Metabolism as a determinant of species susceptibility to 2,3,5-(triglutathion-S-yl)hydroquinone-mediated nephrotoxicity. The role of N-acetylation and N-deacetylation.
2,3,5-(Triglutathion-S-yl)hydroquinone [2,3,5-(triGSyl)HQ] is a potent nephrotoxicant when administered to male rats. We now report that significant species differences exist in susceptibility to 2,3,5-(triGSyl)HQ-mediated nephrotoxicity. Metabolism of glutathione conjugates involves cleavage of teh glutamate and glycine moieties by gamma-glutamyltranspeptidase (gamma-GT) and dipeptidases, respectively, and the nephrotoxicity of 2,3,5-(triGSyl)HQ can be prevented by the inhibition of renal gamma-GT. The resulting cysteine conjugate exhibits a balance between N-acetylation, and N-deacetylation of the mercapturic acid biosynthesis in various species contribute to species susceptibility to 2,3,5-(triGSyl)HQ. Renal gamma-GT activity toward 2,3,5-(triGSyl)HQ was highest in the rat (Fischer 344 and Sprague-Dawley) and consistent with the sensitivity of this species to 2,3,5-(triGSyl)HQ (20 micromol/kg iv)-mediated nephrotoxicity. The gamma-GT-mediated hydrolysis of 2,3,5-(triGSyl)HQ was similar in B6C3F1 and BALB/c mice and guinea pigs. In these species, the gamma-GT activity ranged between 30-45% of the activity measured in rats. Although, the activity of gamma-GT was similar in mice and guinea pigs, only guinea pigs were susceptible to 2,3,5-(triGSyl)HQ (200 micromol/kg iv)-induced renal necrosis. The gamma-GT-mediated hydrolysis of 2,3,5-(triGSyl)HQ was lowest in the hamster, and this species were not susceptible to the renal toxicity of this conjugate. Thus, factors in addition to gamma-GT activity probably contribute to species susceptibility to 2,3,5-(triGSyl)HQ nephrotoxicity. The kinetics of the AT-125-mediated inhibition of gamma-GT differed between species, indicative of potential differences in the regulation of gamma-GT. Consistent with this view, the ratio between the hydrolysis and transpeptidation of 2,3,5-(triGSyl)HQ varied 10-fold between the species examined, and was highest in the guinea pig (0.48) and lowest in the hamster (0.05). Guinea pigs also exhibited the highest renal cytosolic N-deacetylase activity and the lowest N-acetylase activity. The ratios of N-deacetylation to N-acetylation in guinea pigs, BALB/c mice, B6C3F1 mice, hamsters, Fischer 344 rats, and Sprague-Dawley rats were 4.57, 0.16, 0.14, 0.04, 0.03, and 0.02, respectively. Because quinol-cysteine conjugates seem to undergo oxidation more readily than the corresponding mercapturates, the balance of N-deacetylase and N-acetylase in the guinea pig may contribute to the susceptibility of this species to 2,3,5-(triGSyl)HQ nephrotoxicity.


Lymphocyte cAMP and ageing: significance of subset composition, plasma noradrenaline, regular physical training and long-term smoking.
1. We studied 37 healthy men at rest in the supine. position to examine the effect of ageing, smoking and physical training of beta 2-adrenoceptor function, plasma catecholamines and the proportions of various lymphocyte subsets. 2. In 14 young subjects the proportion of natural killer cells was correlated with cAMP production in lymphocytes and inversely correlated with plasma noradrenaline level. 3. In 16 elderly non-smokers plasma noradrenaline was negatively correlated with the natural killer cell subset CD3-CD16+. Lymphocyte cAMP responses did not differ between young and elderly non-smokers, whereas plasma noradrenaline increased slightly but significantly with age. Physical training did not influence either plasma noradrenaline or adrenaline at rest or cAMP in lymphocytes. 4. In seven elderly long-term smokers cAMP production and the viability of lymphocytes were reduced. Plasma noradrenaline attained its highest values in long-term smokers. 5. It is concluded that cAMP production and plasma noradrenaline are related to lymphocyte subset composition. The greater the proportion of natural killer cells and related subsets, the higher is cAMP production and the lower is plasma noradrenaline. Thus, the inverse correlation between lymphocyte cAMP and plasma noradrenaline is indirect and most likely mediated by variability in lymphocyte subset composition. In elderly subjects, reduced cAMP production was observed in long-term smokers, and this abnormality was probably due to a reduced viability of lymphocytes and especially of natural killer cells. The negative correlation between the proportion of natural killer cells and plasma noradrenaline at rest contracts with a well-known mobilizing effect of adrenaline on natural killer cells.


A simple sequence repeat- and single-nucleotide polymorphism-based genetic linkage map of the brown planthopper, Nilaparvata lugens.
In this study, we developed the first genetic linkage map for the major rice insect pest, the brown planthopper (BPH, Nilaparvata lugens). The linkage map was constructed by integrating linkage data from two backcross populations derived from three inbred BPH strains. The consensus map consists of 474 simple sequence repeats, 43 single-nucleotide polymorphisms, and 1 sequence-tagged site, for a total of 518 markers at 472 unique positions in 17 linkage groups. The linkage groups cover 1093.9 cM, with an average distance of 2.3 cM between loci. The average number of marker loci per linkage group was 27.8. The sex-linkage group was identified by exploiting X-linked and Y-specific markers. Our linkage map and the newly developed markers used to create it constitute an essential resource and a useful framework for future genetic analyses in BPH.


Evaluation of the appropriate root pressure for maintaining heartbeat during an aortic cross-clamp for primary repair of the aortic arch in premature infants with associated cardiac anomalies.
We developed a new cardiopulmonary bypass (CPB) method to minimize myocardial damage during aortic arch reconstruction. In this method, coronary flow and heartbeat were stabilized by maintaining the aortic root pressure with an adjusted preload of the ventricle during aortic cross-clamping. This study was performed to determine the appropriate root pressure to maintain the heartbeat without causing deterioration of ventricular function. Study 1. Under partial CPB, the ascending aorta was cross-clamped in 6 pigs (group 1). Experimental data at various systolic aortic root pressures was analysed to determine the appropriate root pressure. Study 2. In group 2 (control, n=6), the aorta was not clamped, while in group 3 (n=6), the aorta was cross-clamped for 60 min and the systolic aortic root pressure was maintained at the pressure determined in study 1. Study 1. The diastolic coronary flow was stabilized at values comparable to that before initiation of CPB (6.6±1.4 ml/beat) when the systolic aortic root pressure was above 80 mmHg. Intracardiac pressure and the myocardial oxygen consumption (MvO2) seemed to be acceptable when the systolic aortic root pressure was below 100 mmHg. Therefore, 90 mmHg was selected for study 2. Study 2. Perioperative cardiac function did not differ between the groups. We concluded that 90 mmHg was the systolic aortic root pressure appropriate for this method.


Wnt and extraocular muscle sparing in amyotrophic lateral sclerosis.
The extraocular muscles (EOM) and their motor neurons are spared in amyotrophic lateral sclerosis (ALS). In limb muscle, axon retraction from the neuromuscular junctions occurs early in the disease. Wnts, a conserved family of secreted signaling molecules, play a critical role in neuromuscular junction formation. This is the first study to examine Wnt signaling for its potential involvement in maintenance of normal morphology in EOM in ALS. Extraocular muscle and limb muscle axons, neuromuscular junctions, and myofibers from control, aging, and ALS subjects and the SOD1(G93A) mouse model of ALS were quantified for their expression of Wnt1, Wnt3a, Wnt5a, Wnt7a, and β-catenin. All four Wnt isoforms were expressed in most axon profiles in all human EOM. Significantly fewer were positive for Wnt1, Wnt3a, and Wnt7a in the human limb muscles. Similar differential patterns in Wnt myofiber expression were also seen except in the case of Wnt7a, where expression was elevated. In the SOD1(G93A) mouse, all four Wnt isoforms were significantly decreased in the neuromuscular junctions at the terminal stage compared to values in age-matched controls. β-Catenin was activated in a subset of myofibers in EOM and limb muscle in all subjects. The differences in expression of Wnts in EOM and limb muscle, particularly at the neuromuscular junction level, suggest that they play a role in the pathophysiology of ALS. Collectively, the data support a role for signaling of Wnts in the preservation of the EOM in ALS and their dysregulation and the subsequent development of pathology in the ALS limb muscles.


Novel vectors for homologous recombination strategies in mouse embryonic stem cells: an ES cell line expressing EGFP under control of the 5T4 promoter.
The use of gene mutation/knock-out strategies in mouse embryonic stem (ES) cells has revolutionized the study of gene function in ES cells and embryonic development. However, the construction of vectors for homologous recombination strategies requires considerable expertise and time. We describe two novel vectors that can generate site specific knock-out or EGFP knock-in ES cells within 6 weeks from construct design to identification of positive ES cell clones. As proof-of-principle, we have utilized the knock-out targeting vector to modify the NEIL2 locus in ES cells. In addition, using the knock-in vector, we have inserted EGFP downstream of the 5T4 oncofetal antigen promoter in ES cells (5T4-GFP ES cells). Undifferentiated 5T4-GFP ES cells lack EGFP and maintain expression of the pluripotent markers OCT-4 and NANOG. Upon differentiation, EGFP expression is increased in 5T4-GFP ES cells and this correlates with 5T4 transcript expression of the unmodified allele, loss of Nanog and Oct-4 transcripts and upregulation of differentiation-associated transcripts. Furthermore, we demonstrate that fluorescent activated cell sorting of 5T4-GFP ES cells allows isolation of pluripotent or differentiated cells from a heterogeneous population. These vectors provide researchers with a rapid method of modifying specific ES cell genes to study cellular differentiation and embryonic development.


A randomized, controlled, open-label trial evaluating the efficacy and safety of chloroquine in the treatment of giardiasis in children.
Giardia duodenalis is among the commonest protozoan parasites in the intestinal tract of humans and may cause significant morbidity worldwide. Although there are several antigiardial agents, treatment failures have been commonly reported. To compare the efficacy and safety of chloroquine (CQ) versus metronidazole (MTZ) in the treatment of children with confirmed G duodenalis mono-infection. A randomized, controlled, open-label trial was carried out at the Cuban Institute of Gastroenterology. One hundred and twenty-two children were randomly assigned to receive either CQ (10 mg/Kg bodyweight twice a day for five days) or MTZ [15 mg/Kg bodyweight divided in three daily does for five days]. All children were asked to provide three faecal samples on days 3, 5 and 7 after treatment completion. Children were considered to be cured, if no Giardia trophozoites or cysts were found in any of the three post-treatment faecal specimens evaluated by direct wet mounts and/or after Ritchie concentration techniques. The frequency of cure was a little higher for CQ than for MTZ but the difference was not statistically significant. Headache was more common in patients treated with CQ as was bitter taste. Yellowish colouration of the urine was more frequent in the MTZ treated group. Chloroquine, for five days, is as efficacious as the recommended treatment with MTZ in children infected with G duodenalis.


Comparative study of the edema-forming activity of Costa Rican snake venoms and its neutralization by a polyvalent antivenom.
The edema-forming activity of eight Costa Rican crotaline snake venoms and its neutralization by a polyvalent antivenom were studied using the mouse footpad test. All of the venoms induced edema, the highest activity being present in the venoms of Bothrops lateralis and Bothrops picadoi. When experiments were performed with preincubation of venom and antivenom, neutralization of edema was poor. Moreover, it was observed that, with some venoms, edema increased when large doses of antivenom were used. This effect was also observed when some venoms were incubated with coral snake antivenom, suggesting that venoms may release some pharmacologically active component(s) from antivenom, since the latter contains traces of alpha-2 and beta globulins. Based on these findings, an alternative approach to the study of the neutralization of edema was used; in this new method, antivenom was injected i.v. before venom administration, thereby avoiding preincubation. With this technique, a much better neutralization of edema was observed, although with some venoms it was still poor. Venoms contain low molecular weight factors which induce edema, suggesting that lack of immunogenicity of some components may cause a poor neutralization. However, such components are responsible for only a minor portion of the edema induced by crude venoms. It is suggested that experiments in which venom and antivenom are preincubated preincubated in testing the neutralization of edema should be avoided, and that a more adequate approach may be an independent inoculation of venom and antivenom.


Change of conduction velocity by regional myelination yields constant latency irrespective of distance between thalamus and cortex.
The widely spanning sensory cortex receives inputs from the disproportionately smaller nucleus of the thalamus, which results in a wide variety of travelling distance among thalamic afferents. Yet, latency from the thalamus to a cortical cell is remarkably constant across the cortex (typically, approximately 2 ms). Here, we found a mechanism that produces invariability of latency among thalamocortical afferents, irrespective of the variability of travelling distances. The conduction velocity (CV) was calculated from excitatory postsynaptic currents recorded from layer IV cells in mouse thalamocortical slices by stimulating the ventrobasal nucleus of the thalamus (VB) and white matter (WM). In adults, the obtained CV for VB to WM (CV(VB-WM); 3.28 +/- 0.11 ms) was approximately 10 times faster than that of WM to layer IV cells (CV(WM-IV); 0.33 +/- 0.05 ms). The CV(VB-WM) was confirmed by recording antidromic single-unit responses from VB cells by stimulating WM. Exclusion of synaptic delay from CV(WM-IV) did not account for the 10-fold difference of CV. By histochemical staining, it was revealed that VB to WM was heavily myelinated, whereas in the cortex staining became substantially weaker. We also found that such morphological and physiological characteristics developed in parallel and were accomplished around postnatal week 4. Considering that VB to WM is longer and more variable in length among afferents than is the intracortical region, such an enormous difference of CV makes conduction time heavily dependent on the length of intracortical region, which is relatively constant. Our finding may well provide a general strategy of connecting multiple sites irrespective of distances in the brain.


Anti-norovirus therapeutics: a patent review (2010-2015).
Human noroviruses are the primary causative agents of acute gastroenteritis and are a pressing public health burden worldwide. There are currently no vaccines or small molecule therapeutics available for the treatment or prophylaxis of norovirus infections. An improved understanding of norovirus biology, as well as the pathogenic mechanisms underlying the disease, has provided the impetus for a range of intense exploratory drug discovery efforts targeting viral and host factors. An overview of norovirus inhibitors disclosed in the patent literature (2010-present) and Clinicaltrials.gov is presented. The review is further enriched and supplemented by recent literature reports. Seminal discoveries made in recent years, including a better understanding of the pathobiology and life cycle of norovirus, the identification and targeting of multiple viral and host factors, the advent of a replicon system and a small animal model for the preclinical evaluation of lead compounds, and the availability of high resolution X-ray crystal structures that can be utilized in structure-based drug design and lead optimization campaigns, collectively suggest that a small molecule therapeutic and prophylactic for norovirus infection is likely to emerge in the not too distant future.


Cigarette litter: smokers' attitudes and behaviors.
Cigarette butts are consistently the most collected items in litter clean-up efforts, which are a costly burden to local economies. In addition, tobacco waste may be detrimental to our natural environment. The tobacco industry has conducted or funded numerous studies on smokers' littering knowledge and behavior, however, non-industry sponsored research is rare. We sought to examine whether demographics and smokers' knowledge and beliefs toward cigarette waste as litter predicts littering behavior. Smokers aged 18 and older (n = 1,000) were interviewed about their knowledge and beliefs towards cigarette waste as litter. Respondents were members of the Research Now panel, an online panel of over three million respondents in the United States. Multivariate logistic regressions were conducted to determine factors significantly predictive of ever having littered cigarette butts or having littered cigarette butts within the past month (p-value < 0.05). The majority (74.1%) of smokers reported having littered cigarette butts at least once in their life, by disposing of them on the ground or throwing them out of a car window. Over half (55.7%) reported disposing of cigarette butts on the ground, in a sewer/gutter, or down a drain in the past month. Those who did not consider cigarette butts to be litter were over three and half times as likely to report having ever littered cigarette butts (OR = 3.68, 95%CI = 2.04, 6.66) and four times as likely to have littered cigarette butts in the past month (OR = 4.00, 95%CI = 2.53, 6.32). Males were significantly more likely to have littered cigarette butts in the past month compared to females (OR = 1.49, 95%CI = 1.14, 1.94). Holding the belief that cigarette butts are not litter was the only belief in this study that predicted ever or past-month littering of cigarette waste. Messages in anti-cigarette-litter campaigns should emphasize that cigarette butts are not just litter but are toxic waste and are harmful when disposed of improperly.


Disproportionality and learning disabilities: parsing apart race, socioeconomic status, and language.
The disproportionate identification of learning disabilities among certain sociodemographic subgroups, typically groups that are already disadvantaged, is perceived as a persistent problem within the education system. The academic and social experiences of students who are misidentified with a learning disability may be severely restricted, whereas students with a learning disability who are never identified are less likely to receive the accommodations and modifications necessary to learn at their maximum potential. The authors use the Education Longitudinal Study of 2002 to describe national patterns in learning disability identification. Results indicate that sociodemographic characteristics are predictive of identification with a learning disability. Although some conventional areas of disproportionality are confirmed (males and language minorities), differences in socioeconomic status entirely account for African American and Hispanic disproportionality. The discrepancy between the results of bivariate and multivariate analyses confirms the importance of employing multivariate multilevel models in the investigation of disproportionality.


Graded, multidimensional intra- and intergroup variations in primary progressive aphasia and post-stroke aphasia.
Language impairments caused by stroke (post-stroke aphasia, PSA) and neurodegeneration (primary progressive aphasia, PPA) have overlapping symptomatology, nomenclature and are classically divided into categorical subtypes. Surprisingly, PPA and PSA have rarely been directly compared in detail. Rather, previous studies have compared certain subtypes (e.g. semantic variants) or have focused on a specific cognitive/linguistic task (e.g. reading). This study assessed a large range of linguistic and cognitive tasks across the full spectra of PSA and PPA. We applied varimax-rotated principal component analysis to explore the underlying structure of the variance in the assessment scores. Similar phonological, semantic and fluency-related components were found for PSA and PPA. A combined principal component analysis across the two aetiologies revealed graded intra- and intergroup variations on all four extracted components. Classification analysis was used to test, formally, whether there were any categorical boundaries for any subtypes of PPA or PSA. Semantic dementia formed a true diagnostic category (i.e. within group homogeneity and distinct between-group differences), whereas there was considerable overlap and graded variations within and between other subtypes of PPA and PSA. These results suggest that (i) a multidimensional rather than categorical classification system may be a better conceptualization of aphasia from both causes; and (ii) despite the very different types of pathology, these broad classes of aphasia have considerable features in common.


The Type IV Secretion System of ICEAfe1: Formation of a Conjugative Pilus in Acidithiobacillus ferrooxidans.
The dispersal of mobile genetic elements and their gene cargo relies on type IV secretion systems (T4SS). In this work the ICEAfe1 Tra-type T4SS nanomachine, encoded in the publicly available genome of Acidithiobacillus ferrooxidans ATCC 23270TY, was characterized in terms of its organization, conservation, expression and mating bridge formation. Twenty-one conjugative genes grouped in four genetic clusters encode the ICEAfe1 T4SS, containing all the indispensable functions for the formation and stabilization of the pili and for DNA processing. The clusters' organization resembles that of other mobile genetic elements (such as plasmids and integrative and conjugative elements-ICEs). Sequence conservation, genetic organization and distribution of the tra system in the genomes of other sequenced Acidithiobacillus spp. suggests that the ICEAfe1 T4SS could mediate the lateral gene transfer between related bacteria. All ICEAfe1 T4SS genes are transcriptionally active and expressed from four independent operons. The transcriptional levels of selected marker genes increase in response to Mitomycin C treatment, a DNA damage elicitor that has acknowledged stimulatory effects on excision rates and gene expression of other ICEs, including ICEAfe1. Using a tailor-made pilin-antiserum against ICEAfe1 T4SS TraA pilin and epifluorescence microscopy, the presence of the conjugative pili on the cell surface of A. ferrooxidans could be demonstrated. Additionally, immunodetection assays, by immunogold, allowed the identification of pili-like extracellular structures. Together, the results obtained in this work demonstrate that the ICEAfe1 T4SS is phylogenetically conserved within the taxon, is expressed at mRNA and protein levels in vivo in the A. ferrooxidans type strain, and produces a pili-like structure of extracellular and intercellular localization in this model acidophile, supporting its functionality. Additional efforts will be required to prove conjugation of the ICEAfe1 or parts of this element through the cognate T4SS.


Effect of novel and familiar mating partners on the duration of sexual receptivity in the female hamster.
The lordosis duration of the female golden hamster (Mesocricetus auratus) in response to novel and familiar mating partners was examined. In Experiment 1, females mated to the point of sexual satiation with one male hamster, and following its removal, showed renewed receptivity in response to the introduction of a second male. Upon sexual satiation with the second male, females either received a novel third male or were reexposed to the original male. Total lordosis duration in the third bout in the group receiving a novel male was significantly greater than in the group reexposed to the original male. In Experiment 2, the insertion of a 1-h delay between the second and the third mating partner had no effect on the female's responsiveness, regardless of whether the third male was the original male or a novel male. In Experiment 3, removal of the ovaries followed by hormone replacement treatment (40 micrograms estradiol benzoate, 72 h prior to testing, and 500 micrograms progesterone, 4 h prior to testing) failed to alter the females' ability to discriminate between novel and familiar mating partners. These results demonstrate that females make clear discriminations among individual mating partners, that this can affect sexual receptivity and will continue to do so following a delay of at least 1 h, and that the effect is not mediated by the release of ovarian steroids.


Pressure dependence of the electrical transport in granular materials.
We report on systematic measurements of the electrical resistance of one- and three-dimensional (1D and 3D) metallic and oxidized granular materials under uni-axial compression. Whatever the dimension of the packing, the resistance follows a power law versus the pressure ([Formula: see text]), with an exponent [Formula: see text] much larger than the ones expected either with elastic or plastic contact between the grains. A simple model based on a statistical description of the micro-contacts between two grains is proposed. It shows that the strong dependence of the resistance on the pressure applied to the granular media is a consequence of large variabilities and heterogeneities present at the contact surface between two grains. Then, the effect of the three-dimensional structure of the packing is investigated using a renormalization process. This allows to reconcile two extreme approaches of a 3D lattice of widely distributed resistances: the effective medium and the percolation theories.


Women awaiting hysterectomy: a qualitative study of issues involved in decisions about oophorectomy.
Women awaiting abdominal hysterectomy have a further decision to take--whether or not to undergo a prophylactic oophorectomy. Prophylactic oophorectomy (removal of healthy ovaries) is commonly undertaken as an adjunct to hysterectomy to prevent ovarian cancer in women who are already having a hysterectomy for menorrhagia, fibroids or severe menstrual pain. It causes an immediate 'surgical menopause'--women no longer produce endogenous oestrogens and they are advised to take hormone replacement therapy (HRT). This descriptive study was undertaken as part of a larger pilot study. Qualitative interviewing study. Hospital outpatient departments in southern England. Women on waiting lists for hysterectomy for benign conditions. In-depth semi-structured interviews were undertaken with 16 women awaiting hysterectomy to examine their views of prophylactic oophorectomy, menopause and HRT use. Interviews were transcribed. Nud*ist was used to organise the data. Analysis was undertaken by two researchers working independently who then met to agree themes. Women held treatment preferences both about the removal or retention of their ovaries and about subsequent HRT use. Many of the women did not want oophorectomy because of a dislike of a sudden menopause. Those who were planning to have an oophorectomy tended to have more severe symptoms. Some women were fearful of ovarian cancer--one woman described it as a 'silent killer'. A further theme was apparent in the discussion of ovaries as 'redundant' with further discussion of healthy versus problematic ovaries. Many women felt inadequately informed about their treatment options and were unaware of important longer term outcomes of oophorectomy such as an increased risk of osteoporosis or of coronary heart disease. It appears that many women emphasise the 'natural' when considering oophorectomy and the use of HRT. Women were uneasy both about a sudden menopause and about the risks of ovarian cancer. And although these women were all about to undergo surgery, they had experienced an apparent serious lack of information especially about long term risks and benefits on which they might base their decision.


Spatial Variation of the Native Colon Microbiota in Healthy Adults.
The microbiome has been implicated in the development of colorectal cancer and inflammatory bowel diseases. The specific traits of these diseases vary along the axis of the digestive tract. Further, variation in the structure of the gut microbiota has been associated with both diseases. We profiled the microbiota of the healthy proximal and distal mucosa and lumen to better understand how bacterial populations vary along the colon. We used a two-colonoscope approach to sample proximal and distal mucosal and luminal contents from the colons of 20 healthy subjects that had not undergone any bowel preparation procedure. The biopsies and home-collected stool were subjected to 16S rRNA gene sequencing, and random forest classification models were built using taxa abundance and location to identify microbiota specific to each site. The right mucosa and lumen had the most similar community structures of the five sites we considered from each subject. The distal mucosa had higher relative abundance of Finegoldia, Murdochiella, Peptoniphilus, Porphyromonas, and Anaerococcus The proximal mucosa had more of the genera Enterobacteriaceae, Bacteroides, and Pseudomonas The classification model performed well when classifying mucosal samples into proximal or distal sides (AUC = 0.808). Separating proximal and distal luminal samples proved more challenging (AUC = 0.599), and specific microbiota that differentiated the two were hard to identify. By sampling the unprepped colon, we identified distinct bacterial populations native to the proximal and distal sides. Further investigation of these bacteria may elucidate if and how these groups contribute to different disease processes on their respective sides of the colon. Cancer Prev Res; 11(7); 393-402. ©2018 AACR.


Direct evidence for Holliday junction crossover isomerization.
The Holliday junction is a key intermediate in genetic recombination. This is a four-stranded branched DNA structure, whose double-helical arms are stacked in two domains; two of the strands are roughly helical, and the other two cross over between domains. Switching the strands between these two roles is known as crossover isomerization; this postulated reversal is thought to be one of the key transformations that the Holliday junction can undergo, because it can lead to changing the products from patch to splice recombinants. We present direct evidence that this reaction can indeed occur in Holliday junctions in solution. We have constructed a double-crossover molecule containing a branched junction, constrained not to be in its favored conformation. This junction is released from the double-crossover molecule by digestion with restriction endonucleases. We demonstrate by means of hydroxyl radical autofootprinting that the junction changes its crossover isomer spontaneously when released from the double crossover. We control for the possibility that the experimental protocol causes the isomerization. We also exclude dissociation and interaction with other molecules in solution as contributing to the phenomenon. Thus, crossover isomerization is an authentic spontaneous transformation of Holliday junctions.


Transcriptional activation of polycomb-repressed genes by ZRF1.
Covalent modification of histones is fundamental in orchestrating chromatin dynamics and transcription. One example of such an epigenetic mark is the mono-ubiquitination of histones, which mainly occurs at histone H2A and H2B. Ubiquitination of histone H2A has been implicated in polycomb-mediated transcriptional silencing. However, the precise role of the ubiquitin mark during silencing is still elusive. Here we show in human cell lines that ZRF1 (zuotin-related factor 1) is specifically recruited to histone H2A when it is ubiquitinated at Lys 119 by means of a novel ubiquitin-interacting domain that is located in the evolutionarily conserved zuotin domain. At the onset of differentiation, ZRF1 specifically displaces polycomb-repressive complex 1 (PRC1) from chromatin and facilitates transcriptional activation. A genome-wide mapping of ZRF1, RING1B and H2A-ubiquitin targets revealed its involvement in the regulation of a large set of polycomb target genes, emphasizing the key role ZRF1 has in cell fate decisions. We provide here a model of the molecular mechanism of switching polycomb-repressed genes to an active state.


Metapopulation dynamics on ephemeral patches.
A challenge for conservation management is to understand how population and habitat dynamics interact to affect species persistence. In real landscapes, timing and duration of disturbances can vary, and species' responses to habitat changes will depend on how timing of dispersal and reproduction events relate to the landscape temporal structure. For instance, increasing disturbance frequency may promote extinction of species that are unable to appropriately time their reproduction in an ever-changing habitat and favor species that are able to track habitat changes. We developed a mathematical model to compare the effects of pulsed dispersal, initiated by shifts in habitat quality, with temporally continuous dispersal. We tested the effects of habitat (and population) turnover rates on metapopulation establishment, persistence, and long-term patch occupancy. Pulsed dispersal reduced patch occupancy and metapopulation longevity when habitat patches are relatively permanent. In such cases, demographic extinction was the primary form of local extinction. Conversely, when habitat patches are short-lived and new ones are frequently formed, pulsed dispersal promoted rapid colonization, increased occupancy, and prolonged metapopulation persistence. Our results show that species responsiveness to habitat disturbance is critical to metapopulation persistence, having profound implications for the species likely to persist in landscapes with altered disturbance regimes.


Functionalized single-walled carbon nanotubes/polypyrrole composites for amperometric glucose biosensors.
This article reports an amperometric glucose biosensor based on a new type of nanocomposite of polypyrrole (PPY) with p-phenyl sulfonate-functionalized single-walled carbon nanotubes (SWCNTs-PhSO3-). An environmentally friendly functionalization procedure of the SWCNTs in the presence of substituted aniline and an oxidative species was adopted. The nanocomposite-modified electrode exhibited excellent electrocatalytic activities towards the reduction or oxidation of H2O2. This feature allowed us to use it as bioplatform on which glucose oxidase (GOx) was immobilized by entrapment in an electropolymerized PPY/SWCNTs-PhSO3- film for the construction of the glucose biosensor. The amperometric detection of glucose was assayed by applying a constant electrode potential value necessary to oxidize or reduce the enzymatically produced H2O2 with minimal interference from the possible coexisting electroactive compounds. With the introduction of a thin film of Prussian blue (PB) at the substrate electrode surface, the PPY/GOx/SWCNTs-PhSO3-/PB system shows synergy between the PB and functionalized SWCNTs which amplifies greatly the electrode sensitivity when operated at low potentials. The biosensor showed good analytical performances in terms of low detection (0.01 mM), high sensitivity (approximately 6 μA mM-1 cm-2), and wide linear range (0.02 to 6 mM). In addition, the effects of applied potential, the electroactive interference, and the stability of the biosensor were discussed. The facile procedure of immobilizing GOx used in the present work can promote the development of other oxidase-based biosensors which could have a practical application in clinical, food, and environmental analysis.


The development and trial of a medication discontinuation program in the department of forensic psychiatry.
When treating mentally ill criminal offenders, improving medication adherence is essential to achieving goals, such as long-term stabilization of symptoms and the prevention of recidivism. Most subjects who are treated under the Medical Treatment and Supervision Act have schizophrenia, which is considered a particularly difficult disorder for which to improve medication adherence. For such patients, we developed a Medication Discontinuation Program (MDP) that aims to improve medication adherence by discontinuing antipsychotic drugs and monitoring changes in psychiatric symptoms. We examined whether there was any utility for the MDP on a trial basis as well as whether it would be worthwhile to introduce the MDP to psychiatric programs. We conducted the MDP with an intervention group (n = 7) and compared Drug Attitude Inventory-30 (DAI-30) scores before and after implementation of the MDP. We also categorized 30 questions of the DAI-30 into three subscales: "awareness of the need for medication", "awareness of the effects of psychiatric drugs", and "impression of medication", and examined factors affecting improvement in medication adherence. The total DAI-30 score significantly increased after completion of the MDP (P = 0.002). Significant elevations after completion of the MDP were also observed in the scores for three subscales of the DAI-30. Our study suggests that the MDP has a possibility of improving medication adherence, and this program might have multidirectional and stimulatory effects on each factor related to the improvement of medication adherence.


Age-related risk of major adverse cardiac event risk and coronary artery disease extent and severity by coronary CT angiography: results from 15 187 patients from the International Multisite CONFIRM Study.
Prior studies evaluating the prognostic utility of cardiac CT angiography (CCTA) have been largely constrained to an all-cause mortality endpoint, with other cardiac endpoints generally not reported. To this end, we sought to determine the relationship of extent and severity of coronary artery disease (CAD) by CCTA to risk of incident major adverse cardiac events (MACEs) (defined as death, myocardial infarction, and late revascularization). We identified subjects without prior known CAD who underwent CCTA and were followed for MACE. CAD by CCTA was defined as none (0% luminal stenosis), mild (1-49% luminal stenosis), moderate (50-69% luminal stenosis), or severe (≥70% luminal stenosis), and ≥50% luminal stenosis was considered as obstructive. CAD severity was judged on per-patient, per-vessel, and per-segment basis. Time to MACE was estimated using univariable and multivariable Cox proportional hazards models. Among 15 187 patients (57 ± 12 years, 55% male), 595 MACE events (3.9%) occurred at a 2.4 ± 1.2 year follow-up. In multivariable analyses, an increased risk of MACE was observed for both non-obstructive [hazard ratio (HR) 2.43, P < 0.001] and obstructive CAD (HR: 11.21, P < 0.001) when compared with patients with normal CCTA. Risk-adjusted MACE increased in a dose-response relationship based on the number of vessels with obstructive CAD ≥50%, with increasing hazards observed for non-obstructive (HR: 2.54, P < 0.001), obstructive one-vessel (HR: 9.15, P < 0.001), two-vessel (HR: 15.00, P < 0.001), or three-vessel or left main (HR: 24.53, P < 0.001) CAD. Among patients stratified by age <65 vs. ≥65 years, older individuals experienced higher risk-adjusted hazards for MACE for non-obstructive, one-, and two-vessel, with similar event rates for three-vessel or left main (P < 0.001 for all) compared with normal individuals age <65. Finally, there was a dose relationship of CAD findings by CCTA and MACE event rates with each advancing decade of life. Among individuals without known CAD, non-obstructive, and obstructive CAD are associated with higher MACE rates, with different risk profiles based on age.


The association of dietary animal and plant protein with putative risk markers of colorectal cancer in overweight pre-diabetic individuals during a weight-reducing programme: a PREVIEW sub-study.
Diets with increased protein content are popular strategies for body weight regulation, but the effect of such diets for the colonic luminal environment is unclear. We aimed to investigate the associations between putative colorectal cancer-related markers and total protein intake, plant and animal proteins, and protein from red and processed meat in pre-diabetic adults (> 25 years). Analyses were based on clinical and dietary assessments at baseline and after 1 year of intervention. Protein intake was assessed from 4-day dietary records. Putative colorectal cancer-related markers identified from 24-h faecal samples collected over three consecutive days were: concentration of short-chain fatty acids, phenols, ammonia, and pH. In total, 79 participants were included in the analyses. We found a positive association between change in total protein intake (slope: 74.72 ± 28.84 µmol per g faeces/E%, p = 0.01), including animal protein intake (slope: 87.63 ± 32.04 µmol per g faeces/E%, p = 0.009), and change in faecal ammonia concentration. For change in ammonia, there was a dose-response trend from the most negative (lowest tertile) to the most positive (highest tertile) association (p = 0.01): in the high tertile, a change in intake of red meat was positively associated with an increase in ammonia excretion (slope: 2.0 ± 0.5 µmol per g faeces/g/day, p < 0.001), whereas no such association was found in the low and medium tertile groups. Increases in total and animal protein intakes were associated with higher excretion of ammonia in faeces after 1 year in overweight pre-diabetic adults undertaking a weight-loss intervention. An increase in total or relative protein intake, or in the ratio of animal to plant protein, was not associated with an increase in faeces of any of the other putative colorectal cancer risk markers. ClinicalTrials.gov Identifier: NCT01777893.


Insights into the metabolism, lifestyle and putative evolutionary history of the novel archaeal phylum 'Diapherotrites'.
The archaeal phylum 'Diapherotrites' was recently proposed based on phylogenomic analysis of genomes recovered from an underground water seep in an abandoned gold mine (Homestake mine in Lead, SD, USA). Here we present a detailed analysis of the metabolic capabilities and genomic features of three single amplified genomes (SAGs) belonging to the 'Diapherotrites'. The most complete of the SAGs, Candidatus 'Iainarchaeum andersonii' (Cand. IA), had a small genome (∼1.24 Mb), short average gene length (822 bp), one ribosomal RNA operon, high coding density (∼90.4%), high percentage of overlapping genes (27.6%) and low incidence of gene duplication (2.16%). Cand. IA genome possesses limited catabolic capacities that, nevertheless, could theoretically support a free-living lifestyle by channeling a narrow range of substrates such as ribose, polyhydroxybutyrate and several amino acids to acetyl-coenzyme A. On the other hand, Cand. IA possesses relatively well-developed anabolic capabilities, although it remains auxotrophic for several amino acids and cofactors. Phylogenetic analysis suggests that the majority of Cand. IA anabolic genes were acquired from bacterial donors via horizontal gene transfer. We thus propose that members of the 'Diapherotrites' have evolved from an obligate symbiotic ancestor by acquiring anabolic genes from bacteria that enabled independent biosynthesis of biological molecules previously acquired from symbiotic hosts. 'Diapherotrites' 16S rRNA genes exhibit multiple mismatches with the majority of archaeal 16S rRNA primers, a fact that could be responsible for their observed rarity in amplicon-generated data sets. The limited substrate range, complex growth requirements and slow growth rate predicted could be responsible for its refraction to isolation.


The reverse transport of DA, what physiological significance?
It is well established that midbrain dopamine neurons innervating the striatum, release their neurotransmitter through an exocytotic process triggered by the neural firing and involving a transient calcium entry in the terminals. Long ago, it had been proposed, however, that another mechanism of release could co-exist with classical exocytosis, involving the reverse-transport of the cytosolic amine by the carrier, ordinarily responsible for uptake function. This atypical mode of release could be evoked directly at the preterminal level by multiple environmental endogenous factors involving transient alterations of the sodium gradient. It cannot be excluded that this mode of release participates in the firing-induced release. In contrast with the classical exocytosis of a preformed DA pool, the reverse-transport of DA requires simultaneous alterations of intraterminal amine metabolism including synthesis and displacement from storage compartment. The concept of a reverse-transport of dopamine is coming from the observations that releasing substances, such as amphetamine-related molecules, actually induce this type of transport. A large set of arguments advocates that reverse-transport plays a role in the maintenance of basal extracellular DA concentration in striatum. It was also often evoked in physiopathological situations including ischemia, neurodegenerative processes, etc. The most recent studies suggest that this release could occur mainly outside the synapses, and thus could constitute a major feature in the paracrine transmission, sometimes evoked for DA.


Implicit encoding of prior probabilities in optimal neural populations.
Optimal coding provides a guiding principle for understanding the representation of sensory variables in neural populations. Here we consider the influence of a prior probability distribution over sensory variables on the optimal allocation of neurons and spikes in a population. We model the spikes of each cell as samples from an independent Poisson process with rate governed by an associated tuning curve. For this response model, we approximate the Fisher information in terms of the density and amplitude of the tuning curves, under the assumption that tuning width varies inversely with cell density. We consider a family of objective functions based on the expected value, over the sensory prior, of a functional of the Fisher information. This family includes lower bounds on mutual information and perceptual discriminability as special cases. In all cases, we find a closed form expression for the optimum, in which the density and gain of the cells in the population are power law functions of the stimulus prior. This also implies a power law relationship between the prior and perceptual discriminability. We show preliminary evidence that the theory successfully predicts the relationship between empirically measured stimulus priors, physiologically measured neural response properties (cell density, tuning widths, and firing rates), and psychophysically measured discrimination thresholds.


Potentiation of curing by a broad-host-range self-transmissible vector for displacing resistance plasmids to tackle AMR.
Plasmids are potent vehicles for spread of antibiotic resistance genes in bacterial populations and often persist in the absence of selection due to efficient maintenance mechanisms. We previously constructed non-conjugative high copy number plasmid vectors that efficiently displace stable plasmids from enteric bacteria in a laboratory context by blocking their replication and neutralising their addiction systems. Here we assess a low copy number broad-host-range self-transmissible IncP-1 plasmid as a vector for such curing cassettes to displace IncF and IncK plasmids. The wild type plasmid carrying the curing cassette displaces target plasmids poorly but derivatives with deletions near the IncP-1 replication origin that elevate copy number about two-fold are efficient. Verification of this in mini IncP-1 plasmids showed that elevated copy number was not sufficient and that the parB gene, korB, that is central to its partitioning and gene control system, also needs to be included. The resulting vector can displace target plasmids from a laboratory population without selection and demonstrated activity in a mouse model although spread is less efficient and requires additional selection pressure.


Optical transmission through compound gold surface relief slit arrays.
The optical transmissions through compound gold surface relief slit arrays were investigated theoretically by using the finite difference time domain method. The differences of transmission, reflection, and absorption spectra of the bare slit and the surface relief grating are discussed. The transmission spectra influenced by different dielectric constants of medium in the two slits and different slits widths. When the two slits fill different dielectrics, the presence of the medium induces a red-shift of the plasmon resonances. Along with the dielectric constant in one slit increasing, there appear obvious dips in the transmission spectra. Based on the magnetic and electric field distributions, Fabry-Pérot-like resonance and phase resonance mechanisms have been suggested for the physical origins of these observations.


Potential of multispectral imaging technology for rapid and non-destructive determination of the microbiological quality of beef filets during aerobic storage.
The performance of a multispectral imaging system has been evaluated in monitoring aerobically packaged beef filet spoilage at different storage temperatures (0, 4, 8, 12, and 16°C). Spectral data in the visible and short wave near infrared area (405-970nm) were collected from the surface of meat samples and correlated with microbiological data (log counts), for total viable counts (TVCs), Pseudomonas spp., and Brochothrix thermosphacta. Qualitative analysis (PLS-DA) was employed for the discrimination of meat samples in three microbiological quality classes based on the values of total viable counts, namely Class 1 (TVC<5.5log10CFU/g), Class 2 (5.5log10CFU/g<TVC<7.0log10CFU/g), and Class 3 (TVC>7.0log10CFU/g). Furthermore, PLS regression models were developed to provide quantitative estimations of microbial counts during meat storage. In both cases model validation was implemented with independent experiments at intermediate storage temperatures (2 and 10°C) using different batches of meat. Results demonstrated good performance in classifying meat samples with overall correct classification rate for the three quality classes ranging from 91.8% to 80.0% for model calibration and validation, respectively. For quantitative estimation, the calculated regression coefficients between observed and estimated counts ranged within 0.90-0.93 and 0.78-0.86 for model development and validation, respectively, depending on the microorganism. Moreover, the calculated average deviation between observations and estimations was 11.6%, 13.6%, and 16.7% for Pseudomonas spp., B. thermosphacta, and TVC, respectively. The results indicated that multispectral vision technology has significant potential as a rapid and non-destructive technique in assessing the microbiological quality of beef fillets.


Longitudinal natural history in young boys with Duchenne muscular dystrophy.
The aim of this prospective multicentric study was to document disease progression in young boys affected by Duchenne muscular dystrophy (DMD) between age 3 and 6 years (±3 months) using the North Star Ambulatory Assessment scale. One hundred fifty-three DMD boys (573 assessments) younger than 6 years (mean: 4.68, SD: 0.84) with a genetically proven DMD diagnoses were included. Our results showed North Star Ambulatory Assessment scores progressively increased with age. The largest increase was observed between age 3 and 4 years but further increase was steadily observed until age of 6 years. Using a multiple linear regression analysis, we found that both the use of corticosteroids and the site of mutation significantly contributed to the North Star Ambulatory Assessment changes (p < 0.001). At each age point, boys on corticosteroid treatment had higher scores than corticosteroid naïve ones (p < 0.001). Similarly, patients with mutations downstream exon 44, had lower baseline scores and lower magnitude of changes compared to those with mutations located at the 5' end of the gene (p < 0,001). Very few boys achieved the age appropriate maximum score. These results provide useful information for the assessment and counselling of young DMD boys and for the design of clinical trials in this age group.


Interobserver and intraobserver variability in measuring the tortuosity of the thoracic aorta on computed tomography.
The variability in measuring the tortuosity of the thoracic aorta has not been previously studied. This study evaluated the interobserver and intraobserver variability of major methods used for measuring the tortuosity of the thoracic aorta in patients with aortic arch or descending thoracic aortic aneurysm. This retrospective study enrolled 66 patients with aortic arch or descending thoracic aortic aneurysm who had undergone thoracic endovascular aortic repair. Two radiologists used preoperative computed tomography images to measure the tortuosity of the thoracic aorta at multiple segments by using the fitting circle diameter, tortuosity index, and centerline angle methods; these measurements were repeated after an interval of >28 days. The variability of the methods was analyzed for interobserver and intraobserver reliability and agreement. The estimated intraclass correlation coefficient (ICC) was used to analyze the reliability. The Bland-Altman plot was used to analyze the interobserver and intraobserver agreement. The association between aortic characteristics, including calcification, luminal irregularity, shape, and diameter, and the variability of the measurements was also analyzed. The interobserver ICC estimates for the tortuosity index at multiple aortic segments, centerline angle methods at the supra-aortic branch orifices, and fitting circle diameter on the greater and lesser curvature sides were 0.97 to 0.98, 0.39 to 0.75, and 0.82 to 0.84, respectively. The corresponding intraobserver ICC estimates were 0.98 to 1.00, 0.44 to 0.75, and 0.82 to 0.85, respectively. In the agreement analysis, the 95% limits of agreement for the tortuosity index, centerline angle, and fitting circle diameter were -5.5% to 5.6%, -10.9% to 10.9%, and -18.0% to 24.0%, respectively. The tortuosity index had the highest ICC estimate and narrowest 99.5% limits of agreement of the three methods. Aortic characteristics, including calcification, grade of atheroma, aneurysm shape, and diameter, were not associated with the variability of the tortuosity index method in the thoracic aorta. The tortuosity index method has low interobserver and intraobserver variability in measuring the tortuosity of the thoracic aorta in patients with thoracic aortic aneurysm. The characteristics of the aorta and aneurysm are not associated with the interobserver or intraobserver variability of the tortuosity index.


Phosphorylation of FMRP and alterations of FMRP complex underlie enhanced mLTD in adult rats triggered by early life seizures.
Outside of Fragile X syndrome (FXS), the role of Fragile-X Mental Retardation Protein (FMRP) in mediating neuropsychological abnormalities is not clear. FMRP, p70-S6 kinase (S6K) and protein phosphatase 2A (PP2A) are thought to cooperate as a dynamic signaling complex. In our prior work, adult rats have enhanced CA1 hippocampal long-term depression (LTD) following an early life seizure (ELS). We now show that mGluR-mediated LTD (mLTD) is specifically enhanced following ELS, similar to FMRP knock-outs. Total FMRP expression is unchanged but S6K is hyperphosphorylated, consistent with S6K overactivation. We postulated that either disruption of the FMRP-S6K-PP2A complex and/or removal of this complex from synapses could explain our findings. Using subcellular fractionation, we were surprised to find that concentrations of FMRP and PP2A were undisturbed in the synaptosomal compartment but reduced in parallel in the cytosolic compartment. Following ELS FMRP phosphorylation was reduced in the cytosolic compartment and increased in the synaptic compartment, in parallel with the compartmentalization of S6K activation. Furthermore, FMRP and PP2A remain bound following ELS. In contrast, the interaction of S6K with FMRP is reduced by ELS. Blockade of PP2A results in enhanced mLTD; this is occluded by ELS. This suggests a critical role for the location and function of the FMRP-S6K-PP2A signaling complex in limiting the amount of mLTD. Specifically, non-synaptic targeting and the function of the complex may influence the "set-point" for regulating mLTD. Consistent with this, striatal-enriched protein tyrosine phosphatase (STEP), an FMRP "target" which regulates mLTD expression, is specifically increased in the synaptosomal compartment following ELS. Further, we provide behavioral data to suggest that FMRP complex dysfunction may underlie altered socialization, a symptom associated and observed in other rodent models of autism, including FXS.


No difference in markers of adipose tissue inflammation between overweight women with polycystic ovary syndrome and weight-matched controls.
BACKGROUND; Previous studies have indicated that peripheral circulating markers of inflammation are elevated in women with polycystic ovary syndrome (PCOS), but thus far no studies concerning markers of inflammation in adipose tissue have been published. The aim of the study was to investigate whether patients with PCOS display increased expression of inflammatory markers in adipose tissue. Twenty overweight patients with PCOS, 10 lean patients with PCOS and 20 overweight controls had subcutaneous fat biopsies and blood samples taken. Adipose tissue levels of mRNA of inflammatory markers were determined by use of real-time PCR. Overweight patients with PCOS had higher relative adipose tissue chemokine ligand 2 (P < 0.01), and its cognate receptor (P < 0.05), tumour necrosis factor-α (P < 0.001), interleukin (IL)-10 (P < 0.001) and IL-18 (P < 0.001) and the monocyte/macrophage markers CD14 (P < 0.01) and CD163 (P < 0.01) mRNA levels compared with lean women with PCOS. There were no differences between overweight patients with PCOS and overweight control subjects in this respect. Within the PCOS group, markers of adipose tissue inflammation correlated significantly with obesity-related metabolic disturbances, but when data were adjusted for age and BMI, most correlations were lost. Overweight, rather than the PCOS diagnosis per se, appears to be the main explanatory variable for elevated adipose tissue inflammation in patients with PCOS.


Delineating the requirements for spontaneous DNA damage resistance pathways in genome maintenance and viability in Saccharomyces cerevisiae.
Cellular metabolic processes constantly generate reactive species that damage DNA. To counteract this relentless assault, cells have developed multiple pathways to resist damage. The base excision repair (BER) and nucleotide excision repair (NER) pathways remove damage whereas the recombination (REC) and postreplication repair (PRR) pathways bypass the damage, allowing deferred removal. Genetic studies in yeast indicate that these pathways can process a common spontaneous lesion(s), with mutational inactivation of any pathway increasing the burden on the remaining pathways. In this study, we examine the consequences of simultaneously compromising three or more of these pathways. Although the presence of a functional BER pathway alone is able to support haploid growth, retention of the NER, REC, or PRR pathway alone is not, indicating that BER is the key damage resistance pathway in yeast and may be responsible for the removal of the majority of either spontaneous DNA damage or specifically those lesions that are potentially lethal. In the diploid state, functional BER, NER, or REC alone can support growth, while PRR alone is insufficient for growth. In diploids, the presence of PRR alone may confer a lethal mutation load or, alternatively, PRR alone may be insufficient to deal with potentially lethal, replication-blocking lesions.


Urban settlements' vulnerability to flood risks in African cities: A conceptual framework.
In the recent past, the frequency and gravity of large-scale flood disasters have increased globally, resulting in casualties, destruction of property and huge economic loss. The destructive flood disaster devastating Louisiana, USA, is a recent example. Despite the availability of advanced technological capabilities for dealing with floods in developed nations, flood disasters continue to become more rampant and disastrous. Developing countries in Africa such as Benin, Ghana, Nigeria, Senegal and Sudan have recently experienced severe flooding, leaving a considerable number of human casualties and thousands displaced. In African cities, most vulnerable urban residents usually have lesser capacity and fewer resources to recover from the shocks of disaster as a result of the failure of governments to build human security for poor African residents. Many scholars have acknowledged the lack of appropriate vulnerability assessment frameworks and policies, questioning the efficiency and effectiveness of the tested models in Africa. The ability to accurately identify, measure and evaluate the various vulnerabilities of affected people and communities is a right step towards reducing disaster risk. This article aimed at developing a framework for assessing urban settlements' vulnerability to flood risks in Africa. The framework is currently being tested to assess various dimensions of vulnerability drivers in three urban communities in Ibadan metropolis, the third largest city in Nigeria, focusing more on flood risk perceptions and behaviour of the risk bearers. It uses participatory and mixed method approaches to socially construct vulnerability of populations at risk. This model emanates from the evaluation of considerable relevant literature and an array of vulnerability assessment frameworks. It integrates some approaches that are applicable to African cities in a bid to create a versatile tool to assess, identify and mitigate the effects of flood disaster risk and reduce urban poor's vulnerability to natural and human-induced hazards.


Community analysis of the abundance and diversity of biting midge species (Diptera: Ceratopogonidae) in three European countries at different latitudes.
The outbreaks of bluetongue and Schmallenberg disease in Europe have increased efforts to understand the ecology of Culicoides biting midges and their role in pathogen transmission. However, most studies have focused on a specific habitat, region, or country. To facilitate wider comparisons, and to obtain a better understanding of the spread of disease through Europe, the present study focused on monitoring biting midge species diversity in three different habitat types and three countries across Europe. Biting midges were trapped using Onderstepoort Veterinary Institute light traps at a total of 27 locations in Sweden, the Netherlands and Italy, comprising farm, peri-urban and wetland habitats. From July 2014 to June 2015 all locations were sampled monthly, except for during the winter months. Trapped midges were counted and identified morphologically. Indices on species richness, evenness and diversity were calculated. Community compositions were analysed using non-metric multidimensional scaling (NMDS) techniques. A total of 50,085 female midges were trapped during 442 collection nights. More than 88% of these belonged to the Obsoletus group. The highest midge diversity was found in Sweden, while species richness was highest in the Netherlands, and most specimens were trapped in Italy. For habitats within countries, diversity of the trapped midges was lowest for farms in all countries. Differences in biting midge species communities were more distinct across the three countries than the three habitat types. A core midge community could be identified, in which the Obsoletus group was the most abundant. Variations in vector communities across countries imply different patterns of disease spread throughout Europe. How specific species and their associated communities affect disease risk is still unclear. Our results emphasize the importance of midge diversity data at community level, how this differs across large geographic range within Europe, and its implications on assessing risks of midge-borne disease outbreaks.


Central toxic keratopathy: a clinical case series.
To describe clinically, a case series of central toxic keratopathy (CTK), and contribute with one possible aetiology of this entity. A retrospective observational case series, which included 12 eyes diagnosed with CTK after undergoing laser refractive surgery in situ keratomileusis (LASIK). From a total of 28 eyes that underwent LASIK in the same week, 12 develop CTK. The mean time of the diagnostic was 4 days after surgery. The resolution time had an interval between 1, 3 and 9 months, and an average of 4.6 months. The occurrence of CTK cases coincided with the change of the brand of surgical gloves, and no further cases were found after their replacement. The CTK clinical findings are well described, but with a yet unknown origin. It seems clear that it does not require specific treatment. It is possible that there might be a relationship between a substance of the gloves and CTK. Further studies are needed to know all the details of this process.


The role of the Primary Healthcare Network in Iran in hepatitis C virus elimination by 2030.
The World Health Organization (WHO) has estimated that about 71 million people in the world have chronic hepatitis C virus (HCV) and 399,000 die every year due to cirrhosis and cancer caused by HCV. Despite efforts to prevent and control hepatitis, there are still many barriers. In September 2000, during a summit in New York organised by the United Nations, world leaders agreed on a set of achievable targets within a limited timeframe, with the goal of combating poverty, hunger, disease, illiteracy, destruction of the environment and of improving the status of women by 2015; these were known as the Millennium Development Goals. The Primary Health Care Network (PHCN) in Iran has played a very important role in health promotion but its achievements could be enhanced further by improving performance and implementing further changes. The WHO strives to achieve goals of sustainable development by 2030 in many countries, thereby raising awareness, encouraging greater community participation, mobilising resources, using evidence-based policies, and increasing screening services, care and treatment. The concept of the PHCN has been very successful in health promotion in Iran, and has provided a basis for accessible and high-quality services for all. Achieving HCV elimination by 2030 could be achieved with the support of the PHCN, and changes in management with regard to screening, prevention and treatment practices.


Statistical methods for designing and assessing the effectiveness of community-based interventions with small numbers.
Community-based interventions are, by their nature, tailored to the particular needs of the members and stakeholders of the community. In the area of safety promotion, the multifaceted aspects of violence and unintentional injuries necessitate complex intertwining of strategies and approaches. The evaluation of the effectiveness of any 'intervention' programme, either as a whole or of its discrete components, is thus challenging. In addition, standard experimental research designs involving controlled comparisons and the use of randomisation are often not practical. The evaluation is complicated further when the numbers of events in the community that constitute outcomes to be measured, before and after the interventions, are small. This manuscript reviews some of the statistical considerations that impact the evaluation of the effectiveness of such intervention approaches, and proposes a meta-regression methodology to address this complex issue. The application of the proposed methodology to the 'Safe Community' Model is provided as an example.


Antibiotics in the environment.
Molecules with antibiotic properties, produced by various microbes, have been around long before mankind recognized their usefulness in preventing and treating bacterial infections. Bacteria have therefore been exposed to selection pressures from antibiotics for very long times, however, generally only on a micro-scale within the immediate vicinity of the antibiotic-producing organisms. In the twentieth century we began mass-producing antibiotics, mainly synthetic derivatives of naturally produced antibiotic molecules, but also a few entirely synthetic compounds. As a consequence, entire bacterial communities became exposed to unprecedented antibiotic selection pressures, which in turn led to the rapid resistance development we are facing today among many pathogens. We are, rightly, concerned about the direct selection pressures of antibiotics on the microbial communities that reside in or on our bodies. However, other environments, outside of our bodies, may also be exposed to antibiotics through different routes, most often unintentionally. There are concerns that increased selection pressures from antibiotics in the environment can contribute to the recruitment of resistance factors from the environmental resistome to human pathogens. This paper attempts to 1) provide a brief overview of environmental exposure routes of antibiotics, 2) provide some thoughts about our current knowledge of the associated risks for humans as well as ecosystems, and 3) indicate management options to reduce risks.


Structure of Flavobacterium psychrophilum populations infecting farmed rainbow trout Oncorhynchus mykiss .
Flavobacterium psychrophilum isolated from rainbow trout Oncorhynchus mykiss suffering from bacterial cold-water disease (BCWD) can dissociate into 2 morphological colony types, rough (R) and smooth (S). However, the presence of the 2 morphotypes in disease outbreaks has not yet been investigated thoroughly. We examined the occurrence of R and S morphotypes in rainbow trout from BCWD outbreaks and in unfertilized eggs from a hatchery. The isolated colony types were characterized by pulsed field gel electrophoresis (PFGE), plasmid analysis, and oxolinic acid susceptibility testing. From most outbreaks, both morphotypes were isolated, although the S type only was isolated from the majority of individual fish. PFGE analysis showed both diverse and indistinguishable genetic patterns among the concurrent morphotypes. While PFGE patterns common to both fish and egg isolates were identified, this was not always the case. Resistant and sensitive isolates of both colony types were isolated from individual disease outbreaks. The plasmid pattern was partly associated with the colony type, showing identical or completely different patterns for the R and S types isolated from the same outbreak. The study showed that within a BCWD outbreak, F. psychrophilum cells with different morphology, plasmid content, antibiotic susceptibility, and PFGE pattern can be isolated, suggesting that F. psychrophilum populations infecting rainbow trout in farm environments can be diverse and thus complicate the control of the disease.


Mutual inhibition and capacity sharing during parallel preparation of serial eye movements.
Many common activities, like reading, scanning scenes, or searching for an inconspicuous item in a cluttered environment, entail serial movements of the eyes that shift the gaze from one object to another. Previous studies have shown that the primate brain is capable of programming sequential saccadic eye movements in parallel. Given that the onset of saccades directed to a target are unpredictable in individual trials, what prevents a saccade during parallel programming from being executed in the direction of the second target before execution of another saccade in the direction of the first target remains unclear. Using a computational model, here we demonstrate that sequential saccades inhibit each other and share the brain's limited processing resources (capacity) so that the planning of a saccade in the direction of the first target always finishes first. In this framework, the latency of a saccade increases linearly with the fraction of capacity allocated to the other saccade in the sequence, and exponentially with the duration of capacity sharing. Our study establishes a link between the dual-task paradigm and the ramp-to-threshold model of response time to identify a physiologically viable mechanism that preserves the serial order of saccades without compromising the speed of performance.


Anthocyanins inhibit high glucose-induced renal tubular cell apoptosis caused by oxidative stress in db/db mice.
Oxidative stress is an important contributory factor resulting the development of kidney injury in patients with diabetes. Numerous in vitro and in vivo studies have suggested that anthocyanins, natural phenols commonly existing in numerous fruits and vegetables, exhibit important antioxidative, anti‑inflammatory and antihyperlipidemic effects; however, their effects and underlying mechanisms on diabetic nephropathy (DN) have not yet been fully determined. In the present study, the regulation of apoptosis metabolism and antioxidative effects exhibited by anthocyanins [grape seed procyanidin (GSPE) and cyanidin‑3‑O‑β‑glucoside chloride (C3G)] were investigated, and the molecular mechanism underlying this process was investigated in vivo and in vitro. GSPE administration was revealed to suppress renal cell apoptosis, as well as suppress the expression of Bcl‑2 in diabetic mouse kidneys. Furthermore, GSPE administration was demonstrated to suppress the expression of thioredoxin interacting protein (TXNIP), in addition to enhancing p38 mitogen‑activation protein kinase (MAPK) and extracellular signal‑regulated kinase 1/2 (ERK1/2) oxidase activity in diabetic mouse kidneys. In vitro experiments using HK‑2 cells revealed that C3G suppressed the generation of HG‑mediated reactive oxygen species, cellular apoptosis, the expression of cleaved caspase‑3 and the Bax/Bcl‑2 ratio; and enhanced the expression of cytochrome c released from mitochondria. Furthermore, treatment with C3G was revealed to suppress the expression of TXNIP, in addition to the phosphorylation of p38 MAPK and ERK1/2 oxidase activity in HK‑2 cells under HG conditions. In addition, treatment with C3G was revealed to attenuate the HG‑induced suppression of the biological activity of thioredoxin, and to enhance the expression of thioredoxin 2 in HK‑2 cells under HG conditions. In conclusion, the present study demonstrated that anthocyanins may exhibit protective effects against HG‑induced renal injury in DN via antioxidant activity.


Fruit carbohydrate metabolism in an introgression line of tomato with increased fruit soluble solids.
A tomato line (IL9-2-5) of the cultivated species, Lycopersicon esculentum, carrying a 9 cM introgression from the wild species, Lycopersicon pennelli, produces fruit with high soluble solids content (Brix), an important determinant of fruit quality for processing. Two quantitative trait loci (QTLs) relating to fruit soluble solids content have been identified within the introgressed segment. One of these QTLs (PW-9-2-5) is silent under the growth conditions used in this study, while a second (Brix-9-2-5) has been shown to encode a fruit apoplastic invertase (Lin5) with altered kinetic properties. In this study, we have undertaken a detailed biochemical analysis of the introgression line to attempt to gain an understanding of the metabolic changes associated with increased fruit soluble solids. Increased Brix in ripe fruit was shown to be the result of increased sucrose and glucose, with a more minor contribution from aspartate and alanine. The introgression leads to a pronounced increase in apoplastic invertase activity in the columella tissue that extends throughout fruit development. Furthermore, columella tissue from IL9-2-5 fruit has a greater capacity to take up exogenously supplied sucrose, an observation that is consistent with the kinetic properties of the introgressed Lin5 allele. Apart from the increase in mature fruit sugar and increases in some amino acids, metabolite profiling revealed few other metabolic perturbations in fruit from IL9-2-5. The only other major change was a dramatic increase in starch accumulation at earlier stages of fruit metabolism. This occurred without any increase in the activity of the enzymes of sucrose metabolism or starch synthesis and may therefore be driven by increased availability of sucrose. We conclude that the major factor that leads to increased fruit sugar in IL9-2-5 is an increase in the capacity to take up sucrose unloaded from the phloem.


Prevalence, symptoms and management of uterine fibroids: an international internet-based survey of 21,746 women.
In 2009 the Uterine Bleeding and Pain Women's Research Study (UBP-WRS) was conducted interviewing 21,479 women across 8 countries in order to gain patient-based prevalence data on uterine pain and bleeding indications and investigate uterine symptoms and women's treatment experiences. This article shows relevant results of the study for the indication uterine fibroids providing data on self-reported prevalence, symptomatology and management of uterine fibroids. 2,500 women (USA: 4,500 women) in each country (Brazil, Canada, France, Germany, Italy, South Korea, the UK, the USA) completed an online survey. Women included were in their reproductive age (age group 15-49 years; USA: 18-49 years) and had ever experienced menstrual bleedings. Quotas were applied for age, region, level of education and household income of respondents. Variables have been analyzed descriptively and exploratory statistical tests have been performed. The self-reported prevalence of uterine fibroids ranged from 4.5% (UK) to 9.8% (Italy), reaching 9.4% (UK) to 17.8% (Italy) in the age group of 40-49 years. Women with a diagnosis of uterine fibroids reported significantly more often about bleeding symptoms than women without a diagnosis: heavy bleedings (59.8% vs. 37.4%), prolonged bleedings (37.3% vs. 15.6%), bleeding between periods (33.3% vs. 13.5%), frequent periods (28.4% vs. 15.2%), irregular and predictable periods (36.3% vs. 23.9%). Furthermore women with diagnosed uterine fibroids reported significantly more often about the following pain symptoms: pressure on the bladder (32.6% vs. 15.0%), chronic pelvic pain (14.5% vs. 2.9%), painful sexual intercourse (23.5% vs. 9.1%) and pain occurring mid-cycle, after and during menstrual bleeding (31.3%, 16.7%, 59.7%, vs. 17.1%, 6.4%, 52.0%). 53.7% of women reported that their symptoms had a negative impact on their life in the last 12 month, influencing their sexual life (42.9%), performance at work (27.7%) and relationship & family (27.2%). Uterine fibroid is a common concern in women at fertile age causing multiple bleeding and pain symptoms which can have a negative impact on different aspects in women's life.


Fast single image super-resolution using estimated low-frequency k-space data in MRI.
Single image super-resolution (SR) is highly desired in many fields but obtaining it is often technically limited in practice. The purpose of this study was to propose a simple, rapid and robust single image SR method in magnetic resonance (MR) imaging (MRI). The idea is based on the mathematical formulation of the intrinsic link in k-space between a given (modulus) low-resolution (LR) image and the desired SR image. The method consists of two steps: 1) estimating the low-frequency k-space data of the desired SR image from a single LR image; 2) reconstructing the SR image using the estimated low-frequency and zero-filled high-frequency k-space data. The method was evaluated on digital phantom images, physical phantom MR images and real brain MR images, and compared with existing SR methods. The proposed SR method exhibited a good robustness by reaching a clearly higher PSNR (25.77dB) and SSIM (0.991) averaged over different noise levels in comparison with existing edge-guided nonlinear interpolation (EGNI) (PSNR=23.78dB, SSIM=0.983), zero-filling (ZF) (PSNR=24.09dB, SSIM=0.985) and total variation (TV) (PSNR=24.54dB, SSIM=0.987) methods while presenting the same order of computation time as the ZF method but being much faster than the EGNI or TV method. The average PSNR or SSIM over different slice images of the proposed method (PSNR=26.33 dB or SSIM=0.955) was also higher than the EGNI (PSNR=25.07dB or SSIM=0.952), ZF (PSNR=24.97dB or SSIM=0.950) and TV (PSNR=25.70dB or SSIM=0.953) methods, demonstrating its good robustness to variation in anatomical structure of the images. Meanwhile, the proposed method always produced less ringing artifacts than the ZF method, gave a clearer image than the EGNI method, and did not exhibit any blocking effect presented in the TV method. In addition, the proposed method yielded the highest spatial consistency in the inter-slice dimension among the four methods. This study proposed a fast, robust and efficient single image SR method with high spatial consistency in the inter-slice dimension for clinical MR images by estimating the low-frequency k-space data of the desired SR image from a single spatial modulus LR image.


Ginger: history and use.
Ginger is well known in the form of ginger sticks or ginger ale. If these are consumed during travel, the traveler imbibes, albeit subconsciously, a healing plant for motion sickness. The efficacy of ginger rhizome for the prevention of nausea, dizziness, and vomiting as symptoms of motion sickness (kinetosis), as well as for postoperative vomiting and vomiting of pregnancy, has been well documented and proved beyond doubt in numerous high-quality clinical studies. The use of this ancient medicine for gastrointestinal problems (stimulation of digestion) has been given scientific approval. Today, medicinal ginger is used mainly for prevention of the symptoms of travel sickness.


Checkpoint inhibitors for renal cell carcinoma: current landscape and future directions.
Immunotherapy with checkpoint inhibitors has arrived and begun to change the landscape of clinical oncology, including for patients with renal cell carcinoma. Specifically, drugs targeting the programmed death 1 and cytotoxic T-lymphocyte associated antigen pathways have demonstrated remarkable responses for patients in clinical trials. In this article, we review the most recent available data for immune checkpoint inhibitors for patients with renal cell carcinoma. We discuss potential strategies for rational combination therapies in these patients, some of which are currently being studied, and address important future considerations for use of these novel agents in the years to come.


An integrative analysis of non-coding regulatory DNA variations associated with autism spectrum disorder.
A number of genetic studies have identified rare protein-coding DNA variations associated with autism spectrum disorder (ASD), a neurodevelopmental disorder with significant genetic etiology and heterogeneity. In contrast, the contributions of functional, regulatory genetic variations that occur in the extensive non-protein-coding regions of the genome remain poorly understood. Here we developed a genome-wide analysis to identify the rare single nucleotide variants (SNVs) that occur in non-coding regions and determined the regulatory function and evolutionary conservation of these variants. Using publicly available datasets and computational predictions, we identified SNVs within putative regulatory regions in promoters, transcription factor binding sites, and microRNA genes and their target sites. Overall, we found that the regulatory variants in ASD cases were enriched in ASD-risk genes and genes involved in fetal neurodevelopment. As with previously reported coding mutations, we found an enrichment of the regulatory variants associated with dysregulation of neurodevelopmental and synaptic signaling pathways. Among these were several rare inherited SNVs found in the mature sequence of microRNAs predicted to affect the regulation of ASD-risk genes. We show a paternally inherited miR-873-5p variant with altered binding affinity for several risk-genes including NRXN2 and CNTNAP2 putatively overlay maternally inherited loss-of-function coding variations in NRXN1 and CNTNAP2 to likely increase the genetic liability in an idiopathic ASD case. Our analysis pipeline provides a new resource for identifying loss-of-function regulatory DNA variations that may contribute to the genetic etiology of complex disorders.


Surgical versus conservative treatment for high-risk stress fractures of the lower leg (anterior tibial cortex, navicular and fifth metatarsal base): a systematic review.
To compare surgical and conservative treatment for high-risk stress fractures of the anterior tibial cortex, navicular and proximal fifth metatarsal. Systematic searches of CENTRAL, MEDLINE, EMBASE, CINAHL, SPORTDiscus and PEDro were performed to identify relevant prospective and retrospective studies. Two reviewers independently extracted data and assessed methodological quality. Main outcomes were return to sport and complication rate. 18 studies were included (2 anterior tibia (N=31), 8 navicular (N=200) and 8 fifth metatarsal (N=246)). For anterior tibial fracture, no studies on initial surgery were eligible. Conservative treatment resulted in high complication rates and few cases returned to sport. For navicular fracture, a weighted mean return to sport of 22 for conservative and 16 weeks for surgical treatment was found. Six weeks of non-weightbearing cast was mostly used as conservative treatment. Surgical procedures varied widely. For the fifth metatarsal fracture, weighted mean return to sport was 19 for conservative and 14 weeks for surgical treatment. Surgery consisted of intramedullary screw fixation or tension band wiring. For conservative methods, insufficient details were reported. Overall, there was a high risk of bias; sample sizes were small and GRADE level of evidence was low. Strong conclusions for surgical or conservative therapy for these high-risk stress fractures cannot be drawn; quality of evidence is low and subjected to a high risk of bias. However, there are unsatisfying outcomes of conservative therapy in the anterior tibia. The role of initial surgery is unknown. For the navicular, surgery provided an earlier return to sport; and when treated conservatively, weightbearing should be avoided. For the fifth metatarsal, surgery provided the best results. Treatment decision-making would greatly benefit from further prospective research. PROSPERO database of systematic reviews: CRD42013004201.


The genome sequence of African rice (Oryza glaberrima) and evidence for independent domestication.
The cultivation of rice in Africa dates back more than 3,000 years. Interestingly, African rice is not of the same origin as Asian rice (Oryza sativa L.) but rather is an entirely different species (i.e., Oryza glaberrima Steud.). Here we present a high-quality assembly and annotation of the O. glaberrima genome and detailed analyses of its evolutionary history of domestication and selection. Population genomics analyses of 20 O. glaberrima and 94 Oryza barthii accessions support the hypothesis that O. glaberrima was domesticated in a single region along the Niger river as opposed to noncentric domestication events across Africa. We detected evidence for artificial selection at a genome-wide scale, as well as with a set of O. glaberrima genes orthologous to O. sativa genes that are known to be associated with domestication, thus indicating convergent yet independent selection of a common set of genes during two geographically and culturally distinct domestication processes.


Principles of postoperative anterior cruciate ligament rehabilitation.
It is known that anterior cruciate ligament (ACL) reconstruction needs to be combined with detailed postoperative rehabilitation in order for patients to return to their pre-injury activity levels, and that the rehabilitation process is as important as the reconstruction surgery. Literature studies focus on how early in the postoperative ACL rehabilitation period rehabilitation modalities can be initiated. Despite the sheer number of studies on this topic, postoperative ACL rehabilitation protocols have not been standardized yet. Could common, "ossified" knowledge or modalities really prove themselves in the literature? Could questions such as "is postoperative brace use really necessary?", "what are the benefits of early restoration of the range of motion (ROM)?", "to what extent is neuromuscular electrical stimulation (NMES) effective in the protection from muscular atrophy?", "how early can proprioception training and open chain exercises begin?", "should strengthening training start in the immediate postoperative period?" be answered for sure? My aim is to review postoperative brace use, early ROM restoration, NMES, proprioception, open/closed chain exercises and early strengthening, which are common modalities in the very comprehensive theme of postoperative ACL rehabilitation, on the basis of several studies (Level of Evidence 1 and 2) and to present the commonly accepted ways they are presently used. Moreover, I have presented the objectives of postoperative ACL rehabilitation in tables and recent miscellaneous studies in the last chapter of the paper.


Emergency medical services intervals and survival in trauma: assessment of the "golden hour" in a North American prospective cohort.
The first hour after the onset of out-of-hospital traumatic injury is referred to as the "golden hour," yet the relationship between time and outcome remains unclear. We evaluate the association between emergency medical services (EMS) intervals and mortality among trauma patients with field-based physiologic abnormality. This was a secondary analysis of an out-of-hospital, prospective cohort registry of adult (aged > or =15 years) trauma patients transported by 146 EMS agencies to 51 Level I and II trauma hospitals in 10 sites across North America from December 1, 2005, through March 31, 2007. Inclusion criteria were systolic blood pressure less than or equal to 90 mm Hg, respiratory rate less than 10 or greater than 29 breaths/min, Glasgow Coma Scale score less than or equal to 12, or advanced airway intervention. The outcome was in-hospital mortality. We evaluated EMS intervals (activation, response, on-scene, transport, and total time) with logistic regression and 2-step instrumental variable models, adjusted for field-based confounders. There were 3,656 trauma patients available for analysis, of whom 806 (22.0%) died. In multivariable analyses, there was no significant association between time and mortality for any EMS interval: activation (odds ratio [OR] 1.00; 95% confidence interval [CI] 0.95 to 1.05), response (OR 1.00; 95% CI 9.97 to 1.04), on-scene (OR 1.00; 95% CI 0.99 to 1.01), transport (OR 1.00; 95% CI 0.98 to 1.01), or total EMS time (OR 1.00; 95% CI 0.99 to 1.01). Subgroup and instrumental variable analyses did not qualitatively change these findings. In this North American sample, there was no association between EMS intervals and mortality among injured patients with physiologic abnormality in the field.


Palladium and visible-light mediated carbonylative Suzuki-Miyaura coupling of unactivated alkyl halides and aryl boronic acids.
Herein, a simple and efficient method for the palladium-catalyzed carbonylation of aryl boronic acids with unactivated alkyl iodides and bromides under visible-light irradiation, ambient temperature and low CO-pressure is presented. Notably, the procedure uses readily available equipment and an inexpensive palladium catalyst to generate the key alkyl radical intermediate. These mild conditions enabled the synthesis of a range of functionalized aryl alkyl ketones including the antipsychotic drug, melperone.


Should flavours be banned in cigarettes and e-cigarettes? Evidence on adult smokers and recent quitters from a discrete choice experiment.
To provide the policy-relevant estimates of impacts of alternative flavour bans on preferences and demand for cigarettes and e-cigarettes in adult smokers and recent quitters. A best-best discrete choice experiment (DCE) is used to elicit smokers' and recent quitters' preferences for flavours, price, health impact and nicotine level in cigarettes and e-cigarettes. Choice of tobacco products and an opt-out option were examined. An efficient design yielded 36 choice sets. Exploded logit choice models were estimated. Flavour bans are modelled by restricting flavour coefficients in the estimated model. A sample of 2031 adult smokers and recent quitters was recruited to complete an online survey and DCE. Current smokers and recent quitters, on average, prefer cigarettes and menthol cigarettes over flavoured e-cigarettes. However, there is substantial preference heterogeneity by younger adults (ages 18-25), race/ethnicity and respondents with higher education. Our predictions suggest that a ban on menthol cigarettes would produce the greatest reduction in the choice of cigarettes (-5.2%), but with an accompanying increase in e-cigarettes use (3.8%). In contrast, banning flavours in e-cigarettes, while allowing menthol in cigarettes would result in the greatest increase in the selection of cigarettes (8.3%), and a decline in the use of e-cigarettes (-11.1%). A ban on all flavours, but tobacco in both products would increase 'opting-out' the most (5.2%) but would also increase choice of cigarettes (2.7%) and decrease choice of e-cigarettes (-7.9%). A ban on flavoured e-cigarettes alone would likely increase the choice of cigarettes in smokers, arguably the more harmful way of obtaining nicotine, whereas a ban on menthol cigarettes alone would likely be more effective in reducing the choice of cigarettes. A ban on all flavours in both products would likely reduce the smoking/vaping rates, but the use of cigarettes would be higher than in the status quo. Policy-makers should use these results to guide the choice of flavour bans in light of their stance on the potential health impacts both products.


Toward high-resolution NMR spectroscopy of microscopic liquid samples.
A longstanding limitation of high-resolution NMR spectroscopy is the requirement for samples to have macroscopic dimensions. Commercial probes, for example, are designed for volumes of at least 5 μL, in spite of decades of work directed toward the goal of miniaturization. Progress in miniaturizing inductive detectors has been limited by a perceived need to meet two technical requirements: (1) minimal separation between the sample and the detector, which is essential for sensitivity, and (2) near-perfect magnetic-field homogeneity at the sample, which is typically needed for spectral resolution. The first of these requirements is real, but the second can be relaxed, as we demonstrate here. By using pulse sequences that yield high-resolution spectra in an inhomogeneous field, we eliminate the need for near-perfect field homogeneity and the accompanying requirement for susceptibility matching of microfabricated detector components. With this requirement removed, typical imperfections in microfabricated components can be tolerated, and detector dimensions can be matched to those of the sample, even for samples of volume ≪5 μL. Pulse sequences that are robust to field inhomogeneity thus enable small-volume detection with optimal sensitivity. We illustrate the potential of this approach to miniaturization by presenting spectra acquired with a flat-wire detector that can easily be scaled to subnanoliter volumes. In particular, we report high-resolution NMR spectroscopy of an alanine sample of volume 500 pL.


Technologies of trust in epidemic response: openness, reflexivity and accountability during the 2014-2016 Ebola outbreak in West Africa.
Trust is an essential component of successful cooperative endeavours. The global health response to the 2014-2016 West Africa Ebola outbreak confronted historically tenuous regional relationships of trust. Challenging sociopolitical contexts and initially inappropriate communication strategies impeded trustworthy relationships between communities and responders during the epidemic. Social scientists affiliated with the Ebola 100-Institut Pasteur project interviewed approximately 160 local, national and international responders holding a wide variety of roles during the epidemic. Focusing on responder's experiences of communities' trust during the epidemic, this qualitative study identifies and explores social techniques for effective emergency response. The response required individuals with diverse knowledges and experiences. Responders' included on-the-ground social mobilisers, health workers and clinicians, government officials, ambulance drivers, contact tracers and many more. We find that trust was fostered through open, transparent and reflexive communication that was adaptive and accountable to community-led response efforts and to real-time priorities. We expand on these findings to identify 'technologies of trust' that can be used to promote actively legitimate trustworthy relationships. Responders engaged the social technologies of openness (a willingness and genuine effort to incorporate multiple perspectives), reflexivity (flexibly responsive to context and ongoing dialogue) and accountability (taking responsibility for local contexts and consequences) to facilitate relations of trust. Technologies of trust contribute to the development of a framework of practical techniques to improve the acceptance and effectiveness of future emergency response strategies.