Phosphoproteome analysis of isoflurane-protected heart mitochondria: phosphorylation of adenine nucleotide translocator-1 on Tyr194 regulates mitochondrial function.
Reversible phosphorylation of mitochondrial proteins is essential in the regulation of respiratory function, energy metabolism, and mitochondrion-mediated cell death. We hypothesized that mitochondrial protein phosphorylation plays a critical role in cardioprotection during pre and postconditioning, two of the most efficient anti-ischaemic therapies. Using phosphoproteomic approaches, we investigated the profiles of phosphorylated proteins in Wistar rat heart mitochondria protected by pharmacological pre and postconditioning elicited by isoflurane. Sixty-one spots were detected by two-dimensional blue-native gel electrophoresis-coupled Western blotting using a phospho-Ser/Thr/Tyr-specific antibody, and 45 of these spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Eleven protein spots related to oxidative phosphorylation, energy metabolism, chaperone, and carrier functions exhibited significant changes in their phosphorylation state when protected mitochondria were compared with unprotected. Using a phosphopeptide enrichment protocol followed by liquid chromatography-MS/MS, 26 potential phosphorylation sites were identified in 19 proteins. Among these, a novel phosphorylation site was detected in adenine nucleotide translocator-1 (ANT1) at residue Tyr(194). Changes in ANT phosphorylation between protected and unprotected mitochondria were confirmed by immunoprecipitation. The biological significance of ANT phosphorylation at Tyr(194) was further tested with site-directed mutagenesis in yeast. Substitution of Tyr(194) with Phe, mimicking the non-phosphorylated state, resulted in the inhibition of yeast growth on non-fermentable carbon sources, implying a critical role of phosphorylation at this residue in regulating ANT function and cellular respiration. Our analysis emphasizes the regulatory functions of the phosphoproteome in heart mitochondria and reveals a novel, potential link between bioenergetics and cardioprotection.


Incomplete X-inactivation initiated by a hypomorphic Xist allele in the mouse.
X chromosome inactivation (X-inactivation) in female mammals is triggered by differential upregulation of the Xist gene on one of the two X chromosomes and subsequent coating of the X in cis with its non-coding transcripts. Although targeted mutation has clearly shown that Xist is essential for X-inactivation in cis, the molecular mechanism by which Xist RNA induces chromosome silencing is largely unknown. Here, we demonstrate that an Xist mutant generated previously in mouse by gene targeting, Xist(IVS), is unique in that it partially retains the capacity to silence the X chromosome. Although Xist(IVS) is differentially upregulated and its mutated transcript coats the X chromosome in cis in embryonic and extra-embryonic tissues, X-inactivation thus initiated does not seem to be fully established. The state of such incomplete inactivation is probably unstable and the mutated X is apparently reactivated in a subset of extra-embryonic tissues and, perhaps, early epiblastic cells. Xist(IVS), which can be referred to as a partial loss-of-function mutation, would provide an opportunity to dissect the molecular mechanism of Xist RNA-mediated chromosome silencing.


Decreased clinical response to infliximab in ankylosing spondylitis is correlated with anti-infliximab formation.
Correlation of serum trough infliximab levels and antibodies to infliximab (anti-infliximab) with clinical response in ankylosing spondylitis. In accordance with the international ASsessment in Ankylosing Spondylitis (ASAS) consensus statement, patients were treated with infliximab (5 mg/kg) every 6 weeks after a starting regimen. Preinfusion sera were collected at baseline, 24 and 54 weeks. At every visit, the 20% improvement response (ASAS-20) was assessed and laboratory tests performed. 24 of the 38 (63%) patients fulfilled ASAS-20 response criteria after 24 weeks of treatment and 21 (53%) after 54 weeks. After 54 weeks, 11 (29%) patients showed undetectable serum trough infliximab levels and detectable anti-infliximab; six of these patients developed an infusion reaction. Anti-infliximab was found significantly more often (p = 0.04) in ASAS-20 non-responders compared with responders at week 54. Serum trough infliximab levels were significantly (p<0.0001) lower in patients with (mean 0.02 mg/l) than in those without (12.7 mg/l) anti-infliximab. In ankylosing spondylitis, high levels of serum trough infliximab correlated with a good clinical response. Detection of anti-infliximab within 54 weeks is associated with undetectable serum trough infliximab levels, reduced response to treatment and increased risk of developing an infusion reaction.


Recommendations to plan a national burden of disease study.
The InfAct (Information for Action) project is a European Commission Joint Action on Health Information which has promoted the potential role of burden of disease (BoD) approaches to improve the current European Union-Health Information System (EU-HIS). It has done so by raising awareness of the concept, the methods used to calculate estimates and their potential implications and uses in policymaking. The BoD approach is a systematic and scientific effort to quantify and compare the magnitude of health loss due to different diseases, injuries, and risk factors with estimates produced by demographic characteristics and geographies for specific points in time. Not all countries have the resources to undertake such work, and may therefore start with a more restricted objective, e.g., a limited number of diseases, or the use of simple measures of population health such as disease prevalence or life expectancy. The main objective to develop these recommendations was to facilitate those countries planning to start a national burden of disease study. These recommendations could be considered as minimum requirements for those countries planning to start a BoD study and includes following elements: (1) Define the objectives of a burden of disease study within the context of your country, (2) Identify, communicate and secure the benefits of performing national burden of disease studies, (3) Secure access to the minimum required data sources, (4) Ensure the minimum required capacity and capability is available to carry out burden of disease study, (5) Establish a clear governance structure for the burden of disease study and stakeholder engagement/involvement, (6) Choose the appropriate methodological approaches and (7) Knowledge translation. These were guided by the results from our survey performed to identify the needs of European countries for BoD studies, a narrative overview from four European countries (Belgium, Germany, The Netherlands and Scotland) and the summary of a comparative study of country health profiles with national health statistics. These recommendations as minimum requirements would facilitate efforts by those European countries who intend to perform national BoD studies.


Fabrication and characterization of diffractive optical elements in InP for monolithic integration with surface-emitting components.
We report on the fabrication and subsequent characterization of binary diffractive optical elements (DOE's) in InP for operation at 1.3 microm. Fresnel lenses of different focal lengths and a DOE that splits and focuses an incident beam into a 1 x 4 array of spots (optical fan-out) were fabricated. We realized the surface reliefs by patterning resist, using electron-beam lithography and etching with a chemically assisted ion beam, which produced well-defined patterns with smooth sidewalls and little if no surface roughness. The measured efficiency for the lenses was 36%. For the fan-out element the efficiency and the uniformity error were 26% and 30%, respectively. Spot sizes as small as 16 microm were measured.


Metabolism of low-density lipoproteins by cultured hepatocytes from normal and homozygous familial hypercholesterolemic subjects.
The profoundly elevated concentrations of low-density lipoproteins (LDL) present in homozygous familial hypercholesterolemia lead to symptomatic cardiovascular disease and death by early adulthood. Studies conducted in nonhepatic tissues demonstrated defective cellular recognition and metabolism of LDL in these patients. Since mammalian liver removes at least half of the LDL in the circulation, the metabolism of LDL by cultured hepatocytes isolated from familial hypercholesterolemic homozygotes was compared to hepatocytes from normal individuals. Fibroblast studies demonstrated that the familial hypercholesterolemic subjects studied were LDL receptor-negative (less than 1% normal receptor activity) and LDL receptor-defective (18% normal receptor activity). Cholesterol-depleted hepatocytes from normal subjects bound and internalized 125I-labeled LDL (Bmax = 2.2 micrograms LDL/mg cell protein). Preincubation of normal hepatocytes with 200 micrograms/ml LDL reduced binding and internalization by approx. 40%. In contrast, 125I-labeled LDL binding and internalization by receptor-negative familial hypercholesterolemic hepatocytes was unaffected by cholesterol loading and considerably lower than normal. This residual LDL uptake could not be ascribed to fluid phase endocytosis as determined by [14C]sucrose uptake. The residual LDL binding by familial hypercholesterolemia hepatocytes led to a small increase in hepatocyte cholesterol content which was relatively ineffective in reducing hepatocyte 3-hydroxy-3-methylglutaryl-CoA reductase activity. Receptor-defective familial hypercholesterolemia hepatocytes retained some degree of regulatable 125I-labeled LDL uptake, but LDL uptake did not lead to normal hepatocyte cholesterol content or 3-hydroxy-3-methylglutaryl-CoA reductase activity. These combined results indicate that the LDL receptor abnormality present in familial hypercholesterolemia fibroblasts reflects deranged hepatocyte LDL recognition and metabolism. In addition, a low-affinity, nonsaturable uptake process for LDL is present in human liver which does not efficiently modulate hepatocyte cholesterol content or synthesis.


Position of dorsal root ganglia in the lumbosacral region in patients with radiculopathy.
When applying pulsed radiofrequency on dorsal root ganglia for treating chronic lower back pain, maximum efficiency can be expected when a needle is placed 1-2 cm peripheral to the dorsal root ganglion. The object of this study is to analyze images taken after adding contrast to transforaminal epidural injection, categorize root ganglia according to anatomical position, and provide a reference for efficient needle positioning in applying pulsed radiofrequency on dorsal root ganglia. From January 2008 to January 2009, 457 patients who visited our hospital for root pain or radiculopathy were treated with transforaminal epidural injection on the nerve roots based on the dermatome of the painful area. Anteroposterior views were taken after injection of contrast. A virtual line was made by connecting the internal and external parts of the spinal pedicle from the contrast images. Then the dorsal root ganglia were categorized as intraspinal (IS), intraforaminal (IF), or extraforaminal (EF). In the fourth lumbar spine, dorsal root ganglia positions were 48% IF, 41% IS, and 6% EF. In the fifth lumbar spine, dorsal root ganglia positions were 75% IF, 10% IS, and 6% EF. In the first sacral spine, dorsal root ganglia locations were 8% IF and 83% IS. Positional categorization of dorsal root ganglia according to contrast images was proven to be good anatomical references for effective radiofrequency or blocking of dorsal root ganglia.


Substance P-induced long-term blockade of spinal adrenergic analgesia: reversal by morphine and naloxone.
Alpha agonists [noradrenaline (NA) and ST-91] inhibit the release of substance P (SP) from the spinal cord and block the biting, licking, scratching syndrome produced by intrathecal SP suggesting that these agents produce analgesia by an interaction with SP systems. In this study we determined the effect of a desensitizing regimen of SP (15 micrograms X 2 at a 30-min interval) on analgesia produced by intrathecal NA in the rat tail-flick test. When NA was injected immediately after the regimen or after a 90-minute delay, NA analgesia was blocked. This blockade persisted up to 11 days after exposure to SP. Exposure to a single dose of SP (15 or 30 micrograms) also blocked NA acutely, but the long-term blockade did not last as long. An identical effect was observed with ST-91. SP (15 micrograms X 2) potentiated the analgesic action of morphine acutely, but no interaction was observed 4 to 7 days later. Pretreatment with morphine and naloxone prevented the long-term blockade by SP. The effect of naloxone was not reversed by naltrexone suggesting that occupation of opiate receptors rather than an apparent agonist effect of naloxone caused the protection. Pretreatment with clonidine had only a slight effect on long-term blockade, but yohimbine was without effect. The present study describes a new long-term interaction between SP and alpha-2 agonists in the spinal cord. The mechanism(s) of the observed blockade by SP remains to be elucidated. However, there appears to be a functionally significant interaction between opiate and alpha-2 receptors in the spinal cord.


MANAGEMENT OF RETINAL PIGMENT EPITHELIUM TEAR DURING ANTI-VASCULAR ENDOTHELIAL GROWTH FACTOR THERAPY.
This article aims to review current evidence on the development, diagnosis, and management of retinal pigment epithelium (RPE) tear during anti-vascular endothelial growth factor (VEGF) therapy. Literature searches were performed using MEDLINE/PubMed databases (cut-off date: August 2019). Three key recommendations were made based on existing literature and clinical experience: 1) Multimodal imaging with color fundus photography, optical coherence tomography, near-infrared reflectance imaging, fundus autofluorescence imaging, optical coherence tomography-angiography, and/or fluorescein angiography are recommended to diagnose RPE tear and assess risk factors. Retinal pigment epithelium tears can be graded by size and foveal involvement. 2) Patients at high risk of developing RPE tear should be monitored after each anti-VEGF injection. If risk factors worsen, it is not yet definitively known whether anti-VEGF administration should be more frequent, or alternatively stopped in such patients. Prospective research into high-risk characteristics is needed. 3) After RPE tear develops, anti-VEGF treatment should be continued in patients with active disease (as indicated by presence of intraretinal or subretinal fluid), although cessation of therapy should be considered in eyes with multilobular tears. Although evidence to support the assumption that anti-VEGF treatment contributes to development of RPE tear is not definitive, some data suggest this link.


Sandwich hologram interferometry: a new dimension in holographic comparison.
New methods are introduced that utilize interferometric comparison of images of diffusely reflecting objects from different hologram plates that are sandwiched together. To make the plate positions optically and mechanically identical during reconstruction and exposure, they are also exposed in sandwiches. If the two sandwiched hologram plates are separated by a small distance a new method of fringe evaluation can be used. Fringes caused by object tilt between two exposures can be eliminated by an analogous, but much larger, tilt of the sandwich hologram during reconstruction. Even the direction of tilt, forward or backward, is found this way.


Phlorofucofuroeckol A from Edible Brown Alga Ecklonia Cava Enhances Osteoblastogenesis in Bone Marrow-Derived Human Mesenchymal Stem Cells.
The deterioration of bone formation is a leading cause of age-related bone disorders. Lack of bone formation is induced by decreased osteoblastogenesis. In this study, osteoblastogenesis promoting effects of algal phlorotannin, phlorofucofuroeckol A (PFF-A), were evaluated. PFF-A was isolated from brown alga Ecklonia cava. The ability of PFF-A to enhance osteoblast differentiation was observed in murine pre-osteoblast cell line MC3T3-E1 and human bone marrow-derived mesenchymal stem cells (huBM-MSCs). Proliferation and alkaline phosphatase (ALP) activity of osteoblasts during differentiation was assayed following PFF-A treatment along extracellular mineralization. In addition, effect of PFF-A on osteoblast maturation pathways such as Runx2 and Smads was analyzed. Treatment of PFF-A was able to enhance the proliferation of differentiating osteoblasts. Also, ALP activity was observed to be increased. Osteoblasts showed increased extracellular mineralization, observed by Alizarin Red staining, following PFF-A treatment. In addition, expression levels of critical proteins in osteoblastogenesis such as ALP, bone morphogenetic protein-2 (BMP-2), osteocalcin and β-catenin were stimulated after the introduction of PFF-A. In conclusion, PFF-A was suggested to be a potential natural product with osteoblastogenesis enhancing effects which can be utilized against bone-remodeling imbalances and osteoporosis-related complications.


Public and private domains of religiosity and adolescent smoking transitions.
We used data from a nationally representative sample of US adolescents in school grades 7 through 12 to explore the effects of public and private religiosity on initiation, escalation, and cessation of smoking. We found that adolescents' decisions to experiment with smoking are influenced by both their individual practice of their faith and by participation in a larger faith community. However, the effects of private and public religiosity are specific to different decision points on the smoking uptake process. Private religiosity was protective against initiation of regular smoking among nonsmokers. It also was protective against initiation of experimental smoking but only when the young person frequently attended religious services or a religious youth group. Although private religiosity appeared to discourage the uptake of smoking, it was unrelated to reduction or cessation once a young person has become addicted to cigarettes. In contrast, public religiosity did predict reduction and cessation of cigarette use among regular smokers. Taken together, these findings demonstrate that the domains in which religiosity are important extend beyond the individual and include religious institutions.


Single cell RNA sequencing to dissect the molecular heterogeneity in lupus nephritis.
Lupus nephritis is a leading cause of mortality among systemic lupus erythematosus (SLE) patients, and its heterogeneous nature poses a significant challenge to the development of effective diagnostics and treatments. Single cell RNA sequencing (scRNA-seq) offers a potential solution to dissect the heterogeneity of the disease and enables the study of similar cell types distant from the site of renal injury to identify novel biomarkers. We applied scRNA-seq to human renal and skin biopsy tissues and demonstrated that scRNA-seq can be performed on samples obtained during routine care. Chronicity index, IgG deposition, and quantity of proteinuria correlated with a transcriptomic-based score composed of IFN-inducible genes in renal tubular cells. Furthermore, analysis of cumulative expression profiles of single cell keratinocytes dissociated from nonlesional, non-sun-exposed skin of patients with lupus nephritis also revealed upregulation of IFN-inducible genes compared with keratinocytes isolated from healthy controls. This indicates the possible use of scRNA-seq analysis of skin biopsies as a biomarker of renal disease. These data support the potential utility of scRNA-seq to provide new insights into the pathogenesis of lupus nephritis and pave the way for exploiting a readily accessible tissue to reflect injury in the kidney.


Interactive effects of chemical and biological controls on food-web composition in saline prairie lakes.
Salinity is restricting habitatability for many biota in prairie lakes due to limited physiological abilities to cope with increasing osmotic stress. Yet, it remains unclear how salinity effects vary among major taxonomic groups and what role other environmental parameters play in shaping food-web composition. To answer these questions, we sampled fish, zooplankton and littoral macroinvertebrates in 20 prairie lakes (Saskatchewan, Canada) characterized by large gradients in water chemistry and lake morphometry. We showed that salinity thresholds differed among major taxonomic groups, as most fishes were absent above salinities of 2 g L-1, while littoral macroinvertebrates were ubiquitous. Zooplankton occurred over the whole salinity range, but changed taxonomic composition as salinity increased. Subsequently, the complexity of fish community (diversity) was associated with large changes in invertebrate communities. The directional changes in invertebrate communities to smaller taxa indicated that complex fish assemblages resulted in higher predation pressure. Most likely, as the complexity of fish community decreased, controls of invertebrate assemblages shifted from predation to competition and ultimately to productivity in hypersaline lakes. Surprisingly, invertebrate predators did not thrive in the absence of fishes in these systems. Furthermore, the here identified salinity threshold for fishes was too low to be a result of osmotic stress. Hence, winterkill was likely an important factor eliminating fishes in low salinity lakes that had high productivity and shallow water depth. Ultimately, while salinity was crucial, intricate combinations of chemical and biological mechanisms also played a major role in controlling the assemblages of major taxonomic groups in prairie lakes.


Increased circulating IgG levels, myocardial immune cells and IgG deposits support a role for an immune response in pre- and end-stage heart failure.
The chronic inflammatory response plays an important role in adverse cardiac remodelling and the development of heart failure (HF). There is also evidence that in the pathogenesis of several cardiovascular diseases, chronic inflammation is accompanied by antibody and complement deposits in the heart, suggestive of a true autoimmune response. However, the role of antibody-mediated immune responses in HF progression is less clear. We assessed whether immune cell infiltration and immunoglobulin levels are associated with HF type and disease stage, taking sex differences into account. We found IgG deposits and increased infiltration of immune cells in the affected myocardium of patients with end-stage HF with reduced ejection fraction (HFrEF, n = 20). Circulating levels of IgG1 and IgG3 were elevated in these patients. Furthermore, the percentage of transitional/regulatory B cells was decreased (from 6.9% to 2.4%) compared with healthy controls (n = 5). Similarly, increased levels of circulating IgG1 and IgG3 were observed in men with left ventricular diastolic dysfunction (LVDD, n = 5), possibly an early stage of HF with preserved EF (HFpEF). In conclusion, IgG deposits and infiltrates of immune cells are present in end-stage HFrEF. In addition, both LVDD patients and end-stage HFrEF patients show elevated levels of circulating IgG1 and IgG3, suggesting an antibody-mediated immune response upon cardiac remodelling, which in the early phase of remodelling appear to differ between men and women. These immunoglobulin subclasses might be used as marker for pre-stage HF and its progression. Future identification of auto-antigens might open possibilities for new therapeutic interventions.


Computational design of receptor and sensor proteins with novel functions.
The formation of complexes between proteins and ligands is fundamental to biological processes at the molecular level. Manipulation of molecular recognition between ligands and proteins is therefore important for basic biological studies and has many biotechnological applications, including the construction of enzymes, biosensors, genetic circuits, signal transduction pathways and chiral separations. The systematic manipulation of binding sites remains a major challenge. Computational design offers enormous generality for engineering protein structure and function. Here we present a structure-based computational method that can drastically redesign protein ligand-binding specificities. This method was used to construct soluble receptors that bind trinitrotoluene, l-lactate or serotonin with high selectivity and affinity. These engineered receptors can function as biosensors for their new ligands; we also incorporated them into synthetic bacterial signal transduction pathways, regulating gene expression in response to extracellular trinitrotoluene or l-lactate. The use of various ligands and proteins shows that a high degree of control over biomolecular recognition has been established computationally. The biological and biosensing activities of the designed receptors illustrate potential applications of computational design.


Working conditions that contribute to absenteeism among nurses in a provincial hospital in the Limpopo Province.
Absenteeism results in an increased workload for nurses who stand in for colleagues and can lead to situations in which a lack of motivation among nurses and a lowering of the quality of patient care may occur. The researcher observed that certain conditions, such as inflexible working schedules, were given as reasons for the absenteeism in units in a provincial hospital. A non-experimental, descriptive, quantitative study was undertaken. The purpose of the article was to describe the working conditions that contribute to absenteeism among the professional and sub-professional nurses at a provincial hospital in the Limpopo province of South Africa. The sample included 107 professional nurses and 163 sub-professional nurses who voluntarily agreed to participate in the study. A questionnaire was used to collect data, which was analysed by using descriptive and inferential statistics. The findings of this study indicated that personal and managerial characteristics, and organisational and working conditions may lead to absenteeism in the workplace. This article focuses on the working conditions that are constraints, namely inadequate group cohesion, inadequate delegation of autonomy, role ambiguity, ineffective routinisation and the effect of the workload in the workplace. Recommendations are made for improving working conditions to combat absenteeism among nurses. The limitations of this study are highlighted.


Sequential gene targeting to make chimeric tumor models with de novo chromosomal abnormalities.
The discovery of chromosomal translocations in leukemia/lymphoma and sarcomas presaged a widespread discovery in epithelial tumors. With the advent of new-generation whole-genome sequencing, many consistent chromosomal abnormalities have been described together with putative driver and passenger mutations. The multiple genetic changes required in mouse models to assess the interrelationship of abnormalities and other mutations are severe limitations. Here, we show that sequential gene targeting of embryonic stem cells can be used to yield progenitor cells to generate chimeric offspring carrying all the genetic changes needed for cell-specific cancer. Illustrating the technology, we show that MLL-ENL fusion is sufficient for lethal leukocytosis and proof of genome integrity comes from germline transmission of the sequentially targeted alleles. This accelerated technology leads to a reduction in mouse numbers (contributing significantly to the 3Rs), allows fluorescence tagging of cancer-initiating cells, and provides a flexible platform for interrogating the interaction of chromosomal abnormalities with mutations.


Mergers and the new workplace: the effects of a merger of two emergency departments on nursing staff.
Even small mergers have big consequences. This case study demonstrates the complex nature of organizational change found even in small organizational subunits and provides important insight for managers and leaders who have completed or are considering such mergers. The study examined the amalgamation of two emergency departments. Quantitative and qualitative techniques were used to analyze the planning, implementation, and outcomes of this amalgamation on the nursing staff. This case study draws upon the literature pertaining to change survivor's syndrome, prescribed approaches to change, and the difficulties in changing cultures and making complex decisions. Change in hospitals has been shown to have negative consequences for nurses. The goal of this study was to examine in greater detail what aspects of change are associated with these effects. The findings suggest that the planning of the amalgamation was consistent with rational decision-making. The goals of changing to a single emergency unit and cost savings were achieved. However, the nurses experienced stress due to the pace of work and the physical layout of the newly renovated department. The findings also suggest that personal-level problems that the nurses experienced were consistent with the consequences of survivor's syndrome, the psychological contract, and communication issues. It was recommended that the organization undertake initiative to alleviate these stresses.


The stereochemical configuration of flavanols influences the level and metabolism of flavanols in humans and their biological activity in vivo.
Extensive epidemiological and clinical evidence associates diets high in flavanol-containing foods with cardiovascular health benefits in humans. Catechin and epicatechin, the most common flavanols in foods, are present in the diet in different enantiomeric forms. This study investigated the influence of the stereochemical configuration of flavanols on their absorption, metabolism, and biological activity. Healthy adult males were asked to consume equal amounts of the stereochemically pure flavanols (-)-epicatechin, (-)-catechin, (+)-catechin, and (+)-epicatechin (1.5mg/kg bw) in a well-defined cocoa-based, dairy-containing drink matrix, and flavanol levels were subsequently determined in plasma and 24-h urine. The results obtained show that the stereochemical configuration of flavanols has a profound influence on their uptake and metabolism in humans. In addition, we assessed the vasodilatory activity of each flavanol stereoisomer in vivo and found (-)-epicatechin to be the single stereoisomer capable of mediating a significant arterial dilation response. Importantly, this effect was independent of the classic antioxidant properties of flavanols. Overall, these results indicate that the proposed beneficial health effects associated with the consumption of flavanol-containing foods will significantly depend on the stereochemical configuration of the flavanols ingested.


Update of potency factors for asbestos-related lung cancer and mesothelioma.
The most recent update of the U.S. Environmental Protection Agency (EPA) health assessment document for asbestos (Nicholson, 1986, referred to as "the EPA 1986 update") is now 20 years old. That document contains estimates of "potency factors" for asbestos in causing lung cancer (K(L)'s) and mesothelioma (K(M)'s) derived by fitting mathematical models to data from studies of occupational cohorts. The present paper provides a parallel analysis that incorporates data from studies published since the EPA 1986 update. The EPA lung cancer model assumes that the relative risk varies linearly with cumulative exposure lagged 10 years. This implies that the relative risk remains constant after 10 years from last exposure. The EPA mesothelioma model predicts that the mortality rate from mesothelioma increases linearly with the intensity of exposure and, for a given intensity, increases indefinitely after exposure ceases, approximately as the square of time since first exposure lagged 10 years. These assumptions were evaluated using raw data from cohorts where exposures were principally to chrysotile (South Carolina textile workers, Hein et al., 2007; mesothelioma only data from Quebec miners and millers, Liddell et al., 1997) and crocidolite (Wittenoom Gorge, Australia miners and millers, Berry et al., 2004) and using published data from a cohort exposed to amosite (Paterson, NJ, insulation manufacturers, Seidman et al., 1986). Although the linear EPA model generally provided a good description of exposure response for lung cancer, in some cases it did so only by estimating a large background risk relative to the comparison population. Some of these relative risks seem too large to be due to differences in smoking rates and are probably due at least in part to errors in exposure estimates. There was some equivocal evidence that the relative risk decreased with increasing time since last exposure in the Wittenoom cohort, but none either in the South Carolina cohort up to 50 years from last exposure or in the New Jersey cohort up to 35 years from last exposure. The mesothelioma model provided good descriptions of the observed patterns of mortality after exposure ends, with no evidence that risk increases with long times since last exposure at rates that vary from that predicted by the model (i.e., with the square of time). In particular, the model adequately described the mortality rate in Quebec chrysotile miners and millers up through >50 years from last exposure. There was statistically significant evidence in both the Wittenoom and Quebec cohorts that the exposure intensity-response is supralinear(1) rather than linear. The best-fitting models predicted that the mortality rate varies as [intensity](0.47) for Wittenoom and as [intensity](0.19) for Quebec and, in both cases, the exponent was significantly less than 1 (p< .0001). Using the EPA models, K(L)'s and K(M)'s were estimated from the three sets of raw data and also from published data covering a broader range of environments than those originally addressed in the EPA 1986 update. Uncertainty in these estimates was quantified using "uncertainty bounds" that reflect both statistical and nonstatistical uncertainties. Lung cancer potency factors (K(L)'s) were developed from 20 studies from 18 locations, compared to 13 locations covered in the EPA 1986 update. Mesothelioma potency factors (K(M)'s) were developed for 12 locations compared to four locations in the EPA 1986 update. Although the 4 locations used to calculate K(M) in the EPA 1986 update include one location with exposures to amosite and three with exposures to mixed fiber types, the 14 K(M)'s derived in the present analysis also include 6 locations in which exposures were predominantly to chrysotile and 1 where exposures were only to crocidolite. The K(M)'s showed evidence of a trend, with lowest K(M)'s obtained from cohorts exposed predominantly to chrysotile and highest K(M)'s from cohorts exposed only to amphibole asbestos, with K(M)'s from cohorts exposed to mixed fiber types being intermediate between the K(M)'s obtained from chrysotile and amphibole environments. Despite the considerable uncertainty in the K(M) estimates, the K(M) from the Quebec mines and mills was clearly smaller than those from several cohorts exposed to amphibole asbestos or a mixture of amphibole asbestos and chrysotile. For lung cancer, although there is some evidence of larger K(L)'s from amphibole asbestos exposure, there is a good deal of dispersion in the data, and one of the largest K(L)'s is from the South Carolina textile mill where exposures were almost exclusively to chrysotile. This K(L) is clearly inconsistent with the K(L) obtained from the cohort of Quebec chrysotile miners and millers. The K(L)'s and K(M)'s derived herein are defined in terms of concentrations of airborne fibers measured by phase-contrast microscopy (PCM), which only counts all structures longer than 5 microm, thicker than about 0.25 microm, and with an aspect ratio > or =3:1. Moreover, PCM does not distinguish between asbestos and nonasbestos particles. One possible reason for the discrepancies between the K(L)'s and K(M)'s from different studies is that the category of structures included in PCM counts does not correspond closely to biological activity. In the accompanying article (Berman and Crump, 2008) the K(L)'s and K(M)'s and related uncertainty bounds obtained in this article are paired with fiber size distributions from the literature obtained using transmission electron microscopy (TEM). The resulting database is used to define K(L)'s and K(M)'s that depend on both the size (e.g., length and width) and mineralogical type (e.g., chrysotile or crocidolite) of an asbestos structure. An analysis is conducted to determine how well different K(L) and K(M) definitions are able to reconcile the discrepancies observed herein among values obtained from different environments.


Evidence for peptide transport across microsomal membranes.
Antigenic peptides bound to class I molecules of the major histocompatibility complex (MHC) are recognized by T-cell receptors during development of an antiviral immune response. T cells respond to peptides derived from cytoplasmic viral proteins as well as viral membrane proteins, indicating that a pathway exists for the transport of proteins or peptides from the cytosol into the compartment(s) where the MHC class I molecules assemble. To investigate this pathway, we have developed an in vitro assay for the transport of peptides into microsomal vesicles. This assay provides evidence for the transport of chemically synthesized peptides (13-21 amino acids) containing N-linked glycosylation acceptor sequences, which serve as glycosylation substrates. Their transport results in depletion of the pool of available dolichol high-mannose oligosaccharides in the lumen of the microsomal vesicles. We have observed transport of peptides derived from antigenic human immunodeficiency virus gag and influenza B nucleoprotein sequences, but transport of a third randomly selected peptide was not detected, suggesting specificity of the transport process. We were not able to demonstrate ATP dependence of this peptide transport process by using apyrase and an ATPase inhibitor. This result was unexpected in light of the recent identification of MHC-linked genes with homology to ATP-binding cassette transporters, which have been proposed to mediate peptide transport.


Use of the roter interaction analysis system to analyze veterinarian-client-patient communication in companion animal practice.
To identify specific components of veterinarian-client-patient communication during clinical appointments in companion animal practice. Cross-sectional descriptive study. A random sample of 50 companion animal practitioners in southern Ontario and a convenience sample of 300 clients and their pets. For each practitioner, 6 clinical appointments (3 wellness appointments and 3 appointments related to a health problem) were videotaped, and the Roter interaction analysis system (RIAS) was used to analyze the resulting 300 videotapes. Statements made during each appointment were classified by means of a communication framework reflecting the 4 essential tasks of the appointment (ie, data gathering, education and counseling, relationship building, and activation and partnership). 57% of the veterinarians contacted (50/87) and 99% of the clients contacted agreed to participate in the study. Mean duration of the appointments was 13 minutes. Typically, veterinarians contributed 62% of the total conversation and clients contributed 38%. Fifty-four percent of the veterinarian interaction was with the client, and 8% was with the pet. Data gathering constituted 9% of the veterinarian-to-client communication and was primarily accomplished through closed-ended questioning; 48% of veterinarian-to-client communication involved client education and counseling, 30% involved relationship building, and 7% involved activation and partnership (the remaining 6% constituted orientation). Results suggest that the RIAS was a reliable method of assessing the structure, process, and content of veterinarian-client-patient communication and that some veterinarians do not use all the tools needed for effective communication.


The Meiotic Recombination Activator PRDM9 Trimethylates Both H3K36 and H3K4 at Recombination Hotspots In Vivo.
In many mammals, including humans and mice, the zinc finger histone methyltransferase PRDM9 performs the first step in meiotic recombination by specifying the locations of hotspots, the sites of genetic recombination. PRDM9 binds to DNA at hotspots through its zinc finger domain and activates recombination by trimethylating histone H3K4 on adjacent nucleosomes through its PR/SET domain. Recently, the isolated PR/SET domain of PRDM9 was shown capable of also trimethylating H3K36 in vitro, raising the question of whether this reaction occurs in vivo during meiosis, and if so, what its function might be. Here, we show that full-length PRDM9 does trimethylate H3K36 in vivo in mouse spermatocytes. Levels of H3K4me3 and H3K36me3 are highly correlated at hotspots, but mutually exclusive elsewhere. In vitro, we find that although PRDM9 trimethylates H3K36 much more slowly than it does H3K4, PRDM9 is capable of placing both marks on the same histone molecules. In accord with these results, we also show that PRDM9 can trimethylate both K4 and K36 on the same nucleosomes in vivo, but the ratio of K4me3/K36me3 is much higher for the pair of nucleosomes adjacent to the PRDM9 binding site compared to the next pair further away. Importantly, H3K4me3/H3K36me3-double-positive nucleosomes occur only in regions of recombination: hotspots and the pseudoautosomal (PAR) region of the sex chromosomes. These double-positive nucleosomes are dramatically reduced when PRDM9 is absent, showing that this signature is PRDM9-dependent at hotspots; the residual double-positive nucleosomes most likely come from the PRDM9-independent PAR. These results, together with the fact that PRDM9 is the only known mammalian histone methyltransferase with both H3K4 and H3K36 trimethylation activity, suggest that trimethylation of H3K36 plays an important role in the recombination process. Given the known requirement of H3K36me3 for double strand break repair by homologous recombination in somatic cells, we suggest that it may play the same role in meiosis.


Intranasal immunization with a vaccinia virus vaccine vector expressing pre-fusion stabilized SARS-CoV-2 spike fully protected mice against lethal challenge with the heavily mutated mouse-adapted SARS2-N501Y MA30 strain of SARS-CoV-2.
The Omicron SARS-CoV-2 variant has been designated a variant of concern because its spike protein is heavily mutated. In particular, Omicron spike is mutated at 5 positions (K417, N440, E484, Q493 and N501) that have been associated with escape from neutralizing antibodies induced by either infection with or immunization against the early Washington strain of SARS-CoV-2. The mouse-adapted strain of SARS-CoV-2, SARS2-N501Y MA30 , contains a spike that is also heavily mutated, with mutations at 4 of the 5 positions in Omicron spike associated with neutralizing antibody escape (K417, E484, Q493 and N501). In this manuscript we show that intranasal immunization with a pre-fusion stabilized Washington strain spike, expressed from a highly attenuated, replication-competent vaccinia virus construct, NYVAC-KC, fully protected mice against disease and death from SARS2-N501Y MA30 . Similarly, immunization by scarification on the skin fully protected against death, but not from mild disease. This data demonstrates that Washington strain spike, when expressed from a highly attenuated, replication-competent poxvirus, administered without parenteral injection can fully protect against the heavily mutated mouse-adapted SARS2-N501Y MA30 .


Missing the Mark: The Complexity of African American Dementia Family Caregiving.
Alzheimer's disease and related dementias (ADRDs) have a significant impact on families. Family nurses are in an ideal position to address the needs of families affected by ADRD. However, to be most effective, family nurses and researchers need culturally appropriate theories to guide practice and research. On November 17, 2018, five nurse researchers presented findings of their research with African American families at the Gerontological Society of America's annual meeting. The results reported and the lively discussion that ensued suggested that the current paradigms framing research and practice with African American families affected by ADRD may not be adequate. There is a need to consider culturally congruent, family-centered theories to guide research and practice with this population of families.


Ultrasound-guided perineural injection with platelet-rich plasma improved the neurophysiological parameters of carpal tunnel syndrome: A case report.
Carpal tunnel syndrome (CTS) is a common and sometimes challenging condition, which causes pain; paresthesia; tingling of the thumb, index and long fingers; and even thenar weakness in the hands. The effectiveness of corticosteroid injections seems to be transient, and there is a lack of information around long-term effects in previous studies. In recent years, platelet-rich plasma (PRP) has proven to bean alternative as it encourages tissue regeneration. We hereby describe a patient with CTS showing significant improvements in electrophysiological parameters after receiving PRP injections. The results revealed significant improvements in the distal motor and sensory latencies as well as the sensory nerve action potential and compound muscle action potential amplitudes of the both median nerves. In summary, if patients are refractory to conservative treatments-such as splinting, oral medication, and corticosteroid injection-PRP may be used as an alternative before surgical intervention.


Pterostilbene prevents AKT-ERK axis-mediated polymerization of surface fibronectin on suspended lung cancer cells independently of apoptosis and suppresses metastasis.
Polymeric fibronectin (polyFN) assembled on suspended breast cancer cells is required for metastasis. Conceivably, drugs that target such polyFN may fight against cancer metastasis. While stilbene analogs trigger pro-apoptotic effect on attached cancer cells, whether they prevent polyFN assembly and metastasis of suspended cancer cells via an apoptosis-independent manner remains unexplored. We depleted suspended Lewis lung carcinoma (LLC) cells of polyFN by silencing the endogenous FN expression or pterostilbene (PS) to examine whether metastasis of lung cancer cells could thus be suppressed. We investigated whether PS regulates AKT-ERK signaling axis to suppress polyFN assembly in suspended LLC cells independently of apoptosis. We tested the therapeutic effects of orally administered PS against cancer metastasis. Both FN-silencing and PS among the three stilbenoids indeed significantly reduced polyFN assembly and lung metastasis of suspended LLC cells in an apoptosis-independent manner. Mechanistically, PS-induced AKT phosphorylation (pAKT) and suppressed ERK phosphorylation (pERK) in suspended LLC cells, whereas pretreatment with a PI3K inhibitor, LY294002, effectively reduced pAKT, rescued pERK, and consequently reversed the PS-suppressed polyFN assembly on LLC cells; these pretreatment effects were then overturned by the ERK inhibitor U0126. Indeed, PS-suppressed lung metastasis was counteracted by LY294002, which was further overruled with U0126. Finally, we found that PS, when orally administered in experimental metastasis assays, both significantly prevented lung colonization and metastasis of LLC cells and reduced the already established tumor growth in the mouse lungs. PS suppressed AKT/ERK-regulated polyFN assembly on suspended LLC cells and pulmonary metastasis. PS possesses potency in both preventing and treating lung metastasis of lung cancer cells in apoptosis-independent and apoptosis-dependent manners, respectively.


Comparative analysis of imaging configurations and objectives for Fourier microscopy.
Fourier microscopy is becoming an increasingly important tool for the analysis of optical nanostructures and quantum emitters. However, achieving quantitative Fourier space measurements requires a thorough understanding of the impact of aberrations introduced by optical microscopes that have been optimized for conventional real-space imaging. Here we present a detailed framework for analyzing the performance of microscope objectives for several common Fourier imaging configurations. To this end, we model objectives from Nikon, Olympus, and Zeiss using parameters that were inferred from patent literature and confirmed, where possible, by physical disassembly. We then examine the aberrations most relevant to Fourier microscopy, including the alignment tolerances of apodization factors for different objective classes, the effect of magnification on the modulation transfer function, and vignetting-induced reductions of the effective numerical aperture for wide-field measurements. Based on this analysis, we identify an optimal objective class and imaging configuration for Fourier microscopy. In addition, the Zemax files for the objectives and setups used in this analysis have been made publicly available as a resource for future studies.


What many of us are doing or should be doing in clinical pathology: a list of the activities of the pathologist in the clinical laboratory.
Mr. Paul Mango, Chief Operating Officer of a hospital-based clinical laboratory network in Pittsburgh, recently performed a survey of patients presenting for phlebotomy. The survey included the question, "What does a pathologist do?" The results were that 50% of the patients had no idea what a pathologist did, and 30% of the patients stated that pathologists examined dead bodies. It is not surprising that there is a limited understanding by patients of the activities of pathologists because patients do not usually see pathologists. However, beyond autopsy and surgical pathology, the activities of pathologists are also not well known to nonpathologist physicians and hospital administrators. A poor understanding of activities in clinical pathology have placed these clinical responsibilities of the pathologist under particular scrutiny for cost reduction. The quantitation of output from anatomic pathology, in number of slides reviewed or number of autopsies performed, is objective and easily understood. As noted in the list of clinical pathology activities that follows, the responsibilities within the clinical laboratory are highly diverse and, if the pathologist handles them successfully, highly contributory to patient care. Thus, it is timely that a compilation of activities in clinical pathology be issued for review by the pathologist community. I would hope that this list will serve as a starting point for a universally accepted group of activities that describes clinical pathology today and that it will be useful for pathologists to make their significant contributions in the clinical laboratory apparent to administrators, fellow physicians, and patients. The clinical laboratory responsibilities should also be valuable to directors of residency training programs to focus training in clinical pathology toward the development of currently desirable expertise.


Cytokine stimulation of T lymphocytes regulates their capacity to induce monocyte production of tumor necrosis factor-alpha, but not interleukin-10: possible relevance to pathophysiology of rheumatoid arthritis.
Previous studies in the laboratory have shown that the pro-inflammatory cytokine tumor necrosis factor (TNF)-alpha plays a pivotal role in the pathogenesis of rheumatoid arthritis (RA). The mechanisms involved in regulating monocyte/macrophage cytokine production are not yet fully understood, but are thought to involve both soluble factors and cell/cell contact with other cell types. We and others have previously demonstrated that T cells activated through the T cell receptor/CD3 complex induce monocyte TNF-alpha production by contact-mediated signals. In this report, we investigated further whether T cells activated by cytokines in the absence of T cell receptor stimulation also regulate monocyte cytokine production. T cells were activated in an antigen-independent manner using the cytokines interleukin (IL)-15 or IL-2 alone, or in combination with IL-6 and TNF-alpha. Subsequently, T cells were fixed and incubated with monocytes. Fixed, cytokine-stimulated T cells induced monocytes to secrete TNF-alpha in a dose-dependent manner, but did not induce secretion of IL-10, a potent endogenous down-regulator of TNF-alpha and other pro-inflammatory cytokines. Stimulation of monocyte TNF-alpha was markedly inhibited when T cells were physically separated from monocytes within the tissue culture well, confirming that T cell contact is necessary. T cell acquisition of monocyte-activating capacity was shown to be dependent on the period of cytokine stimulation, with T cells activated for 8 days more effective than T cells activated for shorter periods. Addition of interferon-gamma or granulocyte/macrophage colony-stimulating factor to the T cell/monocyte cultures enhanced T cell induction of monocyte TNF-alpha by threefold and ninefold, respectively. The results from this model of cognate interaction suggest that cytokine-stimulated T cells, interacting with macrophages in the rheumatoid synovial membrane, may contribute to the continuous excessive production of TNF-alpha observed in the RA joint, and to the imbalance of pro-inflammatory cytokines over anti-inflammatory cytokines.


FAMM Flap in Reconstructing Postsurgical Nasopharyngeal Airway Stenosis.
Introduction. Postsurgical nasopharyngeal airway stenosis can be a challenge to manage. The stenosis could be as a result of any surgical procedure in the nasopharyngeal region that heals extensive scarring and fibrosis. Objective. To evaluate patients with nasopharyngeal stenosis managed with FAMM flap. Study Design. Prospective study of patients with nasopharyngeal stenosis at the Kenyatta National Hospital between 2010 and 2013 managed with FAMM flap. Materials and Methods. Patients with severe nasopharyngeal airway stenosis were reviewed and managed with FAMM flaps at the Kenyatta National Hospital. Postoperatively they were assessed for symptomatic improvement in respiratory distress, patency of the nasopharyngeal airway, and donor site morbidity. Results. A total of 8 patients were managed by the authors in a duration of 4 years with nasopharyngeal stenosis. Five patients were managed with unilateral FAMM flaps in a two-staged surgical procedure. Four patients had complete relieve of the airway obstruction with a patent airway created. One patient had a patent airway created though with only mild improvement in airway obstruction. Conclusion. FAMM flap provides an alternative in the management of postsurgical severe nasopharyngeal stenosis. It is a reliable flap that is easy to raise and could provide adequate epithelium for the stenosed pharynx.


Hypertension management in patients with chronic kidney disease.
Hypertension is one of the major risk factors for the development and progression of chronic kidney disease. The loss of renal function leads to impaired renal autoregulation and renders the kidney vulnerable to the damaging effects of uncontrolled hypertension. Mounting evidence indicates that angiotensin-converting enzyme inhibitors and angiotensin receptor blockers slow the progression of chronic kidney disease through effects beyond lowering blood pressure. Studies are needed to determine whether high doses of the single agent or combination therapy is most effective in providing renal protection. Urinary protein excretion is a useful tool for monitoring and titrating therapy to maximize renal protection. Changes in the serum creatinine concentration and hyperkalemia are complications of antihypertensive therapy in patients with chronic kidney disease that can be successfully managed to allow continued use of renin-angiotensin blockade.


Rifaximin for the treatment of hepatic encephalopathy.
Previous studies suggest that rifaximin is efficacious in the treatment of hepatic encephalopathy. To evaluate the efficacy and safety of rifaximin in addition to lactulose in improving hospitalization outcomes in patients with hepatic encephalopathy. Hospital records of patients evaluated for liver transplantation at a single center between January 2006 and May 2008 were reviewed. Hospitalizations for hepatic encephalopathy and other conditions and the incidence of spontaneous bacterial peritonitis and adverse events were analyzed. Charts of 65 patients who were treated with rifaximin and lactulose were analyzed. Patients received lactulose (20-120 g/d; lactulose phase) before treatment with rifaximin (400- 1200 mg/d; rifaximin phase). During the rifaximin phase, the risk, number, and duration of hospitalizations for hepatic encephalopathy were reduced compared with the lactulose phase. Treatment, age, and Model for End-Stage Liver Disease score were independent predictors of hospitalizations for hepatic encephalopathy (P < .05). The rifaximin phase had fewer hospitalizations than the lactulose phase (36 vs 47, respectively) and a smaller percentage of patients with repeated hospitalizations than the lactulose phase (5% vs 14%, respectively; P = .006) for conditions other than hepatic encephalopathy. A smaller percentage of patients had evidence of spontaneous bacterial peritonitis during the rifaximin phase than the lactulose phase (2% vs 12%, respectively; P = .02). Rifaximin was better tolerated than lactulose. Addition of rifaximin to lactulose therapy significantly reduced the risk and duration of hospitalizations for hepatic encephalopathy.


Fatigue and work safety behavior in men during early fatherhood.
This study investigated the relationship between fatigue and work safety behavior of fathers with new babies. A total of 241 fathers completed a questionnaire at 6 and 12 weeks postpartum with items on fatigue and safety behavior at work. Results revealed that fathers worked long hours, reported a moderate-to-high physical intensity of work, and experienced interrupted sleep averaging less than 6 hours. Fathers also reported moderate fatigue at both 6 and 12 weeks postbirth, which was inversely related to safety behavior. Both fatigue and sleep history made a small but statistically significant contribution to safety behavior results at 6 and 12 weeks postbirth. Findings suggest that working fathers with babies experience fatigue during early fatherhood and are unable to recover due to interrupted and poor sleep patterns. Managers should consider the potential for fatigue to compromise work safety and develop risk management strategies that target new fathers.


Holistic classification of CT attenuation patterns for interstitial lung diseases via deep convolutional neural networks.
Interstitial lung diseases (ILD) involve several abnormal imaging patterns observed in computed tomography (CT) images. Accurate classification of these patterns plays a significant role in precise clinical decision making of the extent and nature of the diseases. Therefore, it is important for developing automated pulmonary computer-aided detection systems. Conventionally, this task relies on experts' manual identification of regions of interest (ROIs) as a prerequisite to diagnose potential diseases. This protocol is time consuming and inhibits fully automatic assessment. In this paper, we present a new method to classify ILD imaging patterns on CT images. The main difference is that the proposed algorithm uses the entire image as a holistic input. By circumventing the prerequisite of manual input ROIs, our problem set-up is significantly more difficult than previous work but can better address the clinical workflow. Qualitative and quantitative results using a publicly available ILD database demonstrate state-of-the-art classification accuracy under the patch-based classification and shows the potential of predicting the ILD type using holistic image.


Constitutive activation of WASp in X-linked neutropenia renders neutrophils hyperactive.
Congenital neutropenia is characterized by low absolute neutrophil numbers in blood, leading to recurrent bacterial infections, and patients often require life-long granulocyte CSF (G-CSF) support. X-linked neutropenia (XLN) is caused by gain-of-function mutations in the actin regulator Wiskott-Aldrich syndrome protein (WASp). To understand the pathophysiology in XLN and the role of WASp in neutrophils, we here examined XLN patients and 2 XLN mouse models. XLN patients had reduced myelopoiesis and extremely low blood neutrophil number. However, their neutrophils had a hyperactive phenotype and were present in normal numbers in XLN patient saliva. Murine XLN neutrophils were hyperactivated, with increased actin dynamics and migration into tissues. We provide molecular evidence that the hyperactivity of XLN neutrophils is caused by WASp in a constitutively open conformation due to contingent phosphorylation of the critical tyrosine-293 and plasma membrane localization. This renders WASp activity less dependent on regulation by PI3K. Our data show that the amplitude of WASp activity inside a cell could be enhanced by cell-surface receptor signaling even in the context in which WASp is already in an active conformation. Moreover, these data categorize XLN as an atypical congenital neutropenia in which constitutive activation of WASp in tissue neutrophils compensates for reduced myelopoiesis.


Foregut mesenchyme contributes cells to pancreatic acini during embryonic development in a chick-quail chimera model.
To understand causes of developmental abnormalities of the pancreas, it is essential to understand its normal embryonic development. Current understanding of the development of pancreatic exocrine tissue is that it develops solely from embryonic epithelium, while the role of the surrounding mesenchyme is to signal to this epithelium and form connective tissue. Recent work in our laboratory has shown that pancreatic bud mesenchyme can contribute cells to islets during embryonic development. However, no published studies have investigated in detail whether mesenchyme contributes cells to the exocrine structures of the pancreas. The aim of this study was to investigate whether cells from foregut mesenchyme can contribute to pancreatic acini during embryonic development. Chick-quail chimera recombinant organs were constructed using pancreatic epithelium and mesenchyme from either the pancreas (n=12) or stomach (n=25). These were cultured for 7 days in 3-D collagen gels. The resulting specimens were analysed using morphological criteria and fluorescent immunocytochemistry against pancreatic amylase, insulin, and the quail-specific nucleolar antigen QCPN. Two independent observers determined the origins of acini as either solely epithelial, solely mesenchymal, or of mixed origin. Results are expressed as percentages of total acini identified in each group. Statistical analysis was performed using chi(2) tests (P<0.01 was considered statistically significant). Recombinations of pancreatic epithelium and pancreatic mesenchyme yielded 11 acini, of which 45% were derived from epithelium only, 45% from mesenchyme only, and 10% of mixed origin. Recombinations of pancreatic epithelium and stomach mesenchyme yielded 78 acini, of which 40% were derived from epithelium only, 32% from mesenchyme only, and 28% of mixed origin. When acini with any mesenchymal cellular contribution were considered as a group, there was no significant difference between stomach and pancreatic mesenchymal contribution (P=0.72). This is the first study to demonstrate the cellular contribution of mesenchyme to pancreatic exocrine structures. Our data show that mesenchyme contributes cells to pancreatic acini during development in this model and that mesenchyme derived from stomach and pancreatic sources are both able to form acini.


Interrelationships of adolescent physical activity, screen-based sedentary behaviour, and social and psychological health.
To examine how adolescent physical activity (PA) and screen-based media sedentary behaviours (SBM) relate to psychological and social health and identify cross-national differences in these relationships. Associations were examined in five regions using two Health Behaviour in School-Aged Children (HBSC) countries from each. Self-reported psychological and social health indices such as self-image, perceived health status, and Life Satisfaction were positively related to PA in all five regions but, with a few exceptions, negatively related to SBM. Negative health indices such as health complaints and tobacco use were negatively related to PA but, with exceptions, positively related to SBM. Significant regional differences were present. Regional differences in correlates of PA and SBM suggest cultural differences in potential effects of PA and SBM and the need to tailor school and public health efforts to the different meanings of PA and SBM for positive and negative health consequences.


Involvement of a novel genistein-inducible multidrug efflux pump of Bradyrhizobium japonicum early in the interaction with Glycine max (L.) Merr.
The early molecular dialogue between soybean and the bacterium Bradyrhizobium japonicum is crucial for triggering their symbiotic interaction. Here we found a single large genomic locus that is widely separated from the symbiosis island and was conspicuously induced within minutes after the addition of genistein. This locus (named BjG30) contains genes for the multidrug efflux pump, TetR family transcriptional regulator, and polyhydroxybutyrate (PHB) metabolism. The induction of BjG30 by genistein was competitively inhibited by daidzein, although both genistein and daidzein are soybean-derived inducers of nodulation (nod) genes. Such a differential expression pattern is also observed in some legume-derived flavonoids, which structurally differ in the hydroxy/deoxy group at the 5-position. In addition, not only did the induction start far in advance of nodW and nodD1 after the addition of genistein, but the levels showed distinct concentration dependence, indicating that the induction pattern of BjG30 is completely different from that of nod genes. The deletion of genes encoding either the multidrug efflux pump or PHB metabolism, especially the former, resulted in defective nodulation performance and nitrogen-fixing capability. Taken together, these results indicate that BjG30, and especially its multidrug efflux pump, may play a key role in the early stage of symbiosis by balancing the dual functions of genistein as both a nod gene inducer and toxicant.


TLR4 signaling induces TLR2 expression in endothelial cells via neutrophil NADPH oxidase.
Interactions of polymorphonuclear neutrophils (PMNs) with endothelial cells may contribute to the activation of endothelial cell responses involved in innate immunity. We explored a novel function of PMN NADPH oxidase in the mechanism of Toll-like receptor-2 (TLR2) upregulation induced by LPS-TLR4 signaling in endothelial cells. We showed that LPS induced TLR2 up-regulation through TLR4- and MyD88-dependent signaling. In neutropenic mice, the LPS-induced NF-kB activation and TLR2 expression were significantly reduced, and both responses were restored upon repletion by PMN obtained from WT mice but not by PMNs from NADPH oxidase gp91pho(-/-) mice. These findings were recapitulated in mouse lung vascular endothelial cells cocultured with PMNs, indicating that the augmented NF-kB activation and the resultant TLR2 upregulation in endothelial cells were secondary to oxidant signaling generated by PMN NADPH oxidase. The functional relevance of NADPH oxidase in mediating TLR4-induced TLR2 expression in endothelial cells was evident by markedly elevated and stable ICAM-1 expression as well as augmented PMN migration in response to sequential challenge with LPS and peptidoglycan. Thus, PMN NADPH oxidase-derived oxidant signaling is an important determinant of the cross talk between TLR4 and TLR2 and the control of endothelial cell activation.


Relationship Between Sleep Quality and Mood: Ecological Momentary Assessment Study.
Sleep disturbances play an important role in everyday affect and vice versa. However, the causal day-to-day interaction between sleep and mood has not been thoroughly explored, partly because of the lack of daily assessment data. Mobile phones enable us to collect ecological momentary assessment data on a daily basis in a noninvasive manner. This study aimed to investigate the relationship between self-reported daily mood and sleep quality. A total of 208 adult participants were recruited to report mood and sleep patterns daily via their mobile phones for 6 consecutive weeks. Participants were recruited in 4 roughly equal groups: depressed and anxious, depressed only, anxious only, and controls. The effect of daily mood on sleep quality and vice versa were assessed using mixed effects models and propensity score matching. All methods showed a significant effect of sleep quality on mood and vice versa. However, within individuals, the effect of sleep quality on next-day mood was much larger than the effect of previous-day mood on sleep quality. We did not find these effects to be confounded by the participants' past mood and sleep quality or other variables such as stress, physical activity, and weather conditions. We found that daily sleep quality and mood are related, with the effect of sleep quality on mood being significantly larger than the reverse. Correcting for participant fixed effects dramatically affected results. Causal analysis suggests that environmental factors included in the study and sleep and mood history do not mediate the relationship.


A general method for identifying recessive diploid-specific mutations in Saccharomyces cerevisiae, its application to the isolation of mutants blocked at intermediate stages of meiotic prophase and characterization of a new gene SAE2.
We describe a general new approach for identifying recessive mutations that affect diploid strains of yeast Saccharomyces cerevisiae and the application of this method to the identification of mutations that confer an intermediate block in meiotic prophase chromosome metabolism. The method uses a temperature-sensitive conjugation mutation ste7-1 in combination with homothallism. The mutations of interest confer a defect in spore formation that is dependent upon a gene required for initiation of meiotic recombination and development of meiosis-specific chromosome structure (SPO11). Identified in this screen were null mutations of the DMC1 gene, nonnull mutations of RAD50 (rad50S), and mutations in three new genes designed SAE1, SAE2 and SAE3 (Sporulation in the Absence of Spo Eleven). Molecular characterization of the SAE2 gene and characterization of meiotic and mitotic phenotypes of sae2 mutants are also presented. The phenotypes conferred by a sae2 null mutation are virtually indistinguishable from those conferred by the previously identified nonnull mutations of RAD50 (rad50S). Most notably, both mutations confer only weak sensitivity to the radiomimetic agent methyl methane sulfonate (MMS) but completely block resection and turnover of meiosis-specific double-strand breaks. These observations provide further evidence that this constellation of phenotypes identifies a specific molecular function.


Guideline implementation research: exploring the gap between evidence and practice in the CRUSADE Quality Improvement Initiative.
Translating research results into routine clinical practice remains difficult. Guidelines, such as the 2002 American College of Cardiology/American Heart Association Guidelines for the Management of Patients with Unstable Angina and non-ST-segment elevation myocardial infarction, have been developed to provide a streamlined, evidence-based approach to patient care that is of high quality and is reproducible. The Can Rapid Risk Stratification of Unstable Angina Patients Suppress ADverse Outcomes with Early Implementation (CRUSADE) Quality Improvement Initiative was developed as a registry for non-ST-segment elevation acute coronary syndromes to track the use of guideline-based acute and discharge treatments for hospitalized patients, as well as outcomes associated with the use of these treatments. Care for more than 200,000 patients at more than 400 high-volume acute care hospitals in the United States was tracked in CRUSADE, with feedback provided to participating physicians and hospitals regarding their performance over time and compared with similar institutions. Such access to data has proved important in stimulating improvements in non-ST-segment elevation acute coronary syndromes care at participating hospitals for delivery of acute and discharge guideline-based therapy, as well as improving outcomes for patients. Providing quality improvement methods such as protocol order sets, continuing education programs, and a CRUSADE Quality Improvement Initiative toolbox serve to actively stimulate physician providers and institutions to improve care. The CRUSADE Initiative has also proven to be a fertile source of research in translation of treatment guidelines into routine care, resulting in more than 52 published articles and 86 abstracts presented at major emergency medicine and cardiology meetings. The cycle for research of guideline implementation demonstrated by CRUSADE includes four major steps--observation, intervention, investigation, and publication--that serve as the basis for evaluating the impact of any evidence-based guideline on patient care. Due to the success of CRUSADE, the American College of Cardiology combined the CRUSADE Initiative with the National Registry for Myocardial Infarction ST-segment elevation myocardial infarction program to form the National Cardiovascular Data Registry-Acute Coronary Treatment & Intervention Outcomes Network Registry beginning in January 2007.


Regulation of Toll-like receptor (TLR)2 and TLR4 on CD14dimCD16+ monocytes in response to sepsis-related antigens.
Rapid overproduction of proinflammatory cytokines are characteristic of sepsis. CD14(dim)CD16(+) monocytes are thought to be major producers of cytokine and have been shown to be elevated in septic patients. Toll-like receptors (TLR) are pattern recognition receptors important in mediating the innate immune response and their activation can lead to production of cytokines. Using whole blood culture and flow cytometry we have investigated TLR2 and TLR4 regulation after stimulation with sepsis-relevant antigens [lipopolysaccharide (LPS), Staphylococcal enterotoxin B (SEB) and peptidoglycan (PGN)]. The percentage of CD14(dim)CD16(+) monocyte population expanded at 20 h post-stimulation, after a rise in tumour necrosis factor (TNF)-alpha and interleukin (IL)-6 at 2 h. A strong positive correlation between the percentage of CD14(dim)CD16(+) monocytes and secreted TNF-alpha was demonstrated (r = 0.72). Furthermore, we were able to induce expansion of the CD14(dim)CD16(+) population to approximately 35% of all monocytes with the addition of recombinant TNF-alpha to the whole blood culture. TLR4 was found to be expressed 2.5 times higher on CD14(dim)CD16(+) compared to CD14(+) CD16(-) monocytes, while TLR2 expression was similar in both subpopulations. The CD14(dim)CD16(+) and CD14(+) CD16(-) monocyte populations were different in their response to various antigens. LPS down-regulated TLR4 by 4.9 times in CD16(+) monocytes compared to only 2.3 times in CD16(-) monocytes at 2 h. LPS was able to up-regulate TLR2 by 6.2 times after 2 h, with no difference between the subpopulations. LPS further up-regulated TLR2 by 18.4 times after 20 h only in the CD14(+) CD16(-) population. PGN and SEB induced no significant changes in TLR2 or TLR4 expression. We hypothesize that following exposure to bacterial antigens, subsequent TNF-alpha drives a differentiation of monocytes into a CD14(dim)CD16(+) subpopulation.


Upper and lower extremity robotic devices for rehabilitation and for studying motor control.
The successful motor rehabilitation of stroke, traumatic brain-injured and spinal cord-injured patients requires an intensive and task-specific therapy approach. Budget constraints limit a hand-to-hand therapy approach, so that intelligent machines may offer a solution to promote motor recovery and obtain a better understanding of motor control. This new field of automated or robot-assisted motor rehabilitation has emerged since the 1990s. This article will present clinically viable devices for upper and lower extremity rehabilitation. The MIT-Manus and the Mirror-Image Motion Enabler robot, which enable unrestricted unilateral or bilateral shoulder and elbow movement, consistently proved superior on the motor impairment level. The ARM guide, which assisted reaching in a straight-line trajectory, and the Bi-Manu-Track, which enabled the bilateral practice of a forearm and wrist movement, are currently being tested. For gait rehabilitation after stroke, the electromechanical gait trainer, GT I, has proved effective compared with treadmill training with body weight support. The Lokomat, consisting of a treadmill and a powered exoskeleton, lessened the therapeutic effort compared with manually assisted treadmill training in spinal cord-injured patients. Future developments will see more degrees of freedom, improved man-machine interaction and the implementation of virtual reality. Technical possibilities are one aspect, but multi-centre trials and a consideration of the unsubstantiated fears among therapists of being replaced by machines will decide on the successful implementation of this most promising field to the benefit of patients.


On the antioxidant activity of carvedilol in human polymorphonuclear leukocytes in vitro and ex vivo.
In the present study we investigated whether the beta-adrenoceptor antagonist carvedilol (CARV) [Dilatrend] prevented reactive oxygen metabolite (ROM) production in human polymorphonuclear leukocytes (PMNL) or interfered with ROM already generated. To specify the site of action of CARV, we evaluated its effect on extra- and intracellular ROM generation. In addition, we studied the effect of CARV therapy on ROM production in whole blood obtained from patients with congestive heart failure (CHF) combined with type II diabetes mellitus. ROM generation in whole blood and isolated PMNL was determined after phorbol 12-myristate-13-acetate (PMA-0.05 micromol/l) stimulation by luminol/isoluminol-enhanced chemiluminescence (CL) in the microplate luminometer Immunotech LM-01T. CARV significantly decreased CL of human whole blood in the concentrations of 10, 20, 50 and 100 micromol/l, both when applied simultaneously and after stimulation. In isolated PMNL, CARV significantly decreased extracellular CL in the concentrations of 50 and 100 micromol/l, intracellular CL was decreased in the concentrations of 20, 50 and 100 micromol/l. The nonstimulated and PMA stimulated CL of whole blood was increased in patients before therapy in comparison with healthy controls. After CARV therapy, 25 mg/day and 50 mg/day, there was a trend to reduce CL as compared to values before therapy. In human PMNL, CARV interfered in vitro and ex vivo with ROM generation as well as with already generated ROM, suggestive of its both "preventive" and "therapeutic" effect.


A systematic review of the accuracy of ultrasound in the diagnosis of endometriosis.
To evaluate transvaginal and transabdominal ultrasound scanning, with or without Doppler, as a diagnostic test for the accurate diagnosis of pelvic endometriosis. The MEDLINE (1966-2001) and EMBASE (1980-2001) databases were searched for relevant studies, published in English. Only studies fulfilling predefined criteria were selected. An assessment of quality was made for each study, and data were then reanalyzed using likelihood ratios to determine the usefulness of the test. In total, 67 papers were identified using the search strategy, of which 17 described relevant studies. Of these, seven fulfilled the inclusion criteria. All seven related to the use of transvaginal gray-scale imaging in the diagnosis of ovarian endometriomata specifically, rather than endometriosis. The positive likelihood ratios ranged from 7.6 to 29.8, and the negative likelihood ratios ranged from 0.1 to 0.4. Confidence intervals were wide. One paper addressed the use of conventional color Doppler with ultrasound: the positive likelihood ratio was 1.2, with a negative likelihood ratio of 0.4. One paper assessed the use of color Doppler energy imaging, and showed a positive likelihood ratio of 33.5 and a negative likelihood ratio of 0.1. Transvaginal ultrasound appears to be a useful test both to make and to exclude the diagnosis of an ovarian endometrioma.


Oxygen gradient ektacytometry does not predict pain in children with sickle cell anaemia.
The loss of red blood cell (RBC) deformability in sickle cell anaemia (SCA) is considered the primary factor responsible for episodes of acute pain and downstream progressive organ dysfunction. Oxygen gradient ektacytometry (Oxygenscan) is a recently commercialised functional assay that aims to describe the deformability of RBCs in SCA at differing oxygen tensions. So far, the Oxygenscan has been evaluated only by a small number of research groups and the validity and clinical value of Oxygenscan-derived biomarkers have not yet been fully established. In this study we examined RBC deformability measured with the Oxygenscan in 91 children with SCA at King's College Hospital in London. We found a significant correlation between Oxygenscan-derived biomarkers and well-recognised modifiers of disease severity in SCA: haemoglobin F and co-inherited α-thalassaemia. We failed, however, to find any independent predictive value of the Oxygenscan in the clinical outcome measure of pain, as well as other important parameters such as hydroxycarbamide treatment. Although the Oxygenscan remains an intriguing tool for basic research, our results question whether it provides any additional information in predicting the clinical course in children with SCA, beyond measuring known markers of disease severity.


Vancouver and the 2010 Olympic Games: physical activity for all?
The City of Vancouver, British Columbia strategically designed and implemented a municipal health promotion Policy--the Vancouver Active Communities policy--to leverage the 2010 Olympic Games. The goal of the policy was to increase physical activity participation among Vancouver residents by 2010. In this paper, we conduct a critical policy analysis of health promotion policy documents that were available on the City of Vancouver's website. We elaborate on the background to the policy and more specifically we examine its content: the problem definition, policy goals, and policy instruments. Our analysis showed inconsistency within the policy, particularly because the implemented policy instruments were not designed to address needs of the identified target populations in need of health promotion efforts, which were used to legitimize the approval of funding for the policy. Inconsistency across municipal policies, especially in terms of promoting physical activity among low-income residents, was also problematic. If other municipalities seek to leverage health promotion funding related to hosting sport mega-events, the programs and services should be designed to benefit the target populations used to justify the funding. Furthermore, municipalities should clearly indicate how funding will be maintained beyond the life expectancy of the mega-event.


The kaleidoscope of autoimmunity: multiple autoimmune syndromes and familial autoimmunity.
Three related lines of evidence sustain the common origin for autoimmune diseases (ADs). First, the clinical evidence corresponding to the kaleidoscope of autoimmunity, which is the co-occurrence of various ADs within an individual or co-occurrence within members of a nuclear family. Second, the physiopathologic evidence indicating that the pathologic mechanisms might be similar among ADs. Last, the genetic evidence indicating that autoimmune phenotypes could represent pleiotropic outcomes of nonspecific disease genes. The two conditions that better illustrate the kaleidoscope of autoimmunity are multiple autoimmune syndromes and familial autoimmunity. The multiple autoimmune syndromes consist of the presence of three or more well-defined autoimmune conditions in a single patient. The familial autoimmunity is defined as the presence of diverse ADs on multiple members of a nuclear family. Herein, both the multiple autoimmune syndromes and familial autoimmunity are discussed and various epidemiological factors considered in the context of the common genetic background of autoimmunity.


Mammalian gastrointestinal parasites in rainforest remnants of Anamalai Hills, Western Ghats, India.
Habitat fragmentation is postulated to be a major factor influencing infectious disease dynamics in wildlife populations and may also be responsible, at least in part, for the recent spurt in the emergence, or re-emergence, of infectious diseases in humans. The mechanism behind these relationships are poorly understood due to the lack of insights into the interacting local factors and insufficient baseline data in ecological parasitology of wildlife. Here, we studied the gastrointestinal parasites of nonhuman mammalian hosts living in 10 rainforest patches of the Anamalai Tiger Reserve, India. We examined 349 faecal samples of 17 mammalian species and successfully identified 24 gastrointestinal parasite taxa including 1 protozoan, 2 trematode, 3 cestode and 18 nematode taxa. Twenty of these parasites are known parasites of humans. We also found that as much as 73% of all infected samples were infected by multiple parasites. In addition, the smallest and most fragmented forest patches recorded the highest parasite richness; the pattern across fragments, however, seemed to be less straightforward, suggesting potential interplay of local factors.


Cardiotoxicity during invasive pneumococcal disease.
Streptococcus pneumoniae is the leading cause of community-acquired pneumonia and sepsis, with adult hospitalization linked to approximately 19% incidence of an adverse cardiac event (e.g., heart failure, arrhythmia, infarction). Herein, we review the specific host-pathogen interactions that contribute to cardiac dysfunction during invasive pneumococcal disease: (1) cell wall-mediated inhibition of cardiomyocyte contractility; (2) the new observation that S. pneumoniae is capable of translocation into the myocardium and within the heart, forming discrete, nonpurulent, microscopic lesions that are filled with pneumococci; and (3) the bacterial virulence determinants, pneumolysin and hydrogen peroxide, that are most likely responsible for cardiomyocyte cell death. Pneumococcal invasion of heart tissue is dependent on the bacterial adhesin choline-binding protein A that binds to laminin receptor on vascular endothelial cells and binding of phosphorylcholine residues on pneumococcal cell wall to platelet-activating factor receptor. These are the same interactions responsible for pneumococcal translocation across the blood-brain barrier during the development of meningitis. We discuss these interactions and how their neutralization, either with antibody or therapeutic agents that modulate platelet-activating factor receptor expression, may confer protection against cardiac damage and meningitis. Considerable collagen deposition was observed in hearts of mice that had recovered from invasive pneumococcal disease. We discuss the possibility that cardiac scar formation after severe pneumococcal infection may explain why individuals who are hospitalized for pneumonia are at greater risk for sudden death up to 1 year after infection.


Auditory working memory for objects vs. features.
This work considers bases for working memory for non-verbal sounds. Specifically we address whether sounds are represented as integrated objects or individual features in auditory working memory and whether the representational format influences WM capacity. The experiments used sounds in which two different stimulus features, spectral passband and temporal amplitude modulation rate, could be combined to produce different auditory objects. Participants had to memorize sequences of auditory objects of variable length (1-4 items). They either maintained sequences of whole objects or sequences of individual features until recall for one of the items was tested. Memory recall was more accurate when the objects had to be maintained as a whole compared to the individual features alone. This is due to interference between features of the same object. Additionally a feature extraction cost was associated with maintenance and recall of individual features, when extracted from bound object representations. An interpretation of our findings is that, at some stage of processing, sounds might be stored as objects in WM with features bound into coherent wholes. The results have implications for feature-integration theory in the context of WM in the auditory system.


Pregnancy incidence and outcomes among women receiving preexposure prophylaxis for HIV prevention: a randomized clinical trial.
Antiretroviral preexposure prophylaxis (PrEP), using tenofovir disoproxil fumarate (TDF) and combination emtricitabine/tenofovir disoproxil fumarate (FTC+TDF), is efficacious for prevention of human immunodeficiency virus (HIV) acquisition. PrEP could reduce periconception HIV risk, but the effect on pregnancy outcomes is not well defined. To assess pregnancy incidence and outcomes among women using PrEP during the periconception period. Randomized trial among 1785 HIV-serodiscordant heterosexual couples (the Partners PrEP Study) in which the female partner was HIV uninfected that demonstrated that PrEP was efficacious for HIV prevention, conducted between July 2008 and June 2013 at 9 sites in Kenya and Uganda. Daily oral TDF (n = 598), combination FTC+TDF (n = 566), or placebo (n = 621) through July 2011, when PrEP demonstrated efficacy for HIV prevention. Thereafter, participants continued receiving active PrEP without placebo. Pregnancy testing occurred monthly and study medication was discontinued when pregnancy was detected. Pregnancy incidence, birth outcomes (live births, pregnancy loss, preterm birth, congenital anomalies), and infant growth. A total of 431 pregnancies occurred. Pregnancy incidence was 10.0 per 100 person-years among women assigned placebo, 11.9 among those assigned TDF (incidence difference, 1.9; 95% CI, -1.1 to 4.9 [P = .22 vs placebo]), and 8.8 among those assigned FTC+TDF (incidence difference, -1.3; 95% CI, -4.1 to 1.5 [P = .39 vs placebo]). Before discontinuation of the placebo treatment group in July 2011, the occurrence of pregnancy loss (96 of 288 pregnancies) was 42.5% for women receiving FTC+TDF compared with 32.3% for those receiving placebo (difference for FTC+TDF vs placebo, 10.2%; 95% CI, -5.3% to 25.7%; P = .16) and was 27.7% for those receiving TDF alone (difference vs placebo, -4.6%; 95% CI, -18.1% to 8.9%; P = .46). After July 2011, the frequency of pregnancy loss (52 of 143 pregnancies) was 37.5% for FTC+TDF and 36.7% for TDF alone (difference, 0.8%; 95% CI, -16.8% to 18.5%; P = .92). Occurrence of preterm birth, congenital anomalies, and growth throughout the first year of life did not differ significantly for infants born to women who received PrEP vs placebo. Among HIV-serodiscordant heterosexual African couples, differences in pregnancy incidence, birth outcomes, and infant growth were not statistically different for women receiving PrEP with TDF alone or combination FTC+TDF compared with placebo at conception. Given that PrEP was discontinued when pregnancy was detected and that CIs for the birth outcomes were wide, definitive statements about the safety of PrEP in the periconception period cannot be made. These results should be discussed with HIV-uninfected women receiving PrEP who are considering becoming pregnant. clinicaltrials.gov Identifier: NCT00557245.


Moving general practitioner training into primary care with cluster-based learning: a qualitative study in the West Midlands.
This study sought to explore the early implementation of cluster-based learning (CBL) pilots in one part of the UK. A qualitative study using semi-structured one-to-one interviews was conducted with 10 key informants responsible for managing and delivering local training programmes across 14 general practice specialty training schemes. Data were analysed using thematic content analysis using the constant comparative method. Wide variation existed in the running of CBL pilots between different schemes. The principal reasons for trialling CBL were the high numbers of general practitioner trainees, resulting in extra pressure placed on postgraduate education facilities. Training programme directors (TPDs) managed and facilitated the current pilots. Effective facilitation of the groups was considered essential, along with good organisation and planning, and enthusiastic facilitators. Having a mixture of specialist trainees from different stages of training promoted shared learning in the group. The cluster-based approach was also judged to enable the targeting of training to meet the needs of learners and to address specific local needs. Responders considered that CBL should ultimately comprise learner-led sessions facilitated by educational supervisors, with TPDs being responsible for leading the improvement of the quality of educational and clinical supervision. The main reasons why CBL was not implemented were anxieties about the risks of losing the perceived benefits of the current training format in the absence of evidence of the added value of CBL. The reluctance of trainers to get involved was a major barrier - this resulted mainly from a lack of confidence with facilitating small groups and funding for their time.


On the Characterization of the Generation Rate and Size-Dependent Crystalline Silica Content of the Dust from Cutting Fiber Cement Siding.
A laboratory testing system was developed to systematically characterize the dust generation rate and size-dependent crystalline silica content when cutting or shaping silica containing materials. The tests of cutting fiber cement siding in this system verify that it provides high test repeatability, making it suitable for the targeted characterizations. The mass-based size distributions obtained from a gravimetric-based instrument and a direct reading instrument both show bimodal lognormal distributions with a larger mode ~13 µm and another mode <5 µm for the dusts from cutting four different brands of fiber cement siding. The generation rates of respirable dust obtained from the two instruments are comparable, and the results from each instrument are similar for the four brands. The silica content in the airborne dusts, however, strongly depends on the amount of silica used in the respective product. It is also observed that the silica content in the airborne dust from cutting the four brands of fiber cement siding showed the same trend of an increase with the aerodynamic diameter of the dust, approaching the silica content levels found in their respective bulk samples. Combining the results for both the dust size distribution and size-dependent silica content, it is found that most of the respirable crystalline silica (RCS) resides in the dust ~2.5 µm in aerodynamic diameter. These results would help guide the development of specific engineering control measures targeted at lowering workers' exposure to RCS while cutting fiber cement siding. With the high repeatability using the laboratory testing system, the dust generation rate could then be characterized under different operating conditions, and with the deployment of various engineering control measures. This would greatly facilitate the systematic evaluation of the control effectiveness and the selection of the optimal control solutions for field trials.


Inter-population variation for Wolbachia induced reproductive incompatibility in the haplodiploid mite Tetranychus urticae.
Recent studies have revealed diverse patterns of cytoplasmic incompatibility (CI) induced by Wolbachia in the two spotted spider mite (Tetranychus urticae Koch). The mechanism of CI consists of two steps: modification (mod) of sperm of infected males and the rescue (resc) of these chromosomes by Wolbachia in the egg, which results in female embryonic mortality (FM), male development (MD) or no CI. Our study reports that Wolbachia infections were highly prevalent infecting all T. urticae populations from various crops in 14 commercial greenhouses in Korea, with two Wolbachia strains expressing distinctive phenotypic effects on hosts. Analyses for wsp gene sequences obtained from collected mite populations revealed all sequences were categorized into two groups (group W1 and W2) discriminated by three diagnostic nucleotides while all Wolbachia strains belonged to the subgroup Ori in Wolbachia supergroup B. Host plants of each mite population were also generally correlated this grouping. Various mating experiments with two mite populations from each group showed that CI patterns and host plants of the mite populations were completely matched with the grouping; no CI (mod(-)resc(+)) for group W1 and mixed pattern of FM and MD (mod(+)resc(+)) for group W2. No distinct changes in fecundity or sex ratio due to Wolbachia infections were observed in four mite populations regardless of Wolbachia grouping. Our study suggests a potential correlation between phenotypic effect of Wolbachia infection and its genetic diversity associated with host plants in Korean mite populations.