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D2C7-immunotoxin (IT), a dual-specific IT targeting wild-type epidermal growth factor receptor (EGFR) and mutant EGFR variant III (EGFRvIII) proteins, demonstrates encouraging survival outcomes in a subset of patients with glioblastoma. We hypothesized that immunosuppression in glioblastoma limits D2C7-IT efficacy. To improve the response rate and reverse immunosuppression, we combined D2C7-IT tumor cell killing with αCD40 costimulation of antigen-presenting cells. In murine glioma models, a single intratumoral injection of D2C7-IT+αCD40 treatment activated a proinflammatory phenotype in microglia and macrophages, promoted long-term tumor-specific CD8 + T cell immunity, and generated cures. D2C7-IT+αCD40 treatment increased intratumoral Slamf6 + CD8 + T cells with a progenitor phenotype and decreased terminally exhausted CD8 + T cells. D2C7-IT+αCD40 treatment stimulated intratumoral CD8 + T cell proliferation and generated cures in glioma-bearing mice despite FTY720-induced peripheral T cell sequestration. Tumor transcriptome profiling established CD40 up-regulation, pattern recognition receptor, cell senescence, and immune response pathway activation as the drivers of D2C7-IT+αCD40 antitumor responses. To determine potential translation, immunohistochemistry staining confirmed CD40 expression in human GBM tissue sections. These promising preclinical data allowed us to initiate a phase 1 study with D2C7-IT+αhCD40 in patients with malignant glioma (NCT04547777) to further evaluate this treatment in humans.

We investigate the role of generalised trust and relation centrism for corruption as perceived by 16,785 firms in 20 lower- and middle- income countries. The empirical results demonstrate that typically, higher levels of relation centrism and generalised trust are linked with corruption as perceived by firms. The results however show significant regional disparities suggesting that firms will need to be aware of these differences, as they are likely to determine their optimal strategic choices when entering or expanding their operations into new territories. For instance, in regions where increased friend centrism is associated with reduced corruption, firms can use weak ties more tactically but will need to be more careful with such weak ties in regions where increased friend centrism is associated with increased corruption.

Background Myalgic encephalomyelitis / chronic fatigue syndrome (ME/CFS) is a common, long-term condition characterised by post-exertional malaise, often with fatigue that is not significantly relieved by rest. ME/CFS has no confirmed diagnostic test or effective treatment and we lack knowledge of its causes. Identification of genes and cellular processes whose disruption adds to ME/CFS risk is a necessary first step towards development of effective therapy. Methods Here we describe DecodeME, an ongoing study co-produced by people with lived experience of ME/CFS and scientists. Together we designed the study and obtained funding and are now recruiting up to 25,000 people in the UK with a clinical diagnosis of ME/CFS. Those eligible for the study are at least 16 years old, pass international study criteria, and lack any alternative diagnoses that can result in chronic fatigue. These will include 5,000 people whose ME/CFS diagnosis was a consequence of SARS-CoV-2 infection. Questionnaires are completed online or on paper. Participants’ saliva DNA samples are acquired by post, which improves participation by more severely-affected individuals. Digital marketing and social media approaches resulted in 29,000 people with ME/CFS in the UK pre-registering their interest in participating. We will perform a genome-wide association study, comparing participants’ genotypes with those from UK Biobank as controls. This should generate hypotheses regarding the genes, mechanisms and cell types contributing to ME/CFS disease aetiology. Discussion The DecodeME study has been reviewed and given a favourable opinion by the North West – Liverpool Central Research Ethics Committee (21/NW/0169). Relevant documents will be available online (www.decodeme.org.uk). Genetic data will be disseminated as associated variants and genomic intervals, and as summary statistics. Results will be reported on the DecodeME website and via open access publications.

Healthcare workers (HCWs) are known to be at increased risk of infection with SARS-CoV-2, although whether these risks are equal across all roles is uncertain. Here we report a retrospective analysis of a large real-world dataset obtained from 10 March to 6 July 2020 in an NHS Foundation Trust in England with 17,126 employees. 3,338 HCWs underwent symptomatic PCR testing (14.4% positive, 2.8% of all staff) and 11,103 HCWs underwent serological testing for SARS-CoV-2 IgG (8.4% positive, 5.5% of all staff). Seropositivity was lower than other hospital settings in England but higher than community estimates. Increased test positivity rates were observed in HCWs from BAME backgrounds and residents in areas of higher social deprivation. A multiple logistic regression model adjusting for ethnicity and social deprivation confirmed statistically significant increases in the odds of testing positive in certain occupational groups, most notably domestic services staff, nurses, and health-care assistants. PCR testing of symptomatic HCWs appeared to underestimate overall infection levels, probably due to asymptomatic seroconversion. Clinical outcomes were reassuring, with only a small minority of HCWs with COVID-19 requiring hospitalization (2.3%) or ICU management (0.7%) and with no deaths. Despite a relatively low level of HCW infection compared to other UK cohorts, there were nevertheless important differences in test positivity rates between occupational groups, robust to adjustment for demographic factors such as ethnic background and social deprivation. Quantitative and qualitative studies are needed to better understand the factors contributing to this risk. Robust informatics solutions for HCW exposure data are essential to inform occupational monitoring.

Purpose: Physical activity (PA) is increasingly being used in hospice care as a rehabilitation strategy to help patients manage symptoms and improve quality of life. However, little is known about ...

BACKGROUND Intestinal volvulus is a potentially life-threatening condition that occurs when loops of bowel twist around its supporting mesentery and associated vasculature. Clinicians often rely on various radiological investigations for prompt diagnosis to avoid complications such as bowel infarction. This review assesses the clinical reliability of the ultrasonographic whirlpool sign (WS) in the diagnosis of intestinal volvulus. METHODS In adherence with the PRISMA (Preferred Reporting Items for Systematic reviews and Meta-analyses) statement, a systematic search of BMJ Best Practice, Cochrane Database of Systematic Reviews, EMBASE, Google Scholar, PubMed, Scopus and Web of Science databases was performed (August 2017), using relevant search terms. Selected studies were ranked for quality and relevance using the CASP (Critical Appraisal Skills Program) tool. RESULTS Sixteen articles (1640 participants) were assessed. The mean and median sample size was 102.5 (SD ± 192.23) and 28 (range 7-770), respectively. The WS was positive in 212 of 255 (83.1%) patients with intestinal volvulus. Meta-analysis of the studies that provided sufficient data resulted in a pooled sensitivity and specificity of 87.42% (95% confidence interval (CI): 81.05-92.25) and 98.63% (95% CI: 97.88-99.18), respectively, with an estimated summary effect of 5.28 (95% CI: 4.47-6.08, P

The Remote & Rural conference 2019 – Creating a Community of Care was held on 11 November 2019 at the Centre for Health Sciences, Raigmore Hospital, Inverness, UK.

Nerve-binding fluorophores with near-infrared (NIR; 650 to 900 nm) emission could reduce iatrogenic nerve injury rates by providing surgeons precise, real-time visualization of the peripheral nervous system. Unfortunately, current systemically administered nerve contrast agents predominantly emit at visible wavelengths and show nonspecific uptake in surrounding tissues such as adipose, muscle, and facia, thus limiting detection to surgically exposed surface-level nerves. Here, a focused NIR fluorophore library was synthesized and screened through multi-tiered optical and pharmacological assays to identify nerve-binding fluorophore candidates for clinical translation. NIR nerve probes enabled micrometer-scale nerve visualization at the greatest reported tissue depths (~2 to 3 mm), a feat unachievable with previous visibly emissive contrast agents. Laparoscopic fluorescent surgical navigation delineated deep lumbar and iliac nerves in swine, most of which were invisible in conventional white-light endoscopy. Critically, NIR oxazines generated contrast against all key surgical tissue classes (muscle, adipose, vasculature, and fascia) with nerve signal-to-background ratios ranging from ~2 (2- to 3-mm depth) to 25 (exposed nerve). Clinical translation of NIR nerve-specific agents will substantially reduce comorbidities associated with surgical nerve damage.

Social interactions are thought to be a critical driver in the evolution of cognitive ability. Cooperative interactions, such as pair bonding, rather than competitive interactions have been largely implicated in the evolution of increased cognition. This is despite competition traditionally being a very strong driver of trait evolution. Males of many species track changes in their social environment and alter their reproductive strategies in response to anticipated levels of competition. We predict this to be cognitively challenging. Using a Drosophila melanogaster model, we are able to distinguish between the effects of a competitive environment versus generic social contact by exposing flies to same-sex same-species competition versus different species partners, shown to present non-competitive contacts. Males increase olfactory learning/memory and visual memory after exposure to conspecific males only, a pattern echoed by increased expression of synaptic genes and an increased need for sleep. For females, largely not affected by mating competition, the opposite pattern was seen. The results indicate that specific social contacts dependent on sex, not simply generic social stimulation, may be an important evolutionary driver for cognitive ability in fruit flies.

Background: Total hip arthroplasty is an effective surgical procedure commonly used worldwide for patients suffering the disabling effects of osteoarthritis when medical therapy is unsuccessful. Traditionally pre- and postoperative information for patients undergoing a hip arthroplasty has been provided by paper-based methods. Electronic health (eHealth) programs to support individualized patient education on preoperative preparation, in-patient care, and home rehabilitation have the potential to increase patient engagement, enhance patient recovery, and reduce potential postoperative complications. Objective: The aim of this study is to compare the addition of an eHealth program versus standard care for pre- and postoperative education on patient outcomes for primary total hip arthroplasty. Methods: One hundred patients undergoing a primary elective total hip arthroplasty will be recruited from a metropolitan hospital in Western Australia to participate in a 6-month parallel randomized control trial. Participants will be randomized to either the standard care group (n=50) and will be given the education booklet and enrolled to attend a 1-hour education session, or the intervention group (n=50), and will receive the same as the standard care plus access to an eHealth program titled “My Hip Journey.” The eHealth program encourages the patient to log in daily, from 2 weeks prior to surgery to 30 days postsurgery. The information on the platform will be aligned with the patient's individual surgical journey and will include exercises to be completed each day for the duration of the program. The primary outcome measure is the Hip Dysfunction and Osteoarthritis Outcome Score, version LK 2.0. Secondary outcome measures include the EuroQoL EQ-5D-5L, a 5-level 5-dimension quality of life measure, and the Self-Efficacy for Managing Chronic Disease Scale. Data will be collected at pre-admission (presurgery) and at 6 weeks, 3 months, and 6 months postsurgery. A patient satisfaction survey will be completed 6 weeks postsurgery and Web-based analytics will be collected 6 months postsurgery. A cost-effectiveness analysis, using the intention-to-treat principle, will be conducted from the hospital’s perspective. Results: Enrollment in the study commenced in January 2018 with recruitment due for completion towards the end of the year. The first results are expected to be submitted for publication in 2019. Conclusions: The outcomes and cost of using an eHealth program to support a patient’s recovery from a hip arthroplasty will be compared with standard care in this study. If the eHealth program is found to be effective, further implementation across clinical practice could lead to improvement in patient outcomes and other surgical areas could be incorporated. Trial Registration: Australian New Zealand Clinical Trials Registry (ANZCTR) ACTRN12617001433392; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=373657&isReview=true (Archived by WebCite at http://www.webcitation.org/6yzoTuggx). Registered Report Identifier: RR1-10.2196/9654

Purpose: To investigate if final year medical students undertaking an OSCE station at a later stage during examination diet were advantaged over their peers who undertook the same station at an earlier stage, and whether any such effect varies by the student’s relative academic standing.\ud \ud Methods: OSCE data from six consecutive final year cohorts totaling 1505 students was analyzed. Mixed effects logistic regression was used to model factors associated with the probability of passing each individual station (random effects for students and circuits; and fixed effects to assess the association with day of examination, time of day, gender and year).\ud \ud Results: Weaker students were more likely to pass if they took their OSCE later in the examination period. The odds of passing a station increased daily by 20%. Overall, the mean number of stations passed by each student increased over the 5 days.\ud \ud Conclusions: Students undertaking the same OSCE stations later in examination period statistically had higher chances of passing compared to their peers, and the weaker students appear to be particularly advantaged. These findings have major implications for OSCE design, to ensure students are not advantaged by examination timing, and weaker students are not “passing in error”.

Dear SirBefore this study was carried out we did introduce some changes to the differing days, e.g. by changing patients or X-rays for example but a balance is required between reducing the impact ...

Dear SirAlthough rapport, safety and professionalism are assessed to some extent and we are looking to improve on this in the future, these aspects alone are not enough to pass a station. Students ...

This perspective article considers the strategy, design and implementation of an electronic bioanalytical laboratory working to GLP and/or GCP regulations. There are a range of available automated systems and laboratory informatics that could be implemented and integrated to make an electronic laboratory. However, which are the appropriate ones to select and what is realistic and cost-effective for an individual laboratory? The answer is to develop an overall automation strategy that is updated periodically after each system or application has been implemented to assess if the strategy is still valid or needs to be changed. As many laboratory informatics applications have functional overlap or convergence, for example, Laboratory Information Management System, Electronic Laboratory Notebook, and Instrument and Chromatography Data Systems, the decision of which application performs a specific task needs to be carefully considered in the overall strategy. Ensuring data integrity and regulatory compliance, especially in light of a number of recent falsification cases, is a mandatory consideration for the overall strategy for an electronic bioanalytical laboratory submitting data to regulatory authorities.

Intercellular adhesion molecule-1 (ICAM-1) is involved in a range of interactions both within the host and between the host and a number of pathogens. Recently we described a mutation within the coding region of the first N-terminal immunoglobulin-like domain of ICAM-1, present at high frequency within African populations, which increased the risk of cerebral malaria. To understand the mechanism by which such a polymorphism might be maintained despite counter-selection by malaria, we have carried out functional assays using both forms of ICAM-1 as soluble Fc chimeric fusion proteins. ICAM-1Kilifi has reduced avidity for LFA-1 compared with ICAM-1ref and binding to soluble fibrinogen was completely abolished with the Kilifi variant. In Plasmodium falciparum adhesion assays, ITO4-A4u binding to ICAM-1Kilifi was reduced compared with binding to the reference form. These results allow for the possibility of balanced selection between the reference and Kilifi forms of ICAM-1 through modulation of inflammatory responses and indicate the existence of differences within ICAM-1-binding P. falciparum isolates which may be relevant to pathogenesis.

The capacity for dentofacial orthopaedics to modify the growth of the jaws and face in the management of Class II malocclusion has been the subject of much debate over the past century. It is only recently, following the publication of randomized controlled trials, that answers based on high level evidence can be provided. This article aims to review the recent literature in order to provide some clarification on the important issues relating to Class II growth modification, including: immediate treatment effects, long term stability of skeletal changes, the effects of differing orthopaedic appliances and the importance of treatment timing.

Integrins are the major adhesion receptors of leukocytes and platelets. Beta1 and beta2 integrin function on leukocytes is crucial for a successful immune response and the platelet integrin alpha(IIb)beta3 initiates the process of blood clotting through binding fibrinogen. Integrins on circulating cells bind poorly to their ligands but become active after 'inside-out' signaling through other membrane receptors. Subjects with leukocyte adhesion deficiency-1 (LAD-I) do not express beta2 integrins because of mutations in the gene specifying the beta2 subunit, and they suffer recurrent bacterial infections. Mutations affecting alpha(IIb)beta3 integrin cause the bleeding disorder termed Glanzmann's thrombasthenia. Subjects with LAD-III show symptoms of both LAD-I and Glanzmann's thrombasthenia. Their hematopoietically-derived cells express beta1, beta2 and beta3 integrins, but defective inside-out signaling causes immune deficiency and bleeding problems. The LAD-III lesion has been attributed to a C --> A mutation in the gene encoding calcium and diacylglycerol guanine nucleotide exchange factor (CALDAGGEF1; official symbol RASGRP2) specifying the CALDAG-GEF1 protein, but we show that this change is not responsible for the LAD-III disorder. Instead, we identify mutations in the KINDLIN3 (official symbol FERMT3) gene specifying the KINDLIN-3 protein as the cause of LAD-III in Maltese and Turkish subjects. Two independent mutations result in decreased KINDLIN3 messenger RNA levels and loss of protein expression. Notably, transfection of the subjects' lymphocytes with KINDLIN3 complementary DNA but not CALDAGGEF1 cDNA reverses the LAD-III defect, restoring integrin-mediated adhesion and migration.

Reduced oocyte quality has been associated with poor fertility of high-performance dairy cows during peak lactation, due to negative energy balance. We examined the role of nonesterified fatty acids (NEFAs), known to accumulate within follicular fluid during under- and overnutrition scenarios, in causing endoplasmic reticulum (ER) stress of in vitro maturated cattle cumulus-oocyte complexes (COCs). NEFA concentrations were: palmitic acid (150 μM), oleic acid (200 μM), and steric acid (75 μM). Abattoir-derived COCs were randomly matured for 24 h in the presence of NEFAs and/or an ER stress inhibitor, salubrinal. Total and hatched blastocyst yields were negatively impacted by NEFA treatment compared with controls, but this was reversed by salubrinal. ER stress markers, activating transcription factor 4 (Atf4) and heat shock protein 5 (Hspa5), but not Atf6, were significantly up-regulated by NEFA treatment within whole COCs but reversed by coincubation with salubrinal. Likewise, glucose uptake and lactate production, measured in spent medium samples, showed a similar pattern, suggesting that cumulus cell metabolism is sensitive to NEFAs via an ER stress-mediated process. In contrast, while mitochondrial DNA copy number was recovered in NEFA-treated oocytes, oocyte autofluorescence of the respiratory chain cofactor, FAD, was lower following NEFA treatment of COCs, and this was not reversed by salubrinal, suggesting the negative impact was via reduced mitochondrial function. These results reveal the significance of NEFA-induced ER stress on bovine COC developmental competence, revealing a potential therapeutic target for improving oocyte quality during peak lactation.

Objectives/Background Parent knowledge about child sleep may influence parent efforts to support healthy sleep in their children. The present study aimed to describe parent knowledge of child sleep in a hospital cohort and potential correlates including barriers to children's bedtime, information seeking about child sleep, and demographics such as child age, parent education, and household income. Methods In total, 115 parents of children aged 2–12 years attending hospital inpatient wards or day ward were approached individually and invited to complete the questionnaire. Questionnaire items were modified measures in published articles, identified through literature review, or developed from clinical experience. Results Parents on average answered half of the knowledge questions accurately, and knowledge about child sleep was positively correlated with education and household income. Parents who sought information from books or the Internet had greater knowledge about child sleep than parents who did not endorse these sources of information. The accuracy of parent estimations of their child's sleep requirements varied: 55% were within recommendations, while 26% underestimated their child's sleep requirements, 19% overestimated. Parents who correctly estimated their child's sleep requirements had higher education and income on average and were more likely to parent younger children (age 2–5 years). Parents who overestimated or underestimated sleep requirements were more likely to identify greater barriers to their child's bedtime. Conclusion Overall knowledge of child sleep in this sample was poor, consistent with samples from North American communities. Future studies should examine whether knowledge about child sleep and barriers to children's bedtimes are related to actual child sleep behaviors or parent practices to support their children to achieve a healthy sleep.

Glucose concentration during cumulus-oocyte complex (COC) maturation influences several functions, including progression of oocyte meiosis, oocyte developmental competence, and cumulus mucification. Glucosamine (GlcN) is an alternative hexose substrate, specifically metabolized through the hexosamine biosynthesis pathway, which provides the intermediates for extracellular matrix formation during cumulus cell mucification. The aim of this study was to determine the influence of GlcN on meiotic progression and oocyte developmental competence following in vitro maturation (IVM). The presence of GlcN during bovine IVM did not affect the completion of nuclear maturation and early cleavage, but severely perturbed blastocyst development. This effect was subsequently shown to be dose-dependent and was also observed for porcine oocytes matured in vitro. Hexosamine biosynthesis upregulation using GlcN supplementation is well known to increase O-linked glycosylation of many intracellular signaling molecules, the best-characterized being the phosphoinositol-3-kinase (PI3K) signaling pathway. We observed extensive O-linked glycosylation in bovine cumulus cells, but not oocytes, following IVM in either the presence or the absence of GlcN. Inhibition of O-linked glycosylation significantly reversed the effect of GlcN-induced reduction in developmental competence, but inhibition of PI3K signaling had no effect. Our data are the first to link hexosamine biosynthesis, involved in cumulus cell mucification, to oocyte developmental competence during in vitro maturation.

In mice, exposure of the uterus to seminal plasma at mating initiates an inflammatory response within the endometrium, which is characterized by production of cytokines that recruit and activate leukocytes. We hypothesized that this seminal plasma-induced inflammatory response would extend to the ovary, increasing leukocyte abundance within corpora lutea and potentially enhancing progesterone synthesis. Female mice mated to males with their seminal vesicles surgically removed exhibited fewer macrophages within corpora lutea on the day after mating, compared with females mated to vasectomized or normal, intact males. The mean number of F4/80-positive macrophages and major histocompatibility complex (MHC) class II-positive activated macrophages was approximately 2-fold fewer in the absence of seminal vesicle fluid. The effects of seminal plasma on macrophage abundance subsided by Day 4 and were not accompanied by a change in serum progesterone levels during luteinization (Days 1, 2, or 4 after mating) or luteolysis (Days 6 or 9). In vitro secretion of progesterone from corpora lutea cultured with or without LH also did not differ between treatment groups. There was no effect of seminal plasma deficiency in males on the number of ovulated ova or corpora lutea in females. These results imply that seminal plasma exposure of the female reproductive tract at mating augments the macrophage population of newly formed corpora lutea, although these additional macrophages seem not to play a role in steroidogenesis and may instead be involved in tissue remodeling within corpora lutea.

I recently listened to a news item on BBC Radio 4 discussing the perennial NHS bed shortage1 and felt relief, tinged with guilt that I was no longer part of it. I retired from a busy inner city general practice a year ago, just three months short of the chequered flag of …

SsgA-like proteins are a family of actinomycete-specific regulatory proteins that control cell division and spore maturation in streptomycetes. SsgA and SsgB together activate sporulation-specific cell division by controlling the localization of FtsZ. Here we report the identification of novel regulators that control the transcription of the ssgA-like genes. Transcriptional regulators controlling ssg gene expression were identified using a DNA-affinity capture assay. Supporting transcriptional and DNA binding studies showed that the ssgA activator gene ssgR is controlled by the TetR-family regulator AtrA, while the γ-butyrolactone-responsive AdpA (SCO2792) and SlbR (SCO0608) and the metabolic regulator Rok7B7 (SCO6008) were identified as candidate regulators for the cell division genes ssgA, ssgB and ssgG. Transcription of the cell division gene ssgB depended on the sporulation genes whiA and whiH, while ssgR, ssgA and ssgD were transcribed independently of the whi genes. Our work sheds new light on the mechanisms by which sporulation-specific cell division is controlled in Streptomyces. Electronic supplementary material The online version of this article (doi:10.1007/s10482-015-0479-2) contains supplementary material, which is available to authorized users.

OBJECTIVE. The purpose of this study was to conduct a systematic review of the efficacy of ultrasound-guided needle lavage in treating calcific tendinitis. MATERIALS AND METHODS. Two independent assessors searched medical databases and screened studies for eligibility. RESULTS. Eleven articles were included. Heterogeneity among included studies precluded meta-analysis. Results of randomized controlled trials suggested no difference in pain relief between needle lavage and other interventions, but the studies were of low quality. CONCLUSION. Additional high-quality evidence is required to determine the relative efficacy of ultrasound-guided needle lavage in the management of calcific tendinitis of the rotator cuff.

Oocyte in vitro maturation (IVM) is an important assisted reproductive technology and research tool. The adoption of IVM into routine clinical practice has been hindered by its significantly lower success rates compared to conventional in vitro fertilization. Cyclic AMP (cAMP) modulation and follicle-stimulating hormone (FSH), independently, have long been known to improve IVM oocyte developmental competence. This study comprehensively examined the effects of FSH and cAMP/cGMP modulation, alone and in combination, on IVM oocyte metabolism and developmental outcomes. Mouse cumulus-oocyte complexes (COCs) were subjected to a 1 h prematuration phase ± the cAMP modulator forskolin and cAMP/cGMP modulator 3-isobutyl-1-methylxanthine followed by IVM ± FSH. Prematuration with these cyclic nucleotide modulators or IVM with FSH significantly improved oocyte developmental competence and reduced spindle abnormalities compared to spontaneous IVM (no treatment); however, these two treatments in combination endowed even greater developmental competence (improved subsequent blastocyst rates and quality; P < 0.05), albeit blastocyst yield and quality remained significantly lower than that of oocytes matured in vivo. A significant additive effect of combined IVM treatments was evident as increased COC lactate production and oxygen consumption and enhanced oocyte oxidative metabolism, ATP production, ATP:ADP ratio, and glutathione levels (P < 0.05). Nevertheless, IVM increased reactive oxygen species production, particularly as a consequence of FSH addition, relative to in vivo matured oocytes. In conclusion, improvements in the embryo yield following IVM is associated with increased COC oxygen consumption and oocyte oxidative metabolism, but these remain metabolically and developmentally less competent relative to in vivo derived oocytes.

SOX13 is the member of the SOX ( S ry related HMG B OX ) family of transcription factors which encodes the type-1 diabetes autoantigen, ICA12, and is expressed in a number of tissues including pancreatic islets and arterial walls. By fluorescence in situ hybridisation, radiation hybrid mapping and YAC analysis we determined that the human SOX13 gene maps to Chromosome 1q31.3–32.1 near the marker D1S504, a region associated with type-1 diabetes susceptibility and familial dilated cardiomyopathy. Mouse Sox13 maps to the syntenic region near the marker D1Mit57. The human SOX13 gene spans >15.5 kb of genomic DNA and is composed of 14 exons with introns interrupting regions encoding the HMG DNA binding domain and the leucine zipper/glutamine-rich dimerisation domain. Comparison with the mouse Sox13 gene suggests the existence of long and short forms of the SOX13 protein which may arise by differential splicing during different stages in embryogenesis. The high sequence conservation between human SOX13 and mouse, Xenopus and trout orthologues implies a conserved function in vertebrates. SOX13 belongs to SOX Group D members which contain a leucine zipper/glutamine-rich region. Phylogenetic analyses of SOX proteins suggest that such domains were acquired after the initial divergence of groups A to G.

Introduction: anticoagulants and anti-platelet drugs have been shown in randomized trials to reduce the risk of stroke in patients with atrial fibrillation (AF). We therefore investigated their use in patients known to be in AF before a stroke, transient ischaemic attack (either cerebral or ocular) or retinal artery occlusion to assess the influence of trials on clinical practice. Methods: inpatients and outpatients with acute stroke, transient ischaemic attack or retinal artery occlusion were prospectively identified by a stroke physician from 1990 to 1997. The presence or absence of AF before the vascular event, and prior use of anticoagulant and anti-platelet drugs were recorded at the time of the assessment and verified using information from general practitioner and hospital case notes. Results: of 1934 patients with stroke or retinal artery occlusion, 191 (10%) were in AF before their ischaemic event. Anticoagulants had been used in 40 (21%) of these, but only in 32 (2%) of the 1743 patients in sinus rhythm [odds ratio (OR) 14.2, 95% confidence interval (CI) 8.6‐23.2]. Anti-platelet drugs had been used in 62 (32%) of those with AF compared with 500 (30%) of those in sinus rhythm (OR 1.2, 95% CI 0.9‐1.64). Of the 161 patients in AF without contraindications to anticoagulants, only 36 (22%) were taking them. Although there was a statistically significant increase in anticoagulant use from 8% in 1990 to 23% in 1996, this could be explained solely by a fall in the age of the patients referred to our hospital. Conclusion: anticoagulation is probably under-used in AF. We found no conclusive evidence that anticoagulation trials have influenced clinical practice. This raises issues about the dissemination and implementation of trial results.

Oocyte in vitro maturation (IVM) is an assisted reproductive technology that involves the maturation of cumulus-oocyte complexes (COCs) that are then capable of normal development. We have shown that epidermal growth factor (EGF)-like peptide signaling is perturbed in mouse COCs undergoing IVM when matured with follicle-stimulating hormone (FSH) and/or EGF, but supplementation of IVM with EGF-like peptides amphiregulin or epiregulin improves oocyte developmental competence. Here we aimed to determine whether EGF-like peptides regulate COC metabolism. Immature 129/Sv mouse COCs underwent IVM with FSH, EGF, amphiregulin, epiregulin, betacellulin, or no treatment (control). Epiregulin significantly increased intraoocyte flavin adenine dinucleotide (FAD) and REDOX (reduction and oxidation) ratio compared to FSH and control. Amphiregulin and epiregulin significantly increased the proportion of J aggregates (from JC-1) in oocyte mitochondria compared to control, FSH, or EGF, and this coupled with FAD and REDOX measures indicates greater mitochondrial activity. There were no differences in glucose consumption, lactate production, or glycolysis between COCs matured with FSH, EGF, and EGF-like peptides. COCs matured with EGF or EGF-like peptides exhibited significantly higher mRNA expression of the hexosamine biosynthesis pathway (HBP) rate-limiting enzyme gene Gfpt2, Has2 expression, and global beta-O-linked glycosylation of proteins, compared to control or FSH, suggesting greater HBP activity. Our findings suggest that 1) EGF-like peptides, particularly epiregulin, induce more oocyte mitochondrial activity than EGF or FSH and 2) EGF-like peptides and EGF induce greater HBP activity, enabling more hyaluronic acid synthesis and protein beta-O-linked glycosylation. These metabolic alterations may be a mechanism by which EGF-like peptides increase oocyte developmental competence.

The aim of the studies described in the present report was to determine the systemic bioavailability of intranasal fluticasone propionate (FP) in healthy volunteers and in patients with atopic rhinitis. In addition, the effect that any FP that was absorbed may have on the hypothalamic pituitary (HPA) axis was also determined. In six separate studies, subjects were administered FP intranasally either once or twice daily for up to 7.5 days. In studies where FP was given once daily, doses ranged from 0.2 to 1mg. In studies where FP was given twice daily, dosages up to 2.0mg twice daily were given. At intervals following the last dose of FP, blood samples were taken for determination of plasma levels of FP and, in a number of studies, cortisol. In some studies, urine was collected for 24 hours following the last dose of FP for the determination of urinary cortisol excretion. Following repeated administration of FP (up to 1.0mg twice daily for 7.5 days), plasma levels of FP remained below the lower limit of detection (0.05 μg/L) for the 8-hour investigational period after the last dose of FP. When FP (0.2mg) was administered intranasally to patients with allergic rhinitis after challenge with allergen, plasma levels of FP remained below the lower limit of detection for 6 hours after administration with FP. Following administration of single and repeated doses of FP intranasally (up to 1.0mg twice daily for 7 days), no significant reduction in plasma cortisol concentrations could be measured. In the same subjects dexamethasone (2mg orally) reduced plasma cortisol concentrations from 143.0 ± 6.39 μg/L to < 10 μg/L. Neither single (up to l.Omg) nor repeated (up to 2.0mg twice daily for 7 days) administration of FP caused a significant reduction in urinary cortisol excretion. Even at doses 10 times higher than the dose used clinically, absorption of FP from the nasal mucosa was low (< 2%). It is concluded that any FP that is absorbed from the nasal mucosa is insufficient to exert any measurable effect on the HPA axis.

Objective—To determine the prevalence of Mycoplasma bovis infection in the lungs of cattle at various times after arrival at a feedlot, to measure the relationship between clinical disease status and the concentration and genotype of M bovis within the lungs, and to investigate changes in the genotype of M bovis over time. Sample—Bronchoalveolar lavage fluid (BALF) from 328 healthy or pneumonic beef cattle and 20 M bovis isolates obtained from postmortem samples. Procedures—The concentration of M bovis in BALF was determined via real-time PCR assays, and M bovis isolates from BALF were genotyped via amplified fragment length polymorphism (AFLP) analysis. Results—Prevalence of M bovis in BALF was 1 of 60 (1.7%) at arrival to a feedlot and 26 of 36 (72.2%) and 36 of 42 (85.7%) at ≤ 15 days and 55 days after arrival, respectively. Neither the concentration nor the AFLP type of M bovis in BALF was correlated with clinical disease status. The M bovis AFLP type differed between early and later sampling periods in 14 of 17 cattle. Conclusions and Clinical Relevance—The findings implied spread of M bovis among calves and suggested that host factors and copathogens may determine disease outcomes in infected calves. Chronic pulmonary infection with M bovis may represent a dynamic situation of bacterial clearance and reinfection with strains of different AFLP type, rather than continuous infection with a single clone. These findings impact our understanding of why cattle with chronic pneumonia and polyarthritis syndrome inadequately respond to antimicrobial treatment.

Bidirectional communication between cumulus cells and the oocyte is necessary to achieve oocyte developmental competence. The aim of the present study was to examine the effects of recombinant human bone morphogenetic protein 15 (rhBMP15) and follicle-stimulating hormone (FSH) supplementation on bovine cumulus-oocyte complex (COC) metabolism during maturation. Bovine COCs were matured in the presence of absence of FSH, rhBMP15, or both for 23 h. The addition of FSH and rhBMP15 increased blastocyst development (without rhBMP15 and FSH, 28.4% % 6 7.4% %; with FSH and rhBMP15, 51.5% % 6 5.4% %; P , 0.05). Glucose uptake and lactate production was significantly increased by greater than 2-fold with FSH (P , 0.05), whereas rhBM15 supplementation did not increase these levels. rhBMP15 supplementation (regardless of FSH) significantly decreased ADP levels in COCs, leading to an increase in ATP:ADP ratios (P , 0.05). Indicators of mitochondrial activity and cellular REDOX, oxidized flavin adenine dinucleotide (FAD ++ ) and reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), levels within the oocyte of COCs were significantly higher with rhBMP15 alone, whereas the presence of FSH diminished the rhBMP15 effect. Regardless of treatment, no changes in REDOX state (FAD ++ :NAD(P)H). The significant increase in FAD ++ and NAD(P)H in COCs with rhBMP15 was mediated via cumulus cells, because no differences were found in denuded oocytes cultured in the presence or absence of FSH, rhBMP15, or both. The present study demonstrates that a principal metabolic consequence of FSH supplementation of COCs is to alter the glycolytic rate of cumulus cells, whereas that of rhBMP15 is to regulate oxidative phosphorylation in the oocyte, even though it acts via cumulus cells. These effects are tempered when FSH and rhBMP15 are present together but, nonetheless, yield the best oocyte developmental competence.

Perfectionism and drive for thinness have both been described as predictors of eating disorders, but the relationship between these two constructs over time requires further investigation, as does the an understanding of what components of perfectionism are important in this relationship. Using a longitudinal design, a population of 175 young adults was followed up over a 4 month period. Structural equation modelling indicated a unidirectional relationship between evaluative concerns and drive for thinness, with evaluative concerns, measured at Time 1 predicting an increase in drive for thinness at Time 2. This finding has potential implications for understanding psychological symptoms that precede eating disorder symptoms, and may help build models about prevention and treatment. As a first study to prospectively examine this relationship, further research is needed to assess the generalisability of the findings, and to explore additional variables that may mediate the relationship between evaluative concerns and drive for thinness.

When subjects are read a list of stimulus words and are asked to g enerate an associate to each word, recall for associates given to emotionally toned stimulu s words is poorer than recall for associates given to stimulus words of a neutral affective tone. The present study reports the results of 4 experiments designed to investigate this finding. Expe riment 1 replicated the effect and showed that when subjects associate to an emotionally toned stimu lus word, that associate is likely to be a word with a congruent emotional tone. Experiment 2 demonstrated that reducing the number of emotionally toned stimulus words in the word list re sulted in an improvement in recall for associates generated to these words. In the third exp eriment recall for associates generated to emotionally toned stimulus words was poorer if the s timulus words were similar in meaning (eg. worry, anxiety, panic, fear) than if they were diss imilar (eg. anxiety, death, passion, greed). Experiment 4 found that stimulus word properti es other than emotionality could be manipulated to demonstrate a differential recall. The resu lts throw doubt on an explanation involving impaired consolidation of the associate due to the a rousing properties of the emotional stimulus words. Rather, they are best explained in terms of th e inability of the retrieval system to discriminate amongst competing sets of responses which are s emantically and emotionally congruent.Resume Lors de la lecture d'une liste de termes a des sujets, en leur deman dant d'associer a chacun un autre mot, I'evocation de mots associes a des termes de caractere emotionnel est moins vive que celle de mots associes a des termes neut res. Quatre experiences ont evalue ces conclusions. L'experience 1 a demontre que lorsque les sujets associent un mot a un terme de caractere emotionnel, ce mot a tendance a renfermer la meme valeur emotionnelle. Les resultats de l'experience 2 indiquent qu'une diminution du nombre de termes de caractere emotionnel de la liste rehausse l'evocation de mots associes. Au cours de la troisieme experience, I'evocation de mots associes aux termes de caractere emotionnel etait moins vive lorsque ces termes avaient un sens semblable (p. ex., inquietude, angoisse) que lorsqu'ils n'etaient pas semblables (p. ex., angoisse, mort). Lesresultats de l'experienc e 4 ont indique qu'il etait possible d'obtenir differents resultats d'evocation en manip ulant d'autres caracteristiques des termes, a part leur caractere emotionnel. La disc ussion des resultats porte sur l'incapacite du systeme de recuperation de discerner ent re des groupes de reponses en concurrence les uns par rapport aux autres.In an experiment examining recall for self generated associates to stimulus word s, Jones, O'Gorman, and Byrne (1987, Experiment 2) presented subjects with 30 stimulus wor ds: 10 pleasant, 10 unpleasant, and 10 of a neutral affective tone. Subjects were aske d to generate an associate to each word and, following this, were again presented with each stimu lus word and asked to recall the associate previously paired with it. The results showed tha t the recall of associates given to affectively toned stimulus words (both pleasant and unpleasa nt) was poorer than the recall of associates given to affectively neutral stimulus words. Simi lar findings have been reported by other workers (Levinger & Clark, 1961; Parkin, Lewinsohn & Folk ard, 1982; Rossmann, 1984; Sinton, 1981).Levinger and Clark (1961) argued that some inhibitory process involving the emotional words might account for this finding but they did not elaborate on how this migh t occur. Other authors, (Eysenck & Wilson, 1973; Parkin et al., 1982), have suggested that the effect is due to the arousing properties of the emotional words interfering with the consolida tion of the associate. A number of studies have investigated this hypothesis starting with the idea that arousal impairs immediate recall but facilitates long term recall (see Eysenck, 1976, for a review). …

Covering: up to early 2011 Streptomycetes and other actinobacteria are renowned as a rich source of natural products of clinical, agricultural and biotechnological value. They are being mined with renewed vigour, supported by genome sequencing efforts, which have revealed a coding capacity for secondary metabolites in vast excess of expectations that were based on the detection of antibiotic activities under standard laboratory conditions. Here we review what is known about the control of production of so-called secondary metabolites in streptomycetes, with an emphasis on examples where details of the underlying regulatory mechanisms are known. Intriguing links between nutritional regulators, primary and secondary metabolism and morphological development are discussed, and new data are included on the carbon control of development and antibiotic production, and on aspects of the regulation of the biosynthesis of microbial hormones. Given the tide of antibiotic resistance emerging in pathogens, this review is peppered with approaches that may expand the screening of streptomycetes for new antibiotics by awakening expression of cryptic antibiotic biosynthetic genes. New technologies are also described that have potential to greatly further our understanding of gene regulation in what is an area fertile for discovery and exploitation.

Computerized system validation is often viewed as a burden and a waste of time to meet regulatory requirements. This article presents a different approach by looking at validation in a bioanalytical laboratory from the business benefits that computer validation can bring. Ask yourself the question, have you ever bought a computerized system that did not meet your initial expectations? This article will look at understanding the process to be automated, the paper to be eliminated and the records to be signed to meet the requirements of the GLP or GCP and Part 11 regulations. This paper will only consider commercial nonconfigurable and configurable software such as plate readers and LC–MS/MS data systems rather than LIMS or custom applications. Two streamlined life cycle models are presented. The first one consists of a single document for validation of nonconfigurable software. The second is for configurable software and is a five-stage model that avoids the need to write functional and design specifications. Both models are aimed at managing the risk each type of software poses whist reducing the amount of documented evidence required for validation.

The Sickness Impact Profile (SIP) and the Wechsler Memory Scale--Revised (WMS-R) were administered to a small sample of end-stage renal failure patients. The memory test successfully discriminated between patients who were rated by their nurses to be well adjusted or poorly adjusted to dialysis treatment. It is concluded that this instrument may be useful in investigating cognitive function in this patient population.

A new series of non-stoichiometric sulfides Ga1−xGexV4S8−δ (0≤x≤1; δ≤0.23) has been synthesized at high temperatures by heating stoichiometric mixtures of the elements in sealed quartz tubes. The samples have been characterized by powder X-ray diffraction, SQUID magnetometry and electrical transport-property measurements. Structural analysis reveals that a solid solution is formed throughout this composition range, whilst thermogravimetric data reveal sulfur deficiency of up to 2.9% in the quaternary phases. Magnetic measurements suggest that the ferromagnetic behavior of the end-member phase GaV4S8 is retained at x≤0.7; samples in this composition range showing a marked increase in magnetization at low temperatures. By contrast Ga0.25Ge0.75V4S8−δ appears to undergo antiferromagnetic ordering at ca. 15 K. All materials with x≠1 are n-type semiconductors whose resistivity falls by almost six orders of magnitude with decreasing Ga content, whilst the end-member phase GeV4S8−δ is a p-type semiconductor. The results demonstrate that the physical properties are determined principally by the degree of electron filling of narrow-band states arising from intracluster V–V interactions.

Five Gram-stain-positive, motile, aerobic strains were isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. All strains are rod-shaped, spore-forming bacteria, whose spores were resistant to UV radiation up to 1000 J m−2. The spores were subterminally positioned and produced an external layer. A polyphasic taxonomic study including traditional biochemical tests, fatty acid analysis, cell-wall typing, lipid analyses, 16S rRNA gene sequencing and DNA–DNA hybridization studies was performed to characterize these novel strains. 16S rRNA gene sequencing and lipid analyses convincingly grouped these novel strains within the genus Bacillus as a cluster separate from already described species. The similarity of 16S rRNA gene sequences among the novel strains was >99 %, but the similarity was only about 97 % with their nearest neighbours Bacillus pocheonensis, Bacillus firmus and Bacillus bataviensis. DNA–DNA hybridization dissociation values were 15 : 0 and anteiso-C15 : 0 and the cell-wall diamino acid was meso-diaminopimelic acid. Based on phylogenetic and phenotypic results, it is concluded that these strains represent a novel species of the genus Bacillus, for which the name Bacillus horneckiae sp. nov. is proposed. The type strain is 1P01SCT (=NRRL B-59162T =MTCC 9535T).