ATM regulates Mre11-dependent DNA end-degradation and microhomology-mediated end joining.
The human disorder ataxia telangiectasia (AT), which is characterized by genetic instability and neurodegeneration, results from mutation of the ataxia telangiectasia mutated (ATM) kinase. The loss of ATM leads to cell cycle checkpoint deficiencies and other DNA damage signaling defects that do not fully explain all pathologies associated with A-T including neuronal loss. In addressing this enigma, we find here that ATM suppresses DNA double-strand break (DSB) repair by microhomology-mediated end joining (MMEJ). We show that ATM repression of DNA end-degradation is dependent on its kinase activities and that Mre11 is the major nuclease behind increased DNA end-degradation and MMEJ repair in A-T. Assessment of MMEJ by an in vivo reporter assay system reveals decreased levels of MMEJ repair in Mre11-knockdown cells and in cells treated with Mre11-nuclease inhibitor mirin. Structure-based modeling of Mre11 dimer engaging DNA ends suggests the 5' ends of a bridged DSB are juxtaposed such that DNA unwinding and 3'-5' exonuclease activities may collaborate to facilitate simultaneous pairing of extended 5' termini and exonucleolytic degradation of the 3' ends in MMEJ. Together our results provide an integrated understanding of ATM and Mre11 in MMEJ: ATM has a critical regulatory function in controlling DNA end-stability and error-prone DSB repair and Mre11 nuclease plays a major role in initiating MMEJ in mammalian cells. These functions of ATM and Mre11 could be particularly important in neuronal cells, which are post-mitotic and therefore depend on mechanisms other than homologous recombination between sister chromatids to repair DSBs.


Prognostic Implications of Pathological Response to Neoadjuvant Chemoradiation in Pathologic Stage III Rectal Cancer.
To evaluate the independent prognostic ability of the American Joint Committee on Cancer (AJCC) tumor regression scores within pathologic stage II and III rectal cancers. Response to neoadjuvant chemoradiation (nCRT) has been debated as a biologic surrogate for tumor biology and prognosis in rectal cancer. AJCC regression scores have been shown to correlate with prognosis. Patient demographics, tumor characteristics, and AJCC scores (0 = complete response; 1 = isolated tumor cells remaining; 2 = residual cancer outgrown by fibrosis; 3 = extensive residual cancer) were assessed from 545 rectal cancer patients treated by nCRT followed by surgery at a single institution. Patients were classified as responders (score 0-2) or nonresponders (score 3). Survival analyses were performed using Cox proportional hazards models. Of 545 cases, 123 and 182 were pathologic stage II and III, respectively. Median follow-up was 4.9 years. AJCC regression scores were not independently prognostic within stage II cancers. However, AJCC scores were strongly associated with prognosis within stage III cancers (nonresponse 5-year overall survival [OS] 27% vs 67%, P < 0.001). Stage III responders (N = 139, 76.4%) had similar outcomes to stage II (5-year OS 67% vs 74%, P = 0.89). Conversely, stage III nonresponders (N = 43, 23.6%) approached stage IV outcomes (5-year OS 27% vs 18%, P = 0.09). On multivariable analysis, nonresponse (hazard ratio 3.2, 95% confidence interval 1.7-6.2), along with positive margin, abdominoperineal resection, and no adjuvant chemotherapy administration were independently associated with worse OS. AJCC response score after nCRT is a novel prognostic factor in pathologic stage III rectal cancer and may guide surveillance and adjuvant therapy decisions.


Functional Characterization of the Osteoarthritis Genetic Risk Residing at ALDH1A2 Identifies rs12915901 as a Key Target Variant.
To identify the functional single-nucleotide polymorphisms (SNPs) and mechanisms conferring increased risk of hand osteoarthritis (OA) at the ALDH1A2 locus, which is a retinoic acid regulatory gene. Tissue samples from 247 patients with knee, hip, or hand OA who had undergone joint surgery were included. RNA-sequencing analysis was used to investigate differential expression of ALDH1A2 and other retinoic acid signaling pathway genes in cartilage. Expression of ALDH1A2 in joint tissues obtained from multiple sites was quantified using quantitative reverse transcription-polymerase chain reaction. Allelic expression imbalance (AEI) was measured by pyrosequencing. The consequences of ALDH1A2 depletion by RNA interference were assessed in primary human chondrocytes. In silico and in vitro analyses were used to pinpoint which, among 62 highly correlated SNPs, could account for the association at the locus. ALDH1A2 expression was observed across multiple joint tissue samples, including osteochondral tissue from the hand. The expression of ALDH1A2 and of several retinoic acid signaling genes was different in diseased cartilage compared to non-diseased cartilage, with ALDH1A2 showing lower levels in OA cartilage. Experimental depletion of ALDH1A2 resulted in changes in the expression levels of a number of chondrogenic markers, including SOX9. In addition, reduced expression of the OA risk-conferring allele was witnessed in a number of joint tissues, with the strongest effect in cartilage. The intronic SNP rs12915901 recapitulated the AEI observed in patient tissues, while the Ets transcription factors were identified as potential mediators of this effect. The ALDH1A2 locus seems to increase the risk of hand OA through decreased expression of ALDH1A2 in joint tissues, with the effect dependent on rs12915901. These findings indicate a mechanism that may now be targeted to modulate OA risk.


Phylogenomic analysis of proteins that are distinctive of Archaea and its main subgroups and the origin of methanogenesis.
The Archaea are highly diverse in terms of their physiology, metabolism and ecology. Presently, very few molecular characteristics are known that are uniquely shared by either all archaea or the different main groups within archaea. The evolutionary relationships among different groups within the Euryarchaeota branch are also not clearly understood. We have carried out comprehensive analyses on each open reading frame (ORFs) in the genomes of 11 archaea (3 Crenarchaeota--Aeropyrum pernix, Pyrobaculum aerophilum and Sulfolobus acidocaldarius; 8 Euryarchaeota--Pyrococcus abyssi, Methanococcus maripaludis, Methanopyrus kandleri, Methanococcoides burtonii, Halobacterium sp. NCR-1, Haloquadratum walsbyi, Thermoplasma acidophilum and Picrophilus torridus) to search for proteins that are unique to either all Archaea or for its main subgroups. These studies have identified 1448 proteins or ORFs that are distinctive characteristics of Archaea and its various subgroups and whose homologues are not found in other organisms. Six of these proteins are unique to all Archaea, 10 others are only missing in Nanoarchaeum equitans and a large number of other proteins are specific for various main groups within the Archaea (e.g. Crenarchaeota, Euryarchaeota, Sulfolobales and Desulfurococcales, Halobacteriales, Thermococci, Thermoplasmata, all methanogenic archaea or particular groups of methanogens). Of particular importance is the observation that 31 proteins are uniquely present in virtually all methanogens (including M. kandleri) and 10 additional proteins are only found in different methanogens as well as A. fulgidus. In contrast, no protein was exclusively shared by various methanogen and any of the Halobacteriales or Thermoplasmatales. These results strongly indicate that all methanogenic archaea form a monophyletic group exclusive of other archaea and that this lineage likely evolved from Archaeoglobus. In addition, 15 proteins that are uniquely shared by M. kandleri and Methanobacteriales suggest a close evolutionary relationship between them. In contrast to the phylogenomics studies, a monophyletic grouping of archaea is not supported by phylogenetic analyses based on protein sequences. The identified archaea-specific proteins provide novel molecular markers or signature proteins that are distinctive characteristics of Archaea and all of its major subgroups. The species distributions of these proteins provide novel insights into the evolutionary relationships among different groups within Archaea, particularly regarding the origin of methanogenesis. Most of these proteins are of unknown function and further studies should lead to discovery of novel biochemical and physiological characteristics that are unique to either all archaea or its different subgroups.


Stages of use: consideration, initiation, utilization, and outcomes of an internet-mediated intervention.
Attrition, or nonuse of the intervention, is a significant problem in e-health. However, the reasons for this phenomenon are poorly understood. Building on Eysenbach's "Law of Attrition", this study aimed to explore the usage behavior of users of e-health services. We used two theoretical models, Andersen's Behavioral Model of Health Service Utilization and Venkatesh's Unified Theory of Acceptance and Use of Technology, to explore the factors associated with uptake and use of an internet-mediated intervention for caregivers taking care of a family member with dementia. A multiphase, longitudinal design was used to follow a convenience sample of 46 family caregivers who received an e-health intervention. Applying the two theories, usage behavior was conceptualized to form four stages: consideration, initiation, utilization (attrition or continuation), and outcome. The variables and measurement scales were selected based on these theories to measure the sociodemographic context, technology aptitudes, and clinical needs of the caregivers. In the Consideration Stage, caregivers who felt that the information communication technology (ICT)-mediated service was easy to use were more likely to consider participating in the study (p = 0.04). In the Initiation Stage, caregivers who showed greater technology acceptance were more likely to initiate service earlier (p = 0.02). In the Utilization Stage, the frequent users were those who had a more positive attitude toward technology (p = 0.04) and a lower perceived caregiver competence (p = 0.04) compared with nonusers. In the Outcome Stage, frequent users experienced a decline in perceived burden compared with an escalation of perceived burden by nonusers (p = 0.02). We illustrate a methodological framework describing how to develop and expand a theory on attrition. The proposed framework highlighted the importance of conceptualizing e-health "use" and "adoption" as dynamic, continuous, longitudinal processes occurring in different stages, influenced by different factors to predict advancement to the next stage. Although usage behavior was influenced mainly by technological factors in the initial stages, both clinical and technological factors were equally important in the later stages. Frequency of use was associated with positive clinical outcomes. A plausible explanation was that intervention benefits motivated the caregivers to continue the service and regular use led to more positive clinical outcome.


Prospective association of liver function biomarkers with development of hepatobiliary cancers.
Serum liver biomarkers (gamma-glutamyl transferase, GGT; alanine aminotransferase, ALT; aspartate aminotransferase, AST; alkaline phosphatase, ALP; total bilirubin) are used as indicators of liver disease, but there is currently little data on their prospective association with risk of hepatobiliary cancers. A nested-case control study was conducted within the prospective EPIC cohort (>520,000 participants, 10 European countries). After a mean 7.5 mean years of follow-up, 121 hepatocellular carcinoma (HCC), 34 intrahepatic bile duct (IHBC) and 131 gallbladder and biliary tract (GBTC) cases were identified and matched to 2 controls each. Circulating biomarkers were measured in serum taken at recruitment into the cohort, prior to cancer diagnosis. Multivariable adjusted conditional logistic regression was used to calculate odds ratios and 95% confidence intervals (OR; 95%CI). In multivariable models, 1SD increase of each log-transformed biomarker was positively associated with HCC risk (OR(GGT)=4.23, 95%CI:2.72-6.59; OR(ALP)=3.43, 95%CI:2.31-5.10;OR(AST)=3.00, 95%CI:2.04-4.42; OR(ALT)=2.69, 95%CI:1.89-3.84; OR(Bilirubin)=2.25, 95%CI:1.58-3.20). Each liver enzyme (OR(GGT)=4.98; 95%CI:1.75-14.17; OR(AST)=3.10, 95%CI:1.04-9.30; OR(ALT)=2.86, 95%CI:1.26-6.48, OR(ALP)=2.31, 95%CI:1.10-4.86) but not bilirubin (OR(Bilirubin)=1.46,95%CI:0.85-2.51) showed a significant association with IHBC. Only ALP was significantly associated with GBTC risk (OR(ALP)=1.59, 95%CI:1.20-2.09). This study shows positive associations between circulating liver biomarkers in sera collected prior to cancer diagnoses and the risks of developing HCC or IHBC, but not GBTC.


Manipulation of LIPSS orientation on silicon surfaces using orthogonally polarized femtosecond laser double-pulse trains.
Laser-induced periodic surface structures (LIPSS) provide an easy and cost-effective means of fabricating gratings and have been widely studied in recent decades. To overcome the challenge of orientation controllability, we developed a feasible and efficient method for manipulating the orientation of LIPSS in real time. Specifically, we used orthogonally polarized and equal-energy femtosecond laser (50 fs, 800 nm) double-pulse trains with time delay about 1ps, total peak laser fluence about 1.0 J/cm2, laser repetition frequency at 100 Hz and scanning speed at 150 μm/s to manipulate the LIPSS orientation on silicon surfaces perpendicular to the scanning direction, regardless of the scanning paths. The underlying mechanism is attributed to the periodic energy deposition along the direction of surface plasmon polaritons (SPPs), which can be controlled oriented along the scanning direction in orthogonally polarized femtosecond laser double-pulse trains surface scan processing. An application of structural colors presents the functionality of our method.


Dysfunctional voltage-gated K+ channels in pulmonary artery smooth muscle cells of patients with primary pulmonary hypertension.
Primary pulmonary hypertension (PPH) is a rare disease of unknown cause. Although PPH and secondary pulmonary hypertension (SPH) share many clinical and pathological characteristics, their origins may be disparate. In pulmonary artery smooth muscle cells (PASMCs), the activity of voltage-gated K+ (KV) channels governs membrane potential (Em) and regulates cytosolic free Ca2+ concentration ([Ca2+]cyt). A rise in [Ca2+]cyt is a trigger of vasoconstriction and a stimulus of smooth muscle proliferation. Fluorescence microscopy and patch clamp techniques were used to measure [Ca2+]cyt, Em, and KV currents in PASMCs. Mean pulmonary arterial pressures were comparable (46+/-4 and 53+/-4 mm Hg; P=0.30) in SPH and PPH patients. However, PPH-PASMCs had a higher resting [Ca2+]cyt than cells from patients with SPH and nonpulmonary hypertension disease. Consistently, PPH-PASMCs had a more depolarized Em than SPH-PASMCs. Furthermore, KV currents were significantly diminished in PPH-PASMCs. Because of the dysfunctional KV channels, the response of [Ca2+]cyt to the KV channel blocker 4-aminopyridine was significantly attenuated in PPH-PASMCs, whereas the response to 60 mmol/L K+ was comparable to that in SPH-PASMCs. These results indicate that KV channel function in PPH-PASMCs is inhibited compared with SPH-PASMCs. The resulting membrane depolarization and increase in [Ca2+]cyt lead to pulmonary vasoconstriction and PASMC proliferation. Our data suggest that defects in PASMC KV channels in PPH patients may be a unique mechanism involved in initiating and maintaining pulmonary vasoconstriction and appear to play a role in the pathogenesis of PPH.


Long weekly working hours and ischaemic heart disease: a follow-up study among 145 861 randomly selected workers in Denmark.
The aim of the present study was to test if incidences of ischaemic heart disease (IHD) and usage of antihypertensive drugs are independent of weekly working hours (WWH) among full-time employees in Denmark. Data on WWH from participants of the Danish labour force surveys, 1999-2013, were linked on an individual level to national registers with data on socioeconomic status (SES), industry, emigrations, redeemed prescriptions, hospital contacts and deaths. Participants were followed until the end of 2014 (on average 7.7 years). Poisson regression was used to model incidence rates as a function of WWH. The analyses were controlled for calendar time, time passed since start of follow-up, employment in the healthcare industry, age, sex, SES and night work. In total, we found 3635 cases of IHD and 20 648 cases of antihypertensive drug usage. The rate ratio of IHD was 0.95 (95% CI 0.85 to 1.06) for 41-48 compared with 32-40 WWH and 1.07 (0.94 to 1.21) for >48 compared with 32-40 WWH. The corresponding rate ratios for antihypertensive drug usage were 0.99 (0.95 to 1.04) and 1.02 (0.97 to 1.08). No statistically significant interactions between WWH and sex, SES and night work, respectively, were found. In this Danish sample, we did not find any statistically significant association between WWH and IHD or antihypertensive drug usage.


Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity.
Production of a potent urease has been described as a trait common to all Helicobacter pylori so far isolated from humans with gastritis as well as peptic ulceration. The detection of urease activity from genes cloned from H. pylori was made possible by use of a shuttle cosmid vector, allowing replication and movement of cloned DNA sequences in either Escherichia coli or Campylobacter jejuni. With this approach, we cloned a 44-kb portion of H. pylori chromosomal DNA which did not lead to urease activity when introduced into E. coli but permitted, although temporarily, biosynthesis of the urease when transferred by conjugation to C. jejuni. The recombinant cosmid (pILL585) expressing the urease phenotype was mapped and used to subclone an 8.1-kb fragment (pILL590) able to confer the same property to C. jejuni recipient strains. By a series of deletions and subclonings, the urease genes were localized to a 4.2-kb region of DNA and were sequenced by the dideoxy method. Four open reading frames were found, encoding polypeptides with predicted molecular weights of 26,500 (ureA), 61,600 (ureB), 49,200 (ureC), and 15,000 (ureD). The predicted UreA and UreB polypeptides correspond to the two structural subunits of the urease enzyme; they exhibit a high degree of homology with the three structural subunits of Proteus mirabilis (56% exact matches) as well as with the unique structural subunit of jack bean urease (55.5% exact matches). Although the UreD-predicted polypeptide has domains relevant to transmembrane proteins, no precise role could be attributed to this polypeptide or to the UreC polypeptide, which both mapped to a DNA sequence shown to be required to confer urease activity to a C. jejuni recipient strain.


Quality of Life after Intestinal Resection in Patients with Crohn Disease: A Systematic Review.
Most patients with Crohn disease (CD) require surgery within 10 years of diagnosis. Intestinal resection is the most commonly performed operation although the effects on health-related quality of life (HRQOL), particularly long-term, are contentious. We conducted a systematic review evaluating the impact of intestinal resection on the HRQOL of CD patients, predictors of postoperative HRQOL, and patient satisfaction with surgery. Nine studies including 1,108 CD patients undergoing intestinal resection were identified as eligible for inclusion. The median age at surgery was 29-41 years with varying follow-up period (range 30 days-5 years). Ileocolic resection was the most commonly performed operation on an elective basis (range 95-100%). HRQOL improved as early as 2 weeks postoperatively and lasted up to 5 years across both generic and gastrointestinal domains. Gender, smoking, and disease recurrence were potential predictors of postoperative HRQOL. Patient satisfaction is high with regard to surgery, with preference for a laparoscopic approach. Intestinal resection in CD patients improved HRQOL in the short- and long-term and patients describe high satisfaction about their surgery. Further studies are needed to validate potential predictors of postoperative HRQOL in this cohort.


Gray-Matter Volume Estimate Score: A Novel Semi-Automatic Method Measuring Early Ischemic Change on CT.
We developed a novel method named Gray-matter Volume Estimate Score (GRAVES), measuring early ischemic changes on Computed Tomography (CT) semi-automatically by computer software. This study aimed to compare GRAVES and Alberta Stroke Program Early CT Score (ASPECTS) with regards to outcome prediction and inter-rater agreement. This was a retrospective cohort study. Among consecutive patients with ischemic stroke in the anterior circulation who received intra-arterial therapy (IAT), those with a readable pretreatment CT were included. Two stroke neurologists independently measured both the GRAVES and ASPECTS. GRAVES was defined as the percentage of estimated hypodense lesion in the gray matter of the ipsilateral hemisphere. Spearman correlation analysis, receiver operating characteristic (ROC) comparison test, and intra-class correlation coefficient (ICC) comparison tests were performed between GRAVES and ASPECTS. Ninety-four subjects (age: 68.7±10.3; male: 54 [54.9%]) were enrolled. The mean GRAVES was 9.0±8.9 and the median ASPECTS was 8 (interquartile range, 6-9). Correlation between ASPECTS and GRAVES was good (Spearman's rank correlation coefficient, 0.642; P<0.001). ROC comparison analysis showed that the predictive value of GRAVES for favorable outcome was not significantly different from that of ASPECTS (area under curve, 0.765 vs. 0.717; P=0.308). ICC comparison analysis revealed that inter-rater agreement of GRAVES was significantly better than that of ASPECTS (0.978 vs. 0.895; P<0.001). GRAVES had a good correlation with ASPECTS. GRAVES was as good as ASPECTS in predicting a favorable clinical outcome, but was better than ASPECTS regarding inter-rater agreement. GRAVES may be used to predict the outcome of IAT.


Variability, negative evidence, and the acquisition of verb argument constructions.
We present a hierarchical Bayesian framework for modeling the acquisition of verb argument constructions. It embodies a domain-general approach to learning higher-level knowledge in the form of inductive constraints (or overhypotheses), and has been used to explain other aspects of language development such as the shape bias in learning object names. Here, we demonstrate that the same model captures several phenomena in the acquisition of verb constructions. Our model, like adults in a series of artificial language learning experiments, makes inferences about the distributional statistics of verbs on several levels of abstraction simultaneously. It also produces the qualitative learning patterns displayed by children over the time course of acquisition. These results suggest that the patterns of generalization observed in both children and adults could emerge from basic assumptions about the nature of learning. They also provide an example of a broad class of computational approaches that can resolve Baker's Paradox.


An Active Type I-E CRISPR-Cas System Identified in Streptomyces avermitilis.
CRISPR-Cas systems, the small RNA-dependent immune systems, are widely distributed in prokaryotes. However, only a small proportion of CRISPR-Cas systems have been identified to be active in bacteria. In this work, a naturally active type I-E CRISPR-Cas system was found in Streptomyces avermitilis. The system shares many common genetic features with the type I-E system of Escherichia coli, and meanwhile shows unique characteristics. It not only degrades plasmid DNA with target protospacers, but also acquires new spacers from the target plasmid DNA. The naive features of spacer acquisition in the type I-E system of S. avermitilis were investigated and a completely conserved PAM 5'-AAG-3' was identified. Spacer acquisition displayed differential strand bias upstream and downstream of the priming spacer, and irregular integrations of new spacers were observed. In addition, introduction of this system into host conferred phage resistance to some extent. This study will give new insights into adaptation mechanism of the type I-E systems in vivo, and meanwhile provide theoretical foundation for applying this system on the genetic modification of S. avermitilis.


A Stillinger-Weber potential for single-layered black phosphorus, and the importance of cross-pucker interactions for a negative Poisson's ratio and edge stress-induced bending.
The distinguishing structural feature of single-layered black phosphorus is its puckered structure, which leads to many novel physical properties. In this work, we first present a new parameterization of the Stillinger-Weber potential for single-layered black phosphorus. In doing so, we reveal the importance of a cross-pucker interaction term in capturing its unique mechanical properties, such as a negative Poisson's ratio. In particular, we show that the cross-pucker interaction enables the pucker to act as a re-entrant hinge, which expands in the lateral direction when it is stretched in the longitudinal direction. As a consequence, single-layered black phosphorus has a negative Poisson's ratio in the direction perpendicular to the atomic plane. As an additional demonstration of the impact of the cross-pucker interaction, we show that it is also the key factor that enables capturing the edge stress-induced bending of single-layered black phosphorus that has been reported in ab initio calculations.


Efficacy of osteopathic manipulative treatment of female patients with migraine: results of a randomized controlled trial.
Migraine is one of the most prevalent neurological disorders in Europe, severely affecting ability to work and quality of life. Medical therapies are considered to be the "gold standard" of treatment. This study addresses osteopathic treatment for acute therapy or prophylactic therapy as an alternative to traditional therapies. Forty-two (42) female patients with migraine were randomized into an intervention group (n = 21) and a control group (n = 21). Outcomes were evaluated with three questionnaires before the treatment (t1) and 6 months later (t2). The intervention group received five 50-minute osteopathic manipulative treatments (OMT) over a 10-week period. The control group did not receive OMT, sham treatment, or physical therapy. Patients of this group only filled the questionnaires. Both groups continued with previously prescribed medication. The Migraine Disability Assessment (MIDAS) and Short Form-36 (SF-36) questionnaires as well as a German "pain questionnaire" were used to assess pain intensity, the impact of migraine on daily life and health-related quality of life (HRQoL), and the number of days subjects suffered from migraine. Three (3) of the eight HRQoL domains of the SF-36 form in the intervention group showed significant improvement (from t1 to t2), with a general betterment exhibited in the other domains. The total MIDAS score, pain intensity, and disturbance in occupation due to migraine as well as number of days of disablements were also significantly reduced. The control group showed insignificant differences in these areas. This study affirms the effects of OMT on migraine headache in regard to decreased pain intensity and the reduction of number of days with migraine as well as working disability, and partly on improvement of HRQoL. Future studies with a larger sample size should reproduce the results with a control group receiving placebo treatment in a long-term follow-up.


Aging induces endothelial dysfunction while sparing arterial thrombosis.
To assess the effects of aging on arterial thrombus formation by comparing 2-year-old with 11-week-old C57Bl6 mice. Aging is a major risk factor for cardiovascular disease. In humans, assessing the direct effects of aging on vascular homeostasis is difficult because it occurs in the presence of other risk factors. Arterial thrombosis is the critical event in cardiovascular diseases; however, it is not known whether aging per se promotes its occurrence. Mice represent an interesting system to address this issue because they age without spontaneously developing other risk factors. Organ chamber experiments confirmed the advanced level of aging of old mice. As previously shown, old mice exhibited endothelial dysfunction; however, arterial thrombosis induced by photochemical injury was unchanged. Arterial tissue factor expression and activity; expressions of tissue factor pathway inhibitor, thrombomodulin, and plasminogen activator inhibitor 1; prothrombin time; partial thromboplastin time; thrombin-antithrombin complex; and platelet activation were comparable in both groups. Although these results cannot be directly extrapolated to humans, this study contributes novel important information on the direct effect of aging on arterial thrombosis and underscores the importance of controlling modifiable risk factors in aged individuals.


GluN2C/GluN2D subunit-selective NMDA receptor potentiator CIQ reverses MK-801-induced impairment in prepulse inhibition and working memory in Y-maze test in mice.
Despite ample evidence supporting the N-methyl-D-aspartate receptor (NMDAR) hypofunction hypothesis of schizophrenia, progress in the development of effective therapeutics based on this hypothesis has been limited. Facilitation of NMDA receptor function by co-agonists (D-serine or glycine) only partially alleviates the symptoms in schizophrenia; other means to facilitate NMDA receptors are required. NMDA receptor sub-types differ in their subunit composition, with varied GluN2 subunits (GluN2A-GluN2D) imparting different physiological, biochemical and pharmacological properties. CIQ is a positive allosteric modulator that is selective for GluN2C/GluN2D-containing NMDA receptors (Mullasseril et al.). The effect of systemic administration of CIQ was tested on impairment in prepulse inhibition (PPI), hyperlocomotion and stereotypy induced by i.p. administration of MK-801 and methamphetamine. The effect of CIQ was also tested on MK-801-induced impairment in working memory in Y-maze spontaneous alternation test. We found that systemic administration of CIQ (20 mg·kg⁻¹, i.p.) in mice reversed MK-801 (0.15 mg·kg⁻¹, i.p.)-induced, but not methamphetamine (3 mg·kg⁻¹, i.p.)-induced, deficit in PPI. MK-801 increased the startle amplitude to pulse alone, which was not reversed by CIQ. In contrast, methamphetamine reduced the startle amplitude to pulse alone, which was reversed by CIQ. CIQ also partially attenuated MK-801- and methamphetamine-induced hyperlocomotion and stereotyped behaviours. Additionally, CIQ reversed the MK-801-induced working memory deficit in spontaneous alternation in a Y-maze. Together, these results suggest that facilitation of GluN2C/GluN2D-containing receptors may serve as an important therapeutic strategy for treating positive and cognitive symptoms in schizophrenia.


Multicolor lattice solitons.
We report on the existence of multicolor solitons supported by periodic lattices made from quadratic nonlinear media. Such lattice solitons bridge the gap between continuous solitons in uniform media and discrete solitons in strongly localized systems and exhibit a wealth of new features. We discovered that, in contrast to uniform media, multipeaked lattice solitons are stable. Thus they open new opportunities for all-optical switching based on soliton packets.


Running Large-Scale Simulations on the Neurorobotics Platform to Understand Vision - The Case of Visual Crowding.
Traditionally, human vision research has focused on specific paradigms and proposed models to explain very specific properties of visual perception. However, the complexity and scope of modern psychophysical paradigms undermine the success of this approach. For example, perception of an element strongly deteriorates when neighboring elements are presented in addition (visual crowding). As it was shown recently, the magnitude of deterioration depends not only on the directly neighboring elements but on almost all elements and their specific configuration. Hence, to fully explain human visual perception, one needs to take large parts of the visual field into account and combine all the aspects of vision that become relevant at such scale. These efforts require sophisticated and collaborative modeling. The Neurorobotics Platform (NRP) of the Human Brain Project offers a unique opportunity to connect models of all sorts of visual functions, even those developed by different research groups, into a coherently functioning system. Here, we describe how we used the NRP to connect and simulate a segmentation model, a retina model, and a saliency model to explain complex results about visual perception. The combination of models highlights the versatility of the NRP and provides novel explanations for inward-outward anisotropy in visual crowding.


Suppression and Contrast Normalization in Motion Processing.
Sensory neurons are activated by a range of stimuli to which they are said to be tuned. Usually, they are also suppressed by another set of stimuli that have little effect when presented in isolation. The interactions between preferred and suppressive stimuli are often quite complex and vary across neurons, even within a single area, making it difficult to infer their collective effect on behavioral responses mediated by activity across populations of neurons. Here, we investigated this issue by measuring, in human subjects (three males), the suppressive effect of static masks on the ocular following responses induced by moving stimuli. We found a wide range of effects, which depend in a nonlinear and nonseparable manner on the spatial frequency, contrast, and spatial location of both stimulus and mask. Under some conditions, the presence of the mask can be seen as scaling the contrast of the driving stimulus. Under other conditions, the effect is more complex, involving also a direct scaling of the behavioral response. All of this complexity at the behavioral level can be captured by a simple model in which stimulus and mask interact nonlinearly at two stages, one monocular and one binocular. The nature of the interactions is compatible with those observed at the level of single neurons in primates, usually broadly described as divisive normalization, without having to invoke any scaling mechanism.SIGNIFICANCE STATEMENT The response of sensory neurons to their preferred stimulus is often modulated by stimuli that are not effective when presented alone. Individual neurons can exhibit multiple modulatory effects, with considerable variability across neurons even in a single area. Such diversity has made it difficult to infer the impact of these modulatory mechanisms on behavioral responses. Here, we report the effects of a stationary mask on the reflexive eye movements induced by a moving stimulus. A model with two stages, each incorporating a divisive modulatory mechanism, reproduces our experimental results and suggests that qualitative variability of masking effects in cortical neurons might arise from differences in the extent to which such effects are inherited from earlier stages.


Assessing agreement between two job-exposure matrices.
This study was undertaken to compare the operating characteristics of the Hoar et al job-exposure matrix and the job-exposure matrix of the National Occupational Hazard Survey (NOHS) of the National Institute for Occupational Safety and Health in the United States. Both matrices were applied to occupations in the metal, paper and wood, and chemical, drug and paints industries, and the extent of agreement between the two was estimated. The agents (exposure entities) selected for the comparison were common to both matrices; altogether 214 agents were included. For the comparison of occupations, the occupational codes of the Hoar et al matrix were converted to those of the NOHS-based matrix. For every occupational group analyzed, each of the 214 exposure entities was classified as to its "presence" or "absence", and a kappa statistic (kappa) was calculated. The kappa(i) ranged from 0.27 to -0.12. In general, most of the agreement between the 2 matrices occurred because they agreed as to the absence of exposure for the exposure entities. Variation in agreement was observed for both the individual exposure entities and for chemical families. These results indicate poor to only fair agreement between the 2 job-exposure matrices, and they underscore the importance of understanding the rules for classifying exposures when any such matrix is used. Investigators also need to be aware of the methods and limitations of the occupational coding systems.


The pregnancy-related dreams of pregnant women.
This study examined the frequency and nature of pregnancy-related dreams in pregnant women in a family practice center and assessed the extent such dreams were discussed with providers of prenatal care. Pregnant women 18 years of age or older who were receiving prenatal care at the University of Missouri-Columbia Family Medical Care Center responded to a two-page self-administered questionnaire. The major variables measured were frequency of pregnancy-related dreams, frequency of frightening dreams, content of dreams, and discussion of dreams. Health professionals providing prenatal care to these women were also surveyed. Eighty-eight pregnant women, 41 family physicians, and 3 nurse practitioners participated in the study. Dreams about their pregnancy or baby were reported by 59 (67 percent) of 88 pregnant women, of whom 22 had experienced at least one frightening dream. Seventeen women reported being upset by a dream. The frequency of dreams increased with advancing gestational age. The content of the most common dream involved conflict with the father of the baby. Most women had talked to another person about their dreams, usually the baby's father. Only 2 women told their physicians about the pregnancy-related dream. One-half of the providers of prenatal care reported discussing pregnancy-related dreams with a pregnant patient at least once during their careers. Dreams about their pregnancy or baby occurred frequently in pregnant women and could be a neglected source of information about the psychological state of the patient.


Transverse wave instability in a solid explosive.
By the use of a metal indentation techinque, microscopic transverse or side-running shock waves can be detected in composition C-4, an explosive consisting largely of cyclonite (RDX) with organic plasticizers. The minimum wave spacing observed is approximately 0.05 millimeters. with a lateral velocity of about 6.5 millimeters per microsecond. The instability was demnostrated in cylindrically divergent detonation waves.


Experimental asynchrony to study self-inflicted lung injury.
Patient self-inflicted lung injury may be associated with worse clinical outcomes and higher mortality. Patient-ventilator asynchrony is associated with increased ventilator days and mortality, and it has been hypothesised as one of the important mechanisms leading to patient self-inflicted lung injury. However, given the observational nature of the key studies in the field so far, the hypothesis that patient-ventilator asynchrony causes patient self-inflicted lung injury has not been supported by evidence yet. Wittenstein and colleagues present a novel approach that enables controlling patient-ventilator asynchrony in a pig model of acute lung injury, to investigate the patient-ventilator asynchrony and patient self-inflicted lung injury causality. Their results suggest that increased patient-ventilator asynchrony associated with poor clinical outcomes reported in observational trials could be a marker, rather than a cause of patient self-inflicted lung injury. These findings on their own are not sufficient to justify a greater tolerance of patient-ventilator asynchrony amongst clinicians, a change for which further experimental work and clinical evidence is needed.


Salivary alpha amylase in on-call from home fire and emergency service personnel.
The effect of working on-call from home on the sympatho-adrenal medullary system activity is currently unknown. This study had two aims, Aim 1: examine salivary alpha amylase awakening response (AAR) and diurnal salivary alpha amylase (sAA) profile in fire and emergency service workers who operate on-call from home following a night on-call with a call (NIGHT-CALL), a night on-call without a call (NO-CALL) and an off-call night (OFF-CALL), and Aim 2: explore whether there was an anticipatory effect of working on-call from home (ON) compared to when there was an off-call (OFF) on the diurnal sAA profile. Participants wore activity monitors, completed sleep and work diaries and collected seven saliva samples a day for one week. AAR area under the curve with respect to ground (AUCG), AAR area under the curve with respect to increase (AUCI), AAR reactivity, diurnal sAA slope, diurnal sAA AUCG and mean 12-h sAA concentrations were calculated. Separate generalised estimating equation models were constructed for each variable of interest for each aim. For Aim 1, there were no differences between NIGHT-CALL or NO-CALL and OFF-CALL for any response variable. For Aim 2, there was no difference between any response variable of interest when ON the following night compared to when OFF the following night (n = 14). These findings suggest that there is no effect of working on-call from home on sAA, but should be interpreted with caution, as overnight data were not collected. Future research, using overnight heart rate monitoring, could help confirm these findings.


Cylindrical micelles of a POSS amphiphilic dendrimer as nano-reactors for polymerization.
A low generation amphiphilic dendrimer, POSS-AD, which has a POSS core and eight amphiphilic arms, was synthesized and used as a nano-reactor to produce well-defined polymer nano-cylinders. Confirmed by small-angle X-ray scattering (SAXS), Raman and NMR spectrometry, monodispersed cylindrical micelles that contain a hydrophilic cavity with a diameter of 2.09 nm and a length of 4.26 nm were produced via co-assembling POSS-AD with hydrophilic liquids, such as H2O and HEMA in hydrophobic solvents. Taking the HEMA/POSS-AD cylindrical micelles as nano-reactors, polymerization of HEMA within the micelles results in polymer nano-cylinders (POSS-ADNPs) with a diameter of 2.24 nm and a length of 5.02 nm. The study confirmed that despite the inability to maintain specific shape in solution, low generation dendrimers form well-defined nano-containers or nano-reactors, which relies on co-assembling with hydrophilic guest molecules. These nano-reactors are robust enough to maintain their shape during the polymerization of the guest molecules. Polymer nano-cylinders with dimensions less than 10 nm can thus be produced from the HEMA/POSS-AD micelles. Since the chemical structure of low-generation dendrimers and the contents of the co-assembled nano-reactors can be easily adjusted, the concept holds the potential for the further developments of low-generation amphiphilic dendrimers.


The geography of measles vaccination in the African Great Lakes region.
Expanded access to measles vaccination was among the most successful public health interventions of recent decades. All WHO regions currently target measles elimination by 2020, yet continued measles circulation makes that goal seem elusive. Using Demographic and Health Surveys with generalized additive models, we quantify spatial patterns of measles vaccination in ten contiguous countries in the African Great Lakes region between 2009-2014. Seven countries have 'coldspots' where vaccine coverage is below the WHO target of 80%. Over 14 million children under 5 years of age live in coldspots across the region, and a total of 8-12 million children are unvaccinated. Spatial patterns of vaccination do not map directly onto sub-national administrative units and transnational coldspots exist. Clustering of low vaccination areas may allow for pockets of susceptibility that sustain circulation despite high overall coverage. Targeting at-risk areas and transnational coordination are likely required to eliminate measles in the region.


MOF and histone H4 acetylation at lysine 16 are critical for DNA damage response and double-strand break repair.
The human MOF gene encodes a protein that specifically acetylates histone H4 at lysine 16 (H4K16ac). Here we show that reduced levels of H4K16ac correlate with a defective DNA damage response (DDR) and double-strand break (DSB) repair to ionizing radiation (IR). The defect, however, is not due to altered expression of proteins involved in DDR. Abrogation of IR-induced DDR by MOF depletion is inhibited by blocking H4K16ac deacetylation. MOF was found to be associated with the DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a protein involved in nonhomologous end-joining (NHEJ) repair. ATM-dependent IR-induced phosphorylation of DNA-PKcs was also abrogated in MOF-depleted cells. Our data indicate that MOF depletion greatly decreased DNA double-strand break repair by both NHEJ and homologous recombination (HR). In addition, MOF activity was associated with general chromatin upon DNA damage and colocalized with the synaptonemal complex in male meiocytes. We propose that MOF, through H4K16ac (histone code), has a critical role at multiple stages in the cellular DNA damage response and DSB repair.


Probiotic Escherichia coli Nissle 1917 inhibits leaky gut by enhancing mucosal integrity.
Probiotics are proposed to positively modulate the intestinal epithelial barrier formed by intestinal epithelial cells (IECs) and intercellular junctions. Disruption of this border alters paracellular permeability and is a key mechanism for the development of enteric infections and inflammatory bowel diseases (IBDs). To study the in vivo effect of probiotic Escherichia coli Nissle 1917 (EcN) on the stabilization of the intestinal barrier under healthy conditions, germfree mice were colonized with EcN or K12 E. coli strain MG1655. IECs were isolated and analyzed for gene and protein expression of the tight junction molecules ZO-1 and ZO-2. Then, in order to analyze beneficial effects of EcN under inflammatory conditions, the probiotic was orally administered to BALB/c mice with acute dextran sodium sulfate (DSS) induced colitis. Colonization of gnotobiotic mice with EcN resulted in an up-regulation of ZO-1 in IECs at both mRNA and protein levels. EcN administration to DSS-treated mice reduced the loss of body weight and colon shortening. In addition, infiltration of the colon with leukocytes was ameliorated in EcN inoculated mice. Acute DSS colitis did not result in an anion secretory defect, but abrogated the sodium absorptive function of the mucosa. Additionally, intestinal barrier function was severely affected as evidenced by a strong increase in the mucosal uptake of Evans blue in vivo. Concomitant administration of EcN to DSS treated animals resulted in a significant protection against intestinal barrier dysfunction and IECs isolated from these mice exhibited a more pronounced expression of ZO-1. This study convincingly demonstrates that probiotic EcN is able to mediate up-regulation of ZO-1 expression in murine IECs and confer protection from the DSS colitis-associated increase in mucosal permeability to luminal substances.


Communication and dissemination strategies to facilitate the use of health-related evidence.
This review examined how to best communicate and disseminate evidence, including uncertain evidence, to inform health care decisions. The review focused on three primary objectives--comparing the effectiveness of: (1) communicating evidence in various contents and formats that increase the likelihood that target audiences will both understand and use the information (KQ 1); (2) a variety of approaches for disseminating evidence from those who develop it to those who are expected to use it (KQ 2); and (3) various ways of communicating uncertainty-associated health-related evidence to different target audiences (KQ 3). A secondary objective was to examine how the effectiveness of communication and dissemination strategies varies across target audiences, including evidence translators, health educators, patients, and clinicians. We searched MEDLINE®, the Cochrane Library, Cochrane Central Trials Registry, PsycINFO®, and the Web of Science. We used a variety of medical subject headings (MeSH terms) and major headings, and used free-text and title and abstract text-word searches. The search was limited to studies on humans published from 2000 to March 15, 2013, for communication and dissemination, given the prior systematic reviews, and from 1966 to March 15, 2013, for communicating uncertainty. We used standard Evidence-based Practice Center methods of dual review of abstracts, full-text articles, and abstractions, and quality ratings and group consensus to resolve disagreements. We used group consensus to grade strength of evidence. The search identified 4,152 articles (after removing duplicates) for all three KQs. After dual review at the title/abstract stage and full-text review stage, we retained 61 articles that directly (i.e., head to head) compared strategies to communicate and disseminate evidence. Across the KQs, many of the comparisons yielded insufficient evidence to draw firm conclusions. For KQ 1, we found that investigators frequently blend more than one communication strategy in interventions. For KQ 2, we found that, compared with single dissemination strategies, multicomponent dissemination strategies are more effective at enhancing clinician behavior, particularly for guideline adherence. Key findings for KQ 3 indicate that evidence on communicating overall strength of recommendation and precision was insufficient, but certain ways of communicating directness and net benefit may be helpful in reducing uncertainty. The lack of comparative research evidence to inform communication and dissemination of evidence, including uncertain evidence, impedes timely clinician, patient, and policymaker awareness, uptake, and use of evidence to improve the quality of care. Expanding investment in communication, dissemination, and implementation research is critical to the identification of strategies to accelerate the translation of comparative effectiveness research into community and clinical practice and the direct benefit of patient care.


Depression and Dyspepsia: An Implication of Islamic Resources.
Depression is a common mental disorder and the leading cause of disability in the worldwide. Based on Islamic resources, indigestion and dyspepsia can lead to depression. In modern medicine, though many somatic diseases have been named as possible causes of depression, the effect of gastrointestinal disorders on depression is still an enigma. Therefore, the focus of this study is to explore the available scientific literature of modern medicine in order to find the footprint of effect of indigestion on depression. In this study, related articles were retrieved from PubMed, Ovid, Proquest and Magiran databases by using the Medical Subject Heading keywords "depression," "psychology," "dyspepsia" and "gastrointestinal diseases." In the next step, studies, which are exactly confirm the Islamic viewpoint, were selected from the retrieved articles. Only one prospective study in 2012 has stated that people with functional gastrointestinal disorders and without elevated levels of anxiety and depression at baseline had significantly higher levels of anxiety and depression at 12-year follow-up. Based on Islamic viewpoint, indigestion can lead to depression, but this aspect approved by only one 12-year prospective population-based study in our review. It seems that it is necessary to conduct complementary studies investigating this hypothesis.


Radiation-sensitive diselenide block co-polymer micellar aggregates: toward the combination of radiotherapy and chemotherapy.
We have developed a potential radiation-sensitive drug-delivery system using active diselenide-containing block co-polymer aggregates in aqueous solution that can load and release anticancer drugs. These aggregates were sensitive to even a low dose of γ-radiation, such as 5 Gy, which is close to the radiation dose received by patients during a single radiotherapy treatment. This line of research may open an avenue for the combination of radiotherapy and chemotherapy.


Calcium ion independent membrane leakage induced by phospholipase-like myotoxins.
The two snake venom myotoxins ammodytin L and myotoxin II, purified respectively from Vipera ammodytes ammodytes and Bothrops asper, have phospholipase-like structures but lack an Asp-49 in the active site and are without normal phospholipase activity. The interaction of these proteins with different types of liposomes indicated that the myotoxins were able to provoke rapid and extensive release of the aqueous content of liposomes. Leakage was measured by two different methods: fluorescence dequenching of liposome-entrapped carboxyfluorescein and ESR measurement of intravesicular TEM-POcholine reduction by external ascorbate. The process was independent of Ca2+ and took place without any detectable phospholipid hydrolysis. Nonmyotoxic phospholipases tested under the same conditions were unable to induce liposome leakage, which could be detected only when Ca2+ was added to the medium and with the concomitant hydrolysis of phospholipids. The kinetics of Ca(2+)-dependent and Ca(2+)-independent leakage were completely different, indicating two different mechanisms of interaction with the lipid bilayer. Studies using diphenylhexatriene as a probe of lipid membrane organization indicated that the myotoxins gave rise to a profound perturbation of the arrangement of the lipid chains in the membrane interior, whereas interaction of Naja naja phospholipase A2 with the membrane surface did not affect lipid organization. On the basis of these results we suggest that a new type of cytolytic reaction mechanism is responsible for the effects of phospholipase-like myotoxins in vivo.


The major 35S-methionine-labeled rapidly transported protein (superprotein) is identical to SNAP-25, a protein of synaptic terminals.
Superprotein is a rapidly axonally transported protein that is conspicuously labeled with 35S-methionine supplied to the cell bodies of retinal ganglion cells. Superprotein candidates are apparent among the rapidly transported proteins of many neurons from the CNS and PNS, including cranial, sympathetic, sensory, and motor neurons from mammals, fish, and amphibians. To determine the identity of Superprotein, we purified it from rabbit visual system and spinal cord and determined the amino acid sequence of seven of its tryptic peptides. The sequence shows that Superprotein is SNAP-25, a protein recently predicted from a cDNA sequence; SNAP-25 has been reported to be concentrated in the synaptic terminals of a selected population of CNS neurons. We measured the amount of radioactivity associated with Superprotein in tissue containing axons (optic tract) and synaptic terminals (superior collicules) of rabbit retinal ganglion cells. Labeled Superprotein disappeared from the superior colliculus more rapidly than another protein (synapsin I-like protein) that is concentrated in synaptic terminals. These results serve to unite the observations on the synthesis, distribution, metabolism, and axonal transport of Superprotein with observations of SNAP-25 and its mRNA.


Pramipexole versus dual release levodopa in restless legs syndrome: a double blind, randomised, cross-over trial.
To compare efficacy and safety of the dopamine agonist pramipexole (PPX) versus reference treatment with dual release levodopa/benserazide (L/B) in de novo patients with restless legs syndrome (RLS). A total of 39 men and women between 25 and 85 years old, fulfilling all clinical criteria for diagnosis of idiopathic RLS, previously untreated, participated. The study was performed as a randomised, double-blind, double-dummy crossover trial with two treatment periods of four weeks and took place in six Swiss certified sleep-centres. Interventions were PPX 0.25-0.75 mg and dual-release L/B 125-375 mg. The primary outcome measure was the frequency of periodic limb movements while in bed (PLM index, PLMI). Secondary endpoints included the changes in patient ratings on the International RLS Study Group Rating Scale (IRLS). Both pramipexole and dual-release L/B were effective in reducing PLM and RLS symptoms. Mean PLMI reduction was -11.5 for PPX and -7.7 for L/B (baseline 21.1 and 21.5), and the mean IRLS score reduction was -7.2 and -4.0 (baseline 20.8 and 21.1). In patients with an IRLS score >20 (38%), a significantly (p = 0.047) higher PLMI reduction for PPX (-8.5), as compared to L/B (-4.3), was found. A higher incidence of "augmentations" and "involuntary movements" for L/B, and "nausea or vomiting" and "hypotension with dizziness" for PPX treatment were noted as adverse effects. This study showed comparable effects of PPX versus dual-release L/B for short-term treatment of de novo patients with mild to moderate RLS.


Retinal arteriolar geometry is associated with cerebral white matter hyperintensities on magnetic resonance imaging.
Cerebral small vessel disease (lacunar stroke and cerebral white matter hyperintensities) is caused by vessel abnormalities of unknown aetiology. Retinal vessels show developmental and pathophysiological similarities to cerebral small vessels and microvessel geometry may influence vascular efficiency. Retinal arteriolar branching angles or coefficients (the ratio of the sum of the cross-sectional areas of the two daughter vessels to the cross-sectional area of the parent vessel at an arteriolar bifurcation) may be associated with cerebral small vessel disease. We performed a cross-sectional observational study in a UK tertiary referral hospital. An experienced stroke physician recruited consecutive patients presenting with lacunar ischaemic stroke with a control group consisting of patients with minor cortical ischaemic stroke. We performed brain magnetic resonance imaging to assess the recent infarct and periventricular and deep white matter hyperintensities. We subtyped stroke with clinical and radiological findings. We took digital retinal photographs to assess retinal arteriolar branching coefficients and branching angles using a semi-automated technique. Two hundred and five patients were recruited (104 lacunar stroke, 101 cortical stroke), mean age 68-years (standard deviation 12). With multivariate analysis, increased branching coefficient was associated with periventricular white matter hyperintensities (P=0.006) and ischaemic heart disease (P<0.001), and decreased branching coefficient with deep white matter hyperintensities (P=0.003), but not with lacunar stroke subtype (P=0.96). We found no associations with retinal branching angles. Retinal arteriolar geometry differs between cerebral small vessel phenotypes. Further research is needed to ascertain the clinical significance of these findings.


Evaluation of administration of West Nile virus vaccine to pregnant broodmares.
To determine whether administration of killed West Nile virus vaccine was associated with pregnancy loss among broodmares. Retrospective cohort study. 595 mares. Records of pregnant mares with known vaccination history from 4 farms were reviewed. Information obtained from 595 mares included mare's identification; farm; age; breed; reproductive status; last breeding date; date last known pregnant; vaccination date; age of conceptus at vaccination; vaccination during the early embryonic, early fetal, and late fetal periods; and whether an early embryonic death (EED), early fetal loss (EFL), or late fetal loss (LFL) occurred. The relationships between the dichotomous outcomes of loss (eg, EED, EFL, LFL) and independent categoric variables (eg, vaccination during the early embryonic, early fetal, or late fetal periods) were examined. Vaccination of pregnant mares during any period of gestation was not associated with increased incidence of pregnancy loss. Many mares are already pregnant at the onset of mosquito season, when mares are more likely to be vaccinated than at other times. Our findings provide evidence that vaccine administration will not compromise pregnancy in horses.


Structural and mechanistic insights into the Artemis endonuclease and strategies for its inhibition.
Artemis (SNM1C/DCLRE1C) is an endonuclease that plays a key role in development of B- and T-lymphocytes and in dsDNA break repair by non-homologous end-joining (NHEJ). Artemis is phosphorylated by DNA-PKcs and acts to open DNA hairpin intermediates generated during V(D)J and class-switch recombination. Artemis deficiency leads to congenital radiosensitive severe acquired immune deficiency (RS-SCID). Artemis belongs to a superfamily of nucleases containing metallo-β-lactamase (MBL) and β-CASP (CPSF-Artemis-SNM1-Pso2) domains. We present crystal structures of the catalytic domain of wildtype and variant forms of Artemis, including one causing RS-SCID Omenn syndrome. The catalytic domain of the Artemis has similar endonuclease activity to the phosphorylated full-length protein. Our structures help explain the predominantly endonucleolytic activity of Artemis, which contrasts with the predominantly exonuclease activity of the closely related SNM1A and SNM1B MBL fold nucleases. The structures reveal a second metal binding site in its β-CASP domain unique to Artemis, which is amenable to inhibition by compounds including ebselen. By combining our structural data with that from a recently reported Artemis structure, we were able model the interaction of Artemis with DNA substrates. The structures, including one of Artemis with the cephalosporin ceftriaxone, will help enable the rational development of selective SNM1 nuclease inhibitors.


Developmental disruption of thyroid hormone: correlations with hearing dysfunction in rats.
A wide variety of environmental contaminants adversely affect thyroid hormone (TH) homeostasis. Hypothyroidism and/or hypothyroxinemia during the early postnatal period in the rat leads to permanent structural damage and loss of function in the cochlea. A major uncertainty in assessing the risks of developmental exposure to thyroid-disrupting chemicals (TDCs) is the lack of a clear characterization of the dose-response relationship, especially in the lower region, between disruption of hormones and adverse consequences. The current work correlated early postnatal hypothyroxinemia with hearing loss in the adult rat. Linear regression was performed on the log transform for total serum thyroxine (T4) concentrations on postnatal day 14 or 21 versus dB(SPL) of hearing loss in adult animals developmentally exposed to TDCs. Regression analyses revealed a highly significant correlation between T4 concentration and hearing loss. In the rat, a 50-60% decrease in circulating T4 was needed to significantly impact hearing function. This correlation suggests that T4 serum concentrations at 14 or 21 days of postnatal age may be a good predictive biomarker in rodents of the adverse consequence of developmental exposure to TDCs.


Rapid HIV-1 testing during labor: a multicenter study.
Timely testing of women in labor with undocumented human immunodeficiency virus (HIV) status could enable immediate provision of antiretroviral prophylaxis. To determine the feasibility and acceptance of rapid HIV testing among women in labor and to assess rapid HIV assay performance. The Mother-Infant Rapid Intervention At Delivery (MIRIAD) study implemented 24-hour counseling and voluntary rapid HIV testing for women in labor at 16 US hospitals from November 16, 2001, through November 15, 2003. A rapid HIV-1 antibody test for whole blood was used. Acceptance of HIV testing; sensitivity, specificity, and predictive value of the rapid test; time from blood collection to patient notification of results. There were 91,707 visits to the labor and delivery units in the study, 7381 of which were by eligible women without documentation of HIV testing. Of these, 5744 (78%) women were approached for rapid HIV testing and 4849 (84%) consented. HIV-1 test results were positive for 34 women (prevalence = 7/1000). Sensitivity and specificity of the rapid test were 100% and 99.9%, respectively; positive predictive value was 90% compared with 76% for enzyme immunoassay (EIA). Factors independently associated with higher test acceptance included younger age, being black or Hispanic, gestational age less than 32 weeks, and having had no prenatal care. Lower acceptance was associated with being admitted between 4 pm and midnight, particularly on Friday nights, but this may be explained in part by fewer available personnel. Median time from blood collection to patient notification of result was 66 minutes (interquartile range, 45-120 minutes), compared with 28 hours for EIA (P<.001). Rapid HIV testing is feasible and delivers accurate and timely test results for women in labor. It provides HIV-positive women prompt access to intrapartum and neonatal antiretroviral prophylaxis, proven to reduce perinatal HIV transmission, and may be particularly applicable to higher-risk populations.


Biophysical feedbacks mediate carbonate chemistry in coastal ecosystems across spatiotemporal gradients.
Ocean acidification (OA) projections are primarily based on open ocean environments, despite the ecological importance of coastal systems in which carbonate dynamics are fundamentally different. Using temperate tide pools as a natural laboratory, we quantified the relative contribution of community composition, ecosystem metabolism, and physical attributes to spatiotemporal variability in carbonate chemistry. We found that biological processes were the primary drivers of local pH conditions. Specifically, non-encrusting producer-dominated systems had the highest and most variable pH environments and the highest production rates, patterns that were consistent across sites spanning 11° of latitude and encompassing multiple gradients of natural variability. Furthermore, we demonstrated a biophysical feedback loop in which net community production increased pH, leading to higher net ecosystem calcification. Extreme spatiotemporal variability in pH is, thus, both impacting and driven by biological processes, indicating that shifts in community composition and ecosystem metabolism are poised to locally buffer or intensify the effects of OA.


Smoking, alcohol consumption, diet and physical activity following stoma formation surgery, stoma-related concerns, and desire for lifestyle advice: a United Kingdom survey.
Adherence to smoking, alcohol consumption, diet and physical activity (PA) guidelines may improve outcomes for people with a stoma. A better understanding of these behaviours following stoma formation surgery and their experiences and attitudes towards receiving lifestyle advice, could help identify specific gaps and inform interventions going forward. The aim of this study was to describe changes in current lifestyle following stoma formation and to explore concerns, desire for lifestyle information, advice and support among people who have or have had a stoma. A sample of adults who currently had or in the past had a stoma for treatment for any medical condition was recruited online through relevant charities and companies, and invited to complete a cross-sectional, online survey. Consenting participants (n = 425) provided demographic information and completed brief, validated questionnaires about their lifestyle, alongside questions around their concerns regarding permanent stoma and experiences of lifestyle information and advice. Responses were summarised using descriptive statistics, and associations between reported concerns about stoma and changes in health behaviours were explored. Most respondents (93%) still had a stoma at the time of completing the survey. The majority (80%) had not consumed at least 5 portions of fruit and vegetables on the previous day and 20% reported they had not participated in at least 30 min of physical activity on any day in the previous week. Most respondents were non-smokers (84%) and did not exceed recommendations for alcohol intake (60%). Most (56%) felt their PA had decreased following stoma formation. Frequencies of concerns about a permanent stoma were high, and appeared to be associated with reported decreases in PA. Of those reporting nausea, 40% felt their diet had worsened since having their stoma. A large proportion of respondents had not received PA (42%) or dietary (30%) advice, and of these > 90% would have liked guidance. Few respondents to this survey were eating the recommended amount of fruit and vegetables, and most reported a decrease in their PA following stoma surgery. Lifestyle advice would be welcomed by this population, which professionals should take into account when addressing stoma- related concerns.


Sequencing-based analysis of the bacterial and fungal composition of kefir grains and milks from multiple sources.
Kefir is a fermented milk-based beverage to which a number of health-promoting properties have been attributed. The microbes responsible for the fermentation of milk to produce kefir consist of a complex association of bacteria and yeasts, bound within a polysaccharide matrix, known as the kefir grain. The consistency of this microbial population, and that present in the resultant beverage, has been the subject of a number of previous, almost exclusively culture-based, studies which have indicated differences depending on geographical location and culture conditions. However, culture-based identification studies are limited by virtue of only detecting species with the ability to grow on the specific medium used and thus culture-independent, molecular-based techniques offer the potential for a more comprehensive analysis of such communities. Here we describe a detailed investigation of the microbial population, both bacterial and fungal, of kefir, using high-throughput sequencing to analyse 25 kefir milks and associated grains sourced from 8 geographically distinct regions. This is the first occasion that this technology has been employed to investigate the fungal component of these populations or to reveal the microbial composition of such an extensive number of kefir grains or milks. As a result several genera and species not previously identified in kefir were revealed. Our analysis shows that the bacterial populations in kefir are dominated by 2 phyla, the Firmicutes and the Proteobacteria. It was also established that the fungal populations of kefir were dominated by the genera Kazachstania, Kluyveromyces and Naumovozyma, but that a variable sub-dominant population also exists.


Spatiotemporal distortions of visual perception at the time of saccades.
Both space and time are grossly distorted during saccades. Here we show that the two distortions are strongly linked, and that both could be a consequence of the transient remapping mechanisms that affect visual neurons perisaccadically. We measured perisaccadic spatial and temporal distortions simultaneously by asking subjects to report both the perceived spatial location of a perisaccadic vertical bar (relative to a remembered ruler), and its perceived timing (relative to two sounds straddling the bar). During fixation and well before or after saccades, bars were localized veridically in space and in time. In different epochs of the perisaccadic interval, temporal perception was subject to different biases. At about the time of the saccadic onset, bars were temporally mislocalized 50-100 ms later than their actual presentation and spatially mislocalized toward the saccadic target. Importantly, the magnitude of the temporal distortions co-varied with the spatial localization bias and the two phenomena had similar dynamics. Within a brief period about 50 ms before saccadic onset, stimuli were perceived with shorter latencies than at other delays relative to saccadic onset, suggesting that the perceived passage of time transiently inverted its direction. Based on this result we could predict the inversion of perceived temporal order for two briefly flashed visual stimuli. We developed a model that simulates the perisaccadic transient change of neuronal receptive fields predicting well the reported temporal distortions. The key aspects of the model are the dynamics of the "remapped" activity and the use of decoder operators that are optimal during fixation, but are not updated perisaccadically.


Hospitals in rural or remote areas: An exploratory review of policies in 8 high-income countries.
Our study reviewed policies in 8 high-income countries (Australia, Canada, United States, Italy, Spain, United Kingdom, Croatia and Estonia) in Europe, Australasia and North America with regard to hospitals in rural or remote areas. We explored whether any specific policies on hospitals in rural or remote areas are in place, and, if not, how countries made sure that the population in remote or rural areas has access to acute inpatient services. We found that only one of the eight countries (Italy) had drawn up a national policy on hospitals in rural or remote areas. In the United States, although there is no singular comprehensive national plan or vision, federal levers have been used to promote access in rural or remote areas and provide context for state and local policy decisions. In Australia and Canada, intermittent policies have been developed at the sub-national level of states and provinces respectively. In those countries where access to hospital services in rural or remote areas is a concern, common challenges can be identified, including the financial sustainability of services, the importance of medical education and telemedicine and the provision of quick transport to more specialized services.


UK National Data Guardian for Health and Care's Review of Data Security: Trust, better security and opt-outs.
Sharing health and social care data is essential to the delivery of high quality health care as well as disease surveillance, public health, and for conducting research. However, these societal benefits may be constrained by privacy and data protection principles. Hence, societies are striving to find a balance between the two competing public interests. Whilst the spread of IT advancements in recent decades has increased the demand for an increased privacy and data protection in many ways health is a special case. UK are adopting guidelines, codes of conduct and regulatory instruments aimed to implement privacy principles into practical settings and enhance public trust. Accordingly, in 2015, the UK National Data Guardian (NDG) requested to conduct a further review of data protection, referred to as Caldicott 3. The scope of this review is to strengthen data security standards and confidentiality. It also proposes a consent system based on an "opt-out" model rather than on "opt-in.Across Europe as well as internationally the privacy-health data sharing balance is not fixed. In Europe enactment of the new EU Data Protection Regulation in 2016 constitute a major breakthrough, which is likely to have a profound effect on European countries and beyond. In Australia and across North America different ways are being sought to balance out these twin requirements of a modern society - to preserve privacy alongside affording high quality health care for an ageing population. Whilst in the UK privacy legal framework remains complex and fragmented into different layers of legislation, which may negatively impact on both the rights to privacy and health the UK is at the forefront in the uptake of international and EU privacy and data protection principles. And, if the privacy regime were reorganised in a more comprehensive manner, it could be used as a sound implementation model for other countries.


Nitrogen comes down to earth: report from the 5th European Nitrogen Fixation Conference.
For four days and four nights, with almost 50 presentations and more than 175 posters, the 5th European Nitrogen Fixation Conference continued a tradition of excellence, bringing scientists from diverse fields such as microbiology, biochemistry, computational genomics, and plant physiology together to address the complex problems associated with biological nitrogen fixation (BNF). The conference was hosted by the John Innes Center and the University of East Anglia in Norwich, England and took place from September 6 through 10, 2002. A diverse range of topics was presented, from the evolution of rhizobial genomes to the plant genes involved in bacterial and fungal symbiosis, to the structure of nitrogenase, and to the means by which nitrogen is shuttled between the symbiotic bacteria and the plant. Additionally, sessions involving broader issues, such as nitrogen fertilizer use and work being done in developing countries, brought home the importance of the research being carried out in BNF around the world.


The H50Q mutation enhances α-synuclein aggregation, secretion, and toxicity.
Over the last two decades, the identification of missense mutations in the α-synuclein (α-Syn) gene SNCA in families with inherited Parkinson disease (PD) has reinforced the central role of α-Syn in PD pathogenesis. Recently, a new missense mutation (H50Q) in α-Syn was described in patients with a familial form of PD and dementia. Here we investigated the effects of this novel mutation on the biophysical properties of α-Syn and the consequences for its cellular function. We found that the H50Q mutation affected neither the structure of free or membrane-bound α-Syn monomer, its interaction with metals, nor its capacity to be phosphorylated in vitro. However, compared with the wild-type (WT) protein, the H50Q mutation accelerated α-Syn fibrillization in vitro. In cell-based models, H50Q mutation did not affect α-Syn subcellular localization or its ability to be phosphorylated by PLK2 and GRK6. Interestingly, H50Q increased α-Syn secretion from SHSY5Y cells into culture medium and induced more mitochondrial fragmentation in hippocampal neurons. Although the transient overexpression of WT or H50Q did not induce toxicity, both species induced significant cell death when added to the culture medium of hippocampal neurons. Strikingly, H50Q exhibited more toxicity, suggesting that the H50Q-related enhancement of α-Syn aggregation and secretion may play a role in the extracellular toxicity of this mutant. Together, our results provide novel insight into the mechanism by which this newly described PD-associated mutation may contribute to the pathogenesis of PD and related disorders.


Separate value comparison and learning mechanisms in macaque medial and lateral orbitofrontal cortex.
Uncertainty about the function of orbitofrontal cortex (OFC) in guiding decision-making may be a result of its medial (mOFC) and lateral (lOFC) divisions having distinct functions. Here we test the hypothesis that the mOFC is more concerned with reward-guided decision making, in contrast with the lOFC's role in reward-guided learning. Macaques performed three-armed bandit tasks and the effects of selective mOFC lesions were contrasted against lOFC lesions. First, we present analyses that make it possible to measure reward-credit assignment--a crucial component of reward-value learning--independently of the decisions animals make. The mOFC lesions do not lead to impairments in reward-credit assignment that are seen after lOFC lesions. Second, we examined how the reward values of choice options were compared. We present three analyses, one of which examines reward-guided decision making independently of reward-value learning. Lesions of the mOFC, but not the lOFC, disrupted reward-guided decision making. Impairments after mOFC lesions were a function of the multiple option contexts in which decisions were made. Contrary to axiomatic assumptions of decision theory, the mOFC-lesioned animals' value comparisons were no longer independent of irrelevant alternatives.


The PPE domain of PPE17 is responsible for its surface localization and can be used to express heterologous proteins on the mycobacterial surface.
PPE represent a peculiar family of mycobacterial proteins characterized by a 180 aminoacids conserved N-terminal domain. Several PPE genes are co-transcribed with a gene encoding for a protein belonging to another family of mycobacterial specific proteins named PE. Only one PE-PPE couple has been extensively characterized so far (PE25-PPE41) and it was shown that these two proteins form a heterodimer and that this interaction is essential for PPE41 stability and translocation through the mycobacterial cell wall. In this study we characterize the PE11-PPE17 couple. In contrast with what was found for PE25-PPE41, we show that PPE17 is not secreted but surface exposed. Moreover, we demonstrate that the presence of PE11 is not necessary for PPE17 stability or for its localization on the mycobacterial surface. Finally, we show that the PPE domain of PPE17 targets the mycobacterial cell wall and that this domain can be used as a fusion partner to expose heterologous proteins on the mycobacterial surface.


Magnitude of benefit of adjuvant chemotherapy for non-small cell lung cancer: meta-analysis of randomized clinical trials.
Several randomized trials investigating the benefit of adjuvant chemotherapy after surgery in non-small cell lung cancer (NSCLC) have provided conflicting results. With over 7000 patients included, we analyzed results of 13 reports over the past 10 years in which patients received either platinum-containing chemotherapy or not. The major endpoint was to assess the magnitude of the benefit of adjuvant chemotherapy in terms of the absolute benefit. All phase III randomized trials and meta-analyses published as peer-reviewed papers or as abstracts from 1994 to 2007 were eligible. A literature-based meta-analysis was performed; event-based overall- and disease-free survival (OS/DFS) and Relative Risks (RRs) with 95% confidence intervals (CIs) were derived. Magnitudes of benefit were evaluated with: absolute benefit and the number of patients treated for one patient to benefit. Seven sub-populations were examined. Combined effect estimation was computed with fixed- and random-effect models; a heterogeneity test was also applied. Twelve trials plus an individual patient meta-analysis (7334 patients) were gathered; the trials were designed to determine if cisplatin- or carboplatin-based chemotherapy improves survival over surgery. When data were pooled and plotted, significant differences in favor of chemotherapy were seen in OS in all seven sub-population, with a relative benefit of 7-12% and an absolute benefit ranging from 2.5% to 4.1%. A more significant trend for chemotherapy was found in DFS. No significant heterogeneity was observed for all outcomes and sub-populations. The absolute benefit of adjuvant chemotherapy remains essentially the same regardless of how data are screened. While significant differences are clearly found in this analysis, the small magnitude of benefit seen with this large population, especially when considering the number of patients needed for one to benefit, raises important issues when weighing risks and benefits of treatment for individual patients.


Metabolism of rabbit angiostatin glycoforms I and II in rabbits: angiostatin-I leaves the intravascular space faster and appears to have greater anti-angiogenic activity than angiostatin-II.
Plasminogen (PLG) exists in the circulation as two glycoforms, I and II. Angiostatin (AST) is a polypeptide that has been cleaved from the kringle region of PLG and has strong anti-angiogenic properties. AST-I and AST-II, which consisted only of kringles 1 through 3, were prepared by the action of urokinase on purified rabbit PLG-I and PLG-II, respectively, in the presence of N-acetyl cysteine, followed by affinity chromatography on lysine-Sepharose. Purified AST-I and AST-II were tested for functional activity with a chick chorioallantoic membrane (CAM) model; when similar amounts were applied to a 6-day CAM, AST-I was substantially more effective than AST-II in decreasing vascular supply to the CAM over a 72-hour period; this activity correlated with a loss of capillaries, probably through apoptosis of endothelial cells. Radiolabeled AST-I and AST-II (iodine 125 and iodine 131) were co-injected intravenously into healthy rabbits to determine their clearances from plasma measured over 3 days. Over a dose range of 0.08 to 2.7 microg/kg, the fractional catabolic rate within the intravascular space (j(3)) indicated that AST-I was cleared 3-fold to 4-fold more rapidly than AST-II (P < .001). The catabolic half-life of AST-I (2.01 +/- 0.19 days) was significantly less than that of AST-II (2.62 +/- 0.20 days). The faster clearance of AST-I from the intravascular space was matched by its more rapid passage than AST-II to the extravascular space of various organs over 60 minutes in vivo. This property of AST-I as compared with AST-II may partially explain its greater anti-angiogenic potential. From the plasma concentrations of PLG-I and PLG-II and their relative behaviors toward rabbit VX-2 lung tumors in vivo, we predict that substantially greater quantities of AST-II than AST-I may be released into the extravascular space of tumors.


Sex determination from the foramen magnum: discriminant function analysis in an eighteenth and nineteenth century British sample.
The successful identification of the deceased is vital to the progress of any forensic investigation. One of the principal biological traits to be established from skeletal remains is the sex of the individual. This becomes more difficult if only parts of a skeleton are found or if the bones are compromised by physical insults such as fire, explosions or violence. The basal region of the occipital bone is covered by a large volume of soft tissue and is therefore in a relatively well-protected anatomical position, and as such, classification of sex using the occipital bone may prove useful in cases of significantly disrupted remains. The aim of this paper is to evaluate manually recorded morphometric variables of the region of the foramen magnum using both discriminant function analysis and linear regression. The skulls utilised in this study were selected from the eighteenth to nineteenth century documented skeletal collection of St. Bride's Church, Fleet Street, London. Adult human skulls n = 158 (male symbol82/female symbol76) were measured to derive statistical functions. The results demonstrated that significant sexual dimorphism is present in the cranial base of the St. Bride's material. The correctly classified crania within this population ranged from 65.8% for univariate functions to 70.3% for multivariate functions within the cranial sample. Males were correctly classified at 70.7% and females at 69.7% using multivariate functions. The linear regression equations predicted sex in the cranial sample correctly for 76% of the males and 70% for the females using different variables; however, overall highest correct prediction percentage was only 68%. Cross-validation brought the percentage down in some cases, but it was concluded that, overall, the expression of sexual dimorphism in the foramen magnum region within the St. Bride's population is significantly demonstrable, and therefore, this area of the skull should be considered useful in the identification of sex.


Lemongrass Extract Possesses Potent Anticancer Activity Against Human Colon Cancers, Inhibits Tumorigenesis, Enhances Efficacy of FOLFOX, and Reduces Its Adverse Effects.
Current chemotherapeutics for metastatic colorectal cancers have limited success and are extremely toxic due to nonselective targeting. Some natural extracts have been traditionally taken and have shown anticancer activity. These extracts have multiple phytochemicals that can target different pathways selectively in cancer cells. We have shown previously that lemongrass (Cymbopogon citratus) extract is effective at inducing cell death in human lymphomas. However, the efficacy of lemongrass extract on human colorectal cancer has not been investigated. Furthermore, its interactions with current chemotherapies for colon cancer is unknown. In this article, we report the anticancer effects of ethanolic lemongrass extract in colorectal cancer models, and importantly, its interactions with FOLFOX and Taxol. Lemongrass extract induced apoptosis in colon cancer cells in a time and dose-dependent manner without harming healthy cells in vitro. Oral administration of lemongrass extract was well tolerated and effective at inhibiting colon cancer xenograft growth in mice. It enhanced the anticancer efficacy of FOLFOX and, interestingly, inhibited FOLFOX-related weight loss in animals given the combination treatment. Furthermore, feeding lemongrass extract to APCmin/+ transgenic mice led to the reduction of intestinal tumors, indicating its preventative potential. Therefore, this natural extract has potential to be developed as a supplemental treatment for colorectal cancer.


Heterogeneity among Mycobacterium ulcerans from French Guiana revealed by multilocus variable number tandem repeat analysis (MLVA).
Buruli ulcer is an emerging and neglected tropical disease caused by Mycobacterium ulcerans. Few cases have been reported so far in the Americas. With 250 cases reported since 1969, French Guiana is the only Buruli ulcer endemic area in the continent. Thus far, no genetic diversity studies of strains of M. ulcerans from French Guiana have been reported. Our goal in the present study was to examine the genetic diversity of M. ulcerans strains in this region by using the Multilocus Variable Number Tandem Repeat Analysis (MLVA) approach. A total of 23 DNA samples were purified from ulcer biopsies or derived from pure cultures. MVLA was used in the study of six previously-described Variable Number of Tandem Repeat (VNTR) markers. A total of three allelic combinations were characterized in our study: genotype I which has been described previously, genotype III which is very similar to genotype I, and genotype II which has distinctly different characteristics in comparison with the other two genotypes. This high degree of genetic diversity appears to be uncommon for M. ulcerans. Further research based on complete genome sequencing of strains belonging to genotypes I and II is in progress and should lead soon to a better understanding of genetic specificities of M. ulcerans strains from French Guiana.


Ganoderma lucidum polysaccharide improves rat DSS-induced colitis by altering cecal microbiota and gene expression of colonic epithelial cells.
The effects of β-glucan on colitis mice are contradictory in previous reports. As a result, it is still unclear whether there is an anti-colitis effect in Ganoderma lucidum polysaccharide (GLP), which is mainly composed of β-glucan. Moreover, the association between GLP function and gut microbiota remains to be elucidated. This study aimed to investigate whether GLP consumption improved rat dextran sodium sulfate (DSS)-induced colitis by regulating gut microbiota and altering colonic epithelial expression. The disease activity index (DAI) scores and the cecal short chain fatty acid (SCFA) levels of DSS-induced colitis rats fed with a GLP diet (Group GLP, n = 6) and a control diet (Group Con, n = 6) were investigated and analyzed. Moreover, the profiles of gut microbiota and colonic epithelial expression were analyzed using metagenomics and transcriptomics. GLP consumption significantly lowered animal DAI scores by producing more SCFAs by increasing SCFA-producing bacteria such as Ruminococcus_1 and reducing pathogens such as Escherichia-Shigella in both the small intestine and cecum of rat. Moreover, GLP consumption regulated 11 genes, including six upregulated (Ccl5, Cd3e, Cd8a, Il21r, Lck, and Trbv) and five downregulated (Ccl3, Gro, Il11, Mhc2, and Ptgs) genes enriched in six inflammation-related Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, resulting in enhancement of immunity and reduction of inflammatory response and colonic cancer risk. GLP consumption alleviated DSS-induced colitis and may have potential for ulcerative colitis relief.


A Chimeric EccB-MycP Fusion Protein is Functional and a Stable Component of the ESX-5 Type VII Secretion System Membrane Complex.
The mycosin protease (MycP) is widely conserved in type VII secretion (T7S) systems throughout Actinobacteria. Within the T7S systems of mycobacteria, also known as the ESX systems, MycP is essential for secretion, which is probably linked to its stabilizing effect on the ESX membrane complex. However, it is unknown how this is mediated, as MycP is not a stable component of this complex. In this study, we set out to create a chimeric fusion protein of EccB5 and MycP5, based on a chimeric gene of eccB and mycP in the T7S locus of Bifidobacterium dentium. We show that this fusion protein is functional and capable of complementing ESX-5 secretion in both an eccB5 and a mycP5 knockout in Mycobacterium marinum. To study the ESX complex containing this fusion protein in more detail, we replaced the original eccB5 and mycP5 of the Mycobacterium xenopi esx-5 locus, reconstituted in Mycobacterium smegmatis, with the chimeric gene. The EccB5-MycP5 fusion construct also restored ESX-5 secretion under these double knockout conditions. Subsequent protein pulldowns on the central complex component EccC5 showed that under these conditions, the EccB5-MycP5 fusion was specifically copurified and a stable component of the ESX-5 complex. Based on our results, we can conclude that MycP5 carries out its essential function in secretion in close proximity to EccB5, indicating that EccB5 is the direct interaction partner of MycP5.


Validation of an assay for the determination of levoglucosan and associated monosaccharide anhydrides for the quantification of wood smoke in atmospheric aerosol.
Biomass burning is becoming an increasing contributor to atmospheric particulate matter, and concern is increasing over the detrimental health effects of inhaling such particles. Levoglucosan and related monosaccharide anhydrides (MAs) can be used as tracers of the contribution of wood burning to total particulate matter. An improved gas chromatography-mass spectrometry method to quantify atmospheric levels of MAs has been developed and, for the first-time, fully validated. The method uses an optimised, low-volume methanol extraction, derivitisation by trimethylsilylation and analysis with high-throughput gas chromatography-mass spectrometry (GC-MS). Recovery of approximately 90 % for levoglucosan, and 70 % for the isomers galactosan and mannosan, was achieved using spiked blank filters estimates. The method was extensively validated to ensure that the precision of the method over five experimental replicates on five repeat experimental occasions was within 15 % for low, mid and high concentrations and accuracy between 85 and 115 %. The lower limit of quantification (LLOQ) was 0.21 and 1.05 ng m(-3) for levoglucosan and galactosan/mannosan, respectively, where the assay satisfied precisions of ≤20 % and accuracies 80-120 %. The limit of detection (LOD) for all analytes was 0.105 ng m(-3). The stability of the MAs, once deposited on aerosol filters, was high over the short term (4 weeks) at room temperature and over longer periods (3 months) when stored at -20 °C. The method was applied to determine atmospheric levels of MAs at an urban background site in Leicester (UK) for a month. Mean concentrations of levoglucosan over the month of May were 21.4 ± 18.3 ng m(-3), 7.5 ± 6.1 ng m(-3) mannosan and 1.8 ± 1.3 ng m(-3) galactosan.


A Stereoconvergent Cyclopropanation Reaction of Styrenes.
The first stereoconvergent cyclopropanation reaction by means of photoredox catalysis using diiodomethane as the methylene source is described. This transformation exhibits broad functional group tolerance and it is characterized by an excellent stereocontrol en route to trans-cyclopropanes regardless of whether E- or Z-styrene substrates were utilized.


Histopathology and prognosis of malignant colorectal polyps treated by endoscopic polypectomy.
The histopathological features and results of treatment of malignant polyps removed by endoscopic polypectomy from 60 patients are presented. The patients were followed for a minimum of five years. Forty six patients were treated by polypectomy alone as local excision was judged complete and the invasive carcinoma was well or moderately well differentiated. Thirty seven of these patients are alive and well after five years and the remaining nine have died of other causes up to four years later: there was no evidence of recurrence in any of these cases. Fourteen patients underwent a subsequent major surgical resection and residual tumour was found at the site of polypectomy in two cases but regional lymph nodes were not involved. Only one patient from this group with a high grade tumour developed metastases although the operative specimen was free of tumour. Malignant polyps can be successfully treated by polypectomy alone provided both the laboratory techniques of examination and the histopathological criteria are strictly applied.


A meta-analysis of procedures to change implicit measures.
Using a novel technique known as network meta-analysis, we synthesized evidence from 492 studies (87,418 participants) to investigate the effectiveness of procedures in changing implicit measures, which we define as response biases on implicit tasks. We also evaluated these procedures' effects on explicit and behavioral measures. We found that implicit measures can be changed, but effects are often relatively weak (|ds| < .30). Most studies focused on producing short-term changes with brief, single-session manipulations. Procedures that associate sets of concepts, invoke goals or motivations, or tax mental resources changed implicit measures the most, whereas procedures that induced threat, affirmation, or specific moods/emotions changed implicit measures the least. Bias tests suggested that implicit effects could be inflated relative to their true population values. Procedures changed explicit measures less consistently and to a smaller degree than implicit measures and generally produced trivial changes in behavior. Finally, changes in implicit measures did not mediate changes in explicit measures or behavior. Our findings suggest that changes in implicit measures are possible, but those changes do not necessarily translate into changes in explicit measures or behavior. (PsycINFO Database Record (c) 2019 APA, all rights reserved).