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mTORC1 controls key processes that regulate cell growth, including mRNA translation, ribosome biogenesis, and autophagy. Environmental amino acids activate mTORC1 by promoting its recruitment to the cytosolic surface of the lysosome, where its kinase is activated downstream of growth factor signaling. mTORC1 is brought to the lysosome by the Rag GTPases, which are tethered to the lysosomal membrane by Ragulator, a lysosome-bound scaffold. Here, we identify c17orf59 as a Ragulator-interacting protein that regulates mTORC1 activity through its interaction with Ragulator at the lysosome. The binding of c17orf59 to Ragulator prevents Ragulator interaction with the Rag GTPases, both in cells and in vitro, and decreases Rag GTPase lysosomal localization. Disruption of the Rag-Ragulator interaction by c17orf59 impairs mTORC1 activation by amino acids by preventing mTOR from reaching the lysosome. By disrupting the Rag-Ragulator interaction to inhibit mTORC1, c17orf59 expression may represent another mechanism to modulate nutrient sensing by mTORC1.

This paper presents an advanced engineering evaluation, using nonlinear analysis of hyper elastic material that provides significant improvement to structural silicone glazing (SSG) design in high performance curtain wall systems. Very high cladding wind pressures required in hurricane zones often result in bulky SSG profile dimensions. Architectural desire for aesthetically slender curtain wall framing sight-lines in combination with a desire to reduce aluminium usage led to optimization of silicone material geometry for better stress distribution.To accomplish accurate simulation of predicted behaviour under structural load, robust stress-strain curves of the silicone material are essential. The silicone manufacturer provided physical property testing via a specialized laboratory protocol. A series of rigorous curve fit techniques were then made to closely model test data in the finite element computer analysis that accounts for nonlinear strain of hyper elastic silicone.Comparison of this advanced design technique to traditional SSG design highlights differences in stress distribution contours in the silicone material. Simplified structural engineering per the traditional SSG design method does not provide accurate forecasting of material and stress optimization as shown in the advanced design.Full-scale specimens subject to structural load testing were performed to verify the design capacity, not only for high wind pressure values, but also for debris impact per ASTM E1886 and ASTM E1996. Also, construction of the test specimens allowed development of SSG installation techniques necessitated by the unique geometry of the silicone profile. Finally, correlation of physical test results with theoretical simulations is made, so evaluation of design confidence is possible. This design technique will introduce significant engineering advancement to the curtain wall industry.

Nasogastric (NG) tubes are commonly used for enteral feeding. Complications of feeding tube misplacement include malnutrition, pulmonary aspiration, and even death. We built a Bayesian network (BN) to analyse the risks associated with available bedside tests to verify tube position. Evidence on test validity (sensitivity and specificity) was retrieved from a systematic review. Likelihood ratios were used to select the best tests for detecting tubes misplaced in the lung or oesophagus. Five bedside tests were analysed including magnetic guidance, aspirate pH, auscultation, aspirate appearance, and capnography/colourimetry. Among these, auscultation and appearance are non-diagnostic towards lung or oesophagus placements. Capnography/ colourimetry can confirm but cannot rule out lung placement. Magnetic guidance can rule out both lung and oesophageal placement. However, as a relatively new technology, further validation studies are needed. The pH test with a cut-off at 5.5 or lower can rule out lung intubation. Lowering the cut-off to 4 not only minimises oesophageal intubation but also provides extra safety as the sensitivity of pH measurement is reduced by feeding, antacid medication, or the use of less accurate pH paper. BN is an effective tool for representing and analysing multi-layered uncertainties in test validity and reliability for the verification of NG tube position. Aspirate pH with a cut-off of 4 is the safest bedside method to minimise lung and oesophageal misplacement.

Receptor-mediated oscillations in cytosolic Ca2+ concentration ([Ca2+]i) could originate either directly from an autonomous Ca2+ feedback oscillator at the inositol 1,4,5-trisphosphate (IP3) receptor or as a secondary consequence of IP3 oscillations driven by Ca2+ feedback on IP3 metabolism. It is challenging to discriminate these alternatives, because IP3 fluctuations could drive Ca2+ oscillations or could just be a secondary response to the [Ca2+]i spikes. To investigate this problem, we constructed a recombinant IP3 buffer using type-I IP3 receptor ligand-binding domain fused to GFP (GFP-LBD), which buffers IP3 in the physiological range. This IP3 buffer slows hormone-induced [IP3] dynamics without changing steady-state [IP3]. GFP-LBD perturbed [Ca2+]i oscillations in a dose-dependent manner: it decreased both the rate of [Ca2+]i rise and the speed of Ca2+ wave propagation and, at high levels, abolished [Ca2+]i oscillations completely. These data, together with computational modeling, demonstrate that IP3 dynamics play a fundamental role in generating [Ca2+]i oscillations and waves.

We report the total and differential cross sections for J/ψ photoproduction with the large acceptance GlueX spectrometer for photon beam energies from the threshold at 8.2 GeV up to 11.44 GeV and over the full kinematic range of momentum transfer squared, t. Such coverage facilitates the extrapolation of the differential cross sections to the forward (t=0) point beyond the physical region. The forward cross section is used by many theoretical models and plays an important role in understanding J/ψ photoproduction and its relation to the J/ψ-proton interaction. These measurements of J/ψ photoproduction near threshold are also crucial inputs to theoretical models that are used to study important aspects of the gluon structure of the proton, such as the gluon generalized parton distribution of the proton, the mass radius of the proton, and the trace anomaly contribution to the proton mass. We observe possible structures in the total cross section energy dependence and find evidence for contributions beyond gluon exchange in the differential cross section close to threshold, both of which are consistent with contributions from open-charm intermediate states.

Salmonella enterica is a bacterial species that is a major cause of illness in humans and food-producing animals. S. enterica exhibits considerable inter-serovar diversity, as evidenced by the large number of host adapted serovars that have been identified. The development of methods to assess genome diversity in S. enterica will help to further define the limits of diversity in this foodborne pathogen. Thus, we evaluated a PCR assay, which targets prophage integrase genes, as a rapid method to investigate S. enterica genome diversity. To evaluate the PCR prophage integrase assay, 49 isolates of S. enterica were selected, including 19 clinical isolates from clonal serovars (Enteritidis and Heidelberg) that commonly cause human illness, and 30 isolates from food-associated Salmonella serovars that rarely cause human illness. The number of integrase genes identified by the PCR assay was compared to the number of integrase genes within intact prophages identified by whole genome sequencing and phage finding program PHASTER. The PCR assay identified a total of 147 prophage integrase genes within the 49 S. enterica genomes (79 integrase genes in the food-associated Salmonella isolates, 50 integrase genes in S. Enteritidis, and 18 integrase genes in S. Heidelberg). In comparison, whole genome sequencing and PHASTER identified a total of 75 prophage integrase genes within 102 intact prophages in the 49 S. enterica genomes (44 integrase genes in the food-associated Salmonella isolates, 21 integrase genes in S. Enteritidis, and 9 integrase genes in S. Heidelberg). Collectively, both the PCR assay and PHASTER identified the presence of a large diversity of prophage integrase genes in the food-associated isolates compared to the clinical isolates, thus indicating a high degree of diversity in the food-associated isolates, and confirming the clonal nature of S. Enteritidis and S. Heidelberg. Moreover, PHASTER revealed a diversity of 29 different types of prophages and 23 different integrase genes within the food-associated isolates, but only identified four different phages and integrase genes within clonal isolates of S. Enteritidis and S. Heidelberg. These results demonstrate the potential usefulness of PCR based detection of prophage integrase genes as a rapid indicator of genome diversity in S. enterica.

Foodborne illnesses continue to have an economic impact on global health care systems. There is a growing concern regarding the increasing frequency of antibiotic resistance in foodborne bacterial pathogens and how such resistance may affect treatment outcomes. In an effort to better understand how to reduce the spread of resistance, many research studies have been conducted regarding the methods by which antibiotic resistance genes are mobilized and spread between bacteria. Transduction by bacteriophages (phages) is one of many horizontal gene transfer mechanisms, and recent findings have shown phage-mediated transduction to be a significant contributor to dissemination of antibiotic resistance genes. Here, we review the viability of transduction as a contributing factor to the dissemination of antibiotic resistance genes in foodborne pathogens of the Enterobacteriaceae family, including non-typhoidal Salmonella and Shiga toxin-producing Escherichia coli, as well as environmental factors that increase transduction of antibiotic resistance genes.

[This corrects the article DOI: 10.1371/journal.pmed.1002220.].

BackgroundPerinatal mortality and morbidity continue to be major global health challenges strongly associated with prematurity and reduced fetal growth, an issue of further interest given the mounting evidence that fetal growth in general is linked to degrees of risk of common noncommunicable diseases in adulthood. Against this background, WHO made it a high priority to provide the present fetal growth charts for estimated fetal weight (EFW) and common ultrasound biometric measurements intended for worldwide use.Methods and findingsWe conducted a multinational prospective observational longitudinal study of fetal growth in low-risk singleton pregnancies of women of high or middle socioeconomic status and without known environmental constraints on fetal growth. Centers in ten countries (Argentina, Brazil, Democratic Republic of the Congo, Denmark, Egypt, France, Germany, India, Norway, and Thailand) recruited participants who had reliable information on last menstrual period and gestational age confirmed by crown-rump length measured at 8-13 wk of gestation. Participants had anthropometric and nutritional assessments and seven scheduled ultrasound examinations during pregnancy. Fifty-two participants withdrew consent, and 1,387 participated in the study. At study entry, median maternal age was 28 y (interquartile range [IQR] 25-31), median height was 162 cm (IQR 157-168), median weight was 61 kg (IQR 55-68), 58% of the women were nulliparous, and median daily caloric intake was 1,840 cal (IQR 1,487-2,222). The median pregnancy duration was 39 wk (IQR 38-40) although there were significant differences between countries, the largest difference being 12 d (95% CI 8-16). The median birthweight was 3,300 g (IQR 2,980-3,615). There were differences in birthweight between countries, e.g., India had significantly smaller neonates than the other countries, even after adjusting for gestational age. Thirty-one women had a miscarriage, and three fetuses had intrauterine death. The 8,203 sets of ultrasound measurements were scrutinized for outliers and leverage points, and those measurements taken at 14 to 40 wk were selected for analysis. A total of 7,924 sets of ultrasound measurements were analyzed by quantile regression to establish longitudinal reference intervals for fetal head circumference, biparietal diameter, humerus length, abdominal circumference, femur length and its ratio with head circumference and with biparietal diameter, and EFW. There was asymmetric distribution of growth of EFW: a slightly wider distribution among the lower percentiles during early weeks shifted to a notably expanded distribution of the higher percentiles in late pregnancy. Male fetuses were larger than female fetuses as measured by EFW, but the disparity was smaller in the lower quantiles of the distribution (3.5%) and larger in the upper quantiles (4.5%). Maternal age and maternal height were associated with a positive effect on EFW, particularly in the lower tail of the distribution, of the order of 2% to 3% for each additional 10 y of age of the mother and 1% to 2% for each additional 10 cm of height. Maternal weight was associated with a small positive effect on EFW, especially in the higher tail of the distribution, of the order of 1.0% to 1.5% for each additional 10 kg of bodyweight of the mother. Parous women had heavier fetuses than nulliparous women, with the disparity being greater in the lower quantiles of the distribution, of the order of 1% to 1.5%, and diminishing in the upper quantiles. There were also significant differences in growth of EFW between countries. In spite of the multinational nature of the study, sample size is a limiting factor for generalization of the charts.ConclusionsThis study provides WHO fetal growth charts for EFW and common ultrasound biometric measurements, and shows variation between different parts of the world.

Extensive research is ongoing that concentrates on finding therapies to enhance CNS regeneration after spinal cord injury (SCI) and to cure paralysis. This review sheds light on the role of the FGFR pathway in the injured spinal cord and discusses various therapies that use FGFR activating ligands to promote regeneration after SCI. We discuss studies that use peripheral nerve grafts or Schwann cell grafts in combination with FGF1 or FGF2 supplementation. Most of these studies show evidence that these therapies successfully enhance axon regeneration into the graft. Further they provide evidence for partial recovery of sensory function shown by electrophysiology and motor activity evidenced by behavioural data. We also present one study that indicates that combination with additional, synergistic factors might further drive the system towards functional regeneration. In essence, this review summarises the potential of nerve and cell grafts combined with FGF1/2 supplementation to improve outcome even after severe spinal cord injury.

The aim of this investigation was to compare serum growth hormone (GH), insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-binding protein-3 (IGFBP-3) in response to a combined sprint and resistance training (CSRT) program in young and middle-aged men.Thirty-eight healthy, moderately trained men participated in this study. Young and middle-aged men were randomly assigned to, a young training group (YT = 10, 21.4±1.2yrs) ora young control group (YC = 9, 21.6±1.8 yrs), a middle-aged training group (MAT = 10, 40.4±2.1 yrs) or a middle-aged control group (MAC = 9, 40.5±1.8 yrs). Participants performed the Wingate Anaerobic Test (WAnT) before and after a 13-week CSRT program (three sessions per week). Blood samples were collected at rest, after warm-up, immediately post-WAnT, and 10 min post-WAnT. CSRT induced increases in GH at rest and in response to the WAnT in YT and MAT (P0.05). In response to the WAnT, amelioration of the age-effect was observed between YT and MAT for IGF-1 and IGF-1/IGFBP-3 ratio following CSRT (P>0.05). These data suggest that CSRT increases the activity of the GH/IGF-1 axis at rest and in response to the WAnT in young and middle-aged men. In addition, CSRT reduces the normal age-related decline of somatotropic hormones in middle-age men.

ABSTRACT Ionic liquid (IL) pretreatment is a promising approach for the conversion of lignocellulose to biofuels. The toxicity of residual IL, however, negatively impacts the performance of industrial enzymes and microorganisms in hydrolysis and fermentation. In this study, a thermophilic microbial community was cultured on switchgrass amended with various levels of the ionic liquid 1-ethyl-3-methylimidazolium acetate. Changes in the microbial community composition and transcription of genes relevant to IL tolerance and lignocellulose hydrolysis were quantified. Increasing the level of IL to 0.1% (wt) led to increased levels of relative abundance and transcription in organisms of the phylum Firmicutes. Interestingly, IL concentrations of up to 1% (wt) also resulted in greater xylanase transcription and enzyme activity as well as increased transcription of endoglucanase, beta-glucosidase, and IL tolerance genes compared to communities without IL. IL levels above 1% (wt) resulted in decreased enzyme activity and transcription of genes involved in lignocellulose hydrolysis. The results indicate that moderate levels of IL select for thermophilic microorganisms that not only tolerate IL but also effectively hydrolyze lignocellulose from switchgrass. Discovery of IL-tolerant organisms and enzymes is critical for the development of biological processes that convert IL-pretreated biomass to biofuels and chemicals. Employing metatranscriptomic analysis of enrichment cultures can facilitate the discovery of microorganisms and enzymes that may be active in the presence of toxic compounds such as ionic liquids. IMPORTANCE Pretreatment using ionic liquids (IL) is a promising approach for the conversion of lignocellulose to biofuels. Because IL can be inhibitory to enzymes and microorganisms involved in downstream hydrolysis and fermentation steps, discovery of IL-tolerant organisms and enzymes is critical for advancing this technology. Employing metatranscriptomics in the analysis of IL-enriched cultures facilitated tracking of dynamic changes in a complex microbial community at the level of gene transcription and doing so with genome resolution. Specific organisms were discovered that could simultaneously tolerate a moderate IL concentration and transcribe a diverse array of cellulolytic enzymes. Gene sequences of cellulolytic enzymes and efflux pumps from those same organisms were also identified, providing important resources for future research on engineering IL-tolerant organisms and enzymes.

The United States Air Force’s 45th Weather Squadron provides wind warnings, including those for downbursts, at the Cape Canaveral Air Force Station and Kennedy Space Center (CCAFS/KSC). This study aims to provide a Random Forest model that classifies thunderstorms’ downburst and null events using a 35-knot wind threshold to separate these two categories. The downburst occurrence was assessed using a dense network of wind observations around CCAFS/KSC. Eight dual-polarization radar signatures that are hypothesized to have physical implications for downbursts at the surface were automatically calculated for 209 storms and ingested into the Random Forest model. The Random Forest model predicted null events more correctly than downburst events, with a True Skill Statistic of 0.40. Strong downburst events were better classified than those with weaker wind magnitudes. The most important radar signatures were found to be the maximum vertically integrated ice and the peak reflectivity. The Random Forest model presented a more reliable performance than an automated prediction method based on thresholds of single radar signatures. Based on these results, the Random Forest method is suggested for continued operational development and testing.

(1) Background: High-intensity interval training (HIIT) exerts effects indicative of improved health in young and older populations. However, prescribing analogous training programmes is inappropriate, as recovery from HIIT is different between young and older individuals. Sprint interval training (SIT) is a derivative of HIIT but with shorter, maximal effort intervals. Prior to prescribing this mode of training, it is imperative to understand the recovery period to prevent residual fatigue affecting subsequent adaptations. (2) Methods: Nine older (6M/3F; mean age of 70 ± 8 years) and nine young (6M/3F; mean age of 24 ± 3 years) participants performed a baseline peak power output (PPO) test. Subsequently, two SIT sessions consisting of three repetitions of 20 s ‘all-out’ stationary cycling bouts interspersed by 3 minutes of self-paced recovery were performed. SIT sessions were followed by 3 days’ rest and 5 days’ rest on two separate occasions, in a randomised crossover design. PPO was measured again to determine whether recovery had been achieved after 3 days or after 5 days. (3) Results: Two-way repeated measure (age (older, young) × 3 time (baseline, 3 days, 5 days)) ANOVA revealed a large effect of age (p = 0.002, n2p = 0.460), with older participants having a lower PPO compared to young participants. A small effect of time (p = 0.702, n2p = 0.022), and a medium interaction between age and time (p = 0.098, n2p = 0.135) was observed. (4) Conclusions: This study demonstrates both young and older adults recover PPO following 3 and 5 days’ rest. As such, both groups could undertake SIT following three days of rest, without a reduction in PPO.

(1) Background: Findings from observational studies of relations between neighborhood environments and health outcomes underscore the importance of both objective and perceived experiences of those environments. A clearer understanding of the factors associated with discrepancies between these two assessment approaches is needed to tailor public health interventions to specific populations. This study examined how individual and neighborhood characteristics affect perceptions of supermarket distance, particularly when perceptions do not match objective measures. (2) Methods: Participants were older adults (n = 880) participating in the Senior Neighborhood Quality of Life Study in the Seattle/King County, WA or Baltimore/Washington, DC regions. Two main analyses were conducted. The primary outcome for Analysis I was participants’ geographic information systems (GIS)-based objective network distance to the closest supermarket. Generalized linear mixed models with block group-level random effects were used to assess associations between objective supermarket distance and individual/neighborhood characteristics. The primary outcome for Analysis II was a categorical “accuracy” variable, based on participants’ perceived distance to the nearest supermarket/grocery store relative to the objective distance, assuming a walking speed of 1.0 m/s. Multivariate log-linear models fit neural networks were used to assess influential covariates. (3) Results: Several significant associations with objective distance to the nearest supermarket were observed, including a negative relationship with body mass index (BMI) (95% CI = −45.56, −0.23), having walked to the supermarket in the last 30 days (−174.86, −59.42), living in a high-walkability neighborhood, and residing in Seattle/King County (−707.69, −353.22). In terms of participants’ distance accuracy, 29% were classified as accurate, 33.9% were “Underestimators”, 24.0% “Overestimators”, and 13.2% responded “Don’t Know”. Compared to Accurate participants, Overestimators were significantly less likely to have walked to the supermarket in the last 30 days, and lived objectively closer to a supermarket; Underestimators perceived significantly higher pedestrian safety and lived objectively further from a supermarket; and Don’t Know were more likely to be women, older, not living independently, and not having recently walked to the supermarket. (4) Conclusions: Both modifiable and nonmodifiable factors influence the accuracy of older adults’ perceptions of their proximity to the nearest supermarket. Recent experience in walking to the closest supermarket, along with personal safety, represent potentially modifiable perceived environmental factors that were related to older adults’ accuracy of perceptions of their neighborhood food environment.

Background: Determining mitotic index by counting mitotic figures (MFs) microscopically from tumor areas with most abundant MF (hotspots [HS]) produces a prognostically useful tumor grading biomarker. However, interobserver concordance identifying MF and HS can be poorly reproducible. Immunolabeling MF, coupled with computer-automated counting by image analysis, can improve reproducibility. A computational system for obtaining MF values across digitized whole-slide images (WSIs) was sought that would minimize impact of artifacts, generate values clinically relatable to counting ten high-power microscopic fields of view typical in conventional microscopy, and that would reproducibly map HS topography. Materials and Methods: Relatively low-resolution WSI scans (0.50 μm/pixel) were imported in grid-tile format for feature-based MF segmentation, from naturally occurring canine melanomas providing a wide range of proliferative activity. MF feature extraction conformed to anti-phospho-histone H3-immunolabeled mitotic (M) phase cells. Computer vision image processing was established to subtract key artifacts, obtain MF counts, and employ rotationally invariant feature extraction to map MF topography. Results: The automated topometric HS (TMHS) algorithm identified mitotic HS and mapped select tissue tiles with greatest MF counts back onto WSI thumbnail images to plot HS topographically. Influence of dye, pigment, and extraneous structure artifacts was minimized. TMHS diagnostic decision support included image overlay graphics of HS topography, as well as a spreadsheet and plot of tile-based MF count values. TMHS performance was validated examining both mitotic HS counting and mapping functions. Significantly correlated TMHS MF mapping and metrics were demonstrated using repeat analysis with WSI in different orientation (R2 = 0.9916) and by agreement with a pathologist (R2 = 0.8605) as well as through assessment of counting function using an independently tuned object counting algorithm (OCA) (R2 = 0.9482). Limits of agreement analysis support method interchangeability. MF counts obtained led to accurate patient survival prediction in all (n = 30) except one case. By contrast, more variable performance was documented when several pathologists examined similar cases using microscopy (pair-wise correlations, rho range = 0.7597–0.9286). Conclusions: Automated TMHS MF segmentation and feature engineering performance were interchangeable with both observer and OCA in digital mode. Moreover, enhanced HS location accuracy and superior method reproducibility were achieved using the automated TMHS algorithm compared to the current practice employing clinical microscopy.