SIK2 inhibition enhances PARP inhibitor activity synergistically in ovarian and triple-negative breast cancers.
Poly(ADP-ribose) polymerase inhibitors (PARP inhibitors) have had an increasing role in the treatment of ovarian and breast cancers. PARP inhibitors are selectively active in cells with homologous recombination DNA repair deficiency caused by mutations in BRCA1/2 and other DNA repair pathway genes. Cancers with homologous recombination DNA repair proficiency respond poorly to PARP inhibitors. Cancers that initially respond to PARP inhibitors eventually develop drug resistance. We have identified salt-inducible kinase 2 (SIK2) inhibitors, ARN3236 and ARN3261, which decreased DNA double-strand break (DSB) repair functions and produced synthetic lethality with multiple PARP inhibitors in both homologous recombination DNA repair deficiency and proficiency cancer cells. SIK2 is required for centrosome splitting and PI3K activation and regulates cancer cell proliferation, metastasis, and sensitivity to chemotherapy. Here, we showed that SIK2 inhibitors sensitized ovarian and triple-negative breast cancer (TNBC) cells and xenografts to PARP inhibitors. SIK2 inhibitors decreased PARP enzyme activity and phosphorylation of class-IIa histone deacetylases (HDAC4/5/7). Furthermore, SIK2 inhibitors abolished class-IIa HDAC4/5/7-associated transcriptional activity of myocyte enhancer factor-2D (MEF2D), decreasing MEF2D binding to regulatory regions with high chromatin accessibility in FANCD2, EXO1, and XRCC4 genes, resulting in repression of their functions in the DNA DSB repair pathway. The combination of PARP inhibitors and SIK2 inhibitors provides a therapeutic strategy to enhance PARP inhibitor sensitivity for ovarian cancer and TNBC.


Propofol prevents autophagic cell death following oxygen and glucose deprivation in PC12 cells and cerebral ischemia-reperfusion injury in rats.
Propofol exerts protective effects on neuronal cells, in part through the inhibition of programmed cell death. Autophagic cell death is a type of programmed cell death that plays elusive roles in controlling neuronal damage and metabolic homeostasis. We therefore studied whether propofol could attenuate the formation of autophagosomes, and if so, whether the inhibition of autophagic cell death mediates the neuroprotective effects observed with propofol. The cell model was established by depriving the cells of oxygen and glucose (OGD) for 6 hours, and the rat model of ischemia was introduced by a transient two-vessel occlusion for 10 minutes. Transmission electron microscopy (TEM) revealed that the formation of autophagosomes and autolysosomes in both neuronal PC12 cells and pyramidal rat hippocampal neurons after respective OGD and ischemia/reperfusion (I/R) insults. A western blot analysis revealed that the autophagy-related proteins, such as microtubule-associated protein 1 light chain 3 (LC3-II), Beclin-1 and class III PI3K, were also increased accordingly, but cytoprotective Bcl-2 protein was decreased. The negative effects of OGD and I/R, including the formation of autophagosomes and autolysosomes, the increase in LC3-II, Beclin-1 and class III PI3K expression and the decline in Bcl-2 production were all inhibited by propofol and specific inhibitors of autophagy, such as 3-methyladenine (3-MA), LY294002 and Bafilomycin A1 (Baf),. Furthermore, in vitro OGD cultures and in vivo I/R rats showed an increase in cell survival following the administration of propofol, as assessed by an MTT assay or histochemical analyses. Our data suggest that propofol can markedly attenuate autophagic processes via the decreased expression of autophagy-related proteins in vitro and in vivo. This inhibition improves cell survival, which provides a novel explanation for the pleiotropic effects of propofol that benefit the nervous system.


Precise alignment of sites required for mu enhancer activation in B cells.
The lymphocyte-specific immunoglobulin mu heavy-chain gene intronic enhancer is regulated by multiple nuclear factors. The previously defined minimal enhancer containing the muA, muE3, and muB sites is transactivated by a combination of the ETS-domain proteins PU.1 and Ets-1 in nonlymphoid cells. The core GGAAs of the muA and muB sites are separated by 30 nucleotides, suggesting that ETS proteins bind to these sites from these same side of the DNA helix. We tested the necessity for appropriate spatial alignment of these elements by using mutated enhancers with altered spacings. A 4- or 10-bp insertion between muE3 and muB inactivated the mu enhancer in S194 plasma cells but did not affect in vitro binding of Ets-1, PU.1, or the muE3-binding protein TFE3, alone or in pairwise combinations. Circular permutation and phasing analyses demonstrated that PU.1 binding but not TFE3 or Ets-1 bends mu enhancer DNA toward the major groove. We propose that the requirement for precise spacing of the muA and muB elements is due in part to a directed DNA bend induced by PU.1.


Post-translational modifications enhance NT-proBNP and BNP production in acute decompensated heart failure.
Increases in plasma B-type natriuretic peptide (BNP) concentrations in those with acutely decompensated heart failure (ADHF) has been mainly attributed to an increase in NPPB gene transcription. Recently, proBNP glycosylation has emerged as a potential regulatory mechanism in the production of amino-terminal (NT)-proBNP and BNP. The aim of the present study was to investigate proBNP glycosylation, and corin and furin activities in ADHF patients. Plasma levels of proBNP, NT-proBNP, BNP, as well as corin and furin concentration and activity were measured in a large cohort of 683 patients presenting with ADHF (n = 468), non-cardiac dyspnoea (non-ADHF: n = 169) and 46 patients with stable chronic heart failure (CHF); the degree of plasma proBNP glycosylation was assessed in a subset of these patients (ADHF: n = 49, non-ADHF: n = 50, CHF: n = 46). Our results showed a decrease in proBNP glycosylation in ADHF patients that paralleled NT-proBNP overproduction (ρ = -0.62, P < 0.001) but less so to BNP. In addition, we observed an increase in furin activity that is positively related to the plasma levels of proBNP, NT-proBNP and BNP overproduction (all P < 0.001, all ρ > 0.88), and negatively related to the degree of proBNP glycosylation (ρ = -0.62, P < 0.001). These comprehensive results provide a paradigm for the post-translational modification of natriuretic peptides in ADHF: as proBNP glycosylation decreases, furin activity increases. This synergistically amplifies the processing of proBNP into BNP and NT-proBNP. http://clinicaltrials.gov/. Identifier: NCT01374880.


Relation between changes in weight parameters and height parameters in prepubertal children: daily weight gain and BMIi changes in relation to linear growth during nutritional rehabilitation of underweight children.
Early detection of abnormal weight loss or gain in childhood may be important for preventive purposes. Variable growth response to nutrition rehabilitation have been reported in children with failure to thrive (FTT) who do not have any chronic disease or systematic illness due to different clinical and nutritional approach in their management. To analyze the association of different body mass index (BMI) and BMI- SDS, to linear growth (Ht-SDS) in different BMI categories of prepubertal children. In addition, we studied the effect of weight changes on linear growth in a randomly selected group of prepubertal underweight children who received nutritional rehabilitation (NR) for 9±2 months. 102 children, between 1 and 9 years, followed at the General Pediatric Clinic, between January 2017 to December 2017, because of abnormal weight gain (decreased or increased) which was not associated with any acute or chronic illness were included in the study. Anthropometric measurements included weigh, height, Ht-SDS, BMI, and BMI-SDS. Children BMI-SDS were categorized into 4 groups: Group 1: BMI-SDS <-2, group 2: BMI-SDS <-1 but >-2, group 3 BMI-SDS >-1 but <2, group 4 BMI-SDS >2. We also evaluated the effects of weight changes on linear growth in a randomly selected group of underweight children who received nutritional counselling and oral nutritional supplementation (n = 51) for 9±months. HT-SDS in children of groups 1 and 2 (underweight and at risk of underweight children) was significantly lower than Ht-SDS of groups 3 and 4 (normal and overweight children). Ht-SDS in children of group 4 was significantly higher than the Ht-SDS of children in group 3. A significant linear correlation was found between BMI-SDS and Ht-SDS in these prepubertal children. After nutritional rehabilitation for a year, 55% of underweight children increased their BMI-SDS and 43% increased their Ht-SDS. Children who had weight gain >7g/d, over the whole period of follow-up, (n =14) increased their BMI-SDS and Ht-SDS significantly after versus before NR. The BMI-SDS and Ht-SDS did not increase significantly in the group of children who had weight gain <7 g/day. 28 children out of 51 improved their BMI-SDS after nutritional rehabilitation (group A) and 23 did not have improvement in their BMI-SDS (Group B). Group A had higher weight gain per day versus group B. Height growth velocity was significantly higher in Group B (7.4±3.6 cm/yr) versus group A (5.7±2.8 cm/yr). Ht-SDS increased significantly in the group of patients who had lower Ht-SDS before NR. Children who had faster linear growth velocity, after nutritional rehabilitation, did not increase their BMI-SDS. Linear regression showed a significant correlation between BMI-SDS and Ht-SDS supporting the notion that proper nutrition and maintaining normal BMI-SDS is essential for adequate gain in height. It appears that calculating the weight gain per day, BMI-SDS and Ht-SDS are clinically useful parameters to detect the effect of weight gain on linear growth and to monitor the nutritional management. Daily weight gain was correlated significantly to height growth rate during nutritional rehabilitation. Based on our findings and literature reports, we suggest an algorithm for follow-up of underweight/ malnourished children based mainly on anthropometric assessment.


Family Planning and the Samburu: A Qualitative Study Exploring the Thoughts of Men on a Population Health and Environment Programme in Rural Kenya.
Population Health and Environment (PHE) strategies are argued to improve ecosystem and human health by addressing family size and its effects on natural resource use, food security, and reproductive health. This study investigates men's views on a PHE family planning (FP) programme delivered among the pastoral Samburu tribe in rural northern Kenya. Three focus group discussions and nine semi-structured interviews were conducted with 27 Samburu men. These discussions revealed support for environmentally-sensitised family planning promotion. Men highlighted their dependency on natural resources and challenges faced in providing for large families and maintaining livestock during drought. These practices were said to lead to natural resource exhaustion, environmental degradation, and wildlife dispersal, undermining key economic benefits of environmental and wildlife conservation. Relating family size to the environment is a compelling strategy to improve support for FP among Samburu men. Kenyan policy-makers should consider integrating community-based PHE strategies among underserved pastoral groups living in fragile ecosystems.


Risk factors associated with occurrence of nematodes in free range pigs in Busia District, Kenya.
Nematode infections are a serious constraint to pig production, especially where free range pig keeping is practiced. This study investigated the epidemiology of nematodes in free range pigs in Busia District, Kenya. Three hundred and six pigs from 135 farms were sampled for faeces that were analysed for nematode eggs per gram (EPG) of faeces using the McMaster technique. The nematode eggs were also identified to genus and species based on morphology. A questionnaire on risk factors was also administered to the pig owners. The overall prevalence and mean nematode EPG were 84.2% and 2,355, respectively. The nematode eggs were identified as those belonging to Oesophagostomum spp. (75%), Strongyloides ransomi (37%), Ascaris suum (18%), Metastrongylus spp. (11%), Trichuris suis (7%) and Physocephalus sexalatus (3%). The prevalence of nematodes was positively correlated (p < 0.05) with the amount of rainfall in the division of the pigs' origin (all nematodes except S. ransomi). The prevalence of nematodes was also associated with the age of the pigs. A lower burden of nematodes was associated (p < 0.05) with a history of deworming (A. suum) and the provision of night housing (S. ransomi and Metastrongylus spp.). In conclusion, this study has provided information on nematode infections and the associated risk factors for free range pigs in Busia District, which can be used when implementing integrated control measures.


Recessive dystrophic epidermolysis bullosa results in painful small fibre neuropathy.
Small fibres in the skin are vulnerable to damage in metabolic or toxic conditions such as diabetes mellitus or chemotherapy resulting in small fibre neuropathy and associated neuropathic pain. Whether injury to the most distal portion of sensory small fibres due to a primary dermatological disorder can cause neuropathic pain is still unclear. Recessive dystrophic epidermolysis bullosa (RDEB) is a rare condition in which mutations of proteins of the dermo-epidermal junction lead to cycles of blistering followed by regeneration of the skin. Damage is exclusive to the skin and mucous membranes, with no known direct compromise of the nervous system. It is increasingly recognized that most RDEB patients experience daily pain, the aetiology of which is unclear but may include inflammation (in the wounds), musculoskeletal (due to atrophy and retraction scars limiting movement) or neuropathic pain. In this study we investigated the incidence of neuropathic pain and examined the presence of nerve dysfunction in RDEB patients. Around three quarters of patients presented with pain of neuropathic characteristics, which had a length-dependent distribution. Quantitative sensory testing of the foot revealed striking impairments in thermal detection thresholds combined with an increased mechanical pain sensitivity and wind up ratio (temporal summation of noxious mechanical stimuli). Nerve conduction studies showed normal large fibre sensory and motor nerve conduction; however, skin biopsy showed a significant decrease in intraepidermal nerve fibre density. Autonomic nervous system testing revealed no abnormalities in heart rate and blood pressure variability however the sympathetic skin response of the foot was impaired and sweat gland innervation was reduced. We conclude that chronic cutaneous injury can lead to injury and dysfunction of the most distal part of small sensory fibres in a length-dependent distribution resulting in disabling neuropathic pain. These findings also support the use of neuropathic pain screening tools in these patients and treatment algorithms designed to target neuropathic pain.


Cost Utility Analysis of Lumbar Interlaminar Epidural Injections in the Treatment of Lumbar Disc Herniation, Central Spinal Stenosis, and Axial or Discogenic Low Back Pain.
Cost utility or cost effective analysis continues to take center stage in the United States for defining and measuring the value of treatments in interventional pain management. Appropriate cost utility analysis has been performed for caudal epidural injections, percutaneous adhesiolysis, and spinal cord stimulation. However, the literature pertaining to lumbar interlaminar epidural injections is lacking, specifically in reference to cost utility analysis derived from randomized controlled trials (RCTs) with a pragmatic approach in a practical setting. To assess the cost utility of lumbar interlaminar epidural injections in managing chronic low back and/or lower extremity pain secondary to lumbar disc herniation, spinal stenosis, and axial or discogenic low back pain. Analysis based on 3 previously published randomized trials of effectiveness of lumbar interlaminar epidural injections assessing their role in disc herniation, spinal stenosis, and axial or discogenic pain. A contemporary, private, specialty referral interventional pain management center in the United States. Cost utility of lumbar interlaminar epidural injections with or without steroids in managing lumbar disc herniation, central spinal stenosis, and discogenic or axial low back pain was conducted with data derived from 3 RCTs that included a 2-year follow-up, with inclusion of 360 patients. The primary outcome was significant improvement defined as at least a 50% in pain reduction and disability status. Direct payment data from 2016 was utilized for assessment of procedural costs. Overall costs, including drug costs, were determined by multiplication of direct procedural payment data by a factor of 1.6 to accommodate for indirect payments respectively for disc herniation, spinal stenosis, discogenic pain. The results of 3 RCTs showed direct cost utility for one year of quality-adjusted life year (QALY) of $2,050.87 for disc herniation, $2,112.25 for axial or discogenic pain without disc herniation, and $1,773.28 for spinal stenosis, with an average cost per one year QALY of $1,976.58, with total estimated costs of $3,425, $3,527, $2,961, and $3,301 respectively. The limitation of this cost utility analysis includes that it is a single center evaluation, even though 360 patients were included in this analysis. Further, only the costs of interventional procedures and physician visits were assessed based on the data, with extrapolation of indirect costs presenting the overall total costs. The benefits of returning to work were not assessed. This cost utility analysis of lumbar interlaminar epidural injections in patients nonresponsive to conservative management in the treatment of disc herniation, central spinal stenosis, and axial or discogenic low back pain in the lumbar spine shows the clinical effectiveness and cost utility of these injections of $1,976.58 for direct costs with a total cost of $3,301 per QALY.


High-precision measurements of (33)S and (34)S fractionation during SO2 oxidation reveal causes of seasonality in SO2 and sulfate isotopic composition.
This study presents high-precision isotope ratio-mass spectrometric measurements of isotopic fractionation during oxidation of SO2 by OH radicals in the gas phase and H2O2 and transition metal ion catalysis (TMI-catalysis) in the aqueous phase. Although temperature dependence of fractionation factors was found to be significant for H2O2 and TMI-catalyzed pathways, results from a simple 1D model revealed that changing partitioning between oxidation pathways was the dominant cause of seasonality in the isotopic composition of sulfate relative to SO2. Comparison of modeled seasonality with observations shows the TMI-catalyzed oxidation pathway is underestimated by more than an order of magnitude in all current atmospheric chemistry models. The three reactions showed an approximately mass-dependent relationship between (33)S and (34)S. However, the slope of the mass-dependent line was significantly different to 0.515 for the OH and TMI-catalyzed pathways, reflecting kinetic versus equilibrium control of isotopic fractionation. For the TMI-catalyzed pathway, both temperature dependence and (33)S/(34)S relationship revealed a shift in the rate-limiting reaction step from dissolution at lower temperatures to TMI-sulfite complex formation at higher temperatures. 1D model results showed that although individual reactions could produce Δ(33)S values between -0.15 and +0.2‰, seasonal changes in partitioning between oxidation pathways caused average sulfate Δ(33)S values of 0‰ throughout the year.


The use, misuse, and abuse of design controls.
FDA-mandated design controls for medical devices provide a structured, systematic engineering paradigm that supports human-centered systems engineering. Engineering design controls are nothing more than the fundamental elements of classical systems engineering. The human focus is enabled through the iterative reidentification of stakeholders, reassessment of their NWDs, and reconciliation of their evolving/conflicting NWDs. The implementation of design controls and embedded risk management must begin prior to commercial development to reap the full benefit of the approach; partial approaches dilute or negate the effectiveness and efficiency of this nearly century-old systems engineering paradigm. Properly employing engineering design controls is a strategic business decision. The central value of this proposition is the reduction of economic, technical, and operational risks for both producers and consumers; regulatory compliance is a secondary benefit. Misuse or abuse of design controls only undermines long-term profitability and increases the risks to the consumer.


Port-site and liver metastases after laparoscopic pelvic and para-aortic lymph node dissection for surgical staging of locally advanced cervical cancer.
Recently, we experienced a case of port-site and liver metastases after 75 cases of laparoscopic transperitoneal pelvic lymph node dissection (PLND) and para-aortic lymph node dissection (PALND) for surgical staging of locally advanced cervical cancer. A 45-year-old-woman with stage IIB cervical adenocarcinoma underwent laparoscopic PLND and PALND for surgical staging. There was no intraperitoneal disease and cervical tumor was not manipulated at the time of laparoscopic surgery. Pathologic examination revealed only one micrometastasis in left internal iliac lymph node (LN), measuring 1 mm, of the 60 pelvic and para-aortic LNs removed. She received concurrent chemoradiation therapy and pelvic mass disappeared completely. One month after the completion of therapy, both lateral and umbilical port-site and liver metastases were detected. We conclude that although cases of port-site metastasis have mostly occurred after extensive disease, the possibility of such complication should be kept in mind at laparoscopy of early cancer and laparoscopy which does not manipulate primary tumor.


Cardiac natriuretic peptides act via p38 MAPK to induce the brown fat thermogenic program in mouse and human adipocytes.
The ability of mammals to resist body fat accumulation is linked to their ability to expand the number and activity of "brown adipocytes" within white fat depots. Activation of β-adrenergic receptors (β-ARs) can induce a functional "brown-like" adipocyte phenotype. As cardiac natriuretic peptides (NPs) and β-AR agonists are similarly potent at stimulating lipolysis in human adipocytes, we investigated whether NPs could induce human and mouse adipocytes to acquire brown adipocyte features, including a capacity for thermogenic energy expenditure mediated by uncoupling protein 1 (UCP1). In human adipocytes, atrial NP (ANP) and ventricular NP (BNP) activated PPARγ coactivator-1α (PGC-1α) and UCP1 expression, induced mitochondriogenesis, and increased uncoupled and total respiration. At low concentrations, ANP and β-AR agonists additively enhanced expression of brown fat and mitochondrial markers in a p38 MAPK-dependent manner. Mice exposed to cold temperatures had increased levels of circulating NPs as well as higher expression of NP signaling receptor and lower expression of the NP clearance receptor (Nprc) in brown adipose tissue (BAT) and white adipose tissue (WAT). NPR-C(-/-) mice had markedly smaller WAT and BAT depots but higher expression of thermogenic genes such as Ucp1. Infusion of BNP into mice robustly increased Ucp1 and Pgc-1α expression in WAT and BAT, with corresponding elevation of respiration and energy expenditure. These results suggest that NPs promote "browning" of white adipocytes to increase energy expenditure, defining the heart as a central regulator of adipose tissue biology.


Combined Usage of MDK Inhibitor Augments Interferon-γ Anti-Tumor Activity in the SKOV3 Human Ovarian Cancer Cell Line.
Ovarian cancer (OC) is a particularly lethal disease due to intratumoral heterogeneity, resistance to traditional chemotherapy, and poor response to targeted therapy and immunotherapy. Interferon-γ (IFN-γ) is an attractive therapeutic cytokine, with positive responses achieved in multiple OC clinical trials. However, clinical application of IFN-γ in OC is still hindered, due to the severe toxicity when used at higher levels, as well as the considerable pro-metastatic adverse effect when used at lower levels. Thus, an effective combined intervention is needed to enhance the anti-tumor efficacy of IFN-γ and to suppress the IFN-γ-induced metastasis. Here, we uncovered that OC cells develop an adaptive strategy by upregulating midkine (MDK) to counteract the IFN-γ-induced anti-tumor activity and to fuel IFN-γ-induced metastasis. We showed that MDK is a critical downstream target of IFN-γ in OC, and that this regulation acts in a dose-dependent manner and is mediated by STAT1. Gain-of-function studies showed that MDK overexpression promotes cell proliferation and metastasis in OC, indicating that IFN-γ-activated MDK may antagonize IFN-γ in inhibiting OC proliferation but synergize IFN-γ in promoting OC metastasis. Subsequently, we assessed the influence of MDK inhibition on IFN-γ-induced anti-proliferation and pro-metastasis effects using an MDK inhibitor (iMDK), and we found that MDK inhibition robustly enhanced IFN-γ-induced growth inhibition (all CIs < 0.1) and reversed IFN-γ-driven epithelial-to-mesenchymal transition (EMT) and metastasis in OC in vitro. Collectively, these data identify an IFN-γ responsive protein, MDK, in counteracting anti-proliferation while endowing the pro-metastatic role of IFN-γ in cancer treatment, and we therefore propose the combined utilization of the MDK inhibitor in IFN-γ-based therapies in future OC treatment.


Mobile farm clinic outreach to address health conditions among Latino migrant farmworkers in Georgia.
Agricultural labor involves exposure to many occupational hazards, some of which can lead to chronic health conditions. The purpose of this study was to conduct an occupational health needs assessment of illnesses and work-related injuries among a Latino migrant farmworker population (recruited to harvest Vidalia onions) in South Georgia. Study data included survey responses from 100 farmworkers attending mobile farm clinics in 2010 at their worker housing residences, supplemented by medical diagnoses data from the same clinics collected over 3 years (2009-2011) for 1161 farmworkers at six different farms. From the survey, the main health problems reported were hypertension (25%), eye problems (12%), musculoskeletal problems (11%), diabetes (10%), and depression (7%). In multivariate analyses, depression scores were associated with having a history of musculoskeletal problems (p = .002). According to the mobile farm clinic data, the most common medical diagnoses included back pain (11.8%), hypertension (11.4%), musculoskeletal problems (11.3%), gastrointestinal disorders (8.6%), eye problems (7.2%), dermatitis or rash (7.0%), and tinea or fungal skin infections (5.6%). The study identified eye and musculoskeletal problems as the major occupational health conditions for this population of farmworkers.


A review of the emerging treatment technologies for PFAS contaminated soils.
Contamination of soils with poly- and perfluoroalkyl substances (PFAS) has become a challenging issue due to the adverse effects of these substances on both the environment and public health. PFAS have strong chemical structures and their bonding with soil makes them challenging to eliminate from soil environments. Traditional methods of soil remediation have not been successful in their reduction or removal from the environment. This paper provides a comprehensive evaluation of existing and emerging technologies for remediating PFAS contaminated soils with guidance on which approach to use in different contexts. The functions of all remediation technologies, their suitability, limitations, and the scale applied from laboratory to the field are presented as a baseline for understanding the research need for treatment in soil environments. To date, the immobilization method has been a significant part of the remediation solution for PFAS contaminated soils, although its long-term efficiency still needs further investigation. Soil washing and thermal treatment techniques have been tested at the field scale, but they are expensive and energy-intensive due to the use of a large volume of washing solvent and the high melting point of PFAS, respectively; both methods need a large initial investment for their installation. Other remediation technologies, such as chemical oxidation, ball milling, and electron beams, have been progressed in the laboratory. However, additional research is needed to make them feasible, cost-effective and applicable in the field.


U.S. Physician-Scientist Workforce in the 21st Century: Recommendations to Attract and Sustain the Pipeline.
The U.S. physician-scientist (PS) workforce is invaluable to the nation's biomedical research effort. It is through biomedical research that certain diseases have been eliminated, cures for others have been discovered, and medical procedures and therapies that save lives have been developed. Yet, the U.S. PS workforce has both declined and aged over the last several years. The resulting decreased inflow and outflow to the PS pipeline renders the system vulnerable to collapsing suddenly as the senior workforce retires. In November 2015, the Alliance for Academic Internal Medicine hosted a consensus conference on the PS workforce to address issues impacting academic medical schools, with input from early-career PSs based on their individual experiences and concerns. One of the goals of the conference was to identify current impediments in attracting and supporting PSs and to develop a new set of recommendations for sustaining the PS workforce in 2016 and beyond. This Perspective reports on the opportunities and factors identified at the conference and presents five recommendations designed to increase entry into the PS pipeline and nine recommendations designed to decrease attrition from the PS workflow.


Further assessment of the effects of occupational radiation exposure in the United Kingdom Atomic Energy Authority mortality study.
The United Kingdom Atomic Energy Authority mortality study was designed to investigate the relation between exposure to ionising radiation and mortality among the authority's employees. The present paper describes some of the problems encountered in assessing occupational exposure to low dose radiation and examines whether the study's conclusions about the relation between exposure and mortality could be affected by the methods used. The study covered the years 1946 to 1979 during which time the frequency with which personal film dosimeters were issued changed from weekly to monthly, and the threshold level below which measurements were not made decreased 20-fold. Exposure from "below threshold" readings made an important contribution to total exposure in the early years. Estimates, based on the remeasurement of a sample of old films, indicated that the average whole body exposure before 1961 may have been about double that which was measured. Furthermore, although records were kept of when dosimeters were lost or damaged, the associated exposures were unknown and could only be estimated. Workers whose dosimeter readings were missing for more than 5% of the time during which they were monitored had higher all cause mortality (p = 0.04) and higher mortality from accidents and violence (p = 0.05) than other radiation workers. The results of analyses of mortality in relation to whole body exposure were compared when the exposures included estimates of the below threshold and missing exposures and when these exposures were assumed to be zero. Some of the findings differed, but none changed sufficiently to alter the general conclusions. Although the trend in mortality from all cancers changed from one in which the increase with exposure was far from statistically significant (p = 0.3) when the below threshold and missing values were assumed to be zero to one that approached significance (p = 0.06) after they were estimated, calculations of the annual excess deaths from cancer per unit dose resulted in broadly similar estimates. Studies of workers exposed to ionising radiation usually focus on mortality in relation to whole body exposure. In the present paper its relation to neutron and surface exposure is also examined. Workers with measured neutron exposures had significantly lower all cause mortality than other workers with a radiation record (p = 0.03). Surface exposure was significantly related to mortality from all cancers (p = 0.02) and prostatic cancer (p less than 0.001). Some data on cancer registration are presented but these cannot be readily interpreted because cancer registration details were available only for ex-employees who may not be typical of the workforce as a whole.


The immune tumour microenvironment of neuroendocrine tumours and its implications for immune checkpoint inhibitors.
Immunotherapy in the form of immune checkpoint inhibitors (ICIs) has transformed the treatment landscape in numerous types of advanced cancer. However, the majority of patients do not benefit from this treatment modality. Although data are scarce, in general, patients with low-grade neuroendocrine tumours (NETs) do not benefit from treatment with ICIs in contrast to patients with neuroendocrine carcinoma, in which a small subgroup of patients may benefit. Low- and intermediate-grade NETs predominantly lack factors associated with response to ICIs treatment, like immune cell infiltration, and have an immunosuppressive tumour metabolism and microenvironment. In addition, because of its potential influence on the response to ICIs, major interest has been shown in the tryptophan-degrading enzymes indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO). These enzymes work along the kynurenine pathway that deplete tryptophan in the tumour microenvironment. IDO and TDO are especially of interest in NETs since some tumours produce serotonin but the majority do not, which potentially deplete the precursor tryptophan. In this review, we summarize the current knowledge on the immune tumour microenvironment of neuroendocrine tumours and implications for treatment with immune checkpoint inhibitors. We also discuss (targetable) factors in the NET tumour microenvironment that potentially modulate the anti-cancer immune response.


Expression of Cpgp40/15 in Toxoplasma gondii: a surrogate system for the study of Cryptosporidium glycoprotein antigens.
Cryptosporidium parvum is a waterborne enteric coccidian that causes diarrheal disease in a wide range of hosts. Development of successful therapies is hampered by the inability to culture the parasite and the lack of a transfection system for genetic manipulation. The glycoprotein products of the Cpgp40/15 gene, gp40 and gp15, are involved in C. parvum sporozoite attachment to and invasion of host cells and, as such, may be good targets for anticryptosporidial therapies. However, the function of these antigens appears to be dependent on the presence of multiple O-linked alpha-N-acetylgalactosamine (alpha-GalNAc) determinants. A eukaryotic expression system that would produce proteins bearing glycosylation patterns similar to those found on the native C. parvum glycoproteins would greatly facilitate the molecular and functional characterization of these antigens. As a unique approach to this problem, the Cpgp40/15 gene was transiently expressed in Toxoplasma gondii, and the expressed recombinant glycoproteins were characterized. Antisera to gp40 and gp15 reacted with the surface membranes of tachyzoites expressing the Cpgp40/15 construct, and this reactivity colocalized with that of antiserum to the T. gondii surface protein SAG1. Surface membrane localization was dependent on the presence of the glycophosphatidylinositol anchor attachment site present in the gp15 coding sequence. The presence of terminal O-linked alpha-GalNAc determinants on the T. gondii recombinant gp40 was confirmed by reactivity with Helix pomatia lectin and the monoclonal antibody 4E9, which recognizes alpha-GalNAc residues, and digestion with alpha-N-acetylgalactosaminidase. In addition to appropriate localization and glycosylation, T. gondii apparently processes the gp40/15 precursor into the gp40 and gp15 component glycopolypeptides, albeit inefficiently. These results suggest that a surrogate system using T. gondii for the study of Cryptosporidium biology may be useful.


Infection management and multidrug-resistant organisms in nursing home residents with advanced dementia.
Infection management in advanced dementia has important implications for (1) providing high-quality care to patients near the end of life and (2) minimizing the public health threat posed by the emergence of multidrug-resistant organisms (MDROs). Prospective cohort study of 362 residents with advanced dementia and their health care proxies in 35 Boston area nursing homes for up to 12 months. Data were collected to characterize suspected infections, use of antimicrobial agents (antimicrobials), clinician counseling of proxies about antimicrobials, proxy preference for the goals of care, and colonization with MDROs (methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and multidrug-resistant gram-negative bacteria). Main outcomes were (1) proportion of suspected infections treated with antimicrobials that met minimum clinical criteria to initiate antimicrobial treatment based on consensus guidelines and (2) cumulative incidence of MDRO acquisition among noncolonized residents at baseline. The cohort experienced 496 suspected infections; 72.4% were treated with antimicrobials, most commonly quinolones (39.8%) and third- or fourth-generation cephalosporins (20.6%). At baseline, 94.8% of proxies stated that comfort was the primary goal of care, and 37.8% received counseling from clinicians about antimicrobial use. Minimum clinical criteria supporting antimicrobial treatment initiation were present for 44.0% of treated episodes and were more likely when proxies were counseled about antimicrobial use (adjusted odds ratio, 1.42; 95% CI, 1.08-1.86) and when the infection source was not the urinary tract (referent). Among noncolonized residents at baseline, the cumulative incidence of MDRO acquisition at 1 year was 48%. Acquisition was associated with exposure (>1 day) to quinolones (adjusted hazard ratio [AHR], 1.89; 95% CI, 1.28-2.81) and third- or fourth-generation cephalosporins (AHR, 1.57; 95% CI, 1.04-2.40). Antimicrobials are prescribed for most suspected infections in advanced dementia but often in the absence of minimum clinical criteria to support their use. Colonization with MDROs is extensive in nursing homes and is associated with exposure to quinolones and third- and fourth-generation cephalosporins. A more judicious approach to infection management may reduce unnecessary treatment in these frail patients, who most often have comfort as their primary goal of care, and the public health threat of MDRO emergence.


Irreversible effects of random modulation on coherence properties of partially polarized light.
The effect of the multiplication of two vector electric fields by random Jones matrices on their coherence properties is investigated by analyzing their interference fringes. It is shown that these random modulations cannot increase the maximal visibility obtained when the polarization states of the two analyzed electric fields are optimized. This new result generalizes a standard property of perfectly polarized light to partially polarized light and is representative of the irreversible effects of random modulations.


Distance walked and run as improved metrics over time-based energy estimation in epidemiological studies and prevention; evidence from medication use.
The guideline physical activity levels are prescribed in terms of time, frequency, and intensity (e.g., 30 minutes brisk walking, five days a week or its energy equivalence) and assume that different activities may be combined to meet targeted goals (exchangeability premise). Habitual runners and walkers may quantify exercise in terms of distance (km/day), and for them, the relationship between activity dose and health benefits may be better assessed in terms of distance rather than time. Analyses were therefore performed to test: 1) whether time-based or distance-based estimates of energy expenditure provide the best metric for relating running and walking to hypertensive, high cholesterol, and diabetes medication use (conditions known to be diminished by exercise), and 2) the exchangeability premise. Logistic regression analyses of medication use (dependent variable) vs. metabolic equivalent hours per day (METhr/d) of running, walking and other exercise (independent variables) using cross-sectional data from the National Runners' (17,201 male, 16,173 female) and Walkers' Health Studies (3,434 male, 12,384 female). Estimated METhr/d of running and walking activity were 38% and 31% greater, respectively, when calculated from self-reported time than distance in men, and 43% and 37% greater in women, respectively. Percent reductions in the odds for hypertension and high cholesterol medication use per METhr/d run or per METhr/d walked were ≥ 2-fold greater when estimated from reported distance (km/wk) than from time (hr/wk). The per METhr/d odds reduction was significantly greater for the distance- than the time-based estimate for hypertension (runners: P<10(-5) for males and P=0.003 for females; walkers: P=0.03 for males and P<10(-4) for females), high cholesterol medication use in runners (P<10(-4) for males and P=0.02 for females) and male walkers (P=0.01 for males and P=0.08 for females) and for diabetes medication use in male runners (P<10(-3)). Although causality between greater exercise and lower prevalence of hypertension, high cholesterol and diabetes cannot be inferred from these cross-sectional data, the results do suggest that distance-based estimates of METhr/d run or walked provide superior metrics for epidemiological analyses to their traditional time-based estimates.


Treatment of schistosomiasis mansoni and japonica in baboons with tubercidin given by direct intravenous injection.
Baboons (Papio cyanocephalus and P. anubis), infected with Schistosoma mansoni or S. japonicum, were treated with single doses of tubercidin (7-deazaadenosine; Tu), 1, 3, and 5 mg per kg of body weight, administered by intravenous drip. Crystalline Tu was dissolved in sterile 0.9% NaCl solution (1 mg per ml), and the solution was delivered at a rate of 4 ml per minute. Detectable short-term host toxicity was limited to the 5 mg per kg dose, mainly in the form of reversible mild to moderate kidney damage. Only this 5 mg per kg dose administered to baboons with relatively heavy S. mansoni infections was capable of completely suppressing fecal egg excretion for 6 to 8 weeks, eliminating the female worms, and terminating active disease, as indicated by histopathological findings. Comparable effects were achieved following the administration of the 3 mg per kg dose to baboons with moderate to heavy S. japonicum infections.


Harmine suppresses bladder tumor growth by suppressing vascular endothelial growth factor receptor 2-mediated angiogenesis.
Angiogenesis is a vital step during the process of oncogenesis of a lot of tumors, with no exception in bladder cancer. One of the useful strategies for the development of new drugs against cancer is targeting angiogenesis. In the present study, we found that a small-molecule natural product, which belonged to the β-carboline alkaloid, named harmine, could strongly inhibit tumor angiogenesis thus exhibiting its ideal treatment efficacy in bladder cancer. In vivo study verified that harmine had the effect of inhibition on human bladder tumor xenograft growth. The inhibitory effect of harmine to bladder cancer growth was coordinated by the effects shown on angiogenesis. To further explore the pharmacological activities of harmine, we tested harmine's influence on blood vessel formation and found that harmine effectively blocked the microvessel sprouting in rat aortic ring assay when stimulated by vascular endothelial growth factor (VEGF). Furthermore, harmine inhibited human umbilical vein endothelial cell (HUVEC) proliferation as well as chemotactic motility, and when we treated HUVEC cell with harmine, the formation of capillary-like structures was also restrained. Moreover, harmine induced bladder cancer cell apoptosis through triggering the caspase-dependent apoptotic pathway and the downstream vascular endothelial growth factor receptor 2 (VEGFR2) kinase pathway was down-regulated, thus suppressing tumor development signals. Herein, our study demonstrated that natural product harmine might have potential in curing human bladder tumor because of its pharmacological function on tumor angiogenesis, trigged by VEGFR2 signaling pathways.


Relationships between high-stakes clinical skills exam scores and program director global competency ratings of first-year pediatric residents.
Responding to mandates from the Accreditation Council for Graduate Medical Education (ACGME) and American Osteopathic Association (AOA), residency programs have developed competency-based assessment tools. One such tool is the American College of Osteopathic Pediatricians (ACOP) program directors' annual report. High-stakes clinical skills licensing examinations, such as the Comprehensive Osteopathic Medical Licensing Examination Level 2-Performance Evaluation (COMLEX-USA Level 2-PE), also assess competency in several clinical domains. The purpose of this study is to investigate the relationships between program director competency ratings of first-year osteopathic residents in pediatrics and COMLEX-USA Level 2-PE scores from 2005 to 2009. The sample included all 94 pediatric first-year residents who took COMLEX-USA Level 2-PE and whose training was reviewed by the ACOP for approval of training between 2005 and 2009. Program director competency ratings and COMLEX-USA Level 2-PE scores (domain and component) were merged and analyzed for relationships. Biomedical/biomechanical domain scores were positively correlated with overall program director competency ratings. Humanistic domain scores were not significantly correlated with overall program director competency ratings, but did show moderate correlation with ratings for interpersonal and communication skills. The six ACGME or seven AOA competencies assessed empirically by the ACOP program directors' annual report could not be recovered by principal component analysis; instead, three factors were identified, accounting for 86% of the variance between competency ratings. A few significant correlations were noted between COMLEX-USA Level 2-PE scores and program director competency ratings. Exploring relationships between different clinical skills assessments is inherently difficult because of the heterogeneity of tools used and overlap of constructs within the AOA and ACGME core competencies.


Physical models for moving shadow and object detection in video.
Current moving object detection systems typically detect shadows cast by the moving object as part of the moving object. In this paper, the problem of separating moving cast shadows from the moving objects in an outdoor environment is addressed. Unlike previous work, we present an approach that does not rely on any geometrical assumptions such as camera location and ground surface/object geometry. The approach is based on a new spatio-temporal albedo test and dichromatic reflection model and accounts for both the sun and the sky illuminations. Results are presented for several video sequences representing a variety of ground materials when the shadows are cast on different surface types. These results show that our approach is robust to widely different background and foreground materials, and illuminations.


A view of amphibian embryology during the last century.
Having started working in the field of amphibian embryology over 50 years ago, I make some comments about the changes that seem to me to have taken place in this field over this period. Over the period 1885 to 1960, much of the highly regarded experimental embryology was conducted on amphibian eggs and embryos. Indeed, much of this work was conducted in Germany and Switzerland using eggs and embryos of European newts (salamanders) and frogs of the Rana group. Xenopus started to be used extensively after the 1950s because eggs and embryos could be obtained throughout the year by hormone injection and because sexually mature animals could be raised from an egg within one year. Since the 1960s, publications using Xenopus have exceeded those using other amphibian species by 100-fold. This short commentary highlights some of the major advances attributable to embryological work with Amphibia and exemplifies these advances by reference to those who have made conspicuous contributions in this area.


Genomic code for Sox10 activation reveals a key regulatory enhancer for cranial neural crest.
The neural crest is a multipotent, stem cell-like population that migrates extensively in the embryo and forms a wide array of derivatives, ranging from neurons to melanocytes and cartilage. Analyses of the gene regulatory network driving neural crest development revealed Sox10 as one of the earliest neural crest-specifying genes, cell-autonomously driving delamination and directly regulating numerous downstream effectors and differentiation gene batteries. In search of direct inputs to the neural crest specifier module, we dissected the chick Sox10 genomic region and isolated two downstream regulatory regions with distinct spatiotemporal activity. A unique element, Sox10E2 represents the earliest-acting neural crest cis-regulatory element, critical for initiating Sox10 expression in newly formed cranial, but not vagal and trunk neural crest. A second element, Sox10E1, acts in later migrating vagal and trunk crest cells. Deep characterization of Sox10E2 reveals Sox9, Ets1, and cMyb as direct inputs mediating enhancer activity. ChIP, DNA-pull down, and gel-shift assays demonstrate their direct binding to the Sox10E2 enhancer in vivo, whereas mutation of their corresponding binding sites, or inactivation of the three upstream regulators, abolishes both reporter and endogenous Sox10 expression. Using cis-regulatory analysis as a tool, our study makes critical connections within the neural crest gene regulatory network, thus being unique in establishing a direct link of upstream effectors to a key neural crest specifier.


The Liverpool brief assessment system for communication skills in the making of doctors.
The teaching of clinical communication skills is gaining importance in medical schools. There is a need to design feasible assessments that are credible to faculty, students and the profession. To design, and assess the reliability and validity of a new communication skills assessment system (the Liverpool Communication Skills Assessment Scale-LCSAS, and the Global Simulated Patient Rating Scale-GSPRS) for employment in OSCEs. The first three annual intakes of students to the new medical curriculum at Liverpool Medical School (n = 600). Two further sub samples of 1st year (n = 60) and 2nd year students (n = 80) were included for further validation purposes in separate studies. Intra class correlation coefficients and generalisability coefficients were employed to assess reliability of the LCSAS and GSPRS. Validity was tested by examining predicted relationships using pearson product moment correlation coefficients. The LCSAS and GSPRS showed reasonable evidence for their reliability and validity although further work is recommended.


Feline Pit2 functions as a receptor for subgroup B feline leukemia viruses.
Different subgroups of feline leukemia virus (FeLV) use different host cell receptors for entry. Subgroup A FeLV (FeLV-A) is the virus that is transmitted from cat to cat, suggesting that cells expressing the FeLV-A receptor are important targets at the earliest stages of infection. FeLV-B evolves from FeLV-A in the infected cat through acquisition of cellular sequences that are related to the FeLV envelope gene. FeLV-Bs have been shown to infect cells using the Pit1 receptor, and some variants can infect cells at a lower efficiency using Pit2. Because these observations were made using receptor proteins of human or rodent origin, the role that Pit1 and Pit2 may play in FeLV-B replication in the cat is unclear. In this study, the feline Pit receptors were cloned and tested for their ability to act as receptors for different FeLV-Bs. Some FeLV-Bs infected cells expressing feline Pit2 and feline Pit1 with equal high efficiency. Variable region A (VRA) in the putative receptor-binding domain (RBD) was a critical determinant for both feline Pit1 and feline Pit2 binding, although other domains in the RBD appear to influence how efficiently the FeLV-B surface unit can bind to feline Pit2 and promote entry via this receptor. An arginine residue at position 73 in VRA was found to be important for envelope binding to feline Pit2 but not feline Pit1. Interestingly, this arginine is not found in endogenous FeLV sequences or in recombinant viruses recovered from feline cells infected with FeLV-A. Thus, while FeLV-Bs that are able to use feline Pit2 can evolve by recombination with endogenous sequences, a subsequent point mutation during reverse transcription may be needed to generate a virus that can efficiently enter the cells using the feline Pit2 as its receptor. These studies suggest that cells expressing the feline Pit2 protein are likely to be targets for FeLV-B infection in the cat.


Association Between Immune-Related Adverse Events and Recurrence-Free Survival Among Patients With Stage III Melanoma Randomized to Receive Pembrolizumab or Placebo: A Secondary Analysis of a Randomized Clinical Trial.
Whether immune-related adverse events (irAEs) indicate drug activity in patients treated with immune checkpoint inhibitors remains unknown. To investigate the association between irAEs and recurrence-free survival (RFS) in the double-blind EORTC 1325/KEYNOTE-054 clinical trial comparing pembrolizumab therapy and placebo for the treatment of patients with high-risk stage III melanoma. A total of 1019 adults with stage III melanoma were randomly assigned on a 1:1 ratio to receive treatment with pembrolizumab therapy or placebo. Eligible patients were adults 18 years and older with complete resection of cutaneous melanoma metastatic to lymph nodes, classified with stage IIIA (at least 1 micrometastasis measuring >1 mm in greatest diameter), IIIB, or IIIC (without in-transit metastasis) cancer. Patients were randomized from August 26, 2015, to November 14, 2016. The clinical cutoff for the data set was October 2, 2017. Analyses were then performed on the database, which was locked on November 28, 2017. Participants were scheduled to receive 200 mg of pembrolizumab or placebo every 3 weeks for a total of 18 doses for approximately 1 year or until disease recurrence, unacceptable toxic effects, major protocol violation, or withdrawal of consent. The association between irAEs and RFS was estimated using a Cox model adjusted for sex, age, and AJCC-7 stage, with a time-varying covariate that had a value of 0 before irAE onset and 1 after irAE onset. Of 1011 patients who began treatment with pembrolizumab therapy or placebo, 622 (61.5%) were men and 389 (38.5%) were women; 386 patients (38.2%) were aged 50 to 64 years, 377 (37.3%) were younger than 50 years, and 248 (24.5%) were 65 years and older. Consistent with the reported main analysis in the intent-to-treat population, RFS was longer in the pembrolizumab arm compared with the placebo arm (hazard ratio [HR], 0.56; 98.4% CI, 0.43-0.74) among patients who started treatment. The incidence of irAEs was 190 (37.4%) in the pembrolizumab arm (n = 509) and 45 (9.0%) in the placebo arm (n = 502); in each treatment group, the incidence was similar for men and women. The occurrence of an irAE was associated with a longer RFS in the pembrolizumab arm (HR, 0.61; 95% CI, 0.39-0.95; P = .03) in both men and women. However, in the placebo arm, this association was not significant. Compared with the placebo arm, the reduction in the hazard of recurrence or death in the pembrolizumab arm was greater after the onset of an irAE than without or before an irAE (HR, 0.37; 95% CI, 0.24-0.57 vs HR, 0.61; 95% CI, 0.49-0.77, respectively; P = .03). In this study, the occurrence of an irAE was associated with a longer RFS in the pembrolizumab arm. ClinicalTrials.gov identifier: NCT02362594; EudraCT identifier: 2014-004944-37.


Sulfoethylated nanofibrillated cellulose: Production and properties.
Sulfoethylated nanofibrillated cellulose (NFCSulf) was produced by an industrially relevant process. The properties of the NFCSulf were compared with those of carboxymethylated nanofibrillated cellulose (NFCCarb), which has been identified as an attractive NFC for several industrial applications. The investigations revealed that NFCSulf is characterized by a higher degree of fibrillation and has superior redispersion properties. Furthermore, NFCSulf displays higher stability in varying pH values as compared to NFCCarb. Hence, NFCSulf may be a more attractive alternative than NFCCarb in applications such as rheological modifiers or adsorbing components in personal care products, in which the performance of NFC must remain unaffected in varying ambient conditions. The superior properties of NFCSulf compared to NFCCarb were proposed to be due to the combination of the unique chemical characteristics of the sulfoethylated reagent, and the larger size of the sulfonate group compared to the carboxymethyl group.


First Report of Fire Blight Disease Caused by Erwinia amylovora on Rockspray (Cotoneaster horizontalis) in Turkey.
In the late summer and early winter of 2008 and 2009, leaf and shoot blight and cankers with reddish and brownish necrotic tissue on mature branches of Cotoneaster horizontalis were investigated in landscape areas of Konya province in Turkey. Disease incidence was estimated at 2%. Bacteria were consistently isolated from the lesions on leaves and shoots on nutrient sucrose agar medium. Twelve representative bacterial strains were isolated and characterized as gram-negative, rod-shaped, mucoid, fermentative, yellow-orange on MS medium, positive for levan formation and acetoin production, no growth at 36°C, positive for gelatin hydrolysis, and negative for indole, urease, oxidase, arginine dehydrolase, reduction of nitrate, and acid production from lactose and inositol (2). Two reference strains of Erwinia amylovora (EaP28 and NCPPB 2791) obtained from the culture collection unit of Selcuk University were used as positive controls. All strains induced a hypersensitive response in tobacco (Nicotiana tobaccum cv. White Burley). All strains were identified as E. amylovora on the basis of amplification of a 1 kb DNA fragment with a species-specific primer set, A/B (1) by PCR, and fatty acid methyl ester profiles determined by Sherlock Microbial Identification System software (TSBA 6 v. 6.00; Microbial ID, Newark, DE) with similarity indices ranging from of 83 to 96%. Pathogenicity tests were performed by injecting 20 μl of a bacterial suspension (108 CFU ml-1) into the shoot tips of 3-year-old C. horizontalis seedlings. Leaf and shoot blighting symptoms were observed within 10 to 15 days, but no symptoms were observed on control plants treated with sterile water. The bacterium was reisolated from the lesions on leaves and shoots and identified as described above. To our knowledge, this is the first report of E. amylovora on cotoneaster in Turkey. Control measures are needed to prevent any further spread of the bacterium to new landscape areas. References: (1) S. Bereswill et al. Appl. Environ. Microbiol. 58:3522, 1992. (2) A. L. Jones and K. Geider. Page 40 in: Laboratory Guide for Identification of Plant Pathological Bacteria, 2001.


Genetic control of gene expression at novel and established chronic obstructive pulmonary disease loci.
Genetic risk loci have been identified for a wide range of diseases through genome-wide association studies (GWAS), but the relevant functional mechanisms have been identified for only a small proportion of these GWAS-identified loci. By integrating results from the largest current GWAS of chronic obstructive disease (COPD) with expression quantitative trait locus (eQTL) analysis in whole blood and sputum from 121 subjects with COPD from the ECLIPSE Study, this analysis identifies loci that are simultaneously associated with COPD and the expression of nearby genes (COPD eQTLs). After integrative analysis, 19 COPD eQTLs were identified, including all four previously identified genome-wide significant loci near HHIP, FAM13A, and the 15q25 and 19q13 loci. For each COPD eQTL, fine mapping and colocalization analysis to identify causal shared eQTL and GWAS variants identified a subset of sites with moderate-to-strong evidence of harboring at least one shared variant responsible for both the eQTL and GWAS signals. Transcription factor binding site (TFBS) analysis confirms that multiple COPD eQTL lead SNPs disrupt TFBS, and enhancer enrichment analysis for loci with the strongest colocalization signals showed enrichment for blood-related cell types (CD3 and CD4+ T cells, lymphoblastoid cell lines). In summary, integrative eQTL and GWAS analysis confirms that genetic control of gene expression plays a key role in the genetic architecture of COPD and identifies specific blood-related cell types as likely participants in the functional pathway from GWAS-associated variant to disease phenotype.


High-resolution molecular genomic autopsy reveals complex sudden unexpected death in epilepsy risk profile.
Advanced variant detection in genes underlying risk of sudden unexpected death in epilepsy (SUDEP) can uncover extensive epistatic complexity and improve diagnostic accuracy of epilepsy-related mortality. However, the sensitivity and clinical utility of diagnostic panels based solely on established cardiac arrhythmia genes in the molecular autopsy of SUDEP is unknown. We applied the established clinical diagnostic panels, followed by sequencing and a high density copy number variant (CNV) detection array of an additional 253 related ion channel subunit genes to analyze the overall genomic variation in a SUDEP of the 3-year-old proband with severe myoclonic epilepsy of infancy (SMEI). We uncovered complex combinations of single nucleotide polymorphisms and CNVs in genes expressed in both neurocardiac and respiratory control pathways, including SCN1A, KCNA1, RYR3, and HTR2C. Our findings demonstrate the importance of comprehensive high-resolution variant analysis in the assessment of personally relevant SUDEP risk. In this case, the combination of de novo single nucleotide polymorphisms (SNPs) and CNVs in the SCN1A and KCNA1 genes, respectively, is suspected to be the principal risk factor for both epilepsy and premature death. However, consideration of the overall biologically relevant variant complexity with its extensive functional epistatic interactions reveals potential personal risk more accurately.


Note: Effects of adding a viscosity-increasing 2 nm-size molecule on dielectric relaxation features and the dynamic heterogeneity view.
To investigate the extent to which dynamic heterogeneity determines the relaxation features, we studied the effect of adding a viscosity-increasing, ∼2 nm size polyhederal oligomeric silsesquioxane (POSS) molecule on dielectric relaxation. Addition of POSS increased the relaxation time of the α-process, τ(m,α), and decreased the non-exponential response parameter, β, which is compatible with the dynamic heterogeneity view. But β varied monotonically with the composition, and there was little variation of β with the temperature, which seems incompatible with that view. Effects of adding nm-size particles in simulations may reveal how the size of dynamically heterogeneous regions affects τ(m,α) and β.


Interphase cytogenetics: analysis of numerical chromosome aberrations in isolated cells.
Probes which recognize pericentromeric repetitive sequences can be used to determine numerical chromosome aberrations in interphase nuclei. Under appropriate stringency conditions, these probes decorate only the chromosome from which they were derived. It has been assumed that abnormalities of signal number in interphase nuclei reflect aneuploidy rather than proliferation, but this has not been clearly demonstrated. In this paper, three alphoid probes (D3Z1, D11Z1, and DXZ1) were localized to the appropriate chromosomes and the factors governing the production of reproducible signal distributions from three aneuploid cervical carcinoma-derived epithelial cell lines were investigated. Abnormalities of signal number represent numerical chromosome aberrations rather than changes associated with proliferation. Using four simple rules of interpretation, reproducible results can be obtained with minimal technical variation and selection bias. These results demonstrate that pericentromeric repetitive probes can be used reproducibly to determine numerical chromosome aberrations independent of cell proliferation in interphase nuclei, a necessary prerequisite for the application of this approach to the analysis of human tumours.


Acute ischemic stroke in children versus young adults.
The aim of this study was to compare children and young adults with acute ischemic stroke (AIS) in 2 large registries. We compared clinical characteristics, stroke etiology, workup, and outcome (modified Rankin scale score [mRS] at 3-6 months) in children (1 month-16 years) and young adults (16.1-45 years) with AIS. Data of children were collected prospectively in the nationwide Swiss NeuroPediatric Stroke Registry, young adults in the Bernese stroke database. Outcome (mRS) and stroke severity (pediatric adaptation of the National Institutes of Health stroke scale [PedNIHSS]) in children were calculated retrospectively. From January 2000 to December 2008, 128 children and 199 young adults suffered from an AIS. Children were more likely to be male than young adults (62%/49%, p = 0.023) and less frequently had hypertension (p = 0.001), hypercholesterolemia (p = 0.003), and a family history of stroke (p = 0.048). Stroke severity was similar in children and young adults (median PedNIHSS/NIHSS 5/6; p = 0.102). Stroke etiology (original TOAST classification) was more likely to be "other determined cause" in children than in young adults (51%/29%; p < .001). Cervicocerebral artery dissections were less frequent in children than in young adults (10%/23%; p = 0.005). Outcome at 3 to 6 months did not differ between children and young adults (p = 0.907); 59% of children and 60% of young adults had a favorable outcome (mRS 0-1). Mortality was similar among children and young adults (4%/6%; p = 0.436). In multivariate analysis, low PedNIHSS/NIHSS was the most important predictor of favorable outcome (p < 0.001). Although stroke etiology and risk factors in children and young adults are different, stroke severity and clinical outcome were similar in both groups.


Environmental responsiveness of tubulin glutamylation in sensory cilia is regulated by the p38 MAPK pathway.
Glutamylation is a post-translational modification found on tubulin that can alter the interaction between microtubules (MTs) and associated proteins. The molecular mechanisms regulating tubulin glutamylation in response to the environment are not well understood. Here, we show that in the sensory cilia of Caenorhabditis elegans, tubulin glutamylation is upregulated in response to various signals such as temperature, osmolality, and dietary conditions. Similarly, tubulin glutamylation is modified in mammalian photoreceptor cells following light adaptation. A tubulin glutamate ligase gene ttll-4, which is essential for tubulin glutamylation of axonemal MTs in sensory cilia, is activated by p38 MAPK. Amino acid substitution of TTLL-4 has revealed that a Thr residue (a putative MAPK-phosphorylation site) is required for enhancement of tubulin glutamylation. Intraflagellar transport (IFT), a bidirectional trafficking system specifically observed along axonemal MTs, is required for the formation, maintenance, and function of sensory cilia. Measurement of the velocity of IFT particles revealed that starvation accelerates IFT, which was also dependent on the Thr residue of TTLL-4. Similarly, starvation-induced attenuation of avoidance behaviour from high osmolality conditions was also dependent on ttll-4. Our data suggest that a novel evolutionarily conserved regulatory system exists for tubulin glutamylation in sensory cilia in response to the environment.


Sociodemographic Factors Contribute to Mental Health Disparities and Access to Services Among Young Men Who Have Sex with Men in New York City.
Young men who have sex with men (YMSM) may be at increased risk for mental health problems including depression, post-traumatic stress (PTSD), and suicidality. The overriding goal of the current investigation was to examine mental health and mental health services in a diverse sample of YMSM. We analyzed cross-sectional data from a cohort study of 598 YMSM, including sociodemographics, mental health, and mental health care. We then tested for bivariate associations, and used multivariable modeling to predict depression, PTSD, suicidality and mental health care utilization. Lower socioeconomic status, unstable housing, and school non-enrollment predicted depression and PTSD scores, while unstable housing and school non-enrollment predicted recent suicide attempt(s). These recent suicide attempt(s) also predicted current utilization of counseling or treatment, any history of psychiatric hospitalization, and any history of psychiatric diagnosis. Black and API men were less likely to have ever accessed mental health counseling or treatment. There were significant class-based differences with regard to mental health outcomes, but not mental health services. Further, recent crises (i.e., suicide attempt, hospitalization) were strong predictors of accessing mental health services. Improving the mental health of YMSM requires addressing the underlying structural factors that influence mental health outcomes and service access.


Hospital and geographic variability in thirty-day all-cause mortality following colorectal cancer surgery.
To assess hospital and geographic variability in 30-day mortality after surgery for CRC and examine the extent to which sociodemographic, area-level, clinical, tumor, treatment, and hospital characteristics were associated with increased likelihood of 30-day mortality in a population-based sample of older CRC patients. Linked Surveillance Epidemiology End Results (SEER) and Medicare data from 47,459 CRC patients aged 66 years or older who underwent surgical resection between 2000 and 2005, resided in 13,182 census tracts, and were treated in 1,447 hospitals. An observational study using multilevel logistic regression to identify hospital- and patient-level predictors of and variability in 30-day mortality. We extracted sociodemographic, clinical, tumor, treatment, hospital, and geographic characteristics from Medicare claims, SEER, and census data. Of 47,459 CRC patients, 6.6 percent died within 30 days following surgery. Adjusted variability in 30-day mortality existed across residential census tracts (predicted mortality range: 2.7-12.3 percent) and hospitals (predicted mortality range: 2.5-10.5 percent). Higher risk of death within 30 days was observed for CRC patients age 85+ (12.7 percent), census-tract poverty rate >20 percent (8.0 percent), two or more comorbid conditions (8.8 percent), stage IV at diagnosis (15.1 percent), undifferentiated tumors (11.6 percent), and emergency surgery (12.8 percent). Substantial, but similar variability was observed across census tracts and hospitals in 30-day mortality following surgery for CRC in patients 66 years and older. Risk of 30-day mortality is driven not only by patient and hospital characteristics but also by larger social and economic factors that characterize geographic areas.


Specific abbreviated injury scale values are responsible for the underestimation of mortality in penetrating trauma patients by the injury severity score.
The Injury Severity Score (ISS) is widely used as a method for rating severity of injury. The ISS is the sum of the squares of the three worst Abbreviated Injury Scale (AIS) values from three body regions. Patients with penetrating injuries tend to have higher mortality rates for a given ISS than patients with blunt injuries. This is thought to be secondary to the increased prevalence of multiple severe injuries in the same body region in patients with penetrating injuries, which the ISS does not account for. We hypothesized that the mechanism-based difference in mortality could be attributed to certain ISS ranges and specific AIS values by body region. Outcome and injury scoring data were obtained from transfused patients admitted to 23 Level I trauma centers. ISS values were grouped into categories, and a logistic regression model was created. Mortality for each ISS category was determined and compared with the ISS 1 to 15 group. An interaction term was added to evaluate the effect of mechanism. Additional logistic regression models were created to examine each AIS category individually. There were 2,292 patients in the cohort. An overall interaction between ISS and mechanism was observed (p = 0.049). Mortality rates between blunt and penetrating patients with an ISS between 25 and 40 were significantly different (23.6 vs. 36.1%; p = 0.022). Within this range, the magnitude of the difference in mortality was far higher for penetrating patients with head injuries (75% vs. 37% for blunt) than truncal injuries (26% vs. 17% for blunt). Penetrating trauma patients with an AIS head of 4 or 5, AIS abdomen of 3, or AIS extremity of 3 all had adjusted mortality rates higher than blunt trauma patients with those values. Significant differences in mortality between blunt and penetrating trauma patients exist at certain ISS and AIS category values. The mortality difference is greatest for head injured patients.


An official American Thoracic Society/European Respiratory Society statement: asthma control and exacerbations: standardizing endpoints for clinical asthma trials and clinical practice.
The assessment of asthma control is pivotal to the evaluation of treatment response in individuals and in clinical trials. Previously, asthma control, severity, and exacerbations were defined and assessed in many different ways. The Task Force was established to provide recommendations about standardization of outcomes relating to asthma control, severity, and exacerbations in clinical trials and clinical practice, for adults and children aged 6 years or older. A narrative literature review was conducted to evaluate the measurement properties and strengths/weaknesses of outcome measures relevant to asthma control and exacerbations. The review focused on diary variables, physiologic measurements, composite scores, biomarkers, quality of life questionnaires, and indirect measures. The Task Force developed new definitions for asthma control, severity, and exacerbations, based on current treatment principles and clinical and research relevance. In view of current knowledge about the multiple domains of asthma and asthma control, no single outcome measure can adequately assess asthma control. Its assessment in clinical trials and in clinical practice should include components relevant to both of the goals of asthma treatment, namely achievement of best possible clinical control and reduction of future risk of adverse outcomes. Recommendations are provided for the assessment of asthma control in clinical trials and clinical practice, both at baseline and in the assessment of treatment response. The Task Force recommendations provide a basis for a multicomponent assessment of asthma by clinicians, researchers, and other relevant groups in the design, conduct, and evaluation of clinical trials, and in clinical practice.


A pilot feasibility randomised controlled trial of two behaviour change interventions compared to usual care to reduce substance misuse in looked after children and care leavers aged 12-20 years: The SOLID study.
Young people in state care, often due to abuse or neglect, have a four-fold increased risk of drug and alcohol use compared to their peers. The SOLID study aimed to investigate the feasibility of a definitive randomised controlled trial, comparing two behaviour change interventions to reduce risky substance use (illicit drugs and alcohol), and improve mental health, in young people in care. We recruited young people in care aged 12-20 years, self-reporting substance use within the previous 12 months and residing in 1 of 6 participating local authority sites in the North East of England. Participants were randomised to either i. Motivational Enhancement Therapy (MET), ii. Social Behaviour and Network Therapy (SBNT) or iii. Control (usual care). All interventions were delivered by trained drug and alcohol workers. Follow-up data were collected 12 months post recruitment. Feasibility for trial progression was compared to pre-specified stop: go criteria (recruitment of 60% of eligible participants, 80% of participants attending 60% of offered sessions and retention of 70% of participants at 12 month follow up). Of 1450 eligible participants, 860 (59%) were screened for drug and alcohol use by social workers, 211 (24.5%) met inclusion criteria for the trial and 112 young people (7.7%) consented and were randomised. Sixty of these 112 participants (54%) completed 12-month follow-up questionnaires. Only 15 out of the 76 (20%) participants allocated to an intervention arm attended any of the offered MET or SBNT sessions. By reference to pre-specified stop: go criteria it is not feasible to conduct a definitive trial for SOLID in its current format. Despite co-designing procedures with staff and young people in care, the screening, referral and treatment pathway did not work here. Future work may require dedicated clinically embedded research resource to evaluate effectiveness of new interventions in services.


Pollution problems, resource policy, and the scientist.
Through exponential increase in population, accompanied by rapid industrial and agricultural expansion, we have reached the point where decisions involving the use of natural resources are much more important and also much more difficult than they were even 10 years ago. The same conditions that make decisions more urgent make them more time-consuming. The Cayuga Lake case illustrates many aspects of a present-day resource management controversy.


Provider feedback improves adherence with AHCPR Smoking Cessation Guideline.
This study evaluated the effect of primary care providers' adherence with the AHCPR Smoking Cessation Guideline after receiving a multicomponent intervention. A quasi-experimental study with one intervention and one control team was conducted in a southeastern Veterans Affairs Medical Center primary care setting. During phase I, chart reviews were conducted to measure baseline provider adherence and documentation of the four A's (ask, advise, assist, arrange). In phase II, the intervention team received a single educational session on the AHCPR Guideline, four A's, and tobacco dependence treatment. This was followed by chart reviews of patients seen 4 to 8 weeks after the educational intervention to measure provider adherence and documentation of the four A's. During phase III, the intervention team received individual and team feedback from the chart reviews in phases I and II and booster education on the AHCPR Guideline. Chart reviews were conducted from patient visits 4 to 8 weeks after the feedback and booster education to determine provider adherence and documentation of the four A's. A nested repeated measures two-factor analysis of variance was performed for each of the following outcomes: ask, advise, assist, and arrange. Data analyses revealed that both the control and the intervention teams had 100% compliance in asking the patient about smoking status. There was a prestudy implementation of the vital sign stamp that included smoking status in this setting. Education on tobacco dependence and the AHCPR Guideline had no significant impact on provider performance with the advisement, assistance, and arrangement of follow-up. However, significant improvements occurred in the intervention team in the advisement (P = 0.05), assistance (P = 0.001), and arrangement of follow-up (P = 0.001) phase after individual and team feedback was provided. This research supports the fact that feedback impacts individuals and team performances and facilitated positive system changes to improve provider adherence with the AHCPR recommendations in treating tobacco dependence.


A preliminary investigation of alpha-lipoic acid treatment of antipsychotic drug-induced weight gain in patients with schizophrenia.
Weight gain and other metabolic disturbances have now become discouraging, major side effects of atypical antipsychotic drugs (AAPDs). The novel strategies required to counteract these serious consequences, however, should avoid modulating the activities of the neurotransmitter receptors involved because those receptors are the therapeutic targets of AAPDs. Adenosine monophosphate-activated protein kinase is an enzyme that plays a pivotal role in energy homeostasis. We hypothesized that alpha-lipoic acid (ALA), which is known to modulate adenosine monophosphate-activated protein kinase activity in the hypothalamus and peripheral tissues, would ameliorate AAPD-induced weight gain. We describe the case series of a 12-week ALA trial in schizophrenia patients treated with AAPDs. Two of 7 enrolled subjects were dropped from the study because of noncompliance and demand for new medication to treat depressive symptoms, respectively. The mean (SD) weight loss was 3.16 (3.20) kg (P = 0.043, last observation carried forward; median, 3.03 kg; range, 0-8.85 kg). On average, body mass index showed a significant reduction (P = 0.028) over the 12 weeks. During the same period, a statistically significant reduction was also observed in total cholesterol levels (P = 0.042), and there was a weak trend toward the reduction in insulin resistance (homeostasis model assessment of insulin resistance) (P = 0.080). Three subjects reported increased energy subjectively. The total scores on the Brief Psychiatric Rating Scale and the Montgomery-Asberg Depression Rating Scale did not vary significantly during the study. These preliminary data suggest the possibility that ALA can ameliorate the adverse metabolic effects induced by AAPDs. To confirm the benefits of ALA, more extended study is warranted.


A qualitative analysis of relatives', health professionals' and service users' views on the involvement in care of relatives in Bipolar Disorder.
Relatives of people with bipolar disorder report that services do not meet their own needs, despite clinical recommendations for the development of care plans for relatives, provision of information regarding their statutory entitlements, and formal involvement in decision making meetings. Further, there is now conclusive evidence highlighting the benefits of relatives' involvement in improving outcomes for service users, relatives, and the health system as a whole. This qualitative study explored the views of relatives of people with bipolar disorder, service users and healthcare professionals regarding the barriers and the facilitators to relatives' involvement in care. Thirty five people were interviewed (12 relatives, 11 service users and 12 healthcare professionals). Audio recordings were transcribed verbatim and common themes in participants' narratives emerged using framework analysis. Participants' accounts confirmed the existence of opportunities for relatives to be involved. These, however, were limited and not always accessible. There were three factors identified that influenced accessibility namely: pre-existing worldviews, the quality of relationships and of communication between those involved, and specific structural impediments. These themes are understood as intertwined and dependent on one another. People's thoughts, beliefs, attitudes, cultural identifications and worldviews often underlie the ways by which they communicate and the quality of their relationship. These, however, need to be conceptualised within operational frameworks and policy agendas in health settings that often limit bipolar relatives' accessibility to opportunities for being more formally involved. Involving relatives leads to clear benefits for relatives, service users, healthcare professionals, and the health system as a whole. Successful involvement of relatives, however, depends on a complex network of processes and interactions among all those involved and requires strategic planning from policy makers, operational plans and allocation of resources.


Quantitative trait genetic linkage analysis of body mass index in familial coronary artery disease.
Body mass index (BMI) is one of the most reproducible and commonly used proxies for obesity and is known to be influenced by many environmental causes as well as genetic factors. Identification of susceptibility genes for BMI regulation has been difficult. Reasons for these inconclusive results are both methodological and related to obesity aetiology. A genome-wide linkage analysis was performed to localise Quantitative trait loci influencing BMI in a large cohort collected in the PROCARDIS coronary heart disease study consisting of 1,812 informative families. Multipoint linkage analysis for BMI was conducted using both a variance component approach and a model-free regression method, and the resulting LOD scores were compared. The strongest evidence for linkage was detected on chromosomes 13 (LOD 1.6). Other regions showing a LOD score greater than 1 were observed on chromosomes 3, 5, 11, 12 and 15. These results were mainly confirmed by the three different approaches used in the analysis. Our study did not find any locus with strongly supporting evidence for linkage to BMI even in such a large sample. Our results confirm the substantial genetic heterogeneity influencing BMI regulation that has emerged from the majority of genome scans so far published.


Fouling and long-term durability of an integrated forward osmosis and membrane distillation system.
An integrated forward osmosis (FO) and membrane distillation (MD) system has great potential for sustainable wastewater reuse. However, the fouling and long-term durability of the system remains largely unknown. This study investigates the fouling behaviour and efficiency of cleaning procedures of FO and MD membranes used for treating domestic wastewater. Results showed that a significant decline in flux of both FO and MD membranes were observed during treatment of wastewater with organic foulants. However, shear force generated by the increased cross-flow physically removed the loosely attached foulants from the FO membrane surface and resulted in 86-88% recovery of flux by cleaning with tap water. For the MD membrane, almost no flux recovery was achieved due to adsorption of organic foulants on the hydrophobic membrane surface, thus indicating significant irreversible fouling/wetting, which may not be effectively cleaned even with chemical reagents. Long-term (10 d) tests showed consistent performance of the FO membrane by rejecting the contaminants. However, organic foulants reduced the hydrophobicity of the MD membrane, caused wetting problems and allowed contaminants to pass through. The results demonstrate that combination of the FO and MD processes can effectively reduce irreversible membrane fouling and solve the wetting problem of the MD membrane.


"I like the way you move": how hormonal changes across the menstrual cycle affect female perceptions of gait.
Variations in hormone concentrations across the menstrual cycle affect human female mate preferences. It has been shown that around the time of ovulation human females prefer more masculine male voices, faces, and bodies while simultaneously preferring less faces that are more feminine. They prefer also displays of male dominance, males with more symmetrical faces, and the scent of males with high levels of body symmetry. The aim of the experiments reported here was to investigate whether there are changes in female preferences for walking gaits across the menstrual cycle. Experiment 1 showed female observers could discriminate between point-light walkers with low and high levels of fluctuating asymmetries in their gaits. Female observers were more sensitive to asymmetries in female gaits than they were for asymmetries in male gaits. Experiment 2 showed that level of gait asymmetry did not affect the abilities of observers to discriminate female from male walkers. Experiment 3 showed that female observers did not change their preference for low and high asymmetry walkers across their menstrual cycles. However, females showed a decreased preference for all female walkers at the time during which it was estimated observers were at peak fertility. That same change in preference was not observed for male walkers. These data suggest female observers may not value gait asymmetry, as a mate selection cue, in the same way that they value asymmetries in faces and bodies. While only "average" gaits were used in these experiments, rather than the gaits of individual walkers, the types of asymmetries in gait tested here were not used in the same way as static cues for judging the apparent healthiness of individuals. Females do discriminate well average female gait asymmetries and do change their preferences for those gaits across their menstrual cycle. Doing so may reflect the operation of processes that equip females with an advantage when competing for mates at times of peak fertility.


Antithrombotic and thrombolytic therapy for ischemic stroke: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition).
This article about treatment and prevention of stroke is part of the Antithrombotic and Thrombolytic Therapy: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition). Grade 1 recommendations are strong and indicate that the benefits do, or do not, outweigh risks, burden, and costs. Grade 2 suggests that individual patients' values may lead to different choices (for a full understanding of the grading, see the "Grades of Recommendations" chapter by Guyatt et al, CHEST 2008; 133:123S-131S). Among the key recommendations in this chapter are the following: For patients with acute ischemic stroke, we recommend administration of IV tissue plasminogen activator (tPA) if treatment is initiated within 3 h of clearly defined symptom onset (Grade 1A). For patients with acute ischemic stroke of > 3 h but < 4.5 h, we suggest clinicians do not use IV tPA (Grade 2A). For patients with acute stroke onset of > 4.5 h, we recommend against the use of IV tPA (Grade 1A). For patients with acute ischemic stroke who are not receiving thrombolysis, we recommend early aspirin therapy (Grade 1A). For acute ischemic stroke patients with restricted mobility, we recommend prophylactic low-dose subcutaneous heparin or low-molecular-weight heparins (Grade 1A). For long-term stroke prevention in patients with noncardioembolic stroke or transient ischemic attack (TIA) [ie, atherothrombotic, lacunar, or cryptogenic], we recommend treatment with an antiplatelet agent (Grade 1A), including aspirin (recommended dose, 50-100 mg/d), the combination of aspirin and extended-release dipyridamole (25 mg/200 mg bid), or clopidogrel (75 mg qd). In these patients, we recommend use of the combination of aspirin and extended-release dipyridamole (25/200 mg bid) over aspirin (Grade 1A) and suggest clopidogrel over aspirin (Grade 2B), and recommend avoiding long-term use of the combination of aspirin and clopidogrel (Grade 1B). For patients who are allergic to aspirin, we recommend clopidogrel (Grade 1A). In patients with atrial fibrillation and a recent stroke or TIA, we recommend long-term oral anticoagulation (target international normalized ratio, 2.5; range, 2.0 to 3.0) [Grade 1A]. In patients with venous sinus thrombosis, we recommend unfractionated heparin (Grade 1B) or low-molecular-weight heparin (Grade 1B) over no anticoagulant therapy during the acute phase.


Influences of microwave irradiation on performances of membrane filtration and catalytic degradation of perfluorooctanoic acid (PFOA).
Perfluorooctanoic Acid (PFOA), one of the common per- and poly fluorinated alkylated substances (PFASs), is increasingly detected in the environment due to the diverse industrial applications and high resistance to degradation processes. This study evaluated degradation of PFOA in microwave-assistant catalytic membrane filtration, a process that integrates microwave catalytic reactions into a ceramic membrane filtration. First, water permeation of the pristine and catalyst-coated membranes were examined under the influence of microwave irradiation to analyse the impacts of the coating layer and water temperature increase on permeate flux, which were well interpreted by the Carman-Kozeny and Hagen-Posieulle (non-slipping and slit-like) models. Then, the PFOA removal was first assessed in a continuous filtration mode with and without microwave irradiation. Our results show that PFOA first adsorbed on membrane and catalyst materials, and then fully penetrated the membrane filter after reaching adsorption equilibrium. Under microwave irradiation (7.2 W·cm-2), approximate 65.9% of PFOA (25 μg·L-1) in the feed solution was degraded within a hydraulic time of 2 min (at the permeate flow rate of 43 LMH) due to the microwave-Fenton like reactions. In addition, low flow rates and moderate catalyst coating densities are critical for optimizing PFOA removal. Finally, potential degradation mechanisms of PFOA were proposed through the analysis of degradation by-products (e.g., PFPeA). The findings may provide new insight into the development of reactive membrane-enabled systems for destruction of refractory PFAS.


Use of adeno-associated virus-mediated delivery of mutant huntingtin to study the spreading capacity of the protein in mice and non-human primates.
In order to model various aspects of Huntington's disease (HD) pathology, in particular protein spread, we administered adeno-associated virus (AAV) expressing green fluorescent protein (GFP) or GFP coupled to HTT-Exon1 (19Q or 103Q) to the central nervous system of adult wild-type (WT) mice and non-human primates. All animals underwent behavioral testing and post-mortem analyses to determine the long-term consequences of AAV injection. Both mice and non-human primates demonstrated behavioral changes at 2-3 weeks post-surgery. In mice, these changes were absent after 3 months while in non-human primates, they persisted in the majority of tested animals. Post-mortem analysis revealed that spreading of the aggregates was limited, although the virus did spread between synaptically-connected brain regions. Despite circumscribed spreading, the presence of mHTT generated changes in endogenous huntingtin (HTT) levels in both models. Together, these results suggest that viral expression of mHTTExon1 can induce spreading and seeding of HTT in both mice and non-human primates.


Methamidophos, dichlorvos, O-methoate and diazinon pesticides used in Turkey make a covalent bond with butyrylcholinesterase detected by mass spectrometry.
Organophosphorus pesticides used most commonly in Turkey include methamidophos, dichlorvos, O-methoate and diazinon. These toxic chemicals or their metabolites make a covalent bond with the active site serine of butyrylcholinesterase. Our goal was to identify the adducts that result from the reaction of human butyrylcholinesterase with these pesticides. Highly purified human butyrylcholinesterase was treated with a 20-fold molar excess of pesticide. The protein was denatured by boiling and digested with trypsin. MS and MSMS spectra of HPLC-purified peptides were acquired on a MALDI-TOF-TOF 4800 mass spectrometer. It was found that methamidophos added a mass of +93, consistent with addition of methoxy aminophosphate. A minor amount of adduct with an added mass of +109 was also found. Dichlorvos and O-methoate both made dimethoxyphosphate (+108) and monomethoxyphosphate adducts (+94). Diazinon gave a novel adduct with an added mass of +152 consistent with diethoxythiophosphate. Inhibition of enzyme activity in the presence of diazinon developed slowly (15 h), concomitant with isomerization of diazinon via a thiono-thiolo rearrangement. The isomer of diazinon yielded diethoxyphosphate and monoethoxyphosphate adducts with added masses of +136 and +108. MSMS spectra confirmed that each of the pesticides studied made a covalent bond with serine 198 of butyrylcholinesterase. These results can be used to identify the class of pesticides to which a patient was exposed.