to improve their maternity care practices to support breastfeeding. Limitations of the usefulness of publicly reported measures were reported by interview participants. Publicly reported, hospital-specific breastfeeding measures may prompt hospitals to monitor and improve maternity care practices related to supporting breastfeeding.


Leukotriene C synthase in mouse mastocytoma cells. An enzyme distinct from cytosolic and microsomal glutathione transferases.
Leukotriene C4 synthesis was studied in preparations from mouse mastocytoma cells. Enzymic conjugation of leukotriene A4 with glutathione was catalysed by both the cytosol and the microsomal fraction. The specific activity of the microsomal fraction (7.8 nmol/min per mg of protein) was 17 times that of the cytosol fraction. The cytosol fraction of the mastocytoma cells contained two glutathione transferases, which were purified to homogeneity and characterized. A microsomal glutathione transferase was purified from mouse liver; this enzyme was shown by immunoblot analysis to be present in the mastocytoma microsomal fraction at a concentration one-tenth or less of that in the liver microsomal fraction. Both the cytosolic and the microsomal glutathione transferases in the mastocytoma cells were identified with enzymes previously characterized, by determining specific activities with various substrates, sensitivities to inhibitors, reactions with antibodies, and physical properties. The purified microsomal glutathione transferase from liver was inactive with leukotriene A4 or its methyl ester as substrate. The cytosolic enzymes displayed activity with leukotriene A4, but their specific activities and intracellular concentrations were too low to account for the leukotriene C4 formation in the mastocytoma cells. The microsomal fraction of the cells contained an enzyme distinguishable by various criteria from the previously studied glutathione transferases. This membrane-bound enzyme, leukotriene C synthase (leukotriene A4:glutathione S-leukotrienyltransferase), appears to carry the main responsibility for the biosynthesis of leukotriene C4.


Impact of orthogonal exchange coupling on magnetic anisotropy in antiferromagnetic oxides/ferromagnetic systems.
The influence of interface exchange coupling on magnetic anisotropy in the antiferromagnetic oxide/Ni system is investigated. We show how interfacial exchange coupling can be employed not only to pin the magnetization of the ferromagnetic layer but also to support magnetic anisotropy to orient the easy magnetization axis perpendicular to the film plane. The fact that this effect is only observed below the Néel temperature of all investigated antiferromagnetic oxides with significantly different magnetocrystalline anisotropies gives evidence that antiferromagnetic ordering is a source of the additional contribution to the perpendicular effective magnetic anisotropy.


Metabolic changes and induction of hepatic lipidosis during feed restriction in llamas.
To determine whether feed restriction induces hepatic lipidosis (HL) in llamas and to evaluate the metabolic changes that develop during feed restriction. 8 healthy adult female llamas. Llamas were fed grass hay at a rate of 0.25% of their body weight per day for 13 to 28 days. Llamas were monitored by use of clinical observation, serum biochemical analyses, and ultrasound-guided liver biopsies. All 8 llamas lost weight and mobilized fat. Five llamas developed HL, including 4 that were nursing crias. During the period of feed restriction, mean serum concentration of bile acids and activities of aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), and gamma-glutamyl transferase (GGT) were significantly higher in llamas that developed HL, compared with llamas that did not. Mean insulin-to-cortisol concentration ratios were lower in llamas with HL before and up to 7 days of feed restriction, compared with those that did not develop HL. HL in llamas may be induced by severe feed restriction, particularly in the face of increased energy demand. Llamas with weight loss attributable to inadequate dietary intake may develop biochemical evidence of hepatopathy and HL. Increases in serum concentration of bile acids and activities of GGT, AST, and SDH may indicate the development of HL in llamas and identify affected animals for aggressive therapeutic intervention.


Direct calculation of the binding free energies of FKBP ligands.
Direct calculations of the absolute free energies of binding for eight ligands to FKBP protein were performed using the Fujitsu BioServer massively parallel computer. Using the latest version of the general assisted model building with energy refinement (AMBER) force field for ligand model parameters and the Bennett acceptance ratio for computing free-energy differences, we obtained an excellent linear fit between the calculated and experimental binding free energies. The rms error from a linear fit is 0.4 kcal/mol for eight ligand complexes. In comparison with a previous study of the binding energies of these same eight ligand complexes, these results suggest that the use of improved model parameters can lead to more predictive binding estimates, and that these estimates can be obtained with significantly less computer time than previously thought. These findings make such direct methods more attractive for use in rational drug design.


Nutrition interventions in women in low-income groups in the UK.
In the UK the mental and physical health and well-being of millions of women are influenced by living in poverty. Low educational attainment, unemployment, low pay and poor areas of residence exacerbate the challenges of obtaining optimal food choices, dietary intake and healthy eating patterns. Poorer women are more likely to eat low amounts of fruits and vegetables, whole grains and fish, and higher amounts of sugar and sweetened drinks compared with more affluent women. Diet contributes to the health inequalities evident in high rates of diet-related morbidity (including obesity) and mortality (including IHD and stroke) and in maternal and child health considerations (including breast-feeding and family diet practices). There is a dearth of research on effective interventions undertaken with low-income women, reflecting some of the challenges of engaging and evaluating programmes with this 'hard to reach' subpopulation. Intervention programmes from the USA, including WISEWOMAN, the Women's Health Initiative, the American Special Supplemental Food Program for Women, Infants and Children and the Expanded Food and Nutrition Education Program provide models for changing behaviour amongst women in the UK, although overall effects of such programmes are fairly modest. Lack of evidence does not mean that that policy work should be not be undertaken, but it is essential that policy work should be evaluated for its ability to engage with target groups as well as for the behavioural change and health outcomes.


Influence of midsole hardness of standard cushioned shoes on running-related injury risk.
In this double-blind randomised controlled trial, we tested if leisure-time runners using shoes with less compliant midsoles have a higher running-related injury (RRI) risk. We provided 247 runners with standard running shoes having either a soft study shoes (soft-SS) or a hard study shoes (hard-SS) midsole and followed them prospectively for 5 months regarding RRI. All information about sports practice and injuries was uploaded on a dedicated internet platform and checked for consistency and completeness. RRI was defined as any first-time pain sustained during or as a result of running practice and impeding normal running activity for at least 1 day. Cox proportional hazards regressions were used to identify RRI risk factors. The type of study shoes used for running was not associated with RRIs (HR=0.92; 95% CI 0.57 to 1.48). The hard-SS had a 15% greater overall stiffness in the heel region. The two study groups were similar regarding personal and sports participation characteristics, except for years of running experience, which was higher (p<0.05) in the hard-SS group. Global RRI incidence was 12.1 RRI/1000 h of running. No between-group differences were found regarding injury location, type, severity or category. Nevertheless, the adjusted regression model revealed positive associations with RRI risk for body mass index (HR=1.126; 95% CI 1.033 to 1.227), previous injury (HR=1.735; 95% CI 1.037 to 2.902) and mean session intensity (HR=1.396; 95% CI 1.040 to 1.874). Protective factors were previous regular running activity (HR=0.422; 95% CI 0.228 to 0.779) and weekly volume of other sports activities (HR=0.702; 95% CI 0.561 to 0.879). Midsole hardness of modern cushioned running shoes does not seem to influence RRI risk.


Sarcopenia, obesity, and natural killer cell immune senescence in aging: altered cytokine levels as a common mechanism.
Human aging is characterized by both physical and physiological frailty. A key feature of frailty, sarcopenia is the age-associated decline in skeletal muscle mass, strength, and endurance that characterize even the healthy elderly. Increases in adiposity, particularly in visceral adipose tissue, are almost universal in aging individuals and can contribute to sarcopenia and insulin resistance by increasing levels of inflammatory cytokines known collectively as adipokines. Aging also is associated with declines in adaptive and innate immunity, known as immune senescence, which are risk factors for cancer and all-cause mortality. The cytokine interleukin-15 (IL-15) is highly expressed in skeletal muscle tissue and declines in aging rodent models. IL-15 inhibits fat deposition and insulin resistance, is anabolic for skeletal muscle in certain situations, and is required for the development and survival of natural killer (NK) lymphocytes. We review the effect that adipokines and myokines have on NK cells, with special emphasis on IL-15. We posit that increased adipokine and decreased IL-15 levels during aging constitute a common mechanism for sarcopenia, obesity, and immune senescence.


Motor protein function in skeletal muscle-a multiple scale approach to contractility.
We present an approach to skeletal muscle contractility and its regulation over different scales ranging from biomechanical studies in intact muscle fibers down to the motility and interaction of single motor protein molecules. At each scale, shortening velocities as a measure for weak cross-bridge cycling rates are extracted and compared. Experimental approaches include transmitted light microscopy, second harmonic generation imaging of contracting myofibrils, and fluorescence microscopy of single molecule motility. Each method yields image sequences that are analyzed with automated image processing algorithms to extract the contraction velocity. Using this approach, we show how to isolate the contribution of the motor proteins actin and myosin and their modulation by regulatory proteins from the concerted action of electro-mechanical activation on a more complex cellular scale. The advantage of this approach is that averaged contraction velocities can be determined on the different scales ranging from isolated motor proteins to sarcomere levels in myofibrils and myofibril arrays within the cellular architecture. Our results show that maximum shortening velocities during in situ electrical activation of sarcomere contraction in intact single muscle cells can substantially deviate from sliding velocities obtained in oriented in vitro motility assays of isolated motor proteins showing that biophysical contraction kinetics not simply translate linearly between contractility scales. To adequately resolve the very fast initial mechanical activation kinetics of shortening at each scale, it was necessary to implement high-speed imaging techniques. In the case of intact fibers and single molecule motility, we achieved a major increase in temporal resolution up to frame rates of 200-1000 fps using CMOS image sensor technology. The data we obtained at this unprecedented temporal resolution and the parameters extracted can be used to validate results obtained from computational models of motor protein interaction and skeletal muscle contractility in health and muscle disease. Our approach is feasible to explain the possible underlying mechanisms that contribute to different shortening velocities at different scales and complexities.


Phospholipase A2, oxidative stress, and neurodegeneration in binge ethanol-treated organotypic slice cultures of developing rat brain.
Brain neurodamage from chronic binge ethanol (EtOH) exposure is linked to neuroinflammation and associated oxidative stress. Using rat organotypic hippocampal-entorhinal cortical (HEC) slice cultures of developing brain age, we reported that binge EtOH promotes release of a neuroinflammatory instigator, arachidonic acid (AA), concomitant with neurodegeneration, and that mepacrine, a global inhibitor of phospholipase A2 (PLA2) enzymes mobilizing AA from phospholipids, is neuroprotective. Here, we sought with binge EtOH-treated HEC cultures to establish that PLA2 activity is responsible in part for significant oxidative stress and to ascertain the PLA2 families responsible for AA release and neurodegeneration. HEC slices, prepared from 1-week-old rats and cultured 2 to 2.5 weeks, were exposed to 100 mM EtOH over 6 successive days, with 4 daytime "withdrawals" (no EtOH). Brain 3-nitrotyrosinated (3-NT)- and 4-hydroxy-2-nonenal (4-HNE)-adducted proteins, oxidative stress footprints, were immunoassayed on days 3 through 6, and mepacrine's effect was determined on day 6. The effects of specific PLA2 inhibitors on neurodegeneration (propidium iodide staining) and AA release (ELISA levels in media) in the cultures were then determined. Also, the effect of JZL184, an inhibitor of monoacylglycerol lipase (MAGL) which is reported to mobilize AA from endocannabinoids during neuroinflammatory insults, was examined. 3-NT- and 4-HNE-adducted proteins were significantly increased by the binge EtOH exposure, consistent with oxidative stress, and mepacrine prevented the increases. The PLA2 inhibitor results implicated secretory PLA2 (group II sPLA2) and to some extent Ca(2+) -independent cytosolic PLA2 (group VI iPLA2) in binge EtOH-induced neurotoxicity and in AA release, but surprisingly, Ca(2+) -dependent cytosolic PLA2 (group IV cPLA2) did not appear important. Furthermore, unlike PLA2 inhibition, MAGL inhibition failed to prevent the neurodegeneration. In these developing HEC slice cultures, pro-oxidative signaling via sPLA2 and iPLA2, but not necessarily cPLA2 or MAGL, is involved in EtOH neurotoxicity. This study provides further insights into neuroinflammatory phospholipase signaling and oxidative stress underlying binge EtOH-induced neurodegeneration in developing (adolescent age) brain in vitro.


River transport of mercury from artisanal and small-scale gold mining and risks for dietary mercury exposure in Madre de Dios, Peru.
Artisanal and small-scale gold mining (ASGM) is a major contributor to deforestation and the largest anthropogenic source of atmospheric mercury worldwide. Despite significant information on the direct health impacts of mercury to ASGM miners, the impact of mercury contamination on downstream communities has not been well characterized, particularly in Peru's Madre de Dios region. In this area, ASGM has increased significantly since 2000 and has led to substantial political and social controversy. This research examined the spatial distribution and transport of mercury through the Madre de Dios River with distance from ASGM activity. This study also characterized risks for dietary mercury exposure to local residents who depend on fish from the river. River sediment, suspended solids from the water column, and fish samples were collected in 2013 at 62 sites near 17 communities over a 560 km stretch of the Madre de Dios River and its major tributaries. In areas downstream of known ASGM activity, mercury concentrations in sediment, suspended solids, and fish within the Madre de Dios River were elevated relative to locations upstream of mining. Fish tissue mercury concentrations were observed at levels representing a public health threat, with greater than one-third of carnivorous fish exceeding the international health standard of 0.5 mg kg(-1). This study demonstrates that communities located hundreds of kilometers downstream of ASGM activity, including children and indigenous populations who may not be involved in mining, are at risk of dietary mercury exposure that exceed acceptable body burdens. This report represents the first systematic study of the region to aid policy decision-making related to ASGM activities in Peru.


Launching a new fellowship for medical students: the first years of the Doris Duke Clinical Research Fellowship Program.
As part of its commitment to increasing the pipeline of physicians pursuing careers in clinical research, the Doris Duke Charitable Foundation launched the Doris Duke Clinical Research Fellowship (CRF) Program for medical students in 2000. The program, which is based at 10 US medical schools, provides medical students from any US medical school with the opportunity to spend 1 year obtaining both didactic and "hands-on" mentored clinical research experience. This article describes the program and summarizes the early outcomes collected during the first 3.5 years of the program. Interest in the program among medical students has been robust and widespread, with 35% of CRF program fellows matriculated at non-CRF schools. Exit surveys of the first three classes of fellows totaling 174 fellows indicated that (1) 97% of the fellows felt that participating in the fellowship was a good decision; (2) commitment to a career in clinical research was increased among the 44% of fellows reporting that they were unsure about pursuing a clinical research career when they began their fellowship; (3) there was no difference in satisfaction level between the fellows who remained at the medical schools in which they were matriculated and those who completed their fellowship at a medical school in which they were not matriculated; and (4) the majority of fellows responded that the didactic component of their fellowship was useful.


Selection of controls for clinical research studies in ophthalmology.
Ophthalmologists are becoming increasingly aware of the need for well-designed, controlled studies to investigate etiology and treatment of ocular conditions. While patients with the conditions of interest, or with characteristics that place them at high risk of eventual development of such conditions, may be readily identified in the ophthalmology clinic, identification of appropriate controls to be used for comparison may be quite difficult. In this article, epidemiologic principles for selection of controls for retrospective (case-control) and prospective (cohort) studies are reviewed. Two related studies, a case-control study in which multiple control groups were employed and a 15-year cohort study of all cases and controls, provided an opportunity to compare controls selected at random from an ophthalmology practice with controls selected at random from the general population. Participation rates were higher among office patients selected for study, both in the case-control study and in the cohort study. In the cohort study, office controls were more likely to lose visual acuity and to develop new degenerative eye conditions. These differences between control groups selected from two different sources emphasize the need for careful evaluation of potential groups of controls with respect to biases that each may bring to interpretation of clinical research findings.


A Naturally Occurring Polymorphism in the HIV-1 Tat Basic Domain Inhibits Uptake by Bystander Cells and Leads to Reduced Neuroinflammation.
HIV-1 Tat protein contributes to HIV-neuropathogenesis in several ways including its ability to be taken up by uninfected bystander CNS cells and to activate inflammatory host genes causing synaptic injury. Here, we report that in the globally dominant HIV-1 clade C, Tat displays a naturally occurring polymorphism, R57S, in its basic domain, which mediates cellular uptake. We examined the effect of this polymorphism on Tat uptake and its consequences for cellular gene transactivation. In decapeptides corresponding to the basic domain, a R57S substitution caused up to a 70% reduction in uptake. We also used a transcellular Tat transactivation assay, where we expressed Tat proteins of HIV-1 clade B (Tat-B) or C (Tat-C) or their position 57 variants in HeLa cells. We quantified the secreted Tat proteins and measured their uptake by TZM-bl cells, which provide readout via an HIV-1 Tat-responsive luciferase gene. Transactivation by Tat-B was significantly reduced by R57S substitution, while that of Tat-C was enhanced by the reciprocal S57R substitution. Finally, we exposed microglia to Tat variants and found that R57 is required for maximal neuroinflammation. The R57S substitution dampened this response. Thus, genetic variations can modulate the ability of HIV-1 Tat to systemically disseminate neuroinflammation.


C1- and R-dependent expression of the maize Bz1 gene requires sequences with homology to mammalian myb and myc binding sites.
Tissue-specific expression of the maize anthocyanin Bronze-1 (Bz1) gene is controlled by the products of several regulatory genes. These include C1 or Pl and R or B that share homology to the myb proto-oncogenes and myc-like genes, respectively. Bz1 expression in embryo tissues is dependent on C1 and an R-sc allele of R. Transient expression from mutated and deleted versions of the Bz1 promoter fused to a luciferase reporter gene was measured in C1, Rscm2 embryos after gene transfer by microprojectiles. This analysis revealed that the sequences between -76 base pairs (bp) and -45 bp and a 9-bp AT-rich block between -88 bp and -80 bp were critical for Bz1 expression. The -76 bp to -45 bp region includes two short sequences that are homologous to the consensus binding sites of the myb- and myc-like proteins. Site-specific mutations of these "myb" and "myc" sequences reduced Bz1 expression to 10% and 1% of normal, respectively. Additionally, a trimer of a 38-bp oligonucleotide containing these myb and myc sites increased the expression of a cauliflower mosaic virus 35S minimal promoter by 26-fold. This enhancement was dependent on both C1 and R. Because the sites critical for Bz1 expression are homologous to the myb and myc consensus binding sequences and the C1 and R proteins share homology with the myb and myc products, respectively, we propose that C1 and R interact with the Bz1 promoter at these sites.


A new photoactivatable near-infrared-emitting QCy7 fluorophore for single-molecule super-resolution microscopy.
A new photoactivatable fluorophore based on a quinone-cyanine-7 (QCy7) scaffold was synthesized and spectroscopically characterized. The fluorophore exhibits fluorescence in the near-infrared spectral window and a large Stokes shift. Its application in super-resolution microscopy as an antibody bioconjugate is demonstrated.


Comparative study of Helicobacter pylori eradication rates with 5-day quadruple "concomitant" therapy and 7-day standard triple therapy.
Several studies have shown the superiority of concomitant quadruple therapy containing 3 antibiotics over triple therapy for Helicobacter pylori infection. The aim of this study was to compare concomitant quadruple therapy with standard triple therapy for first-line H. pylori eradication. A total of 270 patients with proven H. pylori infection were randomly assigned to one of 2 regimens: amoxicillin 1000 mg with clarithromycin 500 mg and lansoprazole 30 mg twice daily for 7 days (triple therapy) or amoxicillin 1000 mg with clarithromycin 500 mg, metronidazole 500 mg, and lansoprazole 30 mg twice daily for 5 days (concomitant therapy). The success of eradication was evaluated 4 to 5 weeks after completion of treatment. Eradication rates were 86.1% in the triple therapy and 91.4% in the concomitant therapy (per protocol), but the difference was not statistically significant. Mild adverse events were more frequently reported in the concomitant-therapy group (35.6%) than in the triple-therapy group (25.2%) (P=0.09). Five-day quadruple concomitant therapy eradicated H. pylori in over 90% of patients. Accordingly, concomitant therapy is thought to be a promising alternative to triple therapy as a first-line treatment regimen for H. pylori eradication.


The role of microRNAs in coronary artery disease: From pathophysiology to diagnosis and treatment.
MicroRNAs (miRNAs) are tiny non-coding RNA molecules that regulate gene expression predominantly at the post-transcriptional level. Far from being simple intracellular regulators, miRNAs have recently been involved in intercellular communication and have been shown to circulate in the bloodstream in stable forms. In the past years specific miRNA expression patterns have been linked to the development of atherosclerosis and coronary artery disease, two closely related conditions. The study of miRNAs has promoted our understanding of the processes involved in the pathogenesis of atherosclerosis and innovative diagnostic and therapeutic approaches have emerged. In this review, we present the role of miRNAs in the development of atherosclerosis, on coronary artery disease progression and we assess their role as diagnostic biomarkers. Finally we evaluate the therapeutic and preventive opportunities that arise from the study of miRNAs in coronary artery disease and especially in myocardial infarction.


Canadian Adolescents' Experiences of Dating Violence: Associations with Social Power Imbalances.
A substantial minority of adolescents experience and use dating violence in their sexual and/or romantic relationships. Limited attention has been paid to exploring theory-driven questions about use and experience of adolescent dating violence (ADV), restricting knowledge about promising prevention targets for diverse groups of youth. To address this gap, this paper investigates whether factors tied to power imbalances (bullying, risk of social marginalization) are associated with patterns of ADV victimization and perpetration in a large sample of Canadian mid-adolescents. We used data from the 2017/2018 Health-Behavior in School-Aged Children (HBSC) study, a nationally representative sample of Canadian youth. Our study was comprised of adolescents who were in grades 9 or 10, and who had dated in the past 12 months (N = 3779). We assessed multiple forms of ADV and bullying victimization and perpetration. We also included six variables assessing adolescents' risk of social marginalization: gender, race/ethnicity, immigration status, family structure, food insecurity, and family affluence. We used latent class analysis to explore the ways adolescents experience and use different forms of ADV, and then examined whether factors tied to power imbalances (bullying, social marginalization) were associated with classes of ADV. Three ADV classes emerged in our sample: uninvolved (65.7%), psychological and cyber victimization only (28.9%), and mutual violence (5.4%). Bullying was most strongly associated with the mutual violence class, suggesting a transformation of power from peer to romantic contexts. Social marginalization variables were associated with ADV patterns in different ways, highlighting the need to use a critical and anti-oppressive lens in ADV research and prevention initiatives.


Examining the role of frequency specificity in the enhancement and suppression of human cortical activity by auditory selective attention.
This study examined the neural basis of auditory selective attention using functional magnetic resonance imaging. The main hypothesis stated that attending to a particular sound frequency would significantly enhance the neural response within those tonotopic regions of the auditory cortex sensitive to that frequency. To test this prediction, low- and high-frequency sound sequences were interleaved to produce two concurrent auditory streams. Six normally hearing participants either performed a task which required them to attend to one or the other stream or listened passively to the sounds while functional images were acquired using a high-resolution (1.5 mm x 1.5 mm x 2.5 mm) sequence. Two statistical comparisons identified the attention-specific and general effects of enhancement. The first controlled for task-related processes, while the second did not. Results demonstrated frequency-specific, attention-specific enhancement in the response to the attended frequency, but no response suppression for the unattended frequency. Instead, a general effect of suppression was found in several posterior sites, possibly related to resting-state processes. Furthermore, there was widespread general enhancement across auditory cortex when performing the task compared to passive listening. This enhancement did include frequency-sensitive regions, but was not restricted to them. In conclusion, our results show partial support for frequency-specific enhancement.


The C.elegans MAPK phosphatase LIP-1 is required for the G(2)/M meiotic arrest of developing oocytes.
In the Caenorhabditis elegans hermaphrodite germline, spatially restricted mitogen-activated protein kinase (MAPK) signalling controls the meiotic cell cycle. First, the MAPK signal is necessary for the germ cells to progress through pachytene of meiotic prophase I. As the germ cells exit pachytene and enter diplotene/diakinesis, MAPK is inactivated and the developing oocytes arrest in diakinesis (G(2)/M arrest). During oocyte maturation, a signal from the sperm reactivates MAPK to promote M phase entry. Here, we show that the MAPK phosphatase LIP-1 dephosphorylates MAPK as germ cells exit pachytene in order to maintain MAPK in an inactive state during oocyte development. Germ cells lacking LIP-1 fail to arrest the cell cycle at the G(2)/M boundary, and they enter a mitotic cell cycle without fertilization. LIP-1 thus coordinates oocyte cell cycle progression and maturation with ovulation and fertilization.


Improving natural ventilation in hospital waiting and consulting rooms to reduce nosocomial tuberculosis transmission risk in a low resource setting.
TB transmission in healthcare facilities is an important public health problem, especially in the often-overcrowded settings of HIV treatment scale-up. The problem is compounded by the emergence of drug resistant TB. Natural ventilation is a low-cost environmental control measure for TB infection control where climate permits that is suited to many different areas in healthcare facilities. There are no published data on the effect of simple structural modifications to existing hospital infrastructure to improve natural ventilation and reduce the risk of nosocomial TB transmission. The purpose of this study was to measure the effect of simple architectural modifications to existing hospital waiting and consulting rooms in a low resource setting on (a) improving natural ventilation and (b) reducing modelled TB transmission risk. Room ventilation was measured pre- and post-modification using a carbon dioxide tracer-gas technique in four waiting rooms and two consulting rooms in two hospitals in Lima, Peru. Modifications included additional windows for cross-ventilation (n = 2 rooms); removing glass from unopenable windows (n = 2); creation of an open skylight (n = 1); re-building a waiting-room in the open air (n = 1). Changes in TB transmission risk for waiting patients, or healthcare workers in consulting rooms, were estimated using mathematical modelling. As a result of the infrastructure modifications, room ventilation in the four waiting rooms increased from mean 5.5 to 15; 11 to 16; 10 to 17; and 9 to 66 air-changes/hour respectively; and in the two consulting rooms from mean 3.6 to 17; and 2.7 to 12 air-changes/hour respectively. There was a median 72% reduction (inter-quartile range 51-82%) in calculated TB transmission risk for healthcare workers or waiting patients. The modifications cost <US$75 in four rooms, and US$1000 and US$7000 in the remaining two rooms. Simple modifications to existing hospital infrastructure considerably increased natural ventilation, and greatly reduced modelled TB transmission risk at little cost.


Validity of two simple measures for estimating life-course socio-economic position in cross-sectional postal survey data in an older population: results from the North Staffordshire Osteoarthritis Project (NorStOP).
Since few cohorts encompass the whole life-course, many studies that measure socio-economic position (SEP) across the life-course rely on participant recall of SEP measures from cross-sectional postal or interview surveys. It is also particularly important that SEP measures should be appropriate for the age of the population studied, as the level of missing data has been shown to increase in older people. The aim of this study was to investigate the accuracy of recall of two SEP measures in older adults, age left school and longest job, by examining their validity in a general population postal survey in North Staffordshire, UK. Sets of questions on education and longest job were included in a questionnaire at different stages of the study. All patients aged 50+ registered with three general practices were sent a baseline Health Questionnaire. 6 years later, 3410 responders were mailed a follow-up Health Questionnaire; a sub-sample of these participants took part in independent qualitative interviews. Validity was assessed by: percentage completion; internal percentage agreement within each set of questions; percentage agreement of qualitative and quantitative data for age left school and longest job; comparing recall of age left school with historical change in legal school leaving age; comparing frequency of pottery job titles with those in 1981 Census data for Stoke-on-Trent. The adjusted response to different stages of the study was 71-85%. Completion of questions was 83-98%. Internal agreement was 84-97% (education) and 95-100% (longest job). Comparison of survey and interview data showed 86% agreement (± 1 year) for age left school and 91% agreement for longest job. The change in age left school data concurred with the historical shift in legal school leaving age. 11% of job titles were pottery in NorStOP data and 15% in Stoke-on-Trent Census data. The results from this study provide evidence for the accuracy of recall of two simple measures of SEP (age left school and longest job) in a postal survey of older adults. Consistency with evidence from external datasets indicated the potential validity of these measures for studying life-course SEP in population surveys.


Incidence of seropositive myasthenia gravis in Cape Town and South Africa.
Myasthenia gravis (MG) is a treatable autoimmune disease characterised by fatiguable weakness of skeletal muscles. More than 85% of MG patients have antibodies to the acetylcholine receptor (AChR) at the neuromuscular junction or are seropositive for MG (SPMG). In the developed world the incidence of MG has increased, particularly among older individuals, but no epidemiological studies have been done on SPMG in Africa. To determine the annual incidence rate (IR) of SPMG in the Cape Town (CT) municipality, and the crude annual IR of SPMG for the whole of South Africa (SA). Positive AChR antibody tests were identified between 1 January 2003 and 1 January 2005 for patients living in CT, and the age- and sex-specific incidences were calculated. To determine the national crude annual IR over the same period, positive assays were identified from the laboratories that process AChR assays for SA. National Census 2001 population statistics formed the denominators. There were 65 positive assays in CT, and 230 nationwide. Based on these figures the annual IR for CT was 11.2 per million per year (95% confidence interval (CI) 8.7 - 14.3), and for South Africa 2.6 per million/year (95% CI 2.2 - 2.9). After a questionnaire response from CT neurologists regarding the routine use of the AChR antibody assay, the annual IR for CT was adjusted to 12.6 per million (95% CI 9.9 - 15.9) to incorporate those presumed to have SPMG without a confirmatory test. In CT, the IR in females was 15.3 per million/year (95% CI 11.2 - 20.4), and in males, 6.8 per million/year (95% CI 4.1 - 10.7). The CT IRs for blacks, coloureds and whites were not statistically different after adjusting for age and gender. The IR of SPMG in CT was 6 times greater in those presenting after the age of 50 years than in those with earlier disease onset (95% CI 3.7 - 9.7). The annual IR of SPMG in CT is much the same as rates recorded recently in other developed countries, but the rest of SA has a much lower IR. A preponderance of MG starting after the age of 50 years reflects a worldwide trend, although the CT data showed a relatively lower-than-expected incidence for older males. IRs for SPMG vary widely in different regions in SA; this is likely to be related to differences in regional health care delivery, and underdiagnosis.


Isolation and Identification of Fusarium solani f. sp. glycines from Soil on Modified Nash and Snyder's Medium.
Modified Nash and Snyder's medium (MNSM) has been used to study soil populations of the fungus Fusarium solani f. sp. glycines, the causal agent of sudden death syndrome (SDS) of soybean. However, no studies have been conducted to confirm the accuracy of this technique in enumerating the SDS pathogen. To determine what portion of the colonies enumerated on MNSM are the SDS pathogen, 282 isolates resembling F. solani f. sp. glycines were collected from soil of two fields with a history of SDS in Arkansas using a soil dilution method with MNSM. The colony morphology of these isolates was compared on MNSM and on potato dextrose agar (PDA). Of these isolates, 112 were tested for pathogenicity on soybean in the greenhouse. In addition to fungi that produced colonies typical of F. solani f. sp. glycines on MNSM, 5 isolates each of four common colony types on MNSM were collected from each field for a total of 40 isolates. These isolates were compared to F. solani f. sp. glycines for colony morphology on PDA and for pathogenicity. Isolates that had colonies on MNSM that resembled F. solani f. sp. glycines had colony morphologies on PDA similar to this pathogen and produced SDS-like foliar symptoms in greenhouse inoculations of soybean. There was a significant quadratic relationship between foliar symptom severity and plant fresh weight. None of the other isolates tested resembled F. solani f. sp. glycines on either MNSM or PDA or produced SDS-foliar symptoms in greenhouse inoculations. These non-SDS isolates were F. solani, F. merismoides, a Pythium sp., and a Paecilomyces sp. All of these isolates produced some root rot, but significantly less than the F. solani f. sp. glycines reference isolate. The results of this study established that soil populations of F. solani f. sp. glycines can be accurately enumerated using MNSM. It should be noted that, on both MNSM and PDA, F. solani f. sp. glycines can resemble some isolates of F. solani f. sp. phaseoli, so pathogenicity tests of random isolates may be required to confirm the forma specialis especially in fields that have a history of both Glycines max and Phaseolus vulgaris.


Using 3D printed models for planning and guidance during endovascular intervention: a technical advance.
Three-dimensional (3D) printing applications in medicine have been limited due to high cost and technical difficulty of creating 3D printed objects. It is not known whether patient-specific, hollow, small-caliber vascular models can be manufactured with 3D printing, and used for small vessel endoluminal testing of devices. Manufacture of anatomically accurate, patient-specific, small-caliber arterial models was attempted using data from a patient's CT scan, free open-source software, and low-cost Internet 3D printing services. Prior to endovascular treatment of a patient with multiple splenic artery aneurysms, a 3D printed model was used preoperatively to test catheter equipment and practice the procedure. A second model was used intraoperatively as a reference. Full-scale plastic models were successfully produced. Testing determined the optimal puncture site for catheter positioning. A guide catheter, base catheter, and microcatheter combination selected during testing was used intraoperatively with success, and the need for repeat angiograms to optimize image orientation was minimized. A difficult and unconventional procedure was successful in treating the aneurysms while preserving splenic function. We conclude that creation of small-caliber vascular models with 3D printing is possible. Free software and low-cost printing services make creation of these models affordable and practical. Models are useful in preoperative planning and intraoperative guidance.


Maintaining data integrity in a rural clinical trial.
Clinical trials conducted in rural resource-poor settings face special challenges in ensuring quality of data collection and handling. The variable nature of these challenges, ways to overcome them, and the resulting data quality are rarely reported in the literature. To provide a detailed example of establishing local data handling capacity for a clinical trial conducted in a rural area, highlight challenges and solutions in establishing such capacity, and to report the data quality obtained by the trial. We provide a descriptive case study of a data system for biological samples and questionnaire data, and the problems encountered during its implementation. To determine the quality of data we analyzed test-retest studies using Kappa statistics of inter- and intra-observer agreement on categorical data. We calculated Technical Errors of Measurement of anthropometric measurements, audit trail analysis was done to assess error correction rates, and residual error rates were calculated by database-to-source document comparison. Initial difficulties included the unavailability of experienced research nurses, programmers and data managers in this rural area and the difficulty of designing new software tools and a complex database while making them error-free. National and international collaboration and external monitoring helped ensure good data handling and implementation of good clinical practice. Data collection, fieldwork supervision and query handling depended on streamlined transport over large distances. The involvement of a community advisory board was helpful in addressing cultural issues and establishing community acceptability of data collection methods. Data accessibility for safety monitoring required special attention. Kappa values and Technical Errors of Measurement showed acceptable values. Residual error rates in key variables were low. The article describes the experience of a single-site trial and does not address challenges particular to multi-site trials. Obtaining and maintaining data integrity in rural clinical trials is feasible, can result in acceptable data quality and can be used to develop capacity in developing country sites. It does, however, involve special challenges and requirements.


Transposable element 'roo' attaches to nuclear matrix of the Drosophila melanogaster.
The genome of eukaryotes is organized into structural units of chromatin loops. This higher order organization is supported by a nuclear skeleton called the nuclear matrix. The genomic DNA associated with the nuclear matrix is called the matrix associated region (MAR). Only a few genome-wide screens have been attempted, although many studies have characterized locusspecific MAR DNA sequences. In this study, a MAR DNA library was prepared from the Drosophila melanogaster Meigen (Diptera: Drosophilidae) genome. One of the sequences identified as a MAR was from a long terminal repeat region of 'roo' retrotransposon (roo MAR). Sequence analysis of roo MAR showed its distribution across the D. melanogaster genome. roo MAR also showed high sequence similarity with a previously identified MAR in Drosophila, namely the 'gypsy' retrotransposon. Analysis of the genes flanking roo MAR insertions in the Drosophila genome showed that genes were co-ordinately expressed. The results from the present study in D. melanogaster suggest this sequence plays an important role in genome organization and function. The findings point to an evolutionary role of retrotransposons in shaping the genomic architecture of eukaryotes.


EARLY MICROVASCULAR AND NEURAL CHANGES IN PATIENTS WITH TYPE 1 AND TYPE 2 DIABETES MELLITUS WITHOUT CLINICAL SIGNS OF DIABETIC RETINOPATHY.
To assess and compare early modifications in inner retinal layer thickness and optical coherence tomography angiography parameters in patients with diabetes mellitus (DM) Types 1 and 2 without clinical signs of diabetic retinopathy. Ninety eyes of 90 subjects (24 Type 1 DM, 36 Type 2 DM, and 30 healthy controls) were prospectively evaluated with spectral domain OCT, swept-source OCT angiography, and color fundus photography (on the same day). Retinal nerve fiber layer, ganglion cell layer (GCL+), and nerve fiber layer + GCL+ (GCL++) thickness were automatically determined by the instrument in the 1, 3, and 6 central mm. On OCT angiography, the following parameters were evaluated: area of foveal avascular zone, number of focally dilated endings of the capillaries (detected only on OCT angiography), presence of regular/irregular foveal avascular zone, capillary loss, and capillary network irregularities in the superficial capillary plexus (SCP) and deep capillary plexus (DCP). Ganglion cell layer+ (P = 0.0099) and GCL++ (P = 0.0367) were significantly thicker in DM Type 1 versus DM Type 2 in 1 central mm, after adjustment for age and DM duration. The area of foveal avascular zone was significantly larger in DM Type 1 versus controls in both SCP and DCP and in DM Type 1 versus Type 2 only in DCP (P < 0.05 for all); the number of focally dilated endings of the capillaries was higher in DM Type 1 versus controls in both SCP and DCP (P < 0.01 for all); and in DM Type 2 versus controls only in DCP (P = 0.007). Perifoveal capillary loss in SCP and inner retinal layer thickness had the highest correlation in both DM types. There are specific neural and microvascular modifications even before clinical signs of diabetic retinopathy in DM Types 1 and 2. Perifoveal capillary loss in the SCP is highly correlated with inner retinal layer. These data may help in characterization of patients at the preclinical stage of diabetic retinopathy.


Endoscopic retrograde cholangiopancreaticography with or without stenting in patients with pancreaticobiliary malignancy, prior to surgery.
Postoperative morbidity and mortality are high in patients undergoing pancreatico-duodenectomy for malignant pancreatico-biliary stricture. Different approaches have been tried to improve the outcomes, including pre-surgical biliary stenting with endoscopic retrograde cholangiopancreaticography (ERCP). To assess the beneficial and harmful effects of biliary stenting via ERCP for pancreatico-biliary stricture confirmed or suspected to be malignant, prior to surgery. We identified trials through The Cochrane Hepato-Biliary Group Controlled Trials Register (October 2006), the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library (Issue 2, 2006), MEDLINE (1950 to October 2006), EMBASE (1980 to October 2006), and Science Citation Index Expanded (1945 to October 2006). We also searched the references in the published papers and wrote to stent producers. Randomised trials comparing ERCP with biliary stenting versus ERCP without biliary stenting for pancreatico-biliary malignancy prior to surgery. Two authors independently selected trials for inclusion and extracted data. The primary pre-surgical, post-surgical, and final outcome measures were mortality. The secondary outcomes were complications such as cholangitis, pancreatitis, bleeding, pancreatic fistula, intra-abdominal abscess, improvement in bilirubin, and quality of life. Dichotomous outcomes were reported as odds ratio (OR) with 95% confidence interval (CI) based on fixed- and random-effect models. We identified two randomised trials with 125 patients undergoing pancreatico-duodenectomy; 62 patients underwent ERCP with biliary stenting and 63 had ERCP without biliary stenting prior to surgery. Pre-surgical mortality was not significantly affected by stenting (OR 3.14, 95% CI 0.12 to 79.26), while there were significantly more complications in the stented group (OR 43.75, 95% CI 2.51 to 761.8). Stenting had no significant effect on the post-surgical mortality (OR 0.75, 95% CI 0.25 to 2.24). However, post-surgical complications were significantly less in the stented group (OR 0.45, 95% CI 0.22 to 0.91). Overall mortality (OR 0.81, 95% CI 0.17 to 3.89) and complications (OR 0.50, 95% CI 0.01 to 23.68) were not significantly different in the two groups. We could not find convincing evidence to support or refute endoscopic biliary stenting on the mortality in patients with pancreatico-biliary malignancy. Large randomised trials are needed to settle the question of pre-surgical biliary stenting.


Management of chronic proximal fistulas after sleeve gastrectomy by laparoscopic Roux-limb placement.
Laparoscopic sleeve gastrectomy (LSG) is an increasingly popular bariatric procedure. However, after LSG, leaks may occur that are difficult to treat and that can persist for months. Treatment is controversial and ranges from stent placement to total gastrectomy. We propose an alternative laparoscopic treatment. The aims of our study were to report on the incidence of chronic fistulas after LSG and the outcome of Roux-limb placement on these defects. The setting was a major teaching hospital in Belgium. From January 1, 2002, to December 31, 2011, we performed LSG as a primary weight loss operation (PLSG) in 728 patients and as a corrective operation (CLSG) in 84 patients. A retrospective chart and database review was conducted. When a chronic leak persisted beyond 4 months, we performed a laparoscopic Roux-limb placement on the defect. Leaks occurred in 26 patients (3.6%) after PLSG and in 6 (7.1%) after CLSG. A leak persisted beyond 4 months in 7 patients (26.9%) after PLSG and in 2 patients (33.3%) after CLSG. Two patients with a chronic fistula after PLSG were referred to our hospital. In 11 patients, a Roux limb was laparoscopically sutured to the defect. The mean time for a chronic fistula to heal after Roux-limb placement was 12.5±10.2 days, and the mean length of hospital stay was 19.6± 14.2. Proximal leaks after LSG are relatively rare. However, a significant number become chronic. Laparoscopic placement of a Roux-limb on the defect is a safe treatment with an eventual succes rate of 100%.


Audiological practice and COVID-19: recommendations that audiological centers can use to maintain the safety and quality of service-expert opinion.
Audiology is an essential service for some patient groups and some interventions. This article sets forth experience-based recommendations for how audiological centers can continue to safely and effectively function during COVID-19. The recommendations are the result of panel discussion and are based on the clinical experience of the panelists/authors. The recommendations cover which patient groups and which interventions should be treated when and whether this can be performed in the clinic or remotely; how to maintain the safety of workplace via optimizing patient flow within the clinic and the sanitation of rooms and equipment; and overcoming communication challenges that COVID-19 intensifies. For essential audiological services to continue under COVID-19, safety measures must be implemented and maintained, and treatment and communication strategies must be adapted to offset communication difficulties due to personal protective equipment (PPE) and social distancing and to bolster patient confidence. In short, it is vital that staff feel safe, that patients either feel the clinic is safe enough to visit or that remote treatment may be an option, and that clinics and patients have a broad agreement on the urgency of any needed service. We hope that these recommendations help clinics effectively accomplish these goals.


The Lake Victoria Island Intervention Study on Worms and Allergy-related diseases (LaVIISWA): study protocol for a randomised controlled trial.
The Hygiene Hypothesis proposes that infection exposure protects against inflammatory conditions. Helminths possess allergen-like molecules and may specifically modulate allergy-related immunological pathways to inhibit responses which protect against them. Mass drug administration is recommended for helminth-endemic communities to control helminth-induced pathology, but may also result in increased rates of inflammation-mediated diseases in resource-poor settings. Immunological studies integrated with implementation of helminth control measures may elucidate how helminth elimination contributes to ongoing epidemics of inflammatory diseases. We present the design of the Lake Victoria Island Intervention Study on Worms and Allergy-related diseases (LaVIISWA), a cluster-randomised trial evaluating the risks and benefits of intensive versus standard anthelminthic treatment for allergy-related diseases and other health outcomes. The setting is comprised of island fishing communities in Mukono district, Uganda. Twenty-six communities have been randomised in a 1:1 ratio to receive standard or intensive anthelminthic intervention for a three-year period. Baseline characteristics were collected immediately prior to intervention rollout, commenced in February 2013. Primary outcomes are reported wheeze in the past 12 months and atopy (skin prick test response and allergen-specific immunoglobulin (asIg) E concentration). Secondary outcomes are visible flexural dermatitis, helminth infections, haemoglobin, growth parameters, hepatosplenomegaly, and responses to vaccine antigens. The trial provides a platform for in-depth analysis of clinical and immunological consequences of the contrasting interventions. The baseline survey has been completed successfully in a challenging environment. Baseline characteristics were balanced between trial arms. Prevalence of Schistosoma mansoni, hookworm, Strongyloides stercoralis and Trichuris trichiura was 52%, 23%, 13%, and 12%, respectively; 31% of Schistosoma mansoni infections were heavy (>400 eggs/gram). The prevalence of reported wheeze and positive skin prick test to any allergen was 5% and 20%, respectively. Respectively, 77% and 87% of participants had Dermatophagoides- and German cockroach-specific IgE above 0.35 kUA/L. These characteristics suggest that the LaVIISWA study will provide an excellent framework for investigating beneficial and detrimental effects of worms and their treatment, and the mechanisms of such effects. This trial was registered with Current Controlled Trials (identifier: ISRCTN47196031) on 7 September 2012.


Evaluating the Use of Tissue Oximetry to Decrease Intensive Unit Monitoring for Free Flap Breast Reconstruction.
Postoperative free flap care has historically required intensive monitoring for 24 hours in an intensive care unit. Continuous monitoring with tissue oximetry has allowed earlier detection of vascular compromise, decreasing flap loss and improving salvage. This study aims to identify whether a fast-track postoperative paradigm can be safely used with tissue oximetry to decrease intensive monitoring and costs. All consecutive microsurgical breast reconstructions performed at a single institution were reviewed (2008-2014) and cases requiring return to the operating room were identified. Data evaluated included patient demographics, the take back time course, and complications of flap loss and salvage. A cost-benefit analysis was performed to analyse the utility of a postoperative intensive monitoring setting. There were 900 flaps performed and 32 required an unplanned return to the operating room. There were 16 flaps that required a reexploration within the first 24 hours; the standard length of intensive unit monitoring. After 4 hours, there were 7 flaps (44%) detected by tissue oximetry for reexploration. After 15 hours of intensive monitoring postoperatively, cost analysis revealed that the majority (15/16; 94%) of failing flaps had been identified and the cost of identifying each subsequent failing flap exceeded the cost of another hour of intensive monitoring. The postoperative paradigm for microsurgical flaps has historically required intensive unit monitoring. Using tissue oximetry, a fast-track pathway can reduce time spent in an intensive monitoring setting from 24 to 15 hours with significant cost savings and minimal risk of missing a failing free flap.


Colocalization of the (Pro)renin Receptor/Atp6ap2 with H+-ATPases in Mouse Kidney but Prorenin Does Not Acutely Regulate Intercalated Cell H+-ATPase Activity.
The (Pro)renin receptor (P)RR/Atp6ap2 is a cell surface protein capable of binding and non-proteolytically activate prorenin. Additionally, (P)RR is associated with H(+)-ATPases and alternative functions in H(+)-ATPase regulation as well as in Wnt signalling have been reported. Kidneys express very high levels of H(+)-ATPases which are involved in multiple functions such as endocytosis, membrane protein recycling as well as urinary acidification, bicarbonate reabsorption, and salt absorption. Here, we wanted to localize the (P)RR/Atp6ap2 along the murine nephron, exmaine whether the (P)RR/Atp6ap2 is coregulated with other H(+)-ATPase subunits, and whether acute stimulation of the (P)RR/Atp6ap2 with prorenin regulates H(+)-ATPase activity in intercalated cells in freshly isolated collecting ducts. We localized (P)PR/Atp6ap2 along the murine nephron by qPCR and immunohistochemistry. (P)RR/Atp6ap2 mRNA was detected in all nephron segments with highest levels in the collecting system coinciding with H(+)-ATPases. Further experiments demonstrated expression at the brush border membrane of proximal tubules and in all types of intercalated cells colocalizing with H(+)-ATPases. In mice treated with NH4Cl, NaHCO3, KHCO3, NaCl, or the mineralocorticoid DOCA for 7 days, (P)RR/Atp6ap2 and H(+)-ATPase subunits were regulated but not co-regulated at protein and mRNA levels. Immunolocalization in kidneys from control, NH4Cl or NaHCO3 treated mice demonstrated always colocalization of PRR/Atp6ap2 with H(+)-ATPase subunits at the brush border membrane of proximal tubules, the apical pole of type A intercalated cells, and at basolateral and/or apical membranes of non-type A intercalated cells. Microperfusion of isolated cortical collecting ducts and luminal application of prorenin did not acutely stimulate H(+)-ATPase activity. However, incubation of isolated collecting ducts with prorenin non-significantly increased ERK1/2 phosphorylation. Our results suggest that the PRR/Atp6ap2 may form a complex with H(+)-ATPases in proximal tubule and intercalated cells but that prorenin has no acute effect on H(+)-ATPase activity in intercalated cells.


Model systems for regeneration: zebrafish.
Tissue damage can resolve completely through healing and regeneration, or can produce permanent scarring and loss of function. The response to tissue damage varies across tissues and between species. Determining the natural mechanisms behind regeneration in model organisms that regenerate well can help us develop strategies for tissue recovery in species with poor regenerative capacity (such as humans). The zebrafish (Danio rerio) is one of the most accessible vertebrate models to study regeneration. In this Primer, we highlight the tools available to study regeneration in the zebrafish, provide an overview of the mechanisms underlying regeneration in this system and discuss future perspectives for the field.