Leptin resistance contributes to obesity and hypertension in mouse models of Bardet-Biedl syndrome.
Bardet-Biedl syndrome (BBS) is a heterogeneous genetic disorder characterized by many features, including obesity and cardiovascular disease. We previously developed knockout mouse models of 3 BBS genes: BBS2, BBS4, and BBS6. To dissect the mechanisms involved in the metabolic disorders associated with BBS, we assessed the development of obesity in these mouse models and found that BBS-null mice were hyperphagic, had low locomotor activity, and had elevated circulating levels of the hormone leptin. The effect of exogenous leptin on body weight and food intake was attenuated in BBS mice, which suggests that leptin resistance may contribute to hyperleptinemia. In other mouse models of obesity, leptin resistance may be selective rather than systemic; although mice became resistant to leptin's anorectic effects, the ability to increase renal sympathetic nerve activity (SNA) was preserved. Although all 3 of the BBS mouse models were similarly resistant to leptin, the sensitivity of renal SNA to leptin was maintained in Bbs4 -/- and Bbs6 -/- mice, but not in Bbs2 -/- mice. Consequently, Bbs4 -/- and Bbs6 -/- mice had higher baseline renal SNA and arterial pressure and a greater reduction in arterial pressure in response to ganglionic blockade. Furthermore, we found that BBS mice had a decreased hypothalamic expression of proopiomelanocortin, which suggests that BBS genes play an important role in maintaining leptin sensitivity in proopiomelanocortin neurons.


Skin penetration of epigallocatechin-3-gallate and quercetin from green tea and Ginkgo biloba extracts vehiculated in cosmetic formulations.
Green tea (Camellia sinensis) and Ginkgo biloba extracts in cosmetic formulations have been suggested to protect the skin against UV-induced damage and skin ageing. Thus, it is very important to assess the human skin penetration of their major flavonoids to verify if they penetrate and remain in the skin to exert their proposed effects. The aim of this study was to evaluate the human skin penetration of epigallocatechin-3-gallate (EGCG) and quercetin from green tea and G. biloba extracts vehiculated in cosmetic formulations. This study was conducted with fresh dermatomed human Caucasian skin from abdominal surgery mounted on static Franz diffusion cells. Skin samples were mounted between two diffusion half-cells and 10 mg/cm(2) of formulations supplemented with 6% of green tea or G. biloba extract were applied on the skin surface. The receptor fluid was removed after 6 and 24 h and analyzed by high-performance liquid chromatography for the quantification of the flavonoids. The stratum corneum was removed by tape stripping and immersed in methanol and the epidermis was mechanically separated from the dermis and triturated in methanol to extract EGCG and quercetin. The results showed that the flavonoids under study penetrated into the skin, without reaching the receptor fluid. The majority of EGCG was quantified in the stratum corneum (0.87 microg/cm(2)), which was statistically higher than the EGCG concentrations found in viable epidermis (0.54 microg/cm(2)) and in the dermis (0.38 microg/cm(2)). The majority of quercetin was quantified in the viable epidermis (0.23 microg/cm(2)), which was statistically higher than the EGCG concentration found in the stratum corneum layer (0.17 microg/cm(2)). Finally, it can be concluded that EGCG and quercetin from green tea and G. biloba extracts vehiculated in cosmetic formulations presented good skin penetration and retention, which can favor their skin effects.


Cancer review: Cholangiocarcinoma.
Cholangiocarcinoma (CCA) is the most common biliary tract malignancy. CCA is classified as intrahepatic, perihilar or distal extrahepatic; the individual subtypes differ in their biologic behavior, clinical presentation, and management. Throughout the last decades, CCA incidence rates had significantly increased. In addition to known established risk factors, novel possible risk factors (i.e. obesity, hepatitis C virus) have been identified that are of high importance in developed countries where CCA prevalence rates have been low. CCA tends to develop on the background of inflammation and cholestasis. In recent years, our understanding of the molecular mechanisms of cholangiocarcinogenesis has increased, thereby, providing the basis for molecularly targeted therapies. In its diagnostic evaluation, imaging techniques have improved, and the role of complementary techniques has been defined. There is a need for improved CCA biomarkers as currently used ones are suboptimal. Multiple staging systems have been developed, but none of these is optimal. The prognosis of CCA is considered dismal. However, treatment options have improved throughout the last two decades for carefully selected subgroups of CCA patients. Perihilar CCA can now be treated with orthotopic liver transplantation with neoadjuvant chemoradiation achieving 5-year survival rates of 68%. Classically considered chemotherapy-resistant, the ABC-02 trial has shown the therapeutic benefit of combination therapy with gemcitabine and cisplatin. The benefits of adjuvant treatments for resectable CCA, local ablative therapies and molecularly targeted therapies still need to be defined. In this article, we will provide the reader with an overview over CCA, and discuss the latest developments and controversies.


Diffuse Large B-Cell Lymphoma in Kenya: MYC, BCL2, and the Cell of Origin.
Diffuse large B-cell lymphoma (DLBCL) is the most commonly diagnosed non-Hodgkin lymphoma in adults in Kenya. Cell of origin (COO) and double expression of MYC and BCL2 are two important prognostic factors for DLBCL. A small subset (5% to 10%) of DLBCL cases show positivity for CD5 and are associated with poor prognosis, whereas CD30 antigen, seen in up to 10% of cases, may be a useful target for therapy. We sought to determine the prevalence of MYC/BCL2 double expression, COO, and proportion of Epstein-Barr virus positivity among patients with DLBCL diagnosed at a tertiary referral laboratory in Kenya. All cases of DLBCL diagnosed from 2012 through 2015 in our pathology department were analyzed. Tumor tissue microarray sections were stained with CD20, CD3, CD5, CD30, BCL2, BCL6, CD10, MUM1, MYC, and Ki67, classified for COO on the basis of the Hans algorithm, and subjected to Epstein-Barr virus-encoded small RNAs in situ hybridization. Among 165 DLBCL cases, the median age was 50 years, and there was no sex predilection. Only 18 (10.9%) cases showed double expression for MYC and BCL2. Germinal center B (GCB)-cell type DLBCL accounted for 67 cases (40.6%) and 97 cases (59.4%) were classified as non-GCB. The mean Ki67 proliferation index was significantly higher in the double-expressing (45%) and non-GCB groups (36%) compared with the non-double-expressing group (29%) and GCB group (26%). Sixteen cases (9.7%) were Epstein-Barr virus-encoded small RNAs positive, 12 (75%) of which were non-GCB. DLBCL in Kenya is seen in much younger patients with the poor prognostic non-GCB-type accounting for 59.4% of cases. MYC and BCL2 double expression was seen in fewer tumors than reported in the literature and in significantly older patients.


Experimental heart transplantation: effect of cyclosporine on expression and activity of metzincins.
Metzincins, such as matrix metalloproteases (MMP), and extracellular matrix (ECM) proteins are differentially regulated in inflammation. We hypothesised that metzincins are also dysregulated in experimental acute cardiac allograft rejection. We investigated the Dark Agouti-to-Lewis (DA-to-Lew) rat model of acute cardiac allograft rejection. Cyclosporine (CsA) (7.5 mg/kg/d) was given from transplantation to sacrifice (day +5). At that time, mRNA levels were analysed by Affymetrix genechip and quantitative reverse transcription polymerase chain reaction (qRTPCR). MMP protein and activities were analysed by immunohistology, fluorometry, zymography and Western blots. In untreated rejected DA allografts, mRNA levels of MMP-2/-7/-9/-/12-/14, a disintegrin and metalloprotease (ADAM)-17, tissue inhibitor of metalloprotease (TIMP)-1/-3 were increased, whereas MMP-11/-16/-24 and TIMP-2/-4 were lowered compared to native DA hearts. With respect to these untreated allografts, CsA lowered mRNA levels of MMP-7, TIMP-1/-3 (TIMP-2/-4 remained relatively low) and ADAM17, but augmented mRNA levels of MMP-11/-16/-23 and of many ECM genes. Immunohistology showed increased staining of MMP-2 in acute rejection (AR). Overall MMP activity was augmented in both transplanted groups, but CsA reduced MMP-9 activity and MMP-14 production. Taken together, MMP and TIMP were upregulated during acute AR. CsA ameliorated histology of rejection but showed potential pro-fibrotic effects. Thus, MMP and TIMP may play a role in acute cardiac allograft rejection, and beneficial modification of the MMP-ECM balance requires interventions beyond CsA.


Hydroxyapatite, fluor-hydroxyapatite and fluorapatite produced via the sol-gel method: dissolution behaviour and biological properties after crystallisation.
Hydroxyapatite (HA), fluor-hydroxyapatite (FHA) with varying levels of fluoride ion substitution and fluorapatite (FA) were synthesised by the sol-gel method as possible implant coating or bone-grafting materials. Calcium nitrate and triethyl phosphite were used as precursors under an ethanol-water based solution. Different amounts of ammonium fluoride were incorporated for the preparation of the FHA and FA sol-gels. After heating and powdering the sol-gels, dissolution behaviour was assessed using ion chromatography to measure Ca(2+) and PO4 (3-) ion release. Biological behaviour was assessed using cellular proliferation with human osteosarcoma cells and alamarBlue™ assay. Statistical analysis was performed with a two way analysis of variance and post hoc testing with a Bonferroni correction. Increasing fluoride substitution into an apatite structure decreased the dissolution rate. Increasing the firing temperature of the HA, FHA and FA sol-gels up to 1,000 °C decreased the dissolution rate. There was significantly higher cellular proliferation on highly substituted FHA and FA than on HA or Titanium. The properties of an implant coating or bone grafting material can be tailored to meet specific requirements by altering the amount of fluoride that is incorporated into the original apatite structure. The dissolution behaviour can further be altered by the temperature at which the sol-gel is fired.


Ligation of ICAM-1 molecules inhibits target cell-induced granule exocytosis of IL-12-activated natural killer cells.
The importance of cell adhesion molecules such as ICAM-1 is emphasized in cell-to-cell interactions that are critical in the generation of effective immune reactions. In this study, the involvement of ICAM-1 in natural killer (NK) cell activities was characterized in IL-12-activated human NK cells. To address the question of whether ligation of ICAM-1 molecules can modulate NK cell cytolytic activities, a 4-h (51)Cr-release assay was performed after pretreatment of NK cells with R6.5 mAb (anti-human ICAM-1 mAb). Ligation of membrane ICAM-1 molecules significantly inhibited IL-12-enhanced NK cytotoxicity against K562, and the pretreatment of neutralizing soluble ICAM-1 with R6.5 mAb blocked this inhibitory effect. The involvement of Ca(2+)-dependent granular exocytosis was evaluated. BLT esterase assay demonstrated that the ligation of ICAM-1 molecules inhibited granular exocytosis of NK cells. Additionally, the ICAM-1-mediated inhibition of Ca(2+) flux in NK cells was detected using Fluo-3AM, while the pretreatment of NK cells with R6.5 mAb did not affect conjugate formation between NK and K562 cells. Collectively, these results suggest that the signals transduced from ICAM-1 molecules might be sufficient to induce inhibitory effects on NK cells.


Liquid-phase chemical sensing using lateral mode resonant cantilevers.
Liquid-phase operation of resonant cantilevers vibrating in an out-of-plane flexural mode has to date been limited by the considerable fluid damping and the resulting low quality factors (Q factors). To reduce fluid damping in liquids and to improve the detection limit for liquid-phase sensing applications, resonant cantilever transducers vibrating in their in-plane rather than their out-of-plane flexural resonant mode have been fabricated and shown to have Q factors up to 67 in water (up to 4300 in air). In the present work, resonant cantilevers, thermally excited in an in-plane flexural mode, are investigated and applied as sensors for volatile organic compounds in water. The cantilevers are fabricated using a complementary metal oxide semiconductor (CMOS) compatible fabrication process based on bulk micromachining. The devices were coated with chemically sensitive polymers allowing for analyte sorption into the polymer. Poly(isobutylene) (PIB) and poly(ethylene-co-propylene) (EPCO) were investigated as sensitive layers with seven different analytes screened with PIB and 12 analytes tested with EPCO. Analyte concentrations in the range of 1-100 ppm have been measured in the present experiments, and detection limits in the parts per billion concentration range have been estimated for the polymer-coated cantilevers exposed to volatile organics in water. These results demonstrate significantly improved sensing properties in liquids and indicate the potential of cantilever-type mass-sensitive chemical sensors operating in their in-plane rather than out-of-plane flexural modes.


Somatic cell counts in bovine milk.
Factors which influence somatic cell counts in bovine milk are reviewed and guidelines for their interpretation are presented. It is suggested that the thresholds of 300 000 and 250 000 cells/mL be used to identify infected quarters and cows respectively. However, it is stressed that somatic cell counts are general indicators of udder health which are subject to the influence of many factors. Therefore the evaluation of several successive counts is preferable to the interpretation of an individual count.Relationships between somatic cell counts and both milk production and milk composition are discussed. Subclinical mastitis reduces milk quality and decreases yield although the relationship between production loss and somatic cell count requires clarification. Finally the availability of somatic cell counting programs in Canada is presented.


Phase I and pharmacologic study of intermittently administered 9-nitrocamptothecin in patients with advanced solid tumors.
9-Nitrocamptothecin (9NC) is an oral camptothecin analogue currently administered at 1.5 mg/m(2)/day x 5 days/week in Phase III studies for pancreatic carcinoma. In an effort to increase the dose administered per day and determine whether the daily dose or number of days of treatment influence toxicity, we performed a Phase I study of 9NC using intermittent schedules of administration. On schedule A, 9NC was administered orally daily x 5 days for 2 weeks every 4 weeks (one cycle). On schedule B, 9NC was administered orally daily x 14 days every 4 weeks (one cycle). Dose levels were determined by adaptive dose finding. Serial blood samples were obtained on day 1 of each schedule for pharmacokinetic studies of 9NC and its 9-aminocamptothecin (9AC) metabolite, and lactone forms were measured by high-performance liquid chromatography. The recommended Phase II doses for schedules A and B were 2.43 and 1.70 mg/m(2)/day, respectively, each providing the same dose intensity (i.e., 24 mg/m(2)/cycle). The primary toxicities on schedules A and B were neutropenia, thrombocytopenia, and diarrhea. On schedule A, two patients with gastric cancer and two patients with pancreatic cancer had stable disease for more than six cycles. On schedule B, one patient with pancreatic cancer had stable disease for more than six cycles, and a patient with pancreatic cancer had a partial response. There was significant interpatient variability in the disposition of 9NC and 9AC. Most of the drug remained in the 9NC form with a ratio of 9NC to 9AC of approximately 4 to 1. These studies suggest that 9NC administered on an intermittent schedule is tolerable and may be an active regimen in patients with gastric or pancreatic cancers. Dosing 9NC on a mg/m(2) basis does not reduce pharmacokinetic variability.


PPAR alpha ligand protects during cisplatin-induced acute renal failure by preventing inhibition of renal FAO and PDC activity.
Previous studies demonstrated that during cisplatin-induced acute renal failure, there is a significant reduction in proximal tubule fatty acid oxidation. We now report on the effects of peroxisome proliferator-activated receptor-alpha (PPAR alpha) ligand Wy-14643 (WY) on the abnormalities of medium chain fatty acid oxidation and pyruvate dehydrogenase complex (PDC) activity in kidney tissue of cisplatin-treated mice. Cisplatin causes a significant reduction in mRNA levels and enzyme activity of mitochondrial medium chain acyl-CoA dehydrogenase (MCAD). PPAR alpha ligand WY ameliorated cisplatin-induced acute renal failure and prevented cisplatin-induced reduction of mRNA levels and enzyme activity of MCAD. In contrast, in cisplatin-treated PPAR alpha null mice, WY did not protect kidney function and did not reverse cisplatin-induced decreased expression of MCAD. Cisplatin inhibited renal PDC activity before the development of acute tubular necrosis, and PDC inhibition was reversed by pretreatment with PPAR alpha agonist WY. Cisplatin also induced increased mRNA and protein levels of pyruvate dehydrogenase kinase-4 (PDK4), and PPAR alpha ligand WY prevented cisplatin-induced increased expression of PDK4 protein levels in wild-type mice. We conclude that PPAR alpha agonists have therapeutic potential for cisplatin-induced acute renal failure. Use of PPAR alpha ligands prevents acute tubular necrosis by ameliorating cisplatin-induced inhibition of two distinct metabolic processes, MCAD-mediated fatty acid oxidation and PDC activity.


Molecular characterization of NbCHMP1 encoding a homolog of human CHMP1 in Nicotiana benthamiana.
In humans, CHMP1 encodes a protein of dual function that plays a role in both modification of chromatin structure and endosomal vesicle trafficking. Recently, it was found that sal1, a CHMP1 homolog in maize, is important for the development of the aleurone cell layers in maize endosperm. In this study, we investigated the structure and function of a Nicotiana benthamiana CHMP1 homolog designated NbCHMP1. NbCHMP1 encodes a small protein with a bipartite nuclear localization signal at its N-terminus, and good homology with the corresponding genes from diverse plants and animals. NbCHMP1 mRNA was present at comparable levels in stems, roots, flowers, and leaves. A GFP fusion of the full length NbCHMP1 protein was localized to the cytosol in distinct structures, while a GFP fusion of its N-terminal 80 aa was targeted to the nucleus, suggesting dual-targeting of NbCHMP1 in plants. Overexpression of NbCHMP1 in yeast did not affect its growth and the expressed protein was present in the cytosol in particulate form. Virus-induced gene silencing of NbCHMP1 resulted in subtle alterations of leaf morphology and color, without significantly affecting plant viability or development. Thus the CHMP1 homolog apparently does not play an essential role in the development of the vegetative tissues of N. benthamiana, in contrast to the essential role of sail in formation of the aleurone cell layers during maize endosperm development.


The roles of first phase, second phase insulin secretion, insulin resistance, and glucose effectiveness of having prediabetes in nonobese old Chinese women.
It has been established that prediabetes can causes significant comorbidities, particularly in the elderly. The deterioration of glucose metabolism are generally considered to be results of the impairment of the 4 factors: first, second insulin secretion (FPIS, SPIS, respectively), glucose effectiveness (GE), and insulin resistance. In this study, we enrolled older women to investigate their relationships with prediabetes.Five thousand four hundred eighty-two nonobese, nondiabetic women were included. They were divided into normal glucose tolerance and prediabetes groups. Receiver operating characteristic curve was performed to investigate the effects on whether to have prediabetes for each factors. Two models were built: Model 1: FPIS + SPIS, and Model 2: model 1 + GE. The area under the receiver operating characteristic (aROC) curve was used to determine the predictive power of these models.The aROC curve of GE was significantly higher than the diagonal line followed by SPIS and FPIS accordingly. The aROC curve of Model 1 (0.611) was not different from GE. However, Model 2 improved significantly up to 0.663. Based on this model, an equation was built (-0.003 × GE - 212.6 × SPIS - 17.9 × insulin resistance + 4.8). If the calculated value is equal or higher than 0 (≥0), then the subject has higher chance to have prediabetes (sensitivity = 0.607, specificity = 0.635).Among the 4 factors, GE is the most important contributor for prediabetes in older women. By building a model composed of FPIS, SPIS, and GE, the aROC curve increased significantly. The equation built from this model could predict prediabetes precisely.


Derivation of the acceptance and self-worth adjustment scale.
The original 55-item Nottingham Adjustment Scale (NAS) is a first generation self-report instrument constructed using classical test theory to evaluate adjustment to vision loss. This study assesses the function of the NAS using Rasch analysis in a sample of adults with visual impairment and presents a revised second-generation instrument. Ninety-nine subjects with established vision loss (median onset 5 years) were administered the NAS. Rasch analysis was performed to: (1) determine optimum response scale function, (2) aid item reduction, (3) determine reliability indices and item targeting, (4) assess unidimensionality using Rasch-based principal component analysis, (5) assess differential item functioning (notable defined as >1.0 logit), and (6) formulate person measures to correlate with Geriatric Depression Scale scores and distance visual acuity to indicate convergent and discriminant validity, respectively. Response categories exhibited underutilization, which when repaired improved response scale functioning and ordered structural calibrations. Misfitting items were removed iteratively until all items had mean-square infit and outfit values of 0.70 to 1.30. However, principal component analysis confirmed insufficient unidimensionality (two contrasts identified, eigenvalues 2.4 and 2.3). Removal of these contrasts and two further iterations restored unidimensionality. Despite item mistargeting (1.58 logits), the revised 19-item instrument demonstrated good person (0.85) and item (0.96) reliability coefficients, good convergent and discriminant validity, and no systematic differential item functioning. The resultant 19-item instrument was termed the Acceptance and Self-Worth Adjustment Scale (AS-WAS). In those with established vision loss, the 19-item Acceptance and Self-Worth Adjustment Scale is a reliable and valid instrument that estimates the level of adjustment concerned with acceptance, attitudes, self-esteem, self-efficacy, and locus of control. An additional measure of depression and anxiety is recommended to assess adjustment in a broader sense. Confirmation of item ordering is required if to be used in those with newly acquired vision loss.


Micrococcal nuclease does not substantially bias nucleosome mapping.
We have mapped sequence-directed nucleosome positioning on genomic DNA molecules using high-throughput sequencing. Chromatins, prepared by reconstitution with either chicken or frog histones, were separately digested to mononucleosomes using either micrococcal nuclease (MNase) or caspase-activated DNase (CAD). Both enzymes preferentially cleave internucleosomal (linker) DNA, although they do so by markedly different mechanisms. MNase has hitherto been very widely used to map nucleosomes, although concerns have been raised over its potential to introduce bias. Having identified the locations and quantified the strength of both the chicken or frog histone octamer binding sites on each DNA, the results obtained with the two enzymes were compared using a variety of criteria. Both enzymes displayed sequence specificity in their preferred cleavage sites, although the nature of this selectivity was distinct for the two enzymes. In addition, nucleosomes produced by CAD nuclease are 8-10 bp longer than those produced with MNase, with the CAD cleavage sites tending to be 4-5 bp further out from the nucleosomal dyad than the corresponding MNase cleavage sites. Despite these notable differences in cleavage behaviour, the two nucleases identified essentially equivalent patterns of nucleosome positioning sites on each of the DNAs tested, an observation that was independent of the histone type. These results indicate that biases in nucleosome positioning data collected using MNase are, under our conditions, not significant.


Role of alcohol in cancers of the upper alimentary tract: use of models in risk assessment.
A case-control study of cancers of the upper alimentary tract was conducted in a hospital to assess the role of several risk factors, including alcohol consumption, which is reported here. Male patients from one community with cancers of the oral cavity (n = 278), pharynx (n = 225), and oesophagus (n = 236) formed the case group. Patients diagnosed as not having cancer (n = 215) formed one control group, and a comparable sample of individuals from the general population (n = 177) formed another control group. The risk of regular alcohol consumption along with the two well established risk factors of tobacco smoking and chewing were assessed from the linear logistic model fitted. The process of model fitting has been elaborated. Adjusted odds ratios of alcohol consumption in those under 60 years of age varied from 1.3 to 3.6-fold for developing oral cavity cancer, from 1.9 to 5.4-fold for pharyngeal cancer, and from 1.5 to 2.7-fold for oesophageal cancer, in different age groups. No association was observed between alcohol consumption and cancer in those over 60 years of age. A synergistic effect was observed for the combined habit of alcohol drinking with tobacco smoking and/or chewing. The fact that age is a risk factor independent of habit is also demonstrated.


A qualitative investigation into the effects of brief training in solution-focused therapy in a social work team.
Previous research into the effectiveness of brief training in psychological therapies for non-therapist health and social services workers has found mixed results regarding transferring learning into practice. Very few published studies have assessed the impact of such training in Solution-Focused Brief Therapy (SFBT), despite such training being popular. This study explored the impact of brief SFBT training for a group of community-based social workers. A qualitative interview-based design was used, with the researcher adopting an ethnographic stance to obtain a rich, detailed and focused account of events. Six social workers working with adults with intellectual disabilities took part in the study. All had attended a 2-day workshop in SFBT 9 months previously. The interviews were transcribed and subjected to thematic analysis. Factors affecting skill transfer included being able to practise, peer and organizational support, and perceived conflicts between SFBT and work role. Whilst specific trained techniques were not consistently adopted, most workers changed the style of their interaction with clients in simple ways which they reported resulted in benefits including improved communication, collaboration, and increased feelings of control and self-efficacy for clients and workers. The findings suggest that in addition to peer and organizational support, trainees' perceptions of the fit of the training with their work role may be a key factor in determining transfer. Recommendations are made for developing training for non-therapists that focuses on helping change the style of their interactions rather than on learning to implement specific psychological therapy techniques.


Choking vs. clutch performance: a study of sport performance under pressure.
Choking research in sport has suggested that an athlete's tendency to choke, versus give a better than usual (i.e., "clutch") performance depends on his or her personality, as well as on situational influences, such as a reliance on explicit (versus implicit) knowledge when pressured. The current study integrated these hypotheses and tested a structural equation model (SEM) to predict sport performance under pressure. Two hundred and one participants attempted two sets of 15 basketball free throws, and were videotaped during their second set of shots as a manipulation of pressure. Results of the model suggest that "reinvesting" attention in the task leads to greater anxiety (cognitive and somatic), which then predicts a higher level of self-focus; self-focus, then, did not lead to improved performance under pressure, whereas feelings of self-reported "perceived control" did help performance. Implications for measurement of these constructs, and their relationships with performance, are discussed.


Loss of form vision impairs spatial imagery.
Previous studies have reported inconsistent results when comparing spatial imagery performance in the blind and the sighted, with some, but not all, studies demonstrating deficits in the blind. Here, we investigated the effect of visual status and individual preferences ("cognitive style") on performance of a spatial imagery task. Participants with blindness resulting in the loss of form vision at or after age 6, and age- and gender-matched sighted participants, performed a spatial imagery task requiring memorization of a 4 × 4 lettered matrix and subsequent mental construction of shapes within the matrix from four-letter auditory cues. They also completed the Santa Barbara Sense of Direction Scale (SBSoDS) and a self-evaluation of cognitive style. The sighted participants also completed the Object-Spatial Imagery and Verbal Questionnaire (OSIVQ). Visual status affected performance on the spatial imagery task: the blind performed significantly worse than the sighted, independently of the age at which form vision was completely lost. Visual status did not affect the distribution of preferences based on self-reported cognitive style. Across all participants, self-reported verbalizer scores were significantly negatively correlated with accuracy on the spatial imagery task. There was a positive correlation between the SBSoDS score and accuracy on the spatial imagery task, across all participants, indicating that a better sense of direction is related to a more proficient spatial representation and that the imagery task indexes ecologically relevant spatial abilities. Moreover, the older the participants were, the worse their performance was, indicating a detrimental effect of age on spatial imagery performance. Thus, spatial skills represent an important target for rehabilitative approaches to visual impairment, and individual differences, which can modulate performance, should be taken into account in such approaches.


Qualitative Behavioral Assessment in Juvenile Farmed Atlantic Salmon (Salmo salar): Potential for On-Farm Welfare Assessment.
There is a growing scientific and legislative consensus that fish are sentient, and therefore have the capacity to experience pain and suffering. The assessment of the welfare of farmed fish is challenging due to the aquatic environment and the number of animals housed together. However, with increasing global production and intensification of aquaculture comes greater impetus for developing effective tools which are suitable for the aquatic environment to assess the emotional experience and welfare of farmed fish. This study therefore aimed to investigate the use of Qualitative Behavioral Assessment (QBA), originally developed for terrestrial farmed animals, in farmed salmon and evaluate its potential for use as a welfare monitoring tool. QBA is a "whole animal" approach based on the description and quantification of the expressive qualities of an animal's dynamic style of behaving, using descriptors such as relaxed, agitated, lethargic, or confident. A list of 20 qualitative descriptors was generated by fish farmers after viewing video-footage showing behavior expressions representative of the full repertoire of salmon in this context. A separate, non-experienced group of 10 observers subsequently watched 25 video clips of farmed salmon, and scored the 20 descriptors for each clip using a Visual Analog Scale (VAS). To assess intra-observer reliability each observer viewed the same 25 video clips twice, in two sessions 10 days apart, with the second clip set presented in a different order. The observers were unaware that the two sets of video clips were identical. Data were analyzed using Principal Component (PC) Analysis (correlation matrix, no rotation), revealing four dimensions that together explained 79% of the variation between video clips, with PC1 (Tense/anxious/skittish-Calm/mellow/relaxed) explaining the greatest percentage of variation (56%). PC1 was the only dimension to show acceptable inter- and intra-observer reliability, and mean PC1 scores correlated significantly to durations of slow and erratic physical movements measured for the same 25 video clips. Further refinements to the methodology may be necessary, but this study is the first to provide evidence for the potential of Qualitative Behavioral Assessment to serve as a time-efficient welfare assessment tool for juvenile salmon under farmed conditions.


Standardized microgel beads as elastic cell mechanical probes.
Cell mechanical measurements are gaining increasing interest in biological and biomedical studies. However, there are no standardized calibration particles available that permit the cross-comparison of different measurement techniques operating at different stresses and time-scales. Here we present the rational design, production, and comprehensive characterization of poly-acrylamide (PAAm) microgel beads mimicking size and overall mechanics of biological cells. We produced mono-disperse beads at rates of 20-60 kHz by means of a microfluidic droplet generator, where the pre-gel composition was adjusted to tune the beads' elasticity in the range of cell and tissue relevant mechanical properties. We verified bead homogeneity by optical diffraction tomography and Brillouin microscopy. Consistent elastic behavior of microgel beads at different shear rates was confirmed by AFM-enabled nanoindentation and real-time deformability cytometry (RT-DC). The remaining inherent variability in elastic modulus was rationalized using polymer theory and effectively reduced by sorting based on forward-scattering using conventional flow cytometry. Our results show that PAAm microgel beads can be standardized as mechanical probes, to serve not only for validation and calibration of cell mechanical measurements, but also as cell-scale stress sensors.


Perspective: National Football League Teams Need Chief Diversity Officers.
The National Football League (NFL) and its teams, some of the world's most profitable sporting properties, face challenges with diversity, equity, and inclusion (DEI). With a history of discriminatory work environments, including a recent high-profile lawsuit, the NFL and its teams have a poor reputation on these issues. This perspective piece investigated NFL teams' utilization of organization employees dedicated to DEI. Utilizing a content analysis of publicly available data, this piece investigated DEI employees at NFL team organizations. The study analyzed the position's characteristics including the name of the role, the department in which it was housed, and the reporting structure. The study also examined the demographics and professional background of the employees in the roles. The findings conclude that NFL teams lag behind other American businesses in their adoption of Chief Diversity Officer (CDO) roles. As of 2022, only 31.25% of NFL teams had a dedicated DEI staff person. Three additional teams host diversity councils utilizing employees with other job responsibilities. The employees filling the CDO roles were majority women and majority Black. Though not the only answer to a complex problem, in order to address these challenges and move forward, the NFL teams should create Chief Diversity Officer (CDO) roles. These positions should have appropriate reporting relationships, well-crafted position responsibilities, generous resources, and qualified and experienced employees.


Comparison of cross-sectional HIV incidence assay results from dried blood spots and plasma.
Assays have been developed for cross-sectional HIV incidence estimation using plasma samples. Large scale surveillance programs are planned using dried blood spot (DBS) specimens for incidence assessment. However, limited information exists on the performance of HIV cross-sectional incidence assays using DBS. The assays evaluated were: Maxim HIV-1 Limiting Antigen Avidity EIA (LAg-Avidity), Sedia HIV-1 BED-Capture EIA (BED-CEIA), and CDC modified BioRad HIV-1/2 Plus O Avidity-based Assay (CDC-BioRad Avidity) using pre-determined cutoff values. 100 matched HIV-1 positive plasma and DBS samples, with known duration of infection, from the Consortium for the Evaluation and Performance of HIV Incidence Assays repository were tested. All assays were run in duplicate. To examine the degree of variability within and between results for each sample type, both categorical and continuous results were analyzed. Associations were assessed with Bland Altman, R2 values and Cohen's kappa coefficient (ĸ). Intra-assay variability using the same sample type was similar for all assays (R2 0.96 to 1.00). The R2 values comparing DBS and plasma results for LAg-Avidity, BED-CEIA, and CDC-BioRad Avidity were 0.96, 0.94, and 0.84, respectively. The concordance and ĸ values between DBS and plasma for all three assays were >87% and >0.64, respectively. The Bland-Altman analysis showed significant differences between plasma and DBS samples. For all three assays, a higher number of samples were classified as recent infections using DBS samples. DBS and plasma sample results were highly correlated. However, when compared to plasma, each assay performed somewhat differently in DBS at the lower and higher ends of the dynamic range. DBS samples were more likely to be classified as recently infected by all three assays, which may lead to overestimation of incidence in surveys using performance criteria derived for plasma samples.


Adaptive cellular immunity in aortic aneurysms: cause, consequence, or context?
Abdominal aortic aneurysms are common and life threatening. Although CD4(+) T cells are abundant in aneurysm tissue, their role in disease progression remains unclear. A new study shows that mouse aortic allografts placed in animals lacking IFN-gamma receptors develop a Th2 inflammatory response with aortic aneurysms, whereas Th1 responses promote intimal hyperplasia. It is expected that these surprising findings will stimulate further efforts to clarify whether adaptive cellular immunity in aneurysm disease is detrimental or potentially beneficial.


Extraordinary anisotropy of ultrafast laser writing in glass.
The unusual dependence of femtosecond laser writing on the light polarization and direction of raster scanning is demonstrated in silica and chalcogenide glasses. Two different mechanisms contributing to the observed anisotropy are identified: the chevron-shaped stress induced by the sample movement and the pulse front tilt of ultrashort light pulse. Control of anisotropies associated with the spatio-temporal asymmetry of an ultrashort pulse beam and scanning geometry is crucial in the ultrafast laser machining of transparent materials.


Helping effort increases with relatedness in bell miners, but 'unrelated' helpers of both sexes still provide substantial care.
Indirect fitness benefits from kin selection can explain why non-breeding individuals help raise the young of relatives. However, the evolution of helping by non-relatives requires direct fitness benefits, for example via group augmentation. Here, we examine nest visit rates, load sizes and prey types delivered by breeding pairs and their helpers in the cooperatively breeding bell miner (Manorina melanophrys). In this system, males remain in their natal colony while young females typically disperse, and helpers of both sexes often assist at multiple nests concurrently. We found extremely clear evidence for the expected effect of genetic relatedness on individual helping effort per nest within colonies. This positive incremental effect of kinship was facultative-i.e. largely the result of within-individual variation in helping effort. Surprisingly, no sex differences were detectable in any aspect of helping, and even non-relatives provided substantial aid. Helpers and breeders of both sexes regulated their provisioning effort by responding visit-by-visit to changes in nestling begging. Helping behaviour in bell miners therefore appears consistent with adaptive cooperative investment in the brood, and kin-selected care by relatives. Similar investment by 'unrelated' helpers of both sexes argues against direct fitness benefits, but is perhaps explained by kin selection at the colony level.


RecD plays an essential function during growth at low temperature in the antarctic bacterium Pseudomonas syringae Lz4W.
The Antarctic psychrotrophic bacterium Pseudomonas syringae Lz4W has been used as a model system to identify genes that are required for growth at low temperature. Transposon mutagenesis was carried out to isolate mutant(s) of the bacterium that are defective for growth at 4 degrees but normal at 22 degrees . In one such cold-sensitive mutant (CS1), the transposon-disrupted gene was identified to be a homolog of the recD gene of several bacteria. Trans-complementation and freshly targeted gene disruption studies reconfirmed that the inactivation of the recD gene leads to a cold-sensitive phenotype. We cloned, sequenced, and analyzed approximately 11.2 kbp of DNA from recD and its flanking region from the bacterium. recD was the last gene of a putative recCBD operon. The RecD ORF was 694 amino acids long and 40% identical (52% similar) to the Escherichia coli protein, and it could complement the E. coli recD mutation. The recD gene of E. coli, however, could not complement the cold-sensitive phenotype of the CS1 mutant. Interestingly, the CS1 strain showed greater sensitivity toward the DNA-damaging agents, mitomycin C and UV. The inactivation of recD in P. syringae also led to cell death and accumulation of DNA fragments of approximately 25-30 kbp in size at low temperature (4 degrees ). We propose that during growth at a very low temperature the Antarctic P. syringae is subjected to DNA damage, which requires direct participation of a unique RecD function. Additional results suggest that a truncated recD encoding the N-terminal segment of (1-576) amino acids is sufficient to support growth of P. syringae at low temperature.


Denervation-induced supersensitivity to calcium of chemically skinned smooth muscle of the guinea-pig vas deferens.
Chronic denervation of the smooth muscle of the guinea-pig vas deferens results in postjunctional supersensitivity. A partial membrane depolarization contributes to the enhanced sensitivity. However, there is evidence that supersensitivity results in part from cellular alterations other than membrane potential changes. In the present study we investigated the possibility that changes in the sensitivity of the contractile proteins may contribute to the phenomenon. The approach was to evaluate the concentration-response relationship for calcium of "skinned" preparations of guinea-pig vas deferens. The tissues were chemically skinned with either triton X-100 or saponin. Concentration-response curves for calcium were constructed 1 week after postganglionic denervation of one of a pair of vasa deferentia. The curve for denervated tissues was shifted significantly to the left of control (1.5-fold at the level of the EC50). In addition to supersensitivity, denervated skinned preparations exhibited a faster rate of contraction. The phenothiazine compounds trifluperazine, fluphenazine and chlorpromazine relaxed skinned muscles which were contracted with calcium. The relaxation was concentration-dependent, and was less pronounced in denervated vasa deferentia than in controls. These results suggest that increased sensitivity of the contractile proteins may contribute to denervation-induced supersensitivity of smooth muscle, and further that this may be related to a change in the activity or amount of calmodulin.


The validity of the 16-item version of the Prodromal Questionnaire (PQ-16) to screen for ultra high risk of developing psychosis in the general help-seeking population.
In order to bring about implementation of routine screening for psychosis risk, a brief version of the Prodromal Questionnaire (PQ; Loewy et al., 2005) was developed and tested in a general help-seeking population. We assessed a consecutive patient sample of 3533 young adults who were help-seeking for nonpsychotic disorders at the secondary mental health services in The Hague with the PQ. We performed logistic regression analyses and CHi-squared Automatic Interaction Detector decision tree analysis to shorten the original 92 items. Receiver operating characteristic curves were used to examine the psychometric properties of the PQ-16. In the general help-seeking population, a cutoff score of 6 or more positively answered items on the 16-item version of the PQ produced correct classification of Comprehensive Assessment of At-Risk Mental State (Yung et al., 2005) psychosis risk/clinical psychosis in 44% of the cases, distinguishing Comprehensive Assessment of At-Risk Mental States (CAARMS) diagnosis from no CAARMS diagnosis with high sensitivity (87%) and specificity (87%). These results were comparable to the PQ-92. The PQ-16 is a good self-report screen for use in secondary mental health care services to select subjects for interviewing for psychosis risk. The low number of items makes it quite appropriate for screening large help-seeking populations, thus enhancing the feasibility of detection and treatment of ultra high-risk patients in routine mental health services.


Platelet-Rich Plasma Versus Surgery for the Management of Recalcitrant Greater Trochanteric Pain Syndrome: A Systematic Review.
To perform a systematic review of the outcomes of platelet-rich plasma (PRP) injections as an in-office procedure versus surgical treatment for recalcitrant greater trochanteric pain syndrome (GTPS). The MEDLINE and Embase databases were searched in June 2019 following the Preferred Reporting Items for Systematic Reviews and Meta-analyses statement. Clinical studies on patients with recalcitrant GTPS treated with PRP or surgery were included. Demographic characteristics, patient-reported outcomes (PROs), and complications were compared. A qualitative analysis using the Methodological Index for Non-randomized Studies and Cochrane Risk of Bias Tool scores was performed. A total of 5 PRP and 5 surgery studies met the inclusion criteria, contributing 94 and 185 patients, respectively. The mean follow-up time was shorter for the PRP studies (range, 2-26 months) than with surgery (range, 12-70 months). The mean Methodological Index for Non-randomized Studies scores for the PRP and surgery groups were 11.25 and 11.4, respectively, and the only randomized trial had a low risk of bias. Two studies in the PRP group (n = 56) reported improvements in the modified Harris Hip Score at final follow-up (from 53.8 to 82.6 and from 56.7 to 74.2). The other PRP studies reported improvements using other measures. In the surgery group, 2 studies reported improvements in the Harris Hip Score (from 53.0 to 80 and from 53.3 to 88) whereas 3 used unique PROs (Oxford score, from 20.4 to 37.3; modified Harris Hip Score, from 54.9 to 76.2; and Merle d'Aubigné and Postel score, from 10.9 to 16.7). Although significant improvement was reported in all studies included, PRP showed a large effect size whereas surgery showed a moderate to large effect size. No major complications were associated with PRP treatment; however, the surgery group reported a higher rate of complications including recurrent external snapping hip, retears resulting from falls, trochanteric fracture, venous thrombosis, and wound-related problems. Both PRP and surgical intervention for the treatment of recalcitrant GTPS showed statistically and clinically significant improvements based on PROs. Although not covered by most medical insurance companies, PRP injections for recalcitrant GTPS provides an effective and safe alternative after failed physical therapy. If surgery is indicated, endoscopy is safer than the open technique. Level IV, systematic review of Level I to IV studies.


Schemas and memory consolidation.
Memory encoding occurs rapidly, but the consolidation of memory in the neocortex has long been held to be a more gradual process. We now report, however, that systems consolidation can occur extremely quickly if an associative "schema" into which new information is incorporated has previously been created. In experiments using a hippocampal-dependent paired-associate task for rats, the memory of flavor-place associations became persistent over time as a putative neocortical schema gradually developed. New traces, trained for only one trial, then became assimilated and rapidly hippocampal-independent. Schemas also played a causal role in the creation of lasting associative memory representations during one-trial learning. The concept of neocortical schemas may unite psychological accounts of knowledge structures with neurobiological theories of systems memory consolidation.


Inaccuracy of intraocular lens power prediction for cataract surgery in angle-closure glaucoma.
To assess the accuracy of intraocular lens (IOL) power predictions for cataract surgery in eyes with primary angle-closure glaucoma (ACG). Because of shifting of the capsular bag apparatus and shortening of the axial length, preoperative calculation of IOL power may be inaccurate for eyes with ACG. This retrospective comparative case series comprised of 42 eyes from 42 patients with primary ACG and 45 eyes from 45 subjects with normal open-angles undergoing uneventful cataract surgery. Anterior segment biometry including anterior chamber depth, lens thickness, and axial length were compared. Using the SRK-II formula, the powers of the implanted IOL and the actual postoperative spherical equivalent (SE) refractive errors were compared between the two groups. Also, the absolute values of differences between predicted and residual SE refractive errors were also analyzed for each group. In ACG patients, anterior chamber depth and axial length were shorter and the lens was thicker than normal controls (all p < 0.001). Even though residual SE refractive error was not significantly different (p = 0.290), the absolute value of the difference between predicted and residual SE refractive error was 0.64 +/- 0.50 diopters in AGC patients and 0.39 +/- 0.36 diopters in control subjects (p = 0.012). The number of eyes that resulted in inaccurate IOL power predictions of more than 0.5 diopters were 21 (50.00%) in the ACG group, but only 12 (26.67%) in the control group (p = 0.043). IOL power predictions for cataract surgery in ACG patients can be inaccurate, and it may be associated with their unique anterior segment anatomy.


Multipotent versus differentiated cell fate selection in the developing Drosophila airways.
Developmental potentials of cells are tightly controlled at multiple levels. The embryonic Drosophila airway tree is roughly subdivided into two types of cells with distinct developmental potentials: a proximally located group of multipotent adult precursor cells (P-fate) and a distally located population of more differentiated cells (D-fate). We show that the GATA-family transcription factor (TF) Grain promotes the P-fate and the POU-homeobox TF Ventral veinless (Vvl/Drifter/U-turned) stimulates the D-fate. Hedgehog and receptor tyrosine kinase (RTK) signaling cooperate with Vvl to drive the D-fate at the expense of the P-fate while negative regulators of either of these signaling pathways ensure P-fate specification. Local concentrations of Decapentaplegic/BMP, Wingless/Wnt, and Hedgehog signals differentially regulate the expression of D-factors and P-factors to transform an equipotent primordial field into a concentric pattern of radially different morphogenetic potentials, which gradually gives rise to the distal-proximal organization of distinct cell types in the mature airway.


Genetic or pharmacological depletion of cannabinoid CB1 receptor protects against dopaminergic neurotoxicity induced by methamphetamine in mice.
Accumulating evidence suggests that cannabinoid ligands play delicate roles in cell survival and apoptosis decisions, and that cannabinoid CB1 receptors (CB1R) modulate dopaminergic function. However, the role of CB1R in methamphetamine (MA)-induced dopaminergic neurotoxicity in vivo remains elusive. Multiple high doses of MA increased phospho-ERK and CB1R mRNA expressions in the striatum of CB1R (+/+) mice. These increases were attenuated by CB1R antagonists (i.e., AM251 and rimonabant), an ERK inhibitor (U0126), or dopamine D2R antagonist (sulpiride). In addition, treatment with MA resulted in dopaminergic impairments, which were attenuated by CB1R knockout or CB1R antagonists (i.e., AM251 and rimonabant). Consistently, MA-induced oxidative stresses (i.e., protein oxidation, lipid peroxidation and reactive oxygen species) and pro-apoptotic changes (i.e., increases in Bax, cleaved PKCδ- and cleaved caspase 3-expression and decrease in Bcl-2 expression) were observed in the striatum of CB1R (+/+) mice. These toxic effects were attenuated by CB1R knockout or CB1R antagonists. Consistently, treatment with four high doses of CB1R agonists (i.e., WIN 55,212-2 36mg/kg and ACEA 16mg/kg) also resulted in significant oxidative stresses, pro-apoptotic changes, and dopaminergic impairments. Since CB1R co-immunoprecipitates PKCδ in the presence of MA or CB1R agonists, we applied PKCδ knockout mice to clarify the role of PKCδ in the neurotoxicity elicited by CB1Rs. CB1R agonist-induced toxic effects were significantly attenuated by CB1R knockout, CB1R antagonists or PKCδ knockout. Therefore, our results suggest that interaction between D2R, ERK and CB1R is critical for MA-induced dopaminergic neurotoxicity and that PKCδ mediates dopaminergic damage induced by high-doses of CB1R agonist.


Attention allows the SNARC effect to operate on multiple number lines.
To investigate whether participants can activate only one spatially oriented number line at a time or multiple number lines simultaneously, they were asked to solve a unit magnitude comparison task (unit smaller/larger than 5) and a parity judgment task (even/odd) on two-digit numbers. In both these primary tasks, decades were irrelevant. After some of the primary task trials (randomly), participants were asked to additionally solve a secondary task based on the previously presented number. In Experiment 1, they had to decide whether the two-digit number presented for the primary task was larger or smaller than 50. Thus, for the secondary task decades were relevant. In contrast, in Experiment 2, the secondary task was a color judgment task, which means decades were irrelevant. In Experiment 1, decades' and units' magnitudes influenced the spatial association of numbers separately. In contrast, in Experiment 2, only the units were spatially associated with magnitude. It was concluded that multiple number lines (one for units and one for decades) can be activated if attention is focused on multiple, separate magnitude attributes.


Role of the oxygen-dependent degradation domain in a hypoxia-inducible gene expression system in vascular endothelial growth factor gene therapy.
STUDY DESIGN.: An in vitro neural hypoxia model and rat spinal cord injury (SCI) model were used to assess the regulation effect of a reporter or therapeutic gene expression by an oxygen-dependent degradation (ODD) domain in a hypoxia-inducible gene expression system with or without the erythropoietin (EPO) enhancer. OBJECTIVE.: To increase vascular endothelial growth factor (VEGF) gene expression in SCI lesions but avoid unwanted overexpression of VEGF in normal sites, we developed a hypoxia-inducible gene expression system consisting of the EPO enhancer upstream of the SV promoter and an ODD domain C-terminally fused to VEGF. SUMMARY OF BACKGROUND DATA.: ODD domain plays a major role in the degradation of hypoxia-inducible factor 1alpha and has been used in a hypoxia-specific gene expression system as a post-translational regulatory factor. METHODS.: The hypoxia-inducible luciferase or VEGF plasmid was constructed using the EPO enhancer combined with or without the ODD domain. The constructed plasmid was transfected into mouse Neuro 2a (N2a) neuroblastoma cells by Lipofectamine 2000, followed by a 24-hour incubation in hypoxia or normoxia. For in vivo analysis, the naked plasmid DNA was directly injected into the injured rat spinal cord. The gene expression was evaluated by luciferase activity assay, enzyme-linked immunosorbent assay, reverse transcriptase-polymerase chain reaction, and immunofluorescence staining. RESULTS.: The EPO enhancer/ODD domain-combined hypoxia-inducible gene expression system clearly increased the expression of the reporter luciferase gene and therapeutic VEGF gene specifically under hypoxic conditions and SCI, and quickly downregulated protein expression to a very low level after reoxygenation. CONCLUSION.: These results strongly suggest the potential applicability of this EPO enhancer/ODD domain-based hypoxia-inducible gene expression system in the development of a safer and more effective VEGF gene therapy for SCI.


Treating Rhythmic and Periodic EEG Patterns in Comatose Survivors of Cardiac Arrest.
Whether the treatment of rhythmic and periodic electroencephalographic (EEG) patterns in comatose survivors of cardiac arrest improves outcomes is uncertain. We conducted an open-label trial of suppressing rhythmic and periodic EEG patterns detected on continuous EEG monitoring in comatose survivors of cardiac arrest. Patients were randomly assigned in a 1:1 ratio to a stepwise strategy of antiseizure medications to suppress this activity for at least 48 consecutive hours plus standard care (antiseizure-treatment group) or to standard care alone (control group); standard care included targeted temperature management in both groups. The primary outcome was neurologic outcome according to the score on the Cerebral Performance Category (CPC) scale at 3 months, dichotomized as a good outcome (CPC score indicating no, mild, or moderate disability) or a poor outcome (CPC score indicating severe disability, coma, or death). Secondary outcomes were mortality, length of stay in the intensive care unit (ICU), and duration of mechanical ventilation. We enrolled 172 patients, with 88 assigned to the antiseizure-treatment group and 84 to the control group. Rhythmic or periodic EEG activity was detected a median of 35 hours after cardiac arrest; 98 of 157 patients (62%) with available data had myoclonus. Complete suppression of rhythmic and periodic EEG activity for 48 consecutive hours occurred in 49 of 88 patients (56%) in the antiseizure-treatment group and in 2 of 83 patients (2%) in the control group. At 3 months, 79 of 88 patients (90%) in the antiseizure-treatment group and 77 of 84 patients (92%) in the control group had a poor outcome (difference, 2 percentage points; 95% confidence interval, -7 to 11; P = 0.68). Mortality at 3 months was 80% in the antiseizure-treatment group and 82% in the control group. The mean length of stay in the ICU and mean duration of mechanical ventilation were slightly longer in the antiseizure-treatment group than in the control group. In comatose survivors of cardiac arrest, the incidence of a poor neurologic outcome at 3 months did not differ significantly between a strategy of suppressing rhythmic and periodic EEG activity with the use of antiseizure medication for at least 48 hours plus standard care and standard care alone. (Funded by the Dutch Epilepsy Foundation; TELSTAR ClinicalTrials.gov number, NCT02056236.).


Familial glucocorticoid deficiency in five Arab kindreds with homozygous point mutations of the ACTH receptor (MC2R): genotype and phenotype correlations.
Familial glucocorticoid deficiency type 1 (FGD1) is a rare autosomal-recessive disorder resulting from defective ACTH receptor (melanocortin receptor type 2, MC2R). Individuals with this condition usually present in infancy or early childhood with the signs and symptoms of isolated glucocorticoid deficiency. To date, hypothyroidism has been reported as an associated feature in a few cases. The clinical findings along with MC2R genetic analysis of five Arab kindreds are described. The subjects were children with the clinical and biochemical features of FGD1. Three patients had associated thyroid dysfunction and two patients had associated growth hormone deficiency (GHD). Mutation analysis of MC2R was performed by direct gene sequencing. Analysis of the MC2R gene revealed a homozygous insertion of a cytosine nucleotide between codons 153 and 154 (c.459_460insC) in all of the patients. This mutation would be expected to cause a translation frame shift after codon 154 and a premature termination codon at 248 of the MC2R mRNA (p.I154fsX248). Associated thyroid dysfunction and GHD were clinical features in the Bedouin patients with FGD1 caused by identical homozygous frameshift mutation in the MC2R gene.


Evolutionary dynamics of Enterococcus faecium reveals complex genomic relationships between isolates with independent emergence of vancomycin resistance.
Enterococcus faecium, a major cause of hospital-acquired infections, remains problematic because of its propensity to acquire resistance to vancomycin, which currently is considered first-line therapy. Here, we assess the evolution and resistance acquisition dynamics of E. faecium in a clinical context using a series of 132 bloodstream infection isolates from a single hospital. All isolates, of which 49 (37 %) were vancomycin-resistant, underwent whole-genome sequencing. E. faecium was found to be subject to high rates of recombination with little evidence of sequence importation from outside the local E. faecium population. Apart from disrupting phylogenetic reconstruction, recombination was frequent enough to invalidate MLST typing in the identification of clonal expansion and transmission events, suggesting that, where available, whole-genome sequencing should be used in tracing the epidemiology of E. faecium nosocomial infections and establishing routes of transmission. Several forms of the Tn1549-like element-vanB gene cluster, which was exclusively responsible for vancomycin resistance, appeared and spread within the hospital during the study period. Several transposon gains and losses and instances of in situ evolution were inferred and, although usually chromosomal, the resistance element was also observed on a plasmid background. There was qualitative evidence for clonal expansions of both vancomycin-resistant and vancomycin-susceptible E. faecium with evidence of hospital-specific subclonal expansion. Our data are consistent with continuing evolution of this established hospital pathogen and confirm hospital vancomycin-susceptible and vancomycin-resistant E. faecium patient transmission events, underlining the need for careful consideration before modifying current E. faecium infection control strategies.


A new settling velocity model to describe secondary sedimentation.
Secondary settling tanks (SSTs) are the most hydraulically sensitive unit operations in biological wastewater treatment plants. The maximum permissible inflow to the plant depends on the efficiency of SSTs in separating and thickening the activated sludge. The flow conditions and solids distribution in SSTs can be predicted using computational fluid dynamics (CFD) tools. Despite extensive studies on the compression settling behaviour of activated sludge and the development of advanced settling velocity models for use in SST simulations, these models are not often used, due to the challenges associated with their calibration. In this study, we developed a new settling velocity model, including hindered, transient and compression settling, and showed that it can be calibrated to data from a simple, novel settling column experimental set-up using the Bayesian optimization method DREAM(ZS). In addition, correlations between the Herschel-Bulkley rheological model parameters and sludge concentration were identified with data from batch rheological experiments. A 2-D axisymmetric CFD model of a circular SST containing the new settling velocity and rheological model was validated with full-scale measurements. Finally, it was shown that the representation of compression settling in the CFD model can significantly influence the prediction of sludge distribution in the SSTs under dry- and wet-weather flow conditions.


Fine-mapping and initial characterization of QT interval loci in African Americans.
The QT interval (QT) is heritable and its prolongation is a risk factor for ventricular tachyarrhythmias and sudden death. Most genetic studies of QT have examined European ancestral populations; however, the increased genetic diversity in African Americans provides opportunities to narrow association signals and identify population-specific variants. We therefore evaluated 6,670 SNPs spanning eleven previously identified QT loci in 8,644 African American participants from two Population Architecture using Genomics and Epidemiology (PAGE) studies: the Atherosclerosis Risk in Communities study and Women's Health Initiative Clinical Trial. Of the fifteen known independent QT variants at the eleven previously identified loci, six were significantly associated with QT in African American populations (P≤1.20×10(-4)): ATP1B1, PLN1, KCNQ1, NDRG4, and two NOS1AP independent signals. We also identified three population-specific signals significantly associated with QT in African Americans (P≤1.37×10(-5)): one at NOS1AP and two at ATP1B1. Linkage disequilibrium (LD) patterns in African Americans assisted in narrowing the region likely to contain the functional variants for several loci. For example, African American LD patterns showed that 0 SNPs were in LD with NOS1AP signal rs12143842, compared with European LD patterns that indicated 87 SNPs, which spanned 114.2 Kb, were in LD with rs12143842. Finally, bioinformatic-based characterization of the nine African American signals pointed to functional candidates located exclusively within non-coding regions, including predicted binding sites for transcription factors such as TBX5, which has been implicated in cardiac structure and conductance. In this detailed evaluation of QT loci, we identified several African Americans SNPs that better define the association with QT and successfully narrowed intervals surrounding established loci. These results demonstrate that the same loci influence variation in QT across multiple populations, that novel signals exist in African Americans, and that the SNPs identified as strong candidates for functional evaluation implicate gene regulatory dysfunction in QT prolongation.


Adaptation strategies of horses with induced forelimb lameness walking on a treadmill.
There is a paucity of research describing the gait pattern of lame horses at the walk. To describe the changes in motion pattern and vertical ground reaction forces (GRFz) in horses with induced forelimb lameness at the walk and compare those changes with the changes observed at the trot. Experimental study. In 10 clinically sound Warmblood horses, moderate forelimb lameness was induced using a sole pressure model followed by trot and walk on a treadmill. Kinematic data were collected using 3D optical motion capture (OMC), and GRFz by an instrumented treadmill. Mixed models were used to compare sound baseline versus forelimb lameness (significance was set at P < .05). Lameness induction significantly reduced peak GRFz on the second force peak, and vertical impulse in the lame limb. Stride and stance duration in all limbs were reduced. Lameness significantly affected the vertical movement symmetry of the head and withers. Maximum limb retraction angle, fetlock extension and protraction speed were reduced in the lame limb. Body centre of mass (COM) translation was reduced in the side-to-side direction and increased in the vertical and fore-aft directions. Several compensatory kinetic and kinematic changes were observed in the nonlame limbs. The observed changes in both kinetics and kinematics were generally smaller at walk with fewer variables being affected, compared to the trot. Only one degree and type of orthopaedic pain (sole pressure) was studied. Compensatory strategies of forelimb lameness at the walk include alteration of several kinetic and kinematic parameters and have some specific patterns and inter-individual differences that are not seen at the trot. However, much like at the trot, head movement and forelimb vertical force symmetry seem to be the most useful parameters to detect forelimb lameness at walk.


Strategy as active waiting.
Successful executives who cut their teeth in stable industries or in developed countries often stumble when they face more volatile markets. They falter, in part, because they assume they can gaze deep into the future and develop a long-term strategy that will confer a sustainable competitive advantage. But visibility into the future of volatile markets is sharply limited because so many different variables are in play. Factors such as technological innovation, customers' evolving needs, government policy, and changes in the capital markets interact with one another to create unexpected outcomes. Over the past six years, Donald Sull, an associate professor at London Business School, has led a research project examining some of the world's most volatile markets, from national markets like China and Brazil to industries like enterprise software, telecommunications, and airlines. One of the most striking findings from this research is the importance of taking action during comparative lulls in the storm. Huge business opportunities are relatively rare; they come along only once or twice in a decade. And, for the most part, companies can't manufacture those opportunities; changes in the external environment converge to make them happen. What managers can do is prepare for these golden opportunities by managing smart during the comparative calm of business as usual. During these periods of active waiting, leaders must probe the future and remain alert to anomalies that signal potential threats or opportunities; exercise restraint to preserve their war chests; and maintain discipline to keep the troops battle ready. When a golden opportunity or"sudden death"threat emerges, managers must have the courage to declare the main effort and concentrate resources to seize the moment.


Encapsulated metallic intraocular foreign body of long duration presenting with cystoid macular edema and normal full-field electroretinogram.
The purpose of the study is to report an unusual case of encapsulated iron intraocular foreign body (IOFB) of long duration presenting with cystoid macular edema (CME) and normal full-field electroretinogram (ERG). A 28-year-old male presented with a history of metal injury 5 years back and subsequent visual loss in the left eye for 3 months. Fundus examination of the left eye showed an encapsulated cyst-like structure inferior to the disc with the surrounding pigmentary changes and CME. B-scan ultrasonography and computed tomography scan confirmed metallic IOFB. A full-field ERG performed before surgery showed a normal response. He underwent pars plana vitrectomy with IOFB extraction and silicone oil injection as a tamponade. Two months later, silicone oil removal was combined with intravitreal triamcinolone acetonide (IVTA) injection for persistent edema. Meticulous clinical evaluation, imaging, electrophysiological testing coupled with surgical intervention, and IVTA injection resulted in resolution of CME with minimal visual improvement. In conclusion long-standing encapsulated iron IOFB on the retina can cause vision-threatening complications without siderosis.


Carotenoid analysis of halophilic archaea by resonance Raman spectroscopy.
Recently, halite and sulfate evaporate rocks have been discovered on Mars by the NASA rovers, Spirit and Opportunity. It is reasonable to propose that halophilic microorganisms could have potentially flourished in these settings. If so, biomolecules found in microorganisms adapted to high salinity and basic pH environments on Earth may be reliable biomarkers for detecting life on Mars. Therefore, we investigated the potential of Resonance Raman (RR) spectroscopy to detect biomarkers derived from microorganisms adapted to hypersaline environments. RR spectra were acquired using 488.0 and 514.5 nm excitation from a variety of halophilic archaea, including Halobacterium salinarum NRC-1, Halococcus morrhuae, and Natrinema pallidum. It was clearly demonstrated that RR spectra enhance the chromophore carotenoid molecules in the cell membrane with respect to the various protein and lipid cellular components. RR spectra acquired from all halophilic archaea investigated contained major features at approximately 1000, 1152, and 1505 cm(-1). The bands at 1505 cm(-1) and 1152 cm(-1) are due to in-phase C=C (nu(1) ) and C-C stretching ( nu(2) ) vibrations of the polyene chain in carotenoids. Additionally, in-plane rocking modes of CH(3) groups attached to the polyene chain coupled with C-C bonds occur in the 1000 cm(-1) region. We also investigated the RR spectral differences between bacterioruberin and bacteriorhodopsin as another potential biomarker for hypersaline environments. By comparison, the RR spectrum acquired from bacteriorhodopsin is much more complex and contains modes that can be divided into four groups: the C=C stretches (1600-1500 cm(-1)), the CCH in-plane rocks (1400-1250 cm(-1)), the C-C stretches (1250-1100 cm(-1)), and the hydrogen out-of-plane wags (1000-700 cm(-1)). RR spectroscopy was shown to be a useful tool for the analysis and remote in situ detection of carotenoids from halophilic archaea without the need for large sample sizes and complicated extractions, which are required by analytical techniques such as high performance liquid chromatography and mass spectrometry.


The Intersection between Ocular and Manual Motor Control: Eye-Hand Coordination in Acquired Brain Injury.
Acute and chronic disease processes that lead to cerebral injury can often be clinically challenging diagnostically, prognostically, and therapeutically. Neurodegenerative processes are one such elusive diagnostic group, given their often diffuse and indolent nature, creating difficulties in pinpointing specific structural abnormalities that relate to functional limitations. A number of studies in recent years have focused on eye-hand coordination (EHC) in the setting of acquired brain injury (ABI), highlighting the important set of interconnected functions of the eye and hand and their relevance in neurological conditions. These experiments, which have concentrated on focal lesion-based models, have significantly improved our understanding of neurophysiology and underscored the sensitivity of biomarkers in acute and chronic neurological disease processes, especially when such biomarkers are combined synergistically. To better understand EHC and its connection with ABI, there is a need to clarify its definition and to delineate its neuroanatomical and computational underpinnings. Successful EHC relies on the complex feedback- and prediction-mediated relationship between the visual, ocular motor, and manual motor systems and takes advantage of finely orchestrated synergies between these systems in both the spatial and temporal domains. Interactions of this type are representative of functional sensorimotor control, and their disruption constitutes one of the most frequent deficits secondary to brain injury. The present review describes the visually mediated planning and control of eye movements, hand movements, and their coordination, with a particular focus on deficits that occur following neurovascular, neurotraumatic, and neurodegenerative conditions. Following this review, we also discuss potential future research directions, highlighting objective EHC as a sensitive biomarker complement within acute and chronic neurological disease processes.


Kaposi's sarcoma-associated herpesvirus-encoded latency-associated nuclear antigen modulates K1 expression through its cis-acting elements within the terminal repeats.
K1 is the first open reading frame encoded by Kaposi's sarcoma-associated herpesvirus (KSHV) and lies positionally to the immediate right of the terminal repeats. K1 is a transmembrane glycoprotein having a functional immunoreceptor tyrosine-based activation motif (ITAM) capable of activating B-cell receptor signaling. K1 is expressed mostly during the lytic cycle of the virus and its promoter lies within the terminal repeat which contains the binding sites for latency-associated nuclear antigen (LANA). The K1 promoter (K1p) having LANA binding sites assayed by reporter assay demonstrated that LANA is capable of down-regulating K1 promoter transcriptional activity. However, the KSHV replication transcription activator RTA up-regulates K1p transcriptional activity. The promoter deleted of LANA binding sites showed loss in LANA-mediated down-regulation but was unaffected for RTA-mediated up-regulation. Increasing amounts of RTA rescued LANA-mediated repression of K1p transcriptional activity in cotransfection experiments. Reporter assay data suggest that LANA binding to its cognate sequence is critical for LANA-mediated repression of K1p as a LANA construct lacking the DNA binding domain was unable to repress K1p transcription. Additionally, KSHV primary infection experiments suggest that K1 is expressed during early infection but is repressed on the establishment of latency and so follows an expression profile similar to that of RTA during infection. Analysis of the promoter sequence revealed the presence of Oct-1 transcription factor binding sites within the -116 to +76 region. Mutational analysis of the Oct-1 sites abolished RTA-mediated transcriptional activation, suggesting that RTA up-regulates K1p transcription through binding to this transcription factor.


Nuclear Activation Analysis: Recent developments in the technique and its inherent attributes give promise of wide applicability.
Nuclear activation analysis-whether with instrumental or radiochemical procedures, manual or automatic systems-seems destined for an important place in analytic chemistry. Experience will determine the best choice of activating particles and the best detectors, or it may even lead to the development of entirely new techniques. But it seems evident that nuclear activation analysis will increasingly satisfy the need for precise, sensitive, and rapid elementary analysis, and will do so at relatively low cost.


Untargeted metabolomics-like screening approach for chemical characterization and differentiation of canopic jar and mummy samples from Ancient Egypt using GC-high resolution MS.
In Ancient Egypt it was common practice to embalm corpses and specific internal organs to ensure eternal life. The exact nature of the employed embalming fluids, particularly for organ preservation within the canopic jars, is debated. Therefore, the aim of the current study, was to chemically characterize and differentiate canopic jars (n = 28) and mummies (n = 6) using gas chromatography - high resolution mass spectrometry (GC-HR MS) with a new untargeted metabolomics-like screening approach; as part of a larger minimal-invasive transdisciplinary study on Ancient Egyptian human tissues. Post-analytical data processing included deconvolution, screening against the NIST 14 spectral database as well as a high resolution metabolomics library, and positive peak evaluation. In the majority of samples the presence of a coniferous resin was indicated by the detection of longiborneol in combination with abietadiene acid derivatives and guajacol. Beeswax, proposedly used for symbolic reasons and/or as a binding agent, was detected in 10 samples. Previously not mentioned in the literature, but identified in the current sample set, were medical-related substances like aniseed constituents, salicylic acid, chamazulene and jacobine. By applying an untargeted metabolomics-like approach to archaeological samples for the first time, extensive statistical analysis was made possible (using both identified and non-identified features; adding up to 4381 features), which showed significant differences in the overall chemical composition of canopic jar and mummy samples using principle component analysis (PCA) and partial least square-discriminant analysis (PLS-DA). This emphasizes the necessity for more extensive canopic jar studies in the future in order to interpret findings correctly.


Assessing palliative care needs: views of patients, informal carers and healthcare professionals.
This paper reports a study to assess the palliative care needs of the adult population served by a healthcare provider organization in Northern Ireland from the perspectives of patients, informal carers and healthcare providers. Assessing palliative care need is a key factor for health service planning. Traditionally, palliative care has been associated with end-of-life care and cancer. More recently, the concept has been extended to include care for both cancer and non-cancer populations. Various approaches have been advocated for assessing need, including the exploration of professional provider and user perspectives of need. Semi-structured qualitative interviews were undertaken with a purposive sample of patients and lay carers receiving palliative care services (n = 24). Focus groups were also conducted with multi-professional palliative care providers (n = 52 participants) and face to face interviews were undertaken with key managerial stakeholders in the area (n = 7). The focus groups and interviews concentrated on assessment of palliative care need. All the interviews were transcribed verbatim and analysed using Burnard's framework. Professional providers experienced difficulty in defining the term palliative care. Difficulties in communication and information exchange, and fragmented co-ordination between services were identified. The main areas of need identified by all participants were social and psychological support; financial concerns; and the need for choice and information. All participants considered that there was inequity between palliative care service provision for patients with cancer and non-cancer diseases. All patients, regardless of diagnosis, should be able to access palliative care appropriate to their individual needs. For this to happen in practice, an integrated approach to palliative care is essential. The study methodology confirms the value of developing a comprehensive approach to assessing palliative care need.


Out-of-hospital auricular acupressure in elder patients with hip fracture: a randomized double-blinded trial.
Auricular acupressure is known to decrease the level of anxiety in patients during ambulance transport. The purpose of this randomized, double-blind, sham control study was to determine whether auricular acupressure can decrease not only the level of anxiety but also the level of pain in a group of elder patients with acute hip fracture. With the assistance of the Vienna Red Cross, 38 patients with acute hip fracture were enrolled into this study. Patients were randomized into two study groups: the true intervention group and the sham control group. Subjects in the true intervention group (n = 18) received bilateral auricular acupressure at three auricular acupressure points for hip pain. Patients in the sham group (n = 20) received bilateral auricular acupressure at sham points. Baseline demographic information, anxiety level, pain level, blood pressure, and heart rate were obtained before the administration of the appropriate acupressure intervention. The level of anxiety, level of pain, hemodynamic profiles, and level of satisfaction were reassessed once the patients arrived at the hospital. Patients in the true intervention groups had less pain (F = 28, p = 0.0001) and anxiety (F = 4.3, p = 0.018) and lower heart rate (F = 18, p = 0.0001) on arrival at the hospital than did patients in the sham control group. As a result, the patients in the true intervention group reported higher satisfaction in the care they received during the ride to the hospital. The authors encourage physicians, health care providers, and emergency rescuers to learn this easy, noninvasive, and inexpensive technique for its effects in decreasing anxiety and pain during emergency transportation.


Noise-enhanced heart rate and sympathetic nerve responses to oscillatory lower body negative pressure in humans.
By injecting noise into the carotid sinus baroreceptors, we previously showed that heart rate (HR) responses to weak oscillatory tilt were enhanced via a mechanism known as "stochastic resonance." It remains unclear, however, whether the same responses would be observed when using oscillatory lower body negative pressure (LBNP), which would unload the cardiopulmonary baroreceptors with physically negligible effects on the arterial system. Also, the vasomotor sympathetic activity directly controlling peripheral resistance against hypotensive stimuli was not observed. We therefore investigated the effects of weak (0 to approximately -10 mmHg) oscillatory (0.03 Hz) LBNP on HR and muscle sympathetic nerve activity (MSNA) while adding incremental noise to the carotid sinus baroreceptors via a pneumatic neck chamber. The signal-to-noise ratio of HR, cardiac interbeat interval, and total MSNA were all significantly improved by increasing noise intensity, while there was no significant change in the arterial blood pressure in synchronized with the oscillatory LBNP. We conclude that the stochastic resonance, affecting both HR and MSNA, results from the interaction of noise with the signal in the brain stem, where the neuronal inputs from the arterial and cardiopulmonary baroreceptors first come together in the nucleus tractus solitarius. Also, these results indicate that the noise could induce functional improvement in human blood pressure regulatory system in overcoming given hypotensive stimuli.


Healing actions of essential oils from Citrus aurantium and d-limonene in the gastric mucosa: the roles of VEGF, PCNA, and COX-2 in cell proliferation.
Previous studies have described the gastroprotective effects of essential oils that are derived from Citrus aurantium (OEC) and its main compound d-limonene (LIM) in a model of chemically induced ulcers in rats. However, these studies do not address the compound's healing effects on the gastric mucosa. Thus, the aim of this work was to evaluate the healing activity of OEC and LIM in acetic acid-induced gastric ulcers in rats, a model that reproduces human chronic ulcers. The obtained results demonstrated that lower effective doses of OEC (250 mg/kg) and LIM (245 mg/kg) induced gastric mucosal healing with a cure rate of 44% and 56%, respectively, compared with the control group (P<.05). During the 14 days of OEC or LIM treatment, none of the groups demonstrated toxicity in terms of body and organ weight or serum biochemical parameters. Both OEC and LIM treatment promoted an increase in epithelial healing, as confirmed by immunohistochemistry, which was greater in the animals that were treated with the positive control. In addition, both treatments increased cellular proliferation as measured by proliferating cell nuclear antigen and cyclooxygenase 2 expression in the gastric mucosa, vascular endothelial growth factor-mediated blood vessel formation in the margin of the ulcer, and production of gastric mucus, which fortifies the gastric protective barrier. We concluded that OEC and LIM, two common flavoring agents, promote gastric mucosal healing without any apparent toxic effect, resulting in better gastric epithelial organization in the treated rats.


Effectiveness of Provider and Community Interventions to Improve Treatment of Uncomplicated Malaria in Nigeria: A Cluster Randomized Controlled Trial.
The World Health Organization recommends that malaria be confirmed by parasitological diagnosis before treatment using Artemisinin-based Combination Therapy (ACT). Despite this, many health workers in malaria endemic countries continue to diagnose malaria based on symptoms alone. This study evaluates interventions to help bridge this gap between guidelines and provider practice. A stratified cluster-randomized trial in 42 communities in Enugu state compared 3 scenarios: Rapid Diagnostic Tests (RDTs) with basic instruction (control); RDTs with provider training (provider arm); and RDTs with provider training plus a school-based community intervention (provider-school arm). The primary outcome was the proportion of patients treated according to guidelines, a composite indicator requiring patients to be tested for malaria and given treatment consistent with the test result. The primary outcome was evaluated among 4946 (93%) of the 5311 patients invited to participate. A total of 40 communities (12 in control, 14 per intervention arm) were included in the analysis. There was no evidence of differences between the three arms in terms of our composite indicator (p = 0.36): stratified risk difference was 14% (95% CI -8.3%, 35.8%; p = 0.26) in the provider arm and 1% (95% CI -21.1%, 22.9%; p = 0.19) in the provider-school arm, compared with control. The level of testing was low across all arms (34% in control; 48% provider arm; 37% provider-school arm; p = 0.47). Presumptive treatment of uncomplicated malaria remains an ingrained behaviour that is difficult to change. With or without extensive supporting interventions, levels of testing in this study remained critically low. Governments and researchers must continue to explore alternative ways of encouraging providers to deliver appropriate treatment and avoid the misuse of valuable medicines. ClinicalTrials.gov NCT01350752.


The enhancement of histone H4 and H2A serine 1 phosphorylation during mitosis and S-phase is evolutionarily conserved.
Histone phosphorylation has long been associated with condensed mitotic chromatin; however, the functional roles of these modifications are not yet understood. Histones H1 and H3 are highly phosphorylated from late G2 through telophase in many organisms, and have been implicated in chromatin condensation and sister chromatid segregation. However, mutational analyses in yeast and biochemical experiments with Xenopus extracts have demonstrated that phosphorylation of H1 and H3 is not essential for such processes. In this study, we investigated additional histone phosphorylation events that may have redundant functions to H1 and H3 phosphorylation during mitosis. We developed an antibody to H4 and H2A that are phosphorylated at their respective serine 1 (S1) residues and found that H4S1/H2AS1 are highly phosphorylated in the mitotic chromatin of worm, fly, and mammals. Mitotic H4/H2A phosphorylation has similar timing and localization as H3 phosphorylation, and closely correlates with the chromatin condensation events during mitosis. We also detected a lower level of H4/H2A phosphorylation in 5-bromo-2-deoxyuridine-positive S-phase cells, which corroborates earlier studies that identified H4S1 phosphorylation on newly synthesized histones during S-phase. The evolutionarily conserved phosphorylation of H4/H2A during the cell cycle suggests that they may have a dual purpose in chromatin condensation during mitosis and histone deposition during S-phase.


Sulforaphane enhances irradiation effects in terms of perturbed cell cycle progression and increased DNA damage in pancreatic cancer cells.
Sulforaphane (SFN), an herbal isothiocyanate enriched in cruciferous vegetables like broccoli and cauliflower, has gained popularity for its antitumor effects in cell lines such as pancreatic cancer. Antiproliferative as well as radiosensitizing properties were reported for head and neck cancer but little is known about its effects in pancreatic cancer cells in combination with irradiation (RT). In four established pancreatic cancer cell lines we investigated clonogenic survival, analyzed cell cycle distribution and compared DNA damage via flow cytometry and western blot after treatment with SFN and RT. Both SFN and RT show a strong and dose dependent survival reduction in clonogenic assays, an induction of a G2/M cell cycle arrest and an increase in γH2AX protein level indicating DNA damage. Effects were more pronounced in combined treatment and both cell cycle perturbation and DNA damage persisted for a longer period than after SFN or RT alone. Moreover, SFN induced a loss of DNA repair proteins Ku 70, Ku 80 and XRCC4. Our results suggest that combination of SFN and RT exerts a more distinct DNA damage and growth inhibition than each treatment alone. SFN seems to be a viable option to improve treatment efficacy of chemoradiation with hopefully higher rates of secondary resectability after neoadjuvant treatment for pancreatic cancer.


Meta-Analysis of Randomized Controlled Trials Involving Anterior Shoulder Instability.
Arthroscopic repair is gaining popularity over open repair for the treatment of bankart lesions. The study aims to evaluate the outcome of arthroscopic repair with open repair in randomised controlled trials conducted comparing the two techniques. We searched the Cochrane library, PubMed and EMBASE up to December 2017 for clinical trials comparing the outcomes of arthroscopic bankart repair with open bankart repair. We used fixed or random effects model depending upon heterogenicity. Dichotomous variables were presented as Risk Ratios (RRs) with 95% Confidence Intervals (CIs), and continuous data were measured as measured differences with 95% CIs. Five studies were included, with sample size ranging from 42 to 196. Fixed effect analysis showed that the shoulder was more stable in open repair (RR=0.897, 95% CI: 0.821 to 0.980, P= 0.94) but the loss of external rotation at shoulder joint was also higher in those had open repair (RR=0.325, SMD=-0.411, 95% CI: -1.229 to 0.407). The functional outcome assessed by Rowe score was better in open repair (P=0.325). The operative time was lesser in arthroscopic repair but was not statistically significant (P=0.085). Our meta-analysis showed that the use of arthroscopic repair though offers better shoulder movement but the open repair is superior in terms of shoulder stability.


A composite picture of TcR alpha/beta(+) CD4(-)CD8(-) T Cells (alpha/beta-DNTCs) in humans with autoimmune lymphoproliferative syndrome.
The discovery of an unusual T-cell subset characterized by the expression of the alpha/beta T-cell receptor without expression of either CD4 or CD8 [alpha/beta-double-negative T cells (alpha/beta-DNTCs)] provided critical insights in the evaluation of a "new" lymphoproliferative disorder known as autoimmune lymphoproliferative syndrome (ALPS). ALPS is a disorder of defective Fas-mediated lymphocyte apoptosis, manifested by accumulation of alpha/beta-DNTCs and other lymphocyte subsets, leading to lymphadenopathy and splenomegaly, autoimmunity, and an increased risk of lymphoma. The expanded population of alpha/beta-DNTCs from ALPS patients has a remarkable uniform phenotype that is for the most part similar to alpha/beta-DNTCs from mice with defective Fas (lpr) or Fas ligand (gld). This is in contrast to the minor alpha/beta-DNTC compartment in healthy individuals that contains multiple, immunophenotypically distinct subpopulations. Current data indicate that alpha/beta-DNTCs from ALPS patients are derived from cytotoxic CD8(+) T cells, chronically activated in vivo but anergic in vitro. Their anergic state may be related to persistent modifications of O-linked carbohydrates on cell surface molecules, such as CD43 and CD45, as well as to the increased presence of interleukin-10. Although largely consistent with a model of (linear) CD8(+) cytotoxic T-cell differentiation, the expression patterns of certain surface molecules, such as CD27 and CD28, are not consistent with this model. This may be the result of the perturbed homeostasis of lymphocytes in ALPS, thereby revealing pathways of differentiation and immunophenotypes, including phenotypes pertaining to cell surface glycosylation that are hidden from view in healthy individuals.


Broadening the phenotypic spectrum of pathogenic LARP7 variants: two cases with intellectual disability, variable growth retardation and distinct facial features.
In 2012 Alazami et al. described a novel syndromic cause of primordial dwarfism with distinct facial features and severe intellectual disability. A homozygous frameshift mutation in LARP7, a chaperone of the noncoding RNA 7SK, was discovered in patients from a single consanguineous Saudi family. To date, only one additional patient has recently been described. To further delineate the phenotype associated with LARP7 mutations, we report two additional cases originating from the Netherlands and Saudi Arabia. The patients presented with intellectual disability, distinct facial features and variable short stature. We describe their clinical features and compare them with the previously reported patients. Both cases were identified by diagnostic whole-exome sequencing, which detected two homozygous pathogenic LARP7 variants: c.1091_1094delCGGT in the Dutch case and c.1045_1051dupAAGGATA in the Saudi Arabian case. Both variants are leading to frameshifts with introduction of premature stop codons, suggesting that loss of function is likely the disease mechanism. This study is an independent confirmation of the syndrome due to LARP7 depletion. Our cases broaden the associated clinical features of the syndrome and contribute to the delineation of the phenotypic spectrum of LARP7 mutations.