Platelet activation, inflammatory mediators and hypercholesterolemia.
Atherogenic cofactors, such as altered cholesterol metabolism, may impact locally on inflammatory responses in atherosclerotic lesions. Blood levels of inflammatory markers (e.g., C-reactive protein, fibrinogen) have been associated with hypercholesterolemia and with overt atherothrombotic disorders. More recently. cytokines (e.g., interleukin-6, interleukin-1beta) and soluble adhesion molecules (e.g., selectins, intercellular adhesion molecule-1, vascular cell adhesion molecule-1) have been associated with both hypercholesterolemia and atherosclerotic disease, suggesting their use as potential therapeutic targets for the non-specific "anti-inflammatory" treatment of atherosclerosis. The inflammatory response associated with hypercholesterolemia involves not only the intrinsic cells of the artery wall. but also circulating cells. Platelets participate in this disease process through the release of a wide variety of biologically active substances. An imbalance of the hemostatic system and persistent in vivo platelet activation can be observed in hypercholesterolemia and may have pathophysiological implications in the development and progression of atherosclerotic plaques. Recent findings on the inflammatory actions of platelets have established the potential for a previously unrecognized biologic role for platelets in inflammation and vascular injury, and have opened new perspectives in the comprehension of the pathogenetic mechanism(s) of atherosclerosis. Stimulated platelets actively synthesize proinflammatory cytokines (e.g., CD40L, IL-1beta) and are able to release chemokines (i.e., platelet factor-4, RANTES) which have been all involved in the inflammatory process associated with hypercholesterolemia. This review will summarize the present understanding of the interplay between hypercholesterolemia, inflammation and platelet activation in the development and progression of atherosclerosis, and we also discuss the effects of lipid-lowering treatment on these phenomena.


Second-order resummed thermodynamic perturbation theory for central-force associating potential: multi-patch colloidal models.
We propose a second-order version of the resummed thermodynamic perturbation theory for patchy colloidal models with arbitrary number of multiply bondable patches. The model is represented by the hard-sphere fluid system with several attractive patches on the surface and resummation is carried out to account for blocking effects, i.e., when the bonding of a particle restricts (blocks) its ability to bond with other particles. The theory represents an extension of the earlier proposed first order resummed thermodynamic perturbation theory for central force associating potential and takes into account formation of the rings of the particles. In the limiting case of singly bondable patches (total blockage), the theory reduces to Wertheim thermodynamic perturbation theory for associating fluids. Closed-form expressions for the Helmholtz free energy, pressure, internal energy, and chemical potential of the model with an arbitrary number of equivalent doubly bondable patches are derived. Predictions of the theory for the model with two patches appears to be in a very good agreement with predictions of new NVT and NPT Monte Carlo simulations, including the region of strong association.


Model-based simulation for early neurosurgical learners.
Restrictions on duty hours and shift length by the Accreditation Council for Graduate Medical Education and public pressure to reduce complications and to improve outcomes in the clinical educational environment have enhanced interest in the use of procedural and surgical simulation to train neurosurgical residents. To introduce simple, available, and, when possible, inexpensive model-based simulation for early learners into the initial stages of neurosurgical residency training. Simulation for early-stage trainees in neurological surgery has taken advantage of model-based systems. The Society of Neurological Surgeons postgraduate year 1 courses have served as one paradigm for designing and using model-based simulators for procedural and surgical skill training as part of a purpose-designed overall curriculum. Ongoing surveys of resident and faculty course participants have supported iterative improvements in simulator models and curriculum from year to year. Simulation for basic neurosurgical and intensive care procedures has been undertaken through the use of available materials, surgical technology, and modifications of related existing model simulators. Simulation of common, standard surgical procedures for early learners may be broken into individual surgical skills and maneuvers to prepare trainees for safe practice of these component skills during live procedures under direct supervision appropriate to their training stage. Model-based simulation is particularly effective for early surgical learners as part of a coordinated curriculum. Almost 600 residents have used model-based simulation during the first 3 years of the Society of Neurological Surgeons boot camp courses, with ongoing modification and improvement of individual simulation models.


Validation of a novel milk progesterone-based tool to monitor luteolysis in dairy cows: Timing of the alerts and robustness against missing values.
Automated monitoring of fertility in dairy cows using milk progesterone is based on the accurate and timely identification of luteolysis. In this way, well-adapted insemination advice can be provided to the farmer to further optimize fertility management. To properly evaluate and compare the performance of new and existing data-processing algorithms, a test data set of progesterone time-series that fully covers the desired variability in progesterone profiles is needed. Further, the data should be measured with a high frequency to allow rapid onset events, such as luteolysis, to be precisely determined. Collecting this type of data would require a lot of time, effort, and budget. In the absence of such data, an alternative was developed using simulated progesterone profiles for multiple cows and lactations, in which the different fertility statuses were represented. To these, relevant variability in terms of cycle characteristics and measurement error was added, resulting in a large cost-efficient data set of well-controlled but highly variable and farm-representative profiles. Besides the progesterone profiles, information on (the timing of) luteolysis was extracted from the modeling approach and used as a reference for the evaluation and comparison of the algorithms. In this study, 2 progesterone monitoring tools were compared: a multiprocess Kalman filter combined with a fixed threshold on the smoothed progesterone values to detect luteolysis, and a progesterone monitoring algorithm using synergistic control, PMASC, which uses a mathematical model based on the luteal dynamics and a statistical control chart to detect luteolysis. The timing of the alerts and the robustness against missing values of both algorithms were investigated using 2 different sampling schemes: one sample per cow every 8 h versus 1 sample per day. The alerts for luteolysis of the PMASC algorithm were on average 20 h earlier compared with the ones of the multiprocess Kalman filter, and their timing was less sensitive to missing values. This was shown by the fact that, when 1 sample per day was used, the Kalman filter gave its alerts on average 24 h later, and the variability in timing of the alerts compared with simulated luteolysis increased with 22%. Accordingly, we postulate that implementation of the PMASC system could improve the consistency of luteolysis detection on farm and lower the analysis costs compared with the current state of the art.


Cognitive Function and Kidney Disease: Baseline Data From the Systolic Blood Pressure Intervention Trial (SPRINT).
Chronic kidney disease is common and is associated with cardiovascular disease, cerebrovascular disease, and cognitive function, although the nature of this relationship remains uncertain. Cross-sectional cohort using baseline data from the Systolic Blood Pressure Intervention Trial (SPRINT). Participants in SPRINT, a randomized clinical trial of blood pressure targets in older community-dwelling adults with cardiovascular disease, chronic kidney disease, or high cardiovascular disease risk and without diabetes or known stroke, who underwent detailed neurocognitive testing in the cognition substudy, SPRINT-Memory and Cognition in Decreased Hypertension (SPRINT-MIND). Urine albumin-creatinine ratio (ACR) and estimated glomerular filtration rate (eGFR). Cognitive function, a priori defined as 5 cognitive domains based on 11 cognitive tests using z scores, and abnormal white matter volume quantified by brain magnetic resonance imaging. Of 9,361 SPRINT participants, 2,800 participated in SPRINT-MIND and 2,707 had complete data; 637 had brain imaging. Mean age was 68 years, 37% were women, 30% were black, and 20% had known cardiovascular disease. Mean eGFR was 70.8±20.9mL/min/1.73m2 and median urine ACR was 9.7 (IQR, 5.7-22.5) mg/g. In adjusted analyses, higher ACR was associated with worse global cognitive function, executive function, memory, and attention, such that each doubling of urine ACR had the same association with cognitive performance as being 7, 10, 6, and 14 months older, respectively. Lower eGFR was independently associated with worse global cognitive function and memory. In adjusted models, higher ACR, but not eGFR, was associated with larger abnormal white matter volume. Cross-sectional only, no patients with diabetes were included. In older adults, higher urine ACR and lower eGFR have independent associations with global cognitive performance with different affected domains. Albuminuria concurrently identifies a higher burden of abnormal brain white matter disease, suggesting that vascular disease may mediate these relationships.


Increased AβPP processing in familial Danish dementia patients.
An autosomal dominant mutation in the BRI2/ITM2B gene causes Familial Danish Dementia (FDD). We have generated a mouse model of FDD, called FDDKI, genetically congruous to the human disease. These mice carry one mutant and one wild type Bri2/Itm2b allele, like FDD patients. Analysis of FDDKI mice and samples from human patients has shown that the Danish mutation causes loss of Bri2 protein. FDDKI mice show synaptic plasticity and memory impairments. BRI2 is a physiological interactor of amyloid-β protein precursor (AβPP), a gene associated with Alzheimer's disease, which inhibits processing of AβPP. AβPP/Bri2 complexes are reduced in synaptic membranes of FDDKI mice. Consequently, AβPP metabolites derived from processing of AβPP by β-, α-, and γ-secretases are increased in Danish dementia mice. AβPP haplodeficiency prevents memory and synaptic dysfunctions, consistent with a role for AβPP-metabolites in the pathogenesis of memory and synaptic deficits. This genetic suppression provides compelling evidence that AβPP and BRI2 functionally interact. Here, we have investigated whether AβPP processing is altered in FDD patients' brain samples. We find that the levels of several AβPP metabolites, including Aβ, are significantly increased in the brain sample derived from an FDD patient. Our data are consistent with the findings in FDDKI mice, and support the hypothesis that the neurological effects of the Danish form of BRI2 are caused by toxic AβPP metabolites, suggesting that Familial Danish and Alzheimer's dementias share common pathogenic mechanisms.


Interactions Between Cells of Sulfobacillus thermosulfidooxidans and Leptospirillum ferriphilum During Pyrite Bioleaching.
Sulfobacillus and Leptospirillum occur frequently in leaching systems. Here we investigated the effects of cells of L. ferriphilum on biofilm formation and leaching performance by S. thermosulfidooxidans. The effects were caused by the presence of L. ferriphilum or an addition of pyrite leach liquor from L. ferriphilum. Data show that the number of attached S. thermosulfidooxidans on pyrite increases, if the pyrite had been pre-colonized by living biofilms of L. ferriphilum, while it decreases if the pre-colonized biofilms had been inactivated. Coaggregation between S. thermosulfidooxidans and L. ferriphilum occurs during the dual-species biofilm formation, but different effects on bioleaching were noted, if the preculture of L. ferriphilum had been different. If L. ferriphilum had been pre-colonized on a pyrite, significantly negative effect was shown. However, if the two species were simultaneously inoculated into a sterile leaching system, the bioleaching efficiency was better than that of a pure culture of S. thermosulfidooxidans. The effect might be related to a metabolic preference of S. thermosulfidooxidans. If S. thermosulfidooxidans performed leaching in a filtered pyrite leachate from L. ferriphilum, the cells preferred to oxidize RISCs instead of ferrous ion and the number of attached cells decreased compared with the control. This study gives an indication that in a short-term multi-species leaching system the role of S. thermosufidooxidans may be related to the time of its introduction.


Antithrombotic treatment of atrial fibrillation in a regional hospital in Hong Kong.
To measure the use, appropriateness, and safety of antithrombotic therapy in Hong Kong Chinese patients with atrial fibrillation. Retrospective review. Regional hospital, Hong Kong. Medical records of all patients with atrial fibrillation admitted to acute internal medicine wards in April 2000 and between July and October 2001 were reviewed for details of antithrombotics given, results of international normalised ratio monitoring for patients receiving warfarin, side-effects, and additional risk factors for complications of atrial fibrillation. Statistical analysis was undertaken to assess factors predictive of antithrombotic use. A total of 207 patients with chronic atrial fibrillation were included in the study. Of these, 44.0% of patients with non-valvular atrial fibrillation without contra-indications for warfarin use were receiving warfarin, 34.1% were receiving aspirin, and 22.0% were receiving no antithrombotic therapy. The majority of patients (69.1%) were treated appropriately according to the American College of Chest Physicians guidelines. The major side-effect rates for warfarin and aspirin were 2.14% and 1.72% per patient-year, respectively, which were comparable with western studies of usual clinical practice. The ischaemic stroke rate for patients taking warfarin or aspirin were 1.40% and 6.02% per patient-year, respectively. The median international normalised ratio was 1.96. The median frequency of international normalised ratio measurement was 45.58 days. This study found that antithrombotic use in a Hong Kong regional hospital for patients with atrial fibrillation was similar to that reported from western institutions. Complication and stroke rates were also comparable to the western data relating to usual clinical practice.


Perceptual advertisement by the prey of stalking or ambushing predators.
There has been previous theoretical explorations of the stability of signals by prey that they have detected a stalking or ambush predator, where such perceptual advertisement dissuades the predator from attacking. Here we use a game theoretical model to extend the theory to consider some empirically-motivated complexities: (i) many perceptual advertisement signals appear to have the potential to vary in intensity, (ii) higher intensity signals are likely to be most costly to produce, and (iii) some high-cost signals (such as staring directly at the predator) can only be utilised if the prey is very confident of the existence of a nearby predator (that is, there are reserved or unfakable signals). We demonstrate that these complexities still allow for stable signalling. However, we do not find solutions where prey use a range of signal intensities to signal different degrees of confidence in the proximity of a predator; with prey simply adopting a binary response of not signalling or always signalling at the same fixed level. However this fixed level will not always be the cheapest possible signal, and we predict that prey that require more certainty about proximity of a predator will use higher-cost signals. The availability of reserved signals does not prohibit the stability of signalling based on lower-cost signals, but we also find circumstances where only the reserved signal is used. We discuss the potential to empirically test our model predictions, and to develop theory further to allow perceptual advertisement to be combined with other signalling functions.


Proteomic analysis of the effects of aged garlic extract and its FruArg component on lipopolysaccharide-induced neuroinflammatory response in microglial cells.
Aged garlic extract (AGE) is widely used as a dietary supplement, and is claimed to promote human health through anti-oxidant/anti-inflammatory activities with hypolipidemic, antiplatelet and neuroprotective effects. Prior studies of AGE have mainly focused on its organosulfur compounds, with little attention paid to its carbohydrate derivatives, such as N-α-(1-deoxy-D-fructos-1-yl)-L-arginine (FruArg). The goal of this study is to investigate actions of AGE and FruArg on antioxidative and neuroinflammatory responses in lipopolysaccharide (LPS)-activated murine BV-2 microglial cells using a proteomic approach. Our data show that both AGE and FruArg can significantly inhibit LPS-induced nitric oxide (NO) production in BV-2 cells. Quantitative proteomic analysis by combining two dimensional differential in-gel electrophoresis (2D-DIGE) with mass spectrometry revealed that expressions of 26 proteins were significantly altered upon LPS exposure, while levels of 20 and 21 proteins exhibited significant changes in response to AGE and FruArg treatments, respectively, in LPS-stimulated BV-2 cells. Notably, approximate 78% of the proteins responding to AGE and FruArg treatments are in common, suggesting that FruArg is a major active component of AGE. MULTICOM-PDCN and Ingenuity Pathway Analyses indicate that the proteins differentially affected by treatment with AGE and FruArg are involved in inflammatory responses and the Nrf2-mediated oxidative stress response. Collectively, these results suggest that AGE and FruArg attenuate neuroinflammatory responses and promote resilience in LPS-activated BV-2 cells by suppressing NO production and by regulating expression of multiple protein targets associated with oxidative stress.


Fraction of Inspired Oxygen Delivered by Elisée™ 350 Turbine Transport Ventilator With a Portable Oxygen Concentrator in an Austere Environment.
Management of critically ill patients in austere environments is a logistic challenge. Availability of oxygen cylinders for the mechanically ventilated patient may be difficult in such a context. One solution is to use a ventilator able to function with an oxygen concentrator (OC). We tested two Elisée™ 350 ventilators paired with SeQual Integra 10-OM oxygen concentrators (OC) (Chart Industries, http://www .chartindustries.com) and evaluated the delivered fraction of inspired oxygen (Fio2). Ventilators were connected to a test lung and Fio2 was measured and indicated by the ventilator. Continuous oxygen was generated by the OC from 0.5L/min to 10L/min, and administered by the specific inlet port of the ventilator. Several combinations of ventilator settings were evaluated to determine the factors affecting the delivered Fio2. The Elisée 350 turbine ventilator is able to deliver a high Fio2 when functioning with an OC. However, modifications of the ventilator settings such as an increase in minute ventilation, inspiratory-to-expiratory ratio, and positive end-expiratory pressure affect delivered Fio2 despite steady-state oxygen flow from the concentrator. OCs provide an alternative to oxygen cylinders for delivering high Fio2 with a turbine ventilator. Nevertheless, Fio2 must be monitored continuously, since it decreases when minute ventilation is increased.


Plasma phosphatidylcholine and sphingomyelin concentrations are associated with depression and anxiety symptoms in a Dutch family-based lipidomics study.
The central nervous system has the second highest concentration of lipids after adipose tissue. Alterations in neural membrane phospho- and sphingolipid composition can influence crucial intra- and intercellular signalling and alter the membrane's properties. Recently, the polyunsaturated fatty acids (PUFA) hypothesis for depression suggests that phospho- and sphingolipid metabolism includes potential pathways for the disease. In 742 people from a Dutch family-based study, we assessed the relationships between 148 different plasma phospho- and sphingolipid species and depression/anxiety symptoms as measured by the Hospital Anxiety and Depression Scales (HADS-A and HADS-D) and the Centre for Epidemiological Studies Depression Scale (CES-D). We observed significant differences in plasma sphingomyelins (SPM), particularly the SPM 23:1/SPM 16:0 ratio, which was inversely correlated with depressive symptom scores. We observed a similar trend for plasma phosphatidylcholines (PC), particularly the molar proportion of PC O 36:4 and its ratio to ceramide CER 20:0. Absolute levels of PC O 36:4 were also associated with depression symptoms in an independent replication. To our knowledge this is the first study on depressive symptoms that focuses on specific phospho- and sphingolipid molecules in plasma rather than total PUFA concentrations. The findings of this lipidomic study suggests that plasma sphingomyelins and ether phospholipids should be further studied for their potential as biomarkers and for a better understanding of the underlying mechanisms of this systemic disease.


Outcomes of Liver Transplant for Adults With Wilson's Disease.
Wilson's disease (WD) is a rare genetic disorder with protean manifestations. Even if liver transplantation (LT) could represent an effective therapeutic option for patients with end-stage liver disease, it has remained controversial in the presence of neuropsychiatric involvement. This study aimed to examine the frequency of adult LT for WD in Italy, focusing on the disease phenotype at the time of LT. A retrospective, observational, multicenter study was conducted across Italy exploring the frequency and characteristics of adults transplanted for WD between 2006 and 2016. A total of 29 adult WD patients underwent LT during the study period at 11 Italian LT centers (accounting for 0.4% of all LTs performed), and 27 of them were considered in this analysis (male/female, n = 9/18; age at LT, 29 years [19-60 years]; median Model for End-Stage Liver Disease score at LT, 27 [6-49]). Isolated hepatic phenotype was the indication for LT in 17 (63%) patients, whereas 2 (7%) patients underwent LT for neurological impairment on compensated liver disease. Overall 1- and 5-year patient survival was excellent (88% and 83%, respectively). Neuropsychiatric symptoms early after LT completely recovered in only a few patients. In conclusion, WD remains an uncommon, unusual indication for LT in Italy, displaying good post-LT graft and patient survival. Because isolated neuropsychiatric involvement represents a rare indication to LT, more data are needed to properly assess the value of LT for WD in this subset of patients.


Ultrasound-guided "short" midline catheters for difficult venous access in the emergency department: a retrospective analysis.
Acutely ill patients admitted to the emergency department (ED) constantly require at least one fast and reliable peripheral intravenous (PIV) access. In many conditions (morbid obesity, underweight state, chronic diseases, intravenous drug abuse, adverse local conditions, etc.), PIV placement may be challenging. Ultrasound guidance is a useful tool for obtaining a peripheral intravenous access in the emergency department, particularly when superficial veins are difficult to identify by palpation and direct visualization, though standard peripheral intravenous cannulas are not ideal for this technique of insertion and may have limited duration. Long polyurethane catheters inserted with ultrasound guidance and direct Seldinger technique appear to have several advantages over short cannulas in terms of success of insertion and of duration. A retrospective analysis was conducted on all the ultrasound-guided peripheral venous accesses obtained by insertion of long polyurethane catheters in patients admitted to the emergency department of our university hospital during 1 year. The main indication to the procedure was the urgent need of a peripheral venous access in patients with superficial veins difficult to palpate and/or visualize. All relevant data concerning the insertion and the maintenance of these peripheral lines were collected from the chart. Seventy-six patients were included in this review. The success rate of insertion was 100 %, with an average of 1.57 punctures per each successful cannulation. The mean time needed for the complete procedure was 9.5 min. In 73 % of patients, the catheter was used for more than 1 week; a minority of catheters were removed prematurely for end of use. No major infective or thrombotic complication was reported. In our experience, 8- to 10-cm-long polyurethane catheters may offer a fast and reliable peripheral venous access in the emergency department, if placed by ultrasound guidance and with the Seldinger technique. Further studies with prospective, randomized, and controlled design are warranted to confirm our results.


In situ synchrotron radiation X-ray diffraction studies on molecular aggregation structure of nylon 12 films during bulge testing.
It is desirable to establish a method for evaluating mechanical properties, such as modulus and strength, of micrometer and sub-micrometer thick polymer films. Bulge tests, where bulge deformation is imposed on films by the pressure of an inert gas, are suitable for satisfying this demand. However, very few studies on polymer films exist in the literature. In this study, bulge testing equipment for in situ synchrotron radiation wide-angle X-ray diffraction (WAXD) measurements is designed and used to study the relationship between the molecular aggregation structure and the mechanical properties of a crystalline nylon 12 (Ny12) film during bulge testing. Isothermally crystallized and quenched Ny12 films exhibited stress-strain curves similar to those obtained by conventional uniaxial elongation. In situ WAXD measurements during bulge testing revealed that the lattice extension of the crystallites is clearly dependent on crystallinity. Concretely, crystallites in the isothermally crystallized film show higher elastic properties than those in the quenched one. The results of the molecular aggregation structure, including the crystal structure and the amorphous chain surrounding the crystallites, of the films during bulge deformation firstly obtained in this study must be useful for designing toughened polymer films.


After-hours coverage: national survey of policies and guidelines for primary care physicians.
To determine the prevalence and content of existing or developing policies and guidelines of medical associations and colleges regarding after-hours care by family physicians and general practitioners, especially legal requirements. Telephone survey in fall 2002, updated in fall 2004. Canada. All national and provincial medical associations, Colleges of Family Physicians, Colleges of Physicians and Surgeons, local government offices for the north, and the Canadian Medical Protective Association (CMPA). RESPONSE TO THE QUESTION: "Does your agency have a policy in place regarding after-hours health care coverage by FPs/GPs, or are there active discussions regarding such a policy?" The College of Physicians and Surgeons of British Columbia was the first to institute a policy, in 1995, requiring physicians to make "specific arrangements" for after-hours care of their patients. The College of Physicians and Surgeons of Alberta adopted a similar policy in 1996 along with a guideline to aid implementation. In 2002, the College of Physicians and Surgeons of Nova Scotia approved a guideline on the Availability of Physicians After Hours. The Saskatchewan Medical Association and the College of Physicians and Surgeons of Saskatchewan formulated a joint policy on medical practice coverage that was released in 2003. Many agencies actively discussed the topic. Provincial and national Colleges of Family Physicians did not have any policies in place. The CMPA does not generate guidelines but released in an information letter in May 2000 a section entitled "Reducing your risk when you're not available." There is increasing interest Canada-wide in setting policy for after-hours care. While provincial Colleges of Physicians and Surgeons have traditionally led the way, a trend toward more collaboration between associations was identified. The effect of policy implementation on physicians' coverage of patients is unclear.


Emerging paradigms in anti-infective drug design.
The need for new drugs to treat microbial infections is pressing. The great progress made in the middle part of the twentieth Century was followed by a period of relative inactivity as the medical needs relating to infectious disease in the wealthier nations receded. Growing realisation that anti-infectives are needed in many parts of the world, to treat neglected diseases as well as to combat the burgeoning risk of resistance to existing drugs, has galvanised a new wave of research into anti-microbial drugs. The transfer of knowledge from the Pharmaceutical industry relating to the importance of understanding how to target drugs successfully within the body, and improved understanding of how pathogens interact with their hosts, is driving a series of new paradigms in anti-infective drug design. Here we provide an overview of those processes as an introduction to a series of articles from experts in this area that emerged from a meeting entitled "Emerging Paradigms in Anti-Infective Drug Design" held in London on the 17th and 18th September 2012. The symposium was organised jointly by British Society for Parasitology (BSP) and the Biological & Medicinal Chemistry sector of the Royal Society of Chemistry (RSC) and held at the London School of Hygiene & Tropical Medicine. The symposium set out to cover all aspects of the identification of new therapeutic modalities for the treatment of neglected and tropical diseases. We aimed to bring together leading scientists from all the disciplines working in this field and cover the pharmacology, medicinal chemistry and drug delivery of potential new medicines. Sessions were held on: "Target diseases and targets for drugs", "Target based medicinal chemistry", "Bioavailability and chemistry", "Targeting intracellular microbes", "Alternative approaches and models", and "New anti-infectives - how do we get there?" This symposium was organised by Simon Croft (LSHTM) and Mike Barrett (University of Glasgow) for the BSP, and David Alker (David Alker Associates) and Andrew Stachulski (University of Liverpool) for the Biological & Medicinal Chemistry sector of the RSC.


High-resolution analysis of parent-of-origin allelic expression in the Arabidopsis Endosperm.
Genomic imprinting is an epigenetic phenomenon leading to parent-of-origin specific differential expression of maternally and paternally inherited alleles. In plants, genomic imprinting has mainly been observed in the endosperm, an ephemeral triploid tissue derived after fertilization of the diploid central cell with a haploid sperm cell. In an effort to identify novel imprinted genes in Arabidopsis thaliana, we generated deep sequencing RNA profiles of F1 hybrid seeds derived after reciprocal crosses of Arabidopsis Col-0 and Bur-0 accessions. Using polymorphic sites to quantify allele-specific expression levels, we could identify more than 60 genes with potential parent-of-origin specific expression. By analyzing the distribution of DNA methylation and epigenetic marks established by Polycomb group (PcG) proteins using publicly available datasets, we suggest that for maternally expressed genes (MEGs) repression of the paternally inherited alleles largely depends on DNA methylation or PcG-mediated repression, whereas repression of the maternal alleles of paternally expressed genes (PEGs) predominantly depends on PcG proteins. While maternal alleles of MEGs are also targeted by PcG proteins, such targeting does not cause complete repression. Candidate MEGs and PEGs are enriched for cis-proximal transposons, suggesting that transposons might be a driving force for the evolution of imprinted genes in Arabidopsis. In addition, we find that MEGs and PEGs are significantly faster evolving when compared to other genes in the genome. In contrast to the predominant location of mammalian imprinted genes in clusters, cluster formation was only detected for few MEGs and PEGs, suggesting that clustering is not a major requirement for imprinted gene regulation in Arabidopsis.


Long-term outcome of patients operated for large ventricular septal defects with increased pulmonary vascular resistance.
There is a paucity of data regarding the long-term outcome of patients operated for ventricular septal defect with severe pulmonary arterial hypertension and elevated pulmonary vascular resistance. We evaluated the long-term follow-up results of a selected cohort of patients with nonrestrictive ventricular septal defect and elevated pulmonary vascular resistance (>6 Wood units). Thirty-eight patients, median age 7.5 years (range 6 months-27 years), with nonrestrictive ventricular septal defect with severe pulmonary hypertension were operated between 1985 and 1996 at our institute. Preoperative pulmonary vascular resistance, ratio of pulmonary blood flow to systemic blood flow, and ratio of pulmonary vascular resistance to systemic vascular resistance were 7.63+/-1.8 Wood units, 1.9+/-0.48, and 0.41+/-0.12, respectively. The majority (68.4%) had perimembranous ventricular septal defect. Thirty patients (79%) had a good outcome and were asymptomatic at a mean follow-up of 8.7 years, with significant reduction in pulmonary artery pressures. Eight patients (21%) had a poor outcome, which included 5 immediate postoperative deaths, 1 late death and 2 surviving patients with persistent severe pulmonary arterial hypertension. There was no significant difference regarding hemodynamic parameters at baseline between those who had a good outcome and those who did not. Eleven patients with a preoperative pulmonary blood flow to systemic blood flow ratio of <2:1. who had a good outcome following surgery, underwent repeat catheterization at follow-up. There was a significant reduction in their mean pulmonary vascular resistance (8.03+/-1.4 v. 4.16+/-1.6 Wood units, p=0.001) and pulmonary vascular resistance to systemic vascular resistance ratio (0.41+/-0.12 v. 0.19+/-0.06, p=0.05). The late results of surgery on this selected group of patients with nonrestrictive ventricular septal defect with high pulmonary vascular resistance are encouraging. Operative correction of the ventricular septal defect should be actively considered in all children presenting with nonrestrictive ventricular septal defect with a significant left-to-right shunt, despite moderately elevated pulmonary vascular resistance. Even among older patients with ventricular septal defect and moderately elevated pulmonary vascular resistance, there is a specific group that does well after operation.


Guidelines for postoperative care in cesarean delivery: Enhanced Recovery After Surgery (ERAS) Society recommendations (part 3).
This Enhanced Recovery After Surgery Guideline for postoperative care in cesarean delivery will provide best practice, evidenced-based recommendations for postoperative care with primarily a maternal focus. The pathway process for scheduled and unscheduled cesarean delivery for this Enhanced Recovery After Surgery cesarean delivery guideline will consider time from completion of cesarean delivery until maternal hospital discharge. The literature search (1966-2017) used Embase and PubMed to search medical subject headings that included "Cesarean Section," "Cesarean Delivery," "Cesarean Section Delivery," and all postoperative Enhanced Recovery After Surgery items. Study selection allowed titles and abstracts to be screened by individual reviewers to identify potentially relevant articles. Metaanalyses, systematic reviews, randomized controlled studies, nonrandomized controlled studies, reviews, and case series were considered for each individual topic. Quality assessment and data analyses evaluated the quality of evidence, and recommendations were evaluated according to the Grading of Recommendations, Assessment, Development and Evaluation system as used and described in previous Enhanced Recovery After Surgery Guidelines. The Enhanced Recovery After Surgery cesarean delivery guideline/pathway has created a pathway for postoperative care. Specifics include sham feeding, nausea and vomiting prevention, postoperative analgesia, nutritional care, glucose control, thromboembolism prophylaxis, early mobilization, urinary drainage, and discharge counseling. A number of elements of postoperative care of women who undergo cesarean delivery are recommended, based on the evidence. As the Enhanced Recovery After Surgery cesarean delivery pathway (elements/processes) are studied, implemented, audited, evaluated, and optimized by the maternity care teams, there will be an opportunity for focused and optimized areas of care and recommendations to be further enhanced.


Trastuzumab Sensitizes Ovarian Cancer Cells to EGFR-targeted Therapeutics.
Early studies have demonstrated comparable levels of HER2/ErbB2 expression in both breast and ovarian cancer. Trastuzumab (Herceptin), a therapeutic monoclonal antibody directed against HER2, is FDA-approved for the treatment of both early and late stage breast cancer. However, clinical studies of trastuzumab in epithelial ovarian cancer (EOC) patients have not met the same level of success. Surprisingly, however, no reports have examined either the basis for primary trastuzumab resistance in ovarian cancer or potential ways of salvaging trastuzumab as a potential ovarian cancer therapeutic. An in vitro model of primary trastuzumab-resistant ovarian cancer was created by long-term culture of HER2-positive ovarian carcinoma-derived cell lines with trastuzumab. Trastuzumab treated vs. untreated parental cells were compared for HER receptor expression, trastuzumab sensitivity, and sensitivity to other HER-targeted therapeutics. In contrast to widely held assumptions, here we show that ovarian cancer cells that are not growth inhibited by trastuzumab are still responsive to trastuzumab. Specifically, we show that responsiveness to alternative HER-targeted inhibitors, such as gefitinib and cetuximab, is dramatically potentiated by long-term trastuzumab treatment of ovarian cancer cells. HER2-positive ovarian carcinoma-derived cells are, therefore, not "unresponsive" to trastuzumab as previously assumed, even when they not growth inhibited by this drug. Given the recent success of EGFR-targeted therapeutics for the treatment of other solid tumors, and the well-established safety profile of trastuzumab, results presented here provide a rationale for re-evaluation of trastuzumab as an experimental ovarian cancer therapeutic, either in concert with, or perhaps as a "primer" for EGFR-targeted therapeutics.


Medicaid matters: children's health and Medicaid eligibility expansions.
In the late 1980s, a series of federal laws were enacted which expanded Medicaid eligibility to more of the nation's children. States had a great amount of discretion in how fast and how far these expansions were implemented. As a result, there was great variation among the states in defining who was eligible for the program. This variation provides a rare opportunity to disentangle the effect of Medicaid from a child's socioeconomic status. Using data from the National Health Interview Survey, we address whether the Medicaid expansions improved the health and functional status of children. Econometric models were developed using fixed-effects regressions, and were estimated separately for white, black, and Hispanic children. White children experienced statistically significant reductions in acute health conditions and functional limitations. Black and Hispanic children showed some evidence of improved health conditions and functional status, but this evidence is inconclusive in the study sample. This may be due to differences in their access to appropriate health services or to the smaller sample size of minorities in each geographic area. The findings are also relevant to the implementation of the Children' Health Insurance Program (CHIP), the latest federal effort to expand access to health care to poor and near poor children. In many states, CHIP is being implemented in whole or in part through further Medicaid expansions.


Calculating luminous flux and lighting levels for domesticated mammals and birds.
This paper considers whether photometric calculations using standard human spectral sensitivity data are satisfactory for applications with other species or whether it would be worthwhile to use bespoke spectral sensitivity functions for each species or group of species. Applications include the lighting of interior areas and the design of photometers. Published spectral sensitivity data for a number of domesticated animals (human, turkey, duck, chicken, cat, rat and mouse) were used to calculate lighting levels for each species and compared with those derived from standard CIE human photopic and scotopic functions. Calculations were made for spectral power distributions of daylight, incandescent light and 12 fluorescent sources commonly used to light interiors. The calculated lighting levels showed clear differences between species and the standard human. Assuming that the resulting effects on retinal illuminance determine the overall perception of the level of light, there may be applications where these differences are important. However, evidence is also presented that the magnitude of these inter-species effects are similar to, or smaller than, those arising from other optical, physiological and psychological factors, which are also likely to influence the resulting perception. It is also important to recognise that lighting-related parameters such as the good colour rendering of surfaces, the avoidance of glare from lamps and other factors that may be species related are sometimes of greater importance than the lighting levels. Our results suggest that a judicial choice of three spectral sensitivity functions would satisfy most circumstances. Firstly, where the overall sensitivity is maximal in the medium to long wavelengths, the standard CIE photopic function will suffice, chicken, turkey and duck fall in this category. Secondly, in a small number of cases where the sensitivity centres on the short to medium wavelengths, the CIE scotopic function should be used, e.g. for the scotopic cat, photopic rat and photopic mouse. Finally, where an animal is also sensitive to the UV region of the spectrum and there is a significant component of UV radiation, then an additional measure of the UV response should be included, as for the photopic rat and photopic mouse.


Natural epigenetic variation contributes to heritable flowering divergence in a widespread asexual dandelion lineage.
Epigenetic variation has been proposed to contribute to the success of asexual plants, either as a contributor to phenotypic plasticity or by enabling transient adaptation via selection on transgenerationally stable, but reversible, epialleles. While recent studies in experimental plant populations have shown the potential for epigenetic mechanisms to contribute to adaptive phenotypes, it remains unknown whether heritable variation in ecologically relevant traits is at least partially epigenetically determined in natural populations. Here, we tested the hypothesis that DNA methylation variation contributes to heritable differences in flowering time within a single widespread apomictic clonal lineage of the common dandelion (Taraxacum officinale s. lat.). Apomictic clone members of the same apomictic lineage collected from different field sites showed heritable differences in flowering time, which was correlated with inherited differences in methylation-sensitive AFLP marker profiles. Differences in flowering between apomictic clone members were significantly reduced after in vivo demethylation using the DNA methyltransferase inhibitor zebularine. This synchronization of flowering times suggests that flowering time divergence within an apomictic lineage was mediated by differences in DNA methylation. While the underlying basis of the methylation polymorphism at functional flowering time-affecting loci remains to be demonstrated, our study shows that epigenetic variation contributes to heritable phenotypic divergence in ecologically relevant traits in natural plant populations. This result also suggests that epigenetic mechanisms can facilitate adaptive divergence within genetically uniform asexual lineages.


Convolution Comparison Pattern: An Efficient Local Image Descriptor for Fingerprint Liveness Detection.
We present a new type of local image descriptor which yields binary patterns from small image patches. For the application to fingerprint liveness detection, we achieve rotation invariant image patches by taking the fingerprint segmentation and orientation field into account. We compute the discrete cosine transform (DCT) for these rotation invariant patches and attain binary patterns by comparing pairs of two DCT coefficients. These patterns are summarized into one or more histograms per image. Each histogram comprises the relative frequencies of pattern occurrences. Multiple histograms are concatenated and the resulting feature vector is used for image classification. We name this novel type of descriptor convolution comparison pattern (CCP). Experimental results show the usefulness of the proposed CCP descriptor for fingerprint liveness detection. CCP outperforms other local image descriptors such as LBP, LPQ and WLD on the LivDet 2013 benchmark. The CCP descriptor is a general type of local image descriptor which we expect to prove useful in areas beyond fingerprint liveness detection such as biological and medical image processing, texture recognition, face recognition and iris recognition, liveness detection for face and iris images, and machine vision for surface inspection and material classification.


Network-Based Integration of Multi-Omics Data Identifies the Determinants of miR-491-5p Effects.
The identification of miRNAs' targets and associated regulatory networks might allow the definition of new strategies using drugs whose association mimics a given miRNA's effects. Based on this assumption we devised a multi-omics approach to precisely characterize miRNAs' effects. We combined miR-491-5p target affinity purification, RNA microarray, and mass spectrometry to perform an integrated analysis in ovarian cancer cell lines. We thus constructed an interaction network that highlighted highly connected hubs being either direct or indirect targets of miR-491-5p effects: the already known EGFR and BCL2L1 but also EP300, CTNNB1 and several small-GTPases. By using different combinations of specific inhibitors of these hubs, we could greatly enhance their respective cytotoxicity and mimic the miR-491-5p-induced phenotype. Our methodology thus constitutes an interesting strategy to comprehensively study the effects of a given miRNA. Moreover, we identified targets for which pharmacological inhibitors are already available for a clinical use or in clinical trials. This study might thus enable innovative therapeutic options for ovarian cancer, which remains the leading cause of death from gynecological malignancies in developed countries.


Left atrial appendage: morphology and function in patients with paroxysmal and persistent atrial fibrillation.
The anatomical and functional characteristics of the left atrial appendage (LAA) and its relationships with anatomical remodeling and ischemic stroke in patients with atrial fibrillation (AF) have not been clearly established. The purpose of this study was to determine whether functional and morphological features of the LAA independently predict clinical outcome and stroke in patients with AF who underwent catheter ablation (CA). Two hundred sixty-four patients with AF, including 176 with paroxysmal AF (PAF, 54.0 ± 11.4 years old, M:F = 138:38) and 88 with persistent AF (PeAF, 56.4 ± 9.6 years old, M:F = 74:14) were studied. Of these patients, 31 (11.7 %) had a history of stroke/TIA (transient ischemic attack). The LA and LAA volumes were 124.0 ± 42.4 and 24.9 ± 4.3 ml in PeAF, these values were greater than those in PAF (81.2 ± 24.8 ml and 21.2 ± 5.1 ml, P < 0.001). The AF type (P = 0.016) and AF duration (P = 0.005), and anti-arrhythmic drugs use (P < 0.001) were significant predictors of AF recurrence after CA in all patients. Compared with patients without history of stroke, stroke patients had larger LA volume (106.9 ± 23.0 vs. 94.0 ± 38.9 ml, P = 0.004) and had lower LAA EF (50.0 ± 11.0 vs. 65.7 ± 13.4 %, P < 0.001). The independent predictors of stroke were age (P = 0.002) and LAA EF (P < 0.001) in PAF patients and that was only age (P = 0.001) in PeAF patients. In anatomical and morphological parameters of the LA and LAA, only depressed systolic function of the LAA was significantly related to stroke/TIA and recurrence of AF after CA in paroxysmal AF patients. Further large scaled prospective study is required for validation.


Changes in bowel function following exoskeletal-assisted walking in persons with spinal cord injury: an observational pilot study.
Prospective, observational study. To explore the effects of exoskeletal-assisted walking (EAW) on bowel function in persons with spinal cord injury (SCI). Ambulatory research facility located in a tertiary care hospital. Individuals 18-65 years of age, with thoracic vertebrae one (T1) to T11 motor-complete paraplegia of at least 12 months duration were enrolled. Pre- and post-EAW training, participants were asked to report on various aspects of their bowel function as well as on their overall quality of life (QOL) as related to their bowel function. Ten participants completed 25-63 sessions of EAW over a period of 12-14 weeks, one participant was lost to follow up due to early withdrawal after ten sessions. Due to the small sample size, each participant's results were presented descriptively in a case series format. At least 5/10 participants reported improvements with frequency of bowel evacuations, less time spent on bowel management per bowel day, fewer bowel accidents per month, reduced laxative and/or stool softener use, and improved overall satisfaction with their bowel program post-EAW training. Furthermore, 8/10 reported improved stool consistency and 7/10 reported improved bowel function related QOL. One participant reported worsening of bowel function post-EAW. Between 50 and 80% of the participants studied reported improvements in bowel function and/or management post-EAW training. EAW training appeared to mitigate SCI-related bowel dysfunction and the potential benefits of EAW on bowel function after SCI is worthy or further study.


Clinical utility and patient acceptance of the computerized Composite International Diagnostic Interview.
The study evaluated the utility to clinicians and the acceptability to patients of the self-administered computerized version of the Composite International Diagnostic Interview (CIDI-Auto) in an acute psychiatric setting. Patients admitted to an acute psychiatric unit completed the CIDI-Auto. Reports of CIDI-Auto diagnoses and symptoms were given to treating psychiatrists, who completed a questionnaire evaluating the accuracy and usefulness of the reports. Patients answered a questionnaire about their attitudes toward computers before completing the CIDI-Auto, and after completing it, they answered a questionnaire about their reactions to the interview. Psychiatrists agreed with only 50 percent of CIDI-Auto current diagnoses and indicated that only 22 percent of CIDI-Auto reports provided useful new diagnoses, although 63 percent helped to clarify diagnoses and 58 percent could save clinicians some time. They endorsed the CIDI-Auto as a possible aid to indirect or remote diagnosis where histories would be taken by nonexpert staff. Ninety-four percent of patients liked the computerized interview, and 83 percent understood the questions without difficulty. Sixty percent felt more comfortable with the computerized interview than with a doctor. Education and previous computer experience promoted positive attitudes and satisfaction with the computerized interview. Psychiatrists considered the current CIDI-Auto completed by patients to be of limited value in diagnosis and history taking. Despite patients' acceptance and positive reactions to the computer interview, satisfactory computerized diagnosis is yet to be attained.


Role of endothelial AADC in cardiac synthesis of serotonin and nitrates accumulation.
Serotonin (5-HT) regulates different cardiac functions by acting directly on cardiomyocytes, fibroblasts and endothelial cells. Today, it is widely accepted that activated platelets represent a major source of 5-HT. In contrast, a supposed production of 5-HT in the heart is still controversial. To address this issue, we investigated the expression and localization of 5-HT synthesizing enzyme tryptophan hydroxylase (TPH) and L-aromatic amino acid decarboxylase (AADC) in the heart. We also evaluated their involvement in cardiac production of 5-HT. TPH1 was weakly expressed in mouse and rat heart and appeared restricted to mast cells. Degranulation of mast cells by compound 48/80 did not modify 5-HT cardiac content in mice. Western blots and immunolabelling experiments showed an abundant expression of AADC in the mouse and rat heart and its co-localization with endothelial cells. Incubation of cardiac homogenate with the AADC substrate (5-hydroxy-L-tryptophan) 5-HTP or intraperitoneal injection of 5-HTP in mice significantly increased cardiac 5-HT. These effects were prevented by the AADC inhibitor benserazide. Finally, 5-HTP administration in mice increased phosphorylation of aortic nitric oxide synthase 3 at Ser (1177) as well as accumulation of nitrates in cardiac tissue. This suggests that the increase in 5-HT production by AADC leads to activation of endothelial and cardiac nitric oxide pathway. These data show that endothelial AADC plays an important role in cardiac synthesis of 5-HT and possibly in 5-HT-dependent regulation of nitric oxide generation.


Rapamycin increases lifespan and inhibits spontaneous tumorigenesis in inbred female mice.
The nutrient-sensing TOR (target of rapamycin) pathway is involved in cellular and organismal aging. Rapamycin, an inhibitor of TOR, extends lifespan in yeast, fruit flies and genetically heterogeneous mice. Here, we demonstrate that lifelong administration of rapamycin extends lifespan in female 129/Sv mice characterized by normal mean lifespan of 2 y. Importantly, rapamycin was administrated intermittently (2 weeks per month) starting from the age of 2 mo. Rapamycin inhibited age-related weight gain, decreased aging rate, increased lifespan (especially in the last survivors) and delayed spontaneous cancer. 22.9% of rapamycin-treated mice survived the age of death of the last mouse in control group. Thus we demonstrated for the first time in normal inbred mice that lifespan can be extended by rapamycin. This opens an avenue to develop optimal doses and schedules of rapamycin as an anti-aging modality.


Thinking within the D box: initial identification of Cdh1-APC substrates in the nervous system.
The anaphase-promoting complex (APC) has a well-established role in cell cycle control, but recent exciting evidence has uncovered unexpected neurobiological functions for this complex E3 ubiquitin ligase. With its co-activator Cdh1, APC's effects upon the nervous system range from regulation of axon growth and patterning to development of synapses to neuronal survival. The Cdh1-APC substrates that control these biological processes in neurons are just beginning to be identified. These findings may offer a glimpse of the wide spectrum of neural activities that are orchestrated by Cdh1-APC.


Hypothyroidism is a Risk Factor for Atrial Fibrillation after Coronary Artery Bypass Graft.
Few reports in the world have shown a differential effect of hypothyroidism in relation to morbidity and mortality following cardiac surgery. To determine the association between preoperative hypothyroidism, composite and disaggregated outcomes of mortality and complications in patients undergoing first-time isolated myocardial revascularization surgery. Historical cohort of patients undergoing myocardial revascularization between January 2008 and December 2014, with 626 patients included for evaluation of the composite and disaggregated outcomes of in-hospital mortality and complications (atrial fibrillation, surgical site infection and reoperation due to bleeding). A logistic regression model was used to determine the association between hypothyroidism and the onset of those outcomes. Cohort of 1696 eligible patients for the study, with 1.8 mortality. Median age, female gender and prevalence of arterial hypertension were all significantly higher among hypothyroid patients. No differences were found in other preoperative or intraoperative characteristics. Hypothyroidism was associated with the presence of the composite outcome, RR 1.6 (1.04-2.4) and atrial fibrillation 1.9 (1.05-3.8). No association with mortality, infections or reoperation due to bleeding was found. Hypothyroidism is a disease that affects females predominantly and does not determine the presence of other comorbidities. Hypothyroidism is a risk factor for the onset of postoperative fibrillation in patients undergoing myocardial revascularization surgery. Postoperative care protocols focused on the prevention of these complications in this type of patients must be instituted.


Recombinant bacillus Calmette-Guérin as a potential vector for preventive HIV type 1 vaccines.
In August 1997, the World Health Organization (WHO) and the Joint United Nations Programme on HIV/AIDS (UNAIDS) convened an expert working group to discuss current strategies for the development of HIV type 1 vaccines. Based on the recent findings of investigators from Japan's National Institute of Infectious Diseases (NIID) in Tokyo using recombinant bacillus Calmette-Guérin (rBCG) as a potential vectored vaccine for HIV, a recommendation was made that further work in this area is a priority. As a result, the working group reconvened in September 1998 to discuss the progress to date with this vaccine approach, as well as areas of related research to assess the feasibility of a BCG-vectored HIV vaccine. This report summarizes the discussions addressing the available scientific data on the potential use of rBCG as a vector for preventive HIV vaccines, the work necessary to move such candidate vaccines into Phase 1 clinical trials, and recommendations targeted at facilitating the long-term development of rBCG-vectored HIV vaccines.


Extinction risk and conservation of the world's sharks and rays.
The rapid expansion of human activities threatens ocean-wide biodiversity. Numerous marine animal populations have declined, yet it remains unclear whether these trends are symptomatic of a chronic accumulation of global marine extinction risk. We present the first systematic analysis of threat for a globally distributed lineage of 1,041 chondrichthyan fishes-sharks, rays, and chimaeras. We estimate that one-quarter are threatened according to IUCN Red List criteria due to overfishing (targeted and incidental). Large-bodied, shallow-water species are at greatest risk and five out of the seven most threatened families are rays. Overall chondrichthyan extinction risk is substantially higher than for most other vertebrates, and only one-third of species are considered safe. Population depletion has occurred throughout the world's ice-free waters, but is particularly prevalent in the Indo-Pacific Biodiversity Triangle and Mediterranean Sea. Improved management of fisheries and trade is urgently needed to avoid extinctions and promote population recovery. DOI: http://dx.doi.org/10.7554/eLife.00590.001.


Effect of early childhood malnutrition on tooth eruption in Haitian adolescents.
The objective of this retrospective cohort study was to determine the effects of early childhood protein-energy malnutrition (EC-PEM) and current nutritional status as defined by anthropomorphic measures on the exfoliation and eruption patterns of teeth among adolescents. Oral clinical examinations were conducted in 2005 using World Health Organization (WHO) diagnostic criteria on 498 11- to 13-year-old Haitians for whom early childhood malnutrition data were available. Anthropomorphic records (weight-for-age) from the Haitian Health Foundation computerized database on children from birth through 5-years old were utilized. Current heights and weights were ascertained. Both sets of data were converted to z-scores based on the National Center for Health Statistics (NCHS) referent database. Based upon these z-scores, EC-PEM and current malnutrition categories were developed for this study. The analyses separately regressed the number of primary and permanent teeth on age, gender, EC-PEM status and current nutritional status. Both a delayed exfoliation of primary teeth and a delayed eruption of permanent teeth were associated with EC-PEM and current stunting in adolescence. The observed associations were either direct and statistically significant or indirectly demonstrated by presenting evidence of confounding. The overall interpretation of the models is that malnutrition beginning in the earliest years and extending throughout childhood influences the exfoliation and eruption of teeth. These findings present evidence of an association between tooth exfoliation/eruption patterns and both EC-PEM and nutritional insufficiency (stunting) throughout childhood. This observed delay in the exfoliation of the primary dentition and in the eruption of the permanent dentition has practical significance in interpreting age-specific dental caries data from populations with different malnutrition experiences.


Functional studies of frataxin.
Mitochondria generate adenosine triphosphate (ATP) but also dangerous reactive oxygen species (ROS). One-electron reduction of dioxygen in the early stages of the electron transport chain yields a superoxide radical that is detoxified by mitochondrial superoxide dismutase to give hydrogen peroxide. The hydroxyl radical is derived from decomposition of hydrogen peroxide via the Fenton reaction, catalyzed by Fe2+ ions. Mitochondria require a constant supply of Fe2+ for heme and iron-sulfur cluster biosyntheses and therefore are particularly susceptible to ROS attack. Two main antioxidant defenses are known in mitochondria: enzymes that catalytically remove ROS, e.g. superoxide dismutase and glutathione peroxidase, and low molecular weight agents that scavenge ROS, including coenzyme Q, glutathione, and vitamins E and C. An effective defensive system, however, should also involve means to control the availability of pro-oxidants such as Fe2+ ions. There is increasing evidence that this function may be carried out by the mitochondrial protein frataxin. Frataxin deficiency is the primary cause of Friedreich's ataxia (FRDA), an autosomal recessive degenerative disease. Frataxin is a highly conserved mitochondrial protein that plays a critical role in iron homeostasis. Respiratory deficits, abnormal cellular iron distribution and increased oxidative damage are associated with frataxin defects in yeast and mouse models of FRDA. The mechanism by which frataxin regulates iron metabolism is unknown. The yeast frataxin homologue (mYfh1p) is activated by Fe(II) in the presence of oxygen and assembles stepwise into a 48-subunit multimer (alpha48) that sequesters >2000 atoms of iron in a ferrihydrite mineral core. Assembly of mYfhlp is driven by two sequential iron oxidation reactions: a fast ferroxidase reaction catalyzed by mYfh1p induces the first assembly step (alpha --> alpha3), followed by a slower autoxidation reaction that promotes the assembly of higher order oligomers yielding alpha48. Depending on the ionic environment, stepwise assembly is associated with the sequestration of < or = 50-75 Fe(II)/subunit. This Fe(II) is initially loosely bound to mYfh1p and can be readily mobilized by chelators or made available to the mitochondrial enzyme ferrochelatase to synthesize heme. However, as iron oxidation and mineralization proceed, Fe(III) becomes progressively inaccessible and a stable iron-protein complex is produced. In conclusion, by coupling iron oxidation with stepwise assembly, frataxin can successively function as an iron chaperon or an iron store. Reduced iron availability and solubility and increased oxidative damage may therefore explain the pathogenesis of FRDA.


Infraorbital nerve block for postoperative pain following cleft lip repair in children.
Postoperative pain is a barrier to the quality of paediatric care, the proper management of which is a challenge. Acute postoperative pain often leads to adverse functional and organic consequences that may compromise surgical outcome. Cleft lip is one of the most common craniofacial birth defects and requires surgical correction early in life. As expected after a surgical intervention in such a sensitive and delicate area, the immediate postoperative period of cleft lip repair may be associated with moderate to severe pain. Infraorbital nerve block associated with general anaesthesia has been used to reduce postoperative pain after cleft lip repair. To assess the effects of infraorbital nerve block for postoperative pain following cleft lip repair in children. We searched the following databases: Cochrane Central Register of Controlled Trials (CENTRAL, the Cochrane Library, Issue 6, 2015), MEDLINE, EMBASE, and Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS) from inception to 17 June 2015. There were no language restrictions. We searched for ongoing trials in the following platforms: the metaRegister of Controlled Trials; ClinicalTrials.gov (the US National Institutes of Health Ongoing Trials Register), and the World Health Organization International Clinical Trials Registry Platform (on 17 June 2015). We checked reference lists of the included studies to identify any additional studies. We contacted specialists in the field and authors of the included trials for unpublished data. We included randomised controlled clinical trials that tested perioperative infraorbital nerve block for cleft lip repair in children, compared with other types of analgesia procedure, no intervention, or placebo (sham nerve block). We considered the type of drug, dosage, and route of administration used in each study. For the purposes of this review, the term 'perioperative' refers to the three phases of surgery, that is preoperative, intraoperative, and postoperative, and commonly includes ward admission, anaesthesia, surgery, and recovery. Two review authors (GF and EH) independently identified, screened, and selected the studies, assessed trial quality, and performed data extraction using the Cochrane Pain, Palliative and Supportive Care Review Group criteria. In case of disagreements, a third review author (EMKS) was consulted. We assessed the evidence using Grading of Recommendations, Assessment, Development and Evaluation (GRADE). We included eight studies involving 353 children in the review. These studies reported different types of interventions (lignocaine or bupivacaine), observation times, and forms of measuring and describing the outcomes, making it difficult to conduct meta-analyses. In the comparison of infraorbital nerve block versus placebo, there was a large effect in mean postoperative pain scores (our first primary outcome) favouring the intervention group (standardised mean difference (SMD) -3.54, 95% confidence interval (CI) -6.13 to -0.95; very low-quality evidence; 3 studies; 120 children). Only one study reported the duration of analgesia (in hours) (second primary outcome) with a difference favouring the intervention group (mean difference (MD) 8.26 hours, 95% CI 5.41 to 11.11; very low-quality evidence) and less supplemental analgesic requirements in the intervention group (risk ratio (RR) 0.05, 95% CI 0.01 to 0.18; low-quality evidence). In the comparison of infraorbital nerve block versus intravenous analgesia, there was a difference favouring the intervention group in mean postoperative pain scores (SMD -1.50, 95% CI -2.40 to -0.60; very low-quality evidence; 2 studies; 107 children) and in the time to feeding (MD -9.45 minutes, 95% CI -17.37 to -1.53; moderate-quality evidence; 2 studies; 128 children). No significant adverse events (third primary outcome) were associated with the intervention, although three studies did not report this outcome. Five out of eight studies found no unwanted side effects after the nerve blocks. Overall, the included studies were at low or unclear risk of bias. The reasons for downgrading the quality of the evidence using GRADE related to the lack of information about randomisation methods and allocation concealment in the studies, very small sample sizes, and heterogeneity of outcome reporting. There is low- to very low-quality evidence that infraorbital nerve block with lignocaine or bupivacaine may reduce postoperative pain more than placebo and intravenous analgesia in children undergoing cleft lip repair. Further studies with larger samples are needed. Future studies should standardise the observation time and the instruments used to measure outcomes, and stratify children by age group.


Iron depletion enhances the effect of sorafenib in hepatocarcinoma.
ABSTACT Human hepatocellular carcinoma (HCC) is known to have a poor prognosis. Sorafenib, a molecular targeted drug, is most commonly used for HCC treatment. However, its effect on HCC is limited in clinical use and therefore new strategies regarding sorafenib treatment are required. Iron overload is known to be associated with progression of chronic hepatitis and increased risk of HCC. We previously reported that iron depletion inhibited cancer cell proliferation and conversely induced angiogenesis. Indeed iron depletion therapy including iron chelator needs to be combined with anti-angiogenic drug for its anti-cancer effect. Since sorafenib has an anti-angiogenic effect by its inhibitory targeting VEGFR, we hypothesized that sorafenib could complement the anti-cancer effect of iron depletion. We retrospectively analyzed the relationship between the efficacy of sorafenib and serum iron-related markers in clinical HCC patients. In clinical cases, overall survival was prolonged in total iron binding capacity (TIBC) high- and ferritin low-patients. This result suggested that the low iron-pooled patients, who could have a potential of more angiogenic properties in/around HCC tumors, could be adequate for sorafenib treatment. We determined the effect of sorafenib (Nexavar®) and/or deferasirox (EXJADE®) on cancer cell viability, and on cell signaling of human hepatocarcinoma HepG2 and HLE cells. Both iron depletion by deferasirox and sorafenib revealed insufficient cytotoxic effect by each monotherapy, however, on the basis of increased angiogenesis by iron depletion, the addition of deferasirox enhanced anti-proliferative effect of sorafenib. Deferasirox was confirmed to increase vascular endothelial growth factor (VEGF) secretion into cellular supernatants by ELISA analysis. In in vivo study sorafenib combined with deferasirox also enhanced sorafenib-induced apoptosis. These results suggested that sorafenib combined with deferasirox could be a novel combination chemotherapy for HCC.


Cultural competence and perceptions of community health workers' effectiveness for reducing health care disparities.
Community health worker (CHW) interventions improve health outcomes of patients from underserved communities, but health professionals' perceptions of their effectiveness may impede integration of CHWs into health care delivery systems. Whether health professionals' attitudes and skills, such as those related to cultural competence, influence perceptions of CHWs, is unknown. A questionnaire was administered to providers and clinical staff from 6 primary care practices in Maryland from April to December 2011. We quantified the associations of self-reported cultural competence and preparedness with attitudes toward the effectiveness of CHWs using logistic regression adjusting for respondent age, race, gender, provider/staff status, and years at the practice. We contacted 200 providers and staff, and 119 (60%) participated. Those reporting more cultural motivation had higher odds of perceiving CHWs as helpful for reducing health care disparities (odds ratio [OR] = 9.66, 95% confidence interval [CI] = 3.48-28.80). Those reporting more frequent culturally competent behaviors also had higher odds of believing CHWs would help reduce health disparities (OR = 3.58, 95% CI = 1.61-7.92). Attitudes toward power and assimilation were not associated with perceptions of CHWs. Cultural preparedness was associated with perceived utility of CHWs in reducing health care disparities (OR = 2.33, 95% CI = 1.21-4.51). Providers and staff with greater cultural competence and preparedness have more positive expectations of CHW interventions to reduce healthcare disparities. Cultural competency training may complement the use of CHWs and support their effective integration into primary care clinics that are seeking to reduce disparities.


Responses of group III and IV muscle afferents to distension of the peripheral vascular bed.
This study was undertaken to test the hypothesis that group III and IV afferents with endings in skeletal muscle signal the distension of the peripheral vascular network. The responses of these slowly conducting afferents to pharmacologically induced vasodilation and to acute obstruction of the venous drainage of the hindlimbs were studied in barbiturate-anesthetized cats. Afferent impulses arising from endings in the triceps surae muscles were recorded from the L(7) and S(1) dorsal roots. Fifteen of the 48 group IV and 3 of the 19 group III afferents tested were stimulated by intra-aortic injections of papaverine (2-2.5 mg/kg). Sixty-two percent of the afferents that responded to papaverine also responded to isoproterenol (50 microg/kg). Seven of the 36 group IV and 2 of the 12 group III afferents tested were excited by acute distension of the hindlimb venous system. Four of the seven group IV afferents responding to venous distension also responded to papaverine (57 vs. 13% for the nonresponding). Finally, we observed that most of the group IV afferents that were excited by dynamic contractions of the triceps surae muscles also responded either to venous distension or to vasodilatory agents. These results are consistent with the histological findings that a large number of group IV endings have their receptive fields close to the venules and suggest that they can be stimulated by the deformation of these vascular structures when peripheral conductance increases. Moreover, such a mechanism offers the possibility of encoding both the effects of muscle contraction through intramuscular pressure changes and the distension of the venular system, thereby monitoring the activity of the veno-muscular pump.


Nurse practitioners' information needs and information seeking: implications for practice and education.
This report is an exploratory study of the information needs and information seeking in a sample of nurse practitioners (NPs) approved to practice in North Carolina. A search and review of relevant literature revealed no studies on this topic. In this study, NPs report their most frequent information needs relate to drug therapy, diagnosis, and other therapy. Their most frequently used information resources are physicians, drug reference manuals, and textbooks. They most frequently confer with physicians on diagnosis and other therapy and other NPs on psychosocial issues.


Identification of potential tumour-associated carbonic anhydrase isozyme IX inhibitors: atom-based 3D-QSAR modelling, pharmacophore-based virtual screening and molecular docking studies.
Tumour hypoxia results in dramatic changes in the gene expression, proliferation and survival of tumour cells. The tumour cells shift towards anaerobic glycolysis which results in change of pH in their microenvironment. In response to this stress, over expression of carbonic anhydrase IX (CA IX) genes is observed in many solid tumours. So, selective inhibition of CA IX can be a promising target for anti-cancer drugs. In this work in silico tools like atom-based 3D-QSAR modelling, pharmacophore-based virtual screening and molecular docking were used to identify potential CA IX inhibitors. Based on the training set used in the QSAR model, twenty pharmacophore models were generated. Out of these, HHHR_1, AHHR_1, DHHHR_1, AHHHR_1 model was used to screen a database of 1,50,000 compounds retrieved from ZINC 15 database. R2 and Q2 was 0.9864 and 0.8799, respectively, for the developed QSAR model. 163 compounds showed a phase screen score above 2.4 in which ZINC02260669 was the highest ranked (screen score, 2.852058) compound in all the four models. Built QSAR model was used to predict the activity of all these 163 compounds and ZINC72370966 showed the highest predicted activity with pKi value of 7.649. These compounds were docked against CA IX (human) protein (PDB ID 5FL6) and molecular docking results showed favourable binding interactions for the best ten identified hits. This work gives design insights and some potential scaffolds which can be developed as CA IX inhibitors.Communicated by Ramaswamy H. Sarma.


The N-terminal PIN domain of the exosome subunit Rrp44 harbors endonuclease activity and tethers Rrp44 to the yeast core exosome.
Nuclear and cytoplasmic forms of the yeast exosome share 10 components, of which only Rrp44/Dis3 is believed to possess 3' exonuclease activity. We report that expression only of Rrp44 lacking 3'-exonuclease activity (Rrp44-exo) supports growth in S288c-related strains (BY4741). In BY4741, rrp44-exo was synthetic-lethal with loss of the cytoplasmic 5'-exonuclease Xrn1, indicating block of mRNA turnover, but not with loss of the nuclear 3'-exonuclease Rrp6. The RNA processing phenotype of rrp44-exo was milder than that seen on Rrp44 depletion, indicating that Rrp44-exo retains important functions. Recombinant Rrp44 was shown to possess manganese-dependent endonuclease activity in vitro that was abolished by four point mutations in the putative metal binding residues of its N-terminal PIN domain. Rrp44 lacking both exonuclease and endonuclease activity failed to support growth in strains depleted of endogenous Rrp44. Strains expressing Rrp44-exo and Rrp44-endo-exo exhibited different RNA processing patterns in vivo suggesting Rrp44-dependent endonucleolytic cleavages in the 5'-ETS and ITS2 regions of the pre-rRNA. Finally, the N-terminal PIN domain was shown to be necessary and sufficient for association with the core exosome, indicating its dual function as a nuclease and structural element.


The earliest cognitive change in a person with familial Alzheimer's disease: presymptomatic neuropsychological features in a pedigree with familial Alzheimer's disease confirmed at necropsy.
Comprehensive, longitudinal neuropsychological assessments are reported in a person "at risk" from autosomal dominant, necropsy confirmed familial Alzheimer's disease. The first assessment showed a moderately selective verbal memory deficit in the context of mild general intellectual impairment. Subsequent testing showed the progressive deterioration of visual memory and a mild decline of perceptual and spatial skills. Language and literacy skills, however, remained comparatively intact. The neuropsychological profiles obtained at each assessment are presented in profile maps. These permit direct longitudinal comparison of cognitive function, and may serve in the comparison of different potential cases of familial Alzheimer's disease. This case sought medical attention for memory difficulties 26 months after the first neuropsychological assessment. These results mark the first cognitive manifestations in a pedigree with familial Alzheimer's disease which, in this case, were seen presymptomatically. The findings are discussed in relation to neuropsychological studies of affected cases, and in terms of their reflecting the heterogeneous nature of familial Alzheimer's disease.


Chronological changes in the eighth cranial nerve compound action potential (CAP) in experimental endolymphatic hydrops: the effects of altering the polarity of click sounds.
Using a guinea pig model of experimental endolymphatic hydrops, click sounds of altered polarity showed different latencies and amplitudes in hydropic compared with normal cochleae. Latency changes appeared as early as 1 week after endolymphatic obstruction. This method can help diagnose endolymphatic hydrops. The goal of the study was to develop an objective electrophysiological diagnosis of endolymphatic hydrops. Endolymphatic hydrops were created surgically in guinea pigs. The latency and the amplitude of the eighth cranial nerve compound action potential (CAP) for click sounds of altered polarity were measured up to 8 weeks after the surgery. At early stages after surgery, the latency for condensation clicks became longer, and at later stages the latencies for both condensation and rarefaction became longer. The discrepancy in the latencies for rarefaction and condensation click sounds (rarefaction minus condensation) became larger by the first week after surgery, but no further discrepancy occurred thereafter. Compared with latency changes, amplitude changes in the CAP were rapid and progressive following surgery, suggesting ongoing damage to hair cells.


Resveratrol induces apoptosis in breast cancer cells by E2F1-mediated up-regulation of ASPP1.
Resveratrol is a natural polyphenolic compound with cancer chemopreventive activity. However, our understanding of the molecular mechanism responsible for resveratrol-induced apoptosis is still very limited. Here, we used MCF-7 and MDA-MB231 breast cancer cells as a model to demonstrate that resveratrol induced the expression of ASPP1, a new member of the ASPP (apoptosis stimulation protein of p53) family, which plays an important role in the regulation of apoptosis. Moreover, resveratrol enhanced apoptosis of MCF-7/ASPP1 cells, accompanied by higher expression of bax and p21. In contrast, siRNA-mediated knockdown of ASPP1 inhibited apoptosis in MB231 cells. Furthermore, we found that higher levels of ASPP1 were associated with adenovirus-mediated overexpression of E2F1 while siRNA-mediated E2F1 knockdown led to down-regulation of ASPP1. In conclusion, our results demonstrate that overexpression of ASPP1 rendered MCF-7 and MDA-MB231 breast cancer cells more sensitive to resveratrol-mediated apoptosis via the E2F pathway, thus suggesting that ASPP1 may represent a novel therapeutic target for resveratrol in human breast cancer.


Coal mine dust lung disease in the modern era.
Coal workers' pneumoconiosis (CWP), as part of the spectrum of coal mine dust lung disease (CMDLD), is a preventable but incurable lung disease that can be complicated by respiratory failure and death. Recent increases in coal production from the financial incentive of economic growth lead to higher respirable coal and quartz dust levels, often associated with mechanization of longwall coal mining. In Australia, the observed increase in the number of new CWP diagnoses since the year 2000 has necessitated a review of recommended respirable dust exposure limits, where exposure limits and monitoring protocols should ideally be standardized. Evidence that considers the regulation of engineering dust controls in the mines is lacking even in high-income countries, despite this being the primary preventative measure. Also, it is a global public health priority for at-risk miners to be systemically screened to detect early changes of CWP and to include confirmed patients within a central registry; a task limited by financial constraints in less developed countries. Characteristic X-ray changes are usually categorized using the International Labour Office classification, although future evaluation by low-dose HRCT) chest scanning may allow for CWP detection and thus avoidance of further exposure, at an earlier stage. Preclinical animal and human organoid-based models are required to explore potential re-purposing of anti-fibrotic and related agents with potential efficacy. Epidemiological patterns and the assessment of molecular and genetic biomarkers may further enhance our capacity to identify susceptible individuals to the inhalation of coal dust in the modern era.


A motivational factors assessment instrument for medication management center employees providing medication therapy management services: A Rasch analysis.
Telephonic medication management centers (MMCs) provide medication therapy management services to help alleviate medication-related issues and improve health outcomes. Motivation factors driving the performance of MMC personnel are key components in achieving these goals. Yet, published literature is limited on how motivation affects MMC personnel performance. To assess validity and reliability of the employee motivation questionnaire (EMQ), a 19-item measure of barriers and facilitators to motivation associated with MMC employees' work performance. Pharmacist, nurse, pharmacy technician, and intern employees (N = 534) from 5 telephonic-based U.S. MMCs were invited to complete the electronic EMQ. Rasch analysis was conducted in Winsteps software using a rating scale model. Construct and content validity and reliability were analyzed with employee and item separation index (SI) and reliability coefficient (RC). Linear regression was conducted to test the association of employee characteristics with individual work performance motivation. A total of 319 employees completed the EMQ, 59.7% response rate. Principal components analysis suggested a unidimensional construct. Employee and item infit and outfit mean squared values met recommended fit criteria (0.5-1.5), suggesting that the data fit the model. An item-person map identified items that were easiest (joy of helping patients) and most difficult (motivated to work harder if incentives were tied to goal achievements) to agree with. Mismatch of employee motivation and item difficulty level on the measurement continuum (-1 to 0.92 logits) indicated a need for additional items that employees perceive as more difficult to agree with. The employee RC was 0.81 and the SI was 2.04; whereas, the item RC was 0.97 and the SI was 5.94. None of the variables tested illustrated statistically significant associations with the person motivation measure. The EMQ illustrated reasonable content validity, good construct validity, and reliability evidence when used to measure motivation factors among MMC employees. Consideration of employee motivation factors may help to better meet MMC program goals and improve patient outcomes.


Improved localization of hidden fluorescent objects in highly scattering slab media based on a two-way transmittance determination.
We present a novel procedure for localizing fluorescing-tagged objects embedded in turbid slab media from fluorescent intensity profiles acquired along a surface of interest. Using a numerical model based on a finite element code, we firstly develop a method devoted to lateral detection by varying the laser source position along one face of the tissue slab. Next, we mainly demonstrate the possibility to accurately assess the depth location by alternately changing the position of the source and the detector at the both sides of the slab. The dimensionless depth indicator derived from this procedure remains independent, over a wide range, on both the optical properties of the host tissue and the probe concentration. The overall findings validate the method in situations involving moderate size object-like tumors tagged with a new smart contrast agent (Cy 5.5) that offers high tumor-to-background contrast and great interest in early cancer diagnostic.


Dyskeratosis congenita.
Dyskeratosis congenita is a rare disorder that often leads to early death owing to a variety of complications and associated disorders. Early diagnosis and intervention is important in care for patients affected by this disease. We describe a patient with dyskeratosis congenita (DC) in a child. Our patient had a 3-year history of transaminitis that was felt to be the result of biopsy proven progressive fibrosis of the liver beginning at age 3. He was referred to the dermatology department because of a chronic, evolving eruption with the hope of establishing a unifying diagnosis. Further examination revealed dystrophic nails, numerous dental caries, and blepharitis. Chromosomal analysis on leukocytes showed significant telomere shortening consistent with DC. Early recognition and long term care is important in patients with DC because of their propensity to develop malignancy, hematologic abnormalities, and infection. Better understanding of this disease may lead to insights into other disorders associated with abnormal telomere maintenance.


Anti-smoking parenting practices: recall by and effect on children's risk of smoking after 3 years.
Parent engagement in anti-smoking parenting practices was examined as a predictor of children's recalled exposure to these practices, the presence of pro-smoking risk factors in children's social environments, and children's odds of initiating smoking. 1,032 parents reported level of engagement in a program that promoted anti-smoking parenting practices for 8-year-old children. 1,032 children were surveyed 6 months and 3 years post-intervention; they reported on exposure to anti-smoking parenting practices, pro-smoking risk factors, and initiation of smoking. If parents reported high engagement in anti-smoking socialization, children had significantly greater recall of anti-smoking parenting practices and significantly fewer pro-smoking risk factors up to 3 years post-intervention. If engagement was moderate or low, children had progressively lower odds of recalling anti-smoking parenting practices relative to controls and they were progressively less likely to differ from controls in exposure to pro-smoking risk factors at follow-up. Achieving full program implementation remains a significant challenge to home-based, parent-led approaches to smoking prevention. However, if parents fully engage in anti-smoking parenting practices, children demonstrate protective effects up to 3 years later.


Two Y genes can replace the entire Y chromosome for assisted reproduction in the mouse.
The Y chromosome is thought to be important for male reproduction. We have previously shown that, with the use of assisted reproduction, live offspring can be obtained from mice lacking the entire Y chromosome long arm. Here, we demonstrate that live mouse progeny can also be generated by using germ cells from males with the Y chromosome contribution limited to only two genes, the testis determinant factor Sry and the spermatogonial proliferation factor Eif2s3y. Sry is believed to function primarily in sex determination during fetal life. Eif2s3y may be the only Y chromosome gene required to drive mouse spermatogenesis, allowing formation of haploid germ cells that are functional in assisted reproduction. Our findings are relevant, but not directly translatable, to human male infertility cases.


Cooperation between the Hepatitis C Virus p7 and NS5B Proteins Enhances Virion Infectivity.
The molecular mechanisms that govern hepatitis C virus (HCV) assembly, release, and infectivity are still not yet fully understood. In the present study, we sequenced a genotype 2A strain of HCV (JFH-1) that had been cell culture adapted in Huh-7.5 cells to produce nearly 100-fold-higher viral titers than the parental strain. Sequence analysis identified nine mutations in the genome, present within both the structural and nonstructural genes. The infectious clone of this virus containing all nine culture-adapted mutations had 10-fold-higher levels of RNA replication and RNA release into the supernatant but had nearly 1,000-fold-higher viral titers, resulting in an increased specific infectivity compared to wild-type JFH-1. Two mutations, identified in the p7 polypeptide and NS5B RNA-dependent RNA polymerase, were sufficient to increase the specific infectivity of JFH-1. We found that the culture-adapted mutation in p7 promoted an increase in the size of cellular lipid droplets following transfection of viral RNA. In addition, we found that the culture-adaptive mutations in p7 and NS5B acted synergistically to enhance the specific viral infectivity of JFH-1 by decreasing the level of sphingomyelin in the virion. Overall, these results reveal a genetic interaction between p7 and NS5B that contributes to virion specific infectivity. Furthermore, our results demonstrate a novel role for the RNA-dependent RNA polymerase NS5B in HCV assembly. Hepatitis C virus assembly and release depend on viral interactions with host lipid metabolic pathways. Here, we demonstrate that the viral p7 and NS5B proteins cooperate to promote virion infectivity by decreasing sphingomyelin content in the virion. Our data uncover a new role for the viral RNA-dependent RNA polymerase NS5B and p7 proteins in contributing to virion morphogenesis. Overall, these findings are significant because they reveal a genetic interaction between p7 and NS5B, as well as an interaction with sphingomyelin that regulates virion infectivity. Our data provide new strategies for targeting host lipid-virus interactions as potential targets for therapies against HCV infection.


Health and health systems in the Commonwealth of Independent States.
The countries of the Commonwealth of Independent States differ substantially in their post-Soviet economic development but face many of the same challenges to health and health systems. Life expectancies dropped steeply in the 1990s, and several countries have yet to recover the levels noted before the dissolution of the Soviet Union. Cardiovascular disease is a much bigger killer in the Commonwealth of Independent States than in western Europe because of hazardous alcohol consumption and high smoking rates in men, the breakdown of social safety nets, rising social inequality, and inadequate health services. These former Soviet countries have embarked on reforms to their health systems, often aiming to strengthen primary care, scale back hospital capacities, reform mechanisms for paying providers and pooling funds, and address the overall shortage of public funding for health. However, major challenges remain, such as frequent private out-of-pocket payments for health care and underdeveloped systems for improvement of quality of care.


Molecular and morphological evidence supports the species status of the Mahachai fighter Betta sp. Mahachai and reveals new species of Betta from Thailand.
Two regions of mitochondrial (mt) DNA, cytochrome c oxidase subunit 1 (COI) and 16S rRNA, were sequenced in nine species of Betta from Thailand and Indonesia. Most species showed little intraspecific COI variation (adjusted mean = 0.48%) including the putative species Betta sp. Mahachai, but one species (Betta smaragdina) included three lineages showing much greater divergence (7.03-13.48%) that probably represent overlooked species. These findings were confirmed by maximum likelihood analysis and Bayesian inference, which revealed well-supported corresponding monophyletic clades. Based on these results and morphological differences, the putative species Betta sp. Mahachai from central Thailand is a species distinct from other members of the B. splendens group and represents a new and hitherto undescribed species. Furthermore, this study also demonstrated the probable existence of two overlooked Betta species found in the Khorat plateau basin, illustrating the utility of mitochondrial genetic markers in the revelation of overlooked diversity.


ATP-Sensitive Potassium Channels in Hyperinsulinism and Type 2 Diabetes: Inconvenient Paradox or New Paradigm?
Secretion of insulin from pancreatic β-cells is complex, but physiological glucose-dependent secretion is dominated by electrical activity, in turn controlled by ATP-sensitive potassium (KATP) channel activity. Accordingly, loss-of-function mutations of the KATP channel Kir6.2 (KCNJ11) or SUR1 (ABCC8) subunit increase electrical excitability and secretion, resulting in congenital hyperinsulinism (CHI), whereas gain-of-function mutations cause underexcitability and undersecretion, resulting in neonatal diabetes mellitus (NDM). Thus, diazoxide, which activates KATP channels, and sulfonylureas, which inhibit KATP channels, have dramatically improved therapies for CHI and NDM, respectively. However, key findings do not fit within this simple paradigm: mice with complete absence of β-cell KATP activity are not hyperinsulinemic; instead, they are paradoxically glucose intolerant and prone to diabetes, as are older human CHI patients. Critically, despite these advances, there has been little insight into any role of KATP channel activity changes in the development of type 2 diabetes (T2D). Intriguingly, the CHI progression from hypersecretion to undersecretion actually mirrors the classical response to insulin resistance in the progression of T2D. In seeking to explain the progression of CHI, multiple lines of evidence lead us to propose that underlying mechanisms are also similar and that development of T2D may involve loss of KATP activity.