Next-Generation Genome-Scale Metabolic Modeling through Integration of Regulatory Mechanisms.
Genome-scale metabolic models (GEMs) are powerful tools for understanding metabolism from a systems-level perspective. However, GEMs in their most basic form fail to account for cellular regulation. A diverse set of mechanisms regulate cellular metabolism, enabling organisms to respond to a wide range of conditions. This limitation of GEMs has prompted the development of new methods to integrate regulatory mechanisms, thereby enhancing the predictive capabilities and broadening the scope of GEMs. Here, we cover integrative models encompassing six types of regulatory mechanisms: transcriptional regulatory networks (TRNs), post-translational modifications (PTMs), epigenetics, protein-protein interactions and protein stability (PPIs/PS), allostery, and signaling networks. We discuss 22 integrative GEM modeling methods and how these have been used to simulate metabolic regulation during normal and pathological conditions. While these advances have been remarkable, there remains a need for comprehensive and widespread integration of regulatory constraints into GEMs. We conclude by discussing challenges in constructing GEMs with regulation and highlight areas that need to be addressed for the successful modeling of metabolic regulation. Next-generation integrative GEMs that incorporate multiple regulatory mechanisms and their crosstalk will be invaluable for discovering cell-type and disease-specific metabolic control mechanisms.


Primary care screening for childhood obesity: a population-based analysis.
The prevalence of obesity among children and adolescents in the western world has increased dramatically. To assess the efficacy of routine childhood obesity screening by primary physicians in the pediatric population in Israel and the utilization of health services by overweight children. The electronic medical records of children aged 60-83 months registered in 39 pediatric primary care centers between January 2001 and October 2004 (n=21,799) were reviewed. Those in whom height and weight were documented during a clinic visit (index visit) were classified as overweight, at risk of overweight, or normal weight according to body mass index percentiles. The number of visits to the pediatrician, laboratory tests and health care costs 12 months after the index visit were calculated. Anthropomorphic measurements were performed in 1556 of the 15,364 children (10.1%) who visited the clinic during the study period. Of these, 398 (25.6%) were overweight, 185 (11.9%) were at risk of overweight, and 973 (62.5%) were normal weight. Children in the first two groups visited the clinic slightly more often than the third group, but the differences were not statistically significant (P = 0.12), and they had significantly more laboratory tests than the rest of the children visiting the clinics (P = 0.053). Health care costs were 6.6% higher for the overweight than the normal-weight children. Electronic medical records are a useful tool for population-based health care assessments. Current screening for obesity in children during routine care in Israel is insufficient and additional education of community pediatricians in diagnosis and intervention is urgently needed.


The spatial distribution of known predictors of autism spectrum disorders impacts geographic variability in prevalence in central North Carolina.
The causes of autism spectrum disorders (ASD) remain largely unknown and widely debated; however, evidence increasingly points to the importance of environmental exposures. A growing number of studies use geographic variability in ASD prevalence or exposure patterns to investigate the association between environmental factors and ASD. However, differences in the geographic distribution of established risk and predictive factors for ASD, such as maternal education or age, can interfere with investigations of ASD etiology. We evaluated geographic variability in the prevalence of ASD in central North Carolina and the impact of spatial confounding by known risk and predictive factors. Children meeting a standardized case definition for ASD at 8 years of age were identified through records-based surveillance for 8 counties biennially from 2002 to 2008 (n=532). Vital records were used to identify the underlying cohort (15% random sample of children born in the same years as children with an ASD, n=11,034), and to obtain birth addresses. We used generalized additive models (GAMs) to estimate the prevalence of ASD across the region by smoothing latitude and longitude. GAMs, unlike methods used in previous spatial analyses of ASD, allow for extensive adjustment of individual-level risk factors (e.g. maternal age and education) when evaluating spatial variability of disease prevalence. Unadjusted maps revealed geographic variation in surveillance-recognized ASD. Children born in certain regions of the study area were up to 1.27 times as likely to be recognized as having ASD compared to children born in the study area as a whole (prevalence ratio (PR) range across the study area 0.57-1.27; global P=0.003). However, geographic gradients of ASD prevalence were attenuated after adjusting for spatial confounders (adjusted PR range 0.72-1.12 across the study area; global P=0.052). In these data, spatial variation of ASD in central NC can be explained largely by factors impacting diagnosis, such as maternal education, emphasizing the importance of adjusting for differences in the geographic distribution of known individual-level predictors in spatial analyses of ASD. These results underscore the critical importance of accounting for such factors in studies of environmental exposures that vary across regions.


The mystery of missing female children in the Caucasus: an analysis of sex ratios by birth order.
Official data on sex ratios at birth suggest a rise in sex-selective abortions in some post-Soviet states following the introduction of ultrasonography. However, questions remain about the validity of official data in these nations as well as whether the high sex ratios at birth are a statistical artifact. Trends in sex ratios at birth from 1985 to 2009 for 12 post-Soviet states were examined using vital registration data. For the three countries that had had a Demographic and Health Survey (DHS) in 2005-2010 (Armenia, Azerbaijan and Moldova), survey data were used to calculate sex ratios at birth according to birth order, and vital registration data for 2010 were used to estimate the number of "missing" female births (if any). Official data revealed elevated sex ratios at birth in Armenia (117), Azerbaijan (116) and Georgia (121), but not in other post-Soviet states. According to DHS data, sex ratios were high in Armenia and Azerbaijan for first births (138 and 113, respectively); if the first child was a girl, the sex ratio in Armenia was even higher for the second birth (154). Overall, the number of girls born in these countries in 2010 was 10% lower than expected, consistent with 1,972 sex-selective abortions in Armenia and 8,381 in Azerbaijan. Sex ratios did not vary by birth order in Moldova. Sex-selective abortion appears to be common in Azerbaijan and Armenia. Family planning and legal interventions are needed to address this issue.


Real-time process dynamics monitoring in Anammox reactors.
Process dynamics in Anammox systems were evaluated through continuous monitoring of pH, oxidation reduction potential (ORP) and conductivity in two separate newly started-up sequencing batch reactors, one seeded with an enriched Anammox sludge and the other seeded with mixed activated sludge. The pH and ORP profiles exhibited characteristic patterns depending on the process dynamics during early start-up, start-up and enrichment phases of the operational period of 410 days. The simultaneously continuing processes of the start-up period showed apparent indicative trend lines in pH and ORP profiles. Conductivity profiles were consistent with the process dynamics in all phases. During the enrichment phase, conductivity decreases could quantitatively be related to process removal efficiencies and all real-time profiles exhibited specific break-points which coincided with the end of Anammox in each cycle. The end of Anammox was observed as an 'apex' on pH profiles and a 'valley' on ORP profiles. The 'apex' and 'valley' points exactly coincided with the end point of the linear decrease in the conductivity profiles. The overall findings suggested a great potential in using real-time pH, ORP and conductivity measurements for quick and reliable monitoring of Anammox systems during start-up and enrichment periods.


Normal responsiveness of CTLA-4-deficient anti-viral cytotoxic T cells.
CTLA-4 has been proposed to negatively regulate immune responses, and mice deficient for CTLA-4 expression succumb to a lymphoproliferative disorder within a few weeks after birth. This study assessed the responsiveness of CTLA-4-deficient T cells expressing a class I-restricted TCR specific for lymphocytic choriomeningitis virus (LCMV). The kinetics of T cell proliferation were studied in vitro after stimulation of T cells with full and partial T cell agonists. No gross abnormalities in CTLA-4-deficient T cells could be detected. Using adoptive transfer experiments, T cell responses were also measured in vivo after infection with LCMV. Low dose infection with LCMV leads to strong expansion of specific T cells followed by a reduction in T cells that parallels the elimination of Ag. The kinetics of T cell expansion and elimination after low dose LCMV infection were not affected by the absence of CTLA-4. High dose infection of mice with LCMV leads to a transient expansion of T cells followed by T cell exhaustion, where all specific T cells are eliminated. T cell exhaustion also occurred in the absence of CTLA-4. Thus, surprisingly, the absence of CTLA-4 did not interfere with T cell activation, down-regulation of ongoing T cell responses after the elimination of Ag, or the exhaustion of T cell responses in the presence of excessive amounts of Ag.


Genetic markers associated with resistance to beta-lactam and quinolone antimicrobials in non-typhoidal Salmonella isolates from humans and animals in central Ethiopia.
Beta-lactam and quinolone antimicrobials are commonly used for treatment of infections caused by non-typhoidal Salmonella (NTS) and other pathogens. Resistance to these classes of antimicrobials has increased significantly in the recent years. However, little is known on the genetic basis of resistance to these drugs in Salmonella isolates from Ethiopia. Salmonella isolates with reduced susceptibility to beta-lactams (n = 43) were tested for genes encoding for beta-lactamase enzymes, and those resistant to quinolones (n = 29) for mutations in the quinolone resistance determining region (QRDR) as well as plasmid mediated quinolone resistance (PMQR) genes using PCR and sequencing. Beta-lactamase genes (bla) were detected in 34 (79.1%) of the isolates. The dominant bla gene was blaTEM, recovered from 33 (76.7%) of the isolates, majority being TEM-1 (24, 72.7%) followed by TEM-57, (10, 30.3%). The blaOXA-10 and blaCTX-M-15 were detected only in a single S. Concord human isolate. Double substitutions in gyrA (Ser83-Phe + Asp87-Gly) as well as parC (Thr57-Ser + Ser80-Ile) subunits of the quinolone resistance determining region (QRDR) were detected in all S. Kentucky isolates with high level resistance to both nalidixic acid and ciprofloxacin. Single amino acid substitutions, Ser83-Phe (n = 4) and Ser83-Tyr (n = 1) were also detected in the gyrA gene. An isolate of S. Miami susceptible to nalidixic acid but intermediately resistant to ciprofloxacin had Thr57-Ser and an additional novel mutation (Tyr83-Phe) in the parC gene. Plasmid mediated quinolone resistance (PMQR) genes investigated were not detected in any of the isolates. In some isolates with decreased susceptibility to ciprofloxacin and/or nalidixic acid, no mutations in QRDR or PMQR genes were detected. Over half of the quinolone resistant isolates in the current study 17 (58.6%) were also resistant to at least one of the beta-lactam antimicrobials. Acquisition of blaTEM was the principal beta-lactamase resistance mechanism and mutations within QRDR of gyrA and parC were the primary mechanism for resistance to quinolones. Further study on extended spectrum beta-lactamase and quinolone resistance mechanisms in other gram negative pathogens is recommended.


Increased Thrombopoiesis and Platelet Activation in Hantavirus-Infected Patients.
Thrombocytopenia is a common finding during viral hemorrhagic fever, which includes hemorrhagic fever with renal syndrome (HFRS). The 2 main causes for thrombocytopenia are impaired thrombopoiesis and/or increased peripheral destruction of platelets. In addition, there is an increased intravascular coagulation risk during HFRS, which could be due to platelet activation. Thrombopoiesis was determined by quantification of platelet counts, thrombopoietin, immature platelet fraction, and mean platelet volume during HFRS. The in vivo platelet activation was determined by quantification of soluble P-selectin (sP-selectin) and glycoprotein VI (sGPVI). The function of circulating platelets was determined by ex vivo stimulation followed by flow cytometry analysis of platelet surface-bound fibrinogen and P-selectin exposure. Intravascular coagulation during disease was determined by scoring for disseminated intravascular coagulation (DIC) and recording thromboembolic complications. The levels of thrombopoietin, immature platelet fraction, and mean platelet volume all indicate increased thrombopoiesis during HFRS. Circulating platelets had reduced ex vivo function during disease compared to follow-up. Most interestingly, we observed significantly increased in vivo platelet activation in HFRS patients with intravascular coagulation (DIC and thromboembolic complications) as shown by sP-selectin and sGPVI levels. HFRS patients have increased thrombopoiesis and platelet activation, which contributes to intravascular coagulation.


Paleogenomics. Genomic structure in Europeans dating back at least 36,200 years.
The origin of contemporary Europeans remains contentious. We obtained a genome sequence from Kostenki 14 in European Russia dating from 38,700 to 36,200 years ago, one of the oldest fossils of anatomically modern humans from Europe. We find that Kostenki 14 shares a close ancestry with the 24,000-year-old Mal'ta boy from central Siberia, European Mesolithic hunter-gatherers, some contemporary western Siberians, and many Europeans, but not eastern Asians. Additionally, the Kostenki 14 genome shows evidence of shared ancestry with a population basal to all Eurasians that also relates to later European Neolithic farmers. We find that Kostenki 14 contains more Neandertal DNA that is contained in longer tracts than present Europeans. Our findings reveal the timing of divergence of western Eurasians and East Asians to be more than 36,200 years ago and that European genomic structure today dates back to the Upper Paleolithic and derives from a metapopulation that at times stretched from Europe to central Asia.


Idiopathic intracranial hypertension in pediatric population: case series from India.
Idiopathic intracranial hypertension (IIH) is a well described entity in adults. In pediatric age group the presentation of disease can vary depending on the age of patients and is less frequently reported. The aim of this study is to describe the clinical features, investigations, treatment and outcome of IIH in pediatric population (age <18 years). This retrospective hospital based study was carried out on 25 children with diagnosis of IIH based on modified Dandys criteria. Their clinical, investigation, treatment, outcome and follow-up for 2 year period were analyzed. Out of the 25 children, the youngest child was 4-month-old infant. The commonest symptom was headache (76%) followed by vomiting and papilledema (72%). The mean cerebrospinal fluid (CSF) pressure was 330 mm of H 2 O. In Infants irritability and bulging anterior fontanelle was seen. A total of 24 patients showed a complete resolution of symptom. None of patient had recurrence over a period of 2 years follow-up. IIH can present at any age group. This is the largest series of IIH reported in pediatric population in India. The clinical features are similar to adult patients except in infants. Absence of papilledema does not exclude the diagnosis of IIH. CSF pressure monitoring is needed in suspected cases of IIH. Early and prompt treatment can prevent deficits.


(Reinforcement?) Learning to forage optimally.
Foraging effectively is critical to the survival of all animals and this imperative is thought to have profoundly shaped brain evolution. Decisions made by foraging animals often approximate optimal strategies, but the learning and decision mechanisms generating these choices remain poorly understood. Recent work with laboratory foraging tasks in humans suggest their behaviour is poorly explained by model-free reinforcement learning, with simple heuristic strategies better describing behaviour in some tasks, and in others evidence of prospective prediction of the future state of the environment. We suggest that model-based average reward reinforcement learning may provide a common framework for understanding these apparently divergent foraging strategies.


Endovascular strategy or open repair for ruptured abdominal aortic aneurysm: one-year outcomes from the IMPROVE randomized trial.
To report the longer term outcomes following either a strategy of endovascular repair first or open repair of ruptured abdominal aortic aneurysm, which are necessary for both patient and clinical decision-making. This pragmatic multicentre (29 UK and 1 Canada) trial randomized 613 patients with a clinical diagnosis of ruptured aneurysm; 316 to an endovascular first strategy (if aortic morphology is suitable, open repair if not) and 297 to open repair. The principal 1-year outcome was mortality; secondary outcomes were re-interventions, hospital discharge, health-related quality-of-life (QoL) (EQ-5D), costs, Quality-Adjusted-Life-Years (QALYs), and cost-effectiveness [incremental net benefit (INB)]. At 1 year, all-cause mortality was 41.1% for the endovascular strategy group and 45.1% for the open repair group, odds ratio 0.85 [95% confidence interval (CI) 0.62, 1.17], P = 0.325, with similar re-intervention rates in each group. The endovascular strategy group and open repair groups had average total hospital stays of 17 and 26 days, respectively, P < 0.001. Patients surviving rupture had higher average EQ-5D utility scores in the endovascular strategy vs. open repair groups, mean differences 0.087 (95% CI 0.017, 0.158), 0.068 (95% CI -0.004, 0.140) at 3 and 12 months, respectively. There were indications that QALYs were higher and costs lower for the endovascular first strategy, combining to give an INB of £3877 (95% CI £253, £7408) or €4356 (95% CI €284, €8323). An endovascular first strategy for management of ruptured aneurysms does not offer a survival benefit over 1 year but offers patients faster discharge with better QoL and is cost-effective. ISRCTN 48334791.


S1PR1 drives a feedforward signalling loop to regulate BATF3 and the transcriptional programme of Hodgkin lymphoma cells.
The Hodgkin/Reed-Sternberg cells of classical Hodgkin lymphoma (HL) are characterised by the aberrant activation of multiple signalling pathways. Here we show that a subset of HL displays altered expression of sphingosine-1-phosphate (S1P) receptors (S1PR)s. S1P activates phosphatidylinositide 3-kinase (PI3-K) in these cells that is mediated by the increased expression of S1PR1 and the decreased expression of S1PR2. We also showed that genes regulated by the PI3-K signalling pathway in HL cell lines significantly overlap with the transcriptional programme of primary HRS cells. Genes upregulated by the PI3-K pathway included the basic leucine zipper transcription factor, ATF-like 3 (BATF3), which is normally associated with the development of dendritic cells. Immunohistochemistry confirmed that BATF3 was expressed in HRS cells of most HL cases. In contrast, in normal lymphoid tissues, BATF3 expression was confined to a small fraction of CD30-positive immunoblasts. Knockdown of BATF3 in HL cell lines revealed that BATF3 contributed to the transcriptional programme of primary HRS cells, including the upregulation of S1PR1. Our data suggest that disruption of this potentially oncogenic feedforward S1P signalling loop could provide novel therapeutic opportunities for patients with HL.


Controlling contractile instabilities in the actomyosin cortex.
The actomyosin cell cortex is an active contractile material for driving cell- and tissue morphogenesis. The cortex has a tendency to form a pattern of myosin foci, which is a signature of potentially unstable behavior. How a system that is prone to such instabilities can rveliably drive morphogenesis remains an outstanding question. Here, we report that in the Caenorhabditis elegans zygote, feedback between active RhoA and myosin induces a contractile instability in the cortex. We discover that an independent RhoA pacemaking oscillator controls this instability, generating a pulsatory pattern of myosin foci and preventing the collapse of cortical material into a few dynamic contracting regions. Our work reveals how contractile instabilities that are natural to occur in mechanically active media can be biochemically controlled to robustly drive morphogenetic events.


Membrane cholesterol, protein phosphorylation, and lipid rafts.
The functions of cholesterol and membrane microdomains in transmembrane signaling remain controversial. Edidin discusses the questions surrounding lipid rafts, membrane microdomains that have been biochemically defined but are difficult to visualize in vivo. He also discusses whether experiments showing correlation of changes in plasma membrane cholesterol with differentiation and the formation of adherens junctions in endothelial cells are consistent with a model in which lipid rafts influence the regulation of these processes.


Short-term secondhand smoke exposure decreases heart rate variability and increases arrhythmia susceptibility in mice.
Exposure to secondhand smoke (SHS), a major indoor air pollutant, is linked to increased cardiovascular morbidity and mortality, including cardiac arrhythmias. However, the mechanisms underlying the epidemiological findings are not well understood. Impaired cardiac autonomic function, indexed by reduced heart rate variability (HRV), may represent an underlying cause. The present study takes advantage of well-defined short-term SHS exposure (3 days, 6 h/day) on HRV and the susceptibility to arrhythmia in mice. With the use of electrocardiograph telemetry recordings in conscious mice, HRV parameters in the time domain were measured during the night after each day of exposure and 24 h after 3 days of exposure to either SHS or filtered air. The susceptibility to arrhythmia was determined after 3 days of exposure. Exposure to a low concentration of SHS [total suspended particle (TSP), 2.4 +/- 3.2; and nicotine, 0.3 +/- 0.1 mg/m(3)] had no significant effect on HRV parameters. In contrast, the exposure to a higher but still environmentally relevant concentration of SHS (TSP, 30 +/- 1; and nicotine, 5 +/- 1 mg/m(3)) significantly reduced HRV starting after the first day of exposure and continuing 24 h after the last day of exposure. Moreover, the exposed mice showed a significant increase in ventricular arrhythmia susceptibility and atrioventricular block. The data suggest that SHS exposure decreased HRV beyond the exposure period and was associated with an increase in arrhythmia susceptibility. The data provide insights into possible mechanisms underlying documented increases in cardiovascular morbidity and mortality in humans exposed to SHS.


Anthrax toxin receptor 2a controls mitotic spindle positioning.
Oriented mitosis is essential during tissue morphogenesis. The Wnt/planar cell polarity (Wnt/PCP) pathway orients mitosis in a number of developmental systems, including dorsal epiblast cell divisions along the animal-vegetal (A-V) axis during zebrafish gastrulation. How Wnt signalling orients the mitotic plane is, however, unknown. Here we show that, in dorsal epiblast cells, anthrax toxin receptor 2a (Antxr2a) accumulates in a polarized cortical cap, which is aligned with the embryonic A-V axis and forecasts the division plane. Filamentous actin (F-actin) also forms an A-V polarized cap, which depends on Wnt/PCP and its effectors RhoA and Rock2. Antxr2a is recruited to the cap by interacting with actin. Antxr2a also interacts with RhoA and together they activate the diaphanous-related formin zDia2. Mechanistically, Antxr2a functions as a Wnt-dependent polarized determinant, which, through the action of RhoA and zDia2, exerts torque on the spindle to align it with the A-V axis.


Vascular endothelial growth factor receptors VEGFR-2 and VEGFR-3 are localized primarily to the vasculature in human primary solid cancers.
Vascular endothelial growth factor (VEGF) signaling is key to tumor angiogenesis and is an important target in the development of anticancer drugs. However, VEGF receptor (VEGFR) expression in human cancers, particularly the relative expression of VEGFR-2 and VEGFR-3 in tumor vasculature versus tumor cells, is poorly defined. VEGFR-2- and VEGFR-3-specific antibodies were identified and used in the immunohistochemical analysis of human primary cancers and normal tissue. The relative vascular localization of both receptors in colorectal and breast cancers was determined by coimmunofluorescence with vascular markers. VEGFR-2 and VEGFR-3 were expressed on vascular endothelium but not on malignant cells in 13 common human solid tumor types (n > 400, bladder, breast, colorectal, head and neck, liver, lung, skin, ovarian, pancreatic, prostate, renal, stomach, and thyroid). The signal intensity of both receptors was significantly greater in vessels associated with malignant colorectal, lung, and breast than adjacent nontumor tissue. In colorectal cancers, VEGFR-2 was expressed on both intratumoral blood and lymphatic vessels, whereas VEGFR-3 was found predominantly on lymphatic vessels. In breast cancers, both receptors were localized to and upregulated on blood vessels. VEGFR-2 and VEGFR-3 are primarily localized to, and significantly upregulated on, tumor vasculature (blood and/or lymphatic) supporting the majority of solid cancers. The primary clinical mechanism of action of VEGF signaling inhibitors is likely to be through the targeting of tumor vessels rather than tumor cells. The upregulation of VEGFR-3 on tumor blood vessels indicates a potential additional antiangiogenic effect for dual VEGFR-2/VEGFR-3-targeted therapy.


4D Genome Rewiring during Oncogene-Induced and Replicative Senescence.
To understand the role of the extensive senescence-associated 3D genome reorganization, we generated genome-wide chromatin interaction maps, epigenome, replication-timing, whole-genome bisulfite sequencing, and gene expression profiles from cells entering replicative senescence (RS) or upon oncogene-induced senescence (OIS). We identify senescence-associated heterochromatin domains (SAHDs). Differential intra- versus inter-SAHD interactions lead to the formation of senescence-associated heterochromatin foci (SAHFs) in OIS but not in RS. This OIS-specific configuration brings active genes located in genomic regions adjacent to SAHDs in close spatial proximity and favors their expression. We also identify DNMT1 as a factor that induces SAHFs by promoting HMGA2 expression. Upon DNMT1 depletion, OIS cells transition to a 3D genome conformation akin to that of cells in replicative senescence. These data show how multi-omics and imaging can identify critical features of RS and OIS and discover determinants of acute senescence and SAHF formation.


Impaired S-nitrosylation of the ryanodine receptor caused by xanthine oxidase activity contributes to calcium leak in heart failure.
S-Nitrosylation is a ubiquitous post-translational modification that regulates diverse biologic processes. In skeletal muscle, hypernitrosylation of the ryanodine receptor (RyR) causes sarcoplasmic reticulum (SR) calcium leak, but whether abnormalities of cardiac RyR nitrosylation contribute to dysfunction of cardiac excitation-contraction coupling remains controversial. In this study, we tested the hypothesis that cardiac RyR2 is hyponitrosylated in heart failure, because of nitroso-redox imbalance. We evaluated excitation-contraction coupling and nitroso-redox balance in spontaneously hypertensive heart failure rats with dilated cardiomyopathy and age-matched Wistar-Kyoto rats. Spontaneously hypertensive heart failure myocytes were characterized by depressed contractility, increased diastolic Ca(2+) leak, hyponitrosylation of RyR2, and enhanced xanthine oxidase derived superoxide. Global S-nitrosylation was decreased in failing hearts compared with nonfailing. Xanthine oxidase inhibition restored global and RyR2 nitrosylation and reversed the diastolic SR Ca(2+) leak, improving Ca(2+) handling and contractility. Together these findings demonstrate that nitroso-redox imbalance causes RyR2 oxidation, hyponitrosylation, and SR Ca(2+) leak, a hallmark of cardiac dysfunction. The reversal of this phenotype by inhibition of xanthine oxidase has important pathophysiologic and therapeutic implications.


Nitrogen removal via a single-stage PN-Anammox process in a novel combined biofilm reactor.
This study developed a partial nitrification (PN) and anaerobic ammonia oxidation (Anammox) process for treating high-ammonia wastewater using an innovative biofilm system in which ammonia oxidizing bacteria grew on fluidized Kaldnes (K1) carriers and Anammox bacteria grew on fixed acryl resin carriers. The airlift loop biofilm reactor (ALBR) was stably operated for more than 4 months under the following conditions: 35 ± 2 °C, pH 7.5-8.0 and dissolved oxygen (DO) of 0.5-3.5 mg/L. The results showed that the total nitrogen removal efficiency reached a maximum of 75% and the total nitrogen removal loading rate was above 0.4 kg/(d·m3). DO was the most efficient control parameter in the mixed biofilm system, and values below 1.5 mg/L were observed in the riser zone for the PN reaction, while values below 0.8 mg/L were observed in the downer zone for the Anammox reaction. Scanning electron microscopy and Fluorescence In Situ Hybridization images showed that most of the nitrifying bacteria were distributed on the K1 carriers and most of the Anammox bacteria were distributed within the acryl resin carriers. Therefore, the results indicate that the proposed combined biofilm system is easy to operate and efficient for the treatment of high-ammonia wastewater.


Molecular and Pathogenic Characterization of Fusarium redolens, a New Causal Agent of Fusarium Yellows in Chickpea.
The association of Fusarium redolens with wilting-like symptoms in chickpea in Lebanon, Morocco, Pakistan, and Spain is reported for the first time, together with the molecular and pathogenic characterization of isolates of the pathogen from chickpea of diverse geographic origin. Maximum parsimony analysis of sequences of the translation elongation factor 1α (TEF-1α) gene grouped all F. redolens isolates from chickpea in the same main clade. Pathogenicity assays using three chickpea cultivars and isolates from different geographic origins indicated that F. redolens is mildly virulent on chickpea. Moreover, infection of chickpea by F. redolens induces a disease syndrome similar to that caused by the yellowing pathotype of F. oxysporum f. sp. ciceris, including leaf yellowing and necrosis that develop upward from the stem base, and premature senescence of the plant. In contrast, F. redolens does not cause discoloration of the vascular tissues in chickpea but does cause brown necrotic lesions in the tap root and necrosis of lateral roots. F. redolens is not easily differentiated from F. oxysporum f. sp. ciceris using morphology-based diagnosis, and the two species cause similar symptoms on chickpea; therefore, the use of molecular protocols should help to avoid misdiagnoses of Fusarium yellows in chickpea.


Metformin modifies glutamine metabolism in an in vitro and in vivo model of hepatic encephalopathy.
to analyze the effect of metformin on ammonia production derived from glutamine metabolism in vitro and in vivo. twenty male Wistar rats were studied for 28 days after a porto-caval anastomosis (n = 16) or a sham operation (n = 4). Porto-caval shunted animals were randomized into two groups (n = 8) and either received 30 mg/kg/day of metformin for two weeks or were control animals. Plasma ammonia concentration, Gls gene expression and K-type glutaminase activity were measured in the small intestine, muscle and kidney. Furthermore, Caco2 were grown in different culture media containing glucose/glutamine as the main carbon source and exposed to different concentrations of the drug. The expression of genes implicated in glutamine metabolism were analyzed. metformin was associated with a significant inhibition of glutaminase activity levels in the small intestine of porto-caval shunted rats (0.277 ± 0.07 IU/mg vs 0.142 ± 0.04 IU/mg) and a significant decrease in plasma ammonia (204.3 ± 24.4 µg/dl vs 129.6 ± 16.1 µg/dl). Glucose withdrawal induced the expression of the glutamine transporter SLC1A5 (2.54 ± 0.33 fold change; p < 0.05). Metformin use reduced MYC levels in Caco2 and consequently, SLC1A5 and GLS expression, with a greater effect in cells dependent on glutaminolytic metabolism. metformin regulates ammonia homeostasis by modulating glutamine metabolism in the enterocyte, exerting an indirect control of both the uptake and degradation of glutamine. This entails a reduction in the production of metabolites and energy through this pathway and indirectly causes a decrease in ammonia production that could be related to a decreased risk of HE development.


Coupled effects of treated effluent irrigation and wetting-drying cycles on transport of triazines through unsaturated soil columns.
The physical and chemical parameters controlling the movement of atrazine (6-chloro-N2-ethyl-N4-isopropyl-l,3,5-triazine-2,4-diamine; 98.8%) and prometryn [N,N'-bis(1-methylethyl)-6-(methylthio)-l,3,5triazine-2,4-diamine; 99.5%] were investigated in columns infiltrated with treated effluent under unsaturated transient conditions and subjected to drying events at 22 or 60 degrees C followed by rewetting. Three soils varying in soil pH and texture and three solutions were used. The infiltrating solutions consisted of either a CaCl2 matrix (CC), a swine waste-derived lagoon effluent (SW), or a simulated buffer solution (SB) representative of the element composition and pH of the SW but with no dissolved organic matter. Several parameters were monitored including leachate triazine concentrations, pH, dissolved organic carbon (DOC), inorganic carbon, and flow rates. Compared with CC, application of SW and SB increased column leachate pH, enhanced dissolution of organic carbon and particle dispersion, and decreased average flow rates, which allowed for increased desorption time. The coupled effect of these processes enhanced movement of triazines in some cases, with SW generally having the greatest effect. The individual effect of increased pH was more pronounced for prometryn (pKa=4.05) versus atrazine (pKa=1.66), and most dramatic for the soil with the lowest initial pH. High-temperature drying, which simulated intensive evaporation, further enhanced the dissolution of soil organic matter and the reduction in leachate flow rates with SW and SB applications; however, the net effect under the experimental conditions employed varied with soil type. Relative to low-temperature drying, high-temperature drying in the silty clay loam-packed columns reduced pesticide migration.


Preschool children living in joint physical custody arrangements show less psychological symptoms than those living mostly or only with one parent.
Joint physical custody (JPC), where children spend about equal time in both parent's homes after parental separation, is increasing. The suitability of this practice for preschool children, with a need for predictability and continuity, has been questioned. In this cross-sectional study, we used data on 3656 Swedish children aged three to five years living in intact families, JPC, mostly with one parent or single care. Linear regression analyses were conducted with the Strengths and Difficulties Questionnaire, completed by parents and preschool teachers, as the outcome measure. Children in JPC showed less psychological problems than those living mostly (adjusted B 1.81; 95% CI [0.66 to 2.95]) or only with one parent (adjusted B 1.94; 95% CI [0.75 to 3.13]), in parental reports. In preschool teacher reports, the adjusted Betas were 1.27, 95% CI [0.14 to 2.40] and 1.41, 95% CI [0.24 to 2.58], respectively. In parental reports, children in JPC and those in intact families had similar outcomes, while teachers reported lower unadjusted symptom scores for children in intact families. Joint physical custody arrangements were not associated with more psychological symptoms in children aged 3-5, but longitudinal studies are needed to account for potential preseparation differences.


Sequence variants in SLC16A11 are a common risk factor for type 2 diabetes in Mexico.
Performing genetic studies in multiple human populations can identify disease risk alleles that are common in one population but rare in others, with the potential to illuminate pathophysiology, health disparities, and the population genetic origins of disease alleles. Here we analysed 9.2 million single nucleotide polymorphisms (SNPs) in each of 8,214 Mexicans and other Latin Americans: 3,848 with type 2 diabetes and 4,366 non-diabetic controls. In addition to replicating previous findings, we identified a novel locus associated with type 2 diabetes at genome-wide significance spanning the solute carriers SLC16A11 and SLC16A13 (P = 3.9 × 10(-13); odds ratio (OR) = 1.29). The association was stronger in younger, leaner people with type 2 diabetes, and replicated in independent samples (P = 1.1 × 10(-4); OR = 1.20). The risk haplotype carries four amino acid substitutions, all in SLC16A11; it is present at ~50% frequency in Native American samples and ~10% in east Asian, but is rare in European and African samples. Analysis of an archaic genome sequence indicated that the risk haplotype introgressed into modern humans via admixture with Neanderthals. The SLC16A11 messenger RNA is expressed in liver, and V5-tagged SLC16A11 protein localizes to the endoplasmic reticulum. Expression of SLC16A11 in heterologous cells alters lipid metabolism, most notably causing an increase in intracellular triacylglycerol levels. Despite type 2 diabetes having been well studied by genome-wide association studies in other populations, analysis in Mexican and Latin American individuals identified SLC16A11 as a novel candidate gene for type 2 diabetes with a possible role in triacylglycerol metabolism.


A unified design space of synthetic stripe-forming networks.
Synthetic biology is a promising tool to study the function and properties of gene regulatory networks. Gene circuits with predefined behaviours have been successfully built and modelled, but largely on a case-by-case basis. Here we go beyond individual networks and explore both computationally and synthetically the design space of possible dynamical mechanisms for 3-node stripe-forming networks. First, we computationally test every possible 3-node network for stripe formation in a morphogen gradient. We discover four different dynamical mechanisms to form a stripe and identify the minimal network of each group. Next, with the help of newly established engineering criteria we build these four networks synthetically and show that they indeed operate with four fundamentally distinct mechanisms. Finally, this close match between theory and experiment allows us to infer and subsequently build a 2-node network that represents the archetype of the explored design space.


C. elegans whole-genome sequencing reveals mutational signatures related to carcinogens and DNA repair deficiency.
Mutation is associated with developmental and hereditary disorders, aging, and cancer. While we understand some mutational processes operative in human disease, most remain mysterious. We used Caenorhabditis elegans whole-genome sequencing to model mutational signatures, analyzing 183 worm populations across 17 DNA repair-deficient backgrounds propagated for 20 generations or exposed to carcinogens. The baseline mutation rate in C. elegans was approximately one per genome per generation, not overtly altered across several DNA repair deficiencies over 20 generations. Telomere erosion led to complex chromosomal rearrangements initiated by breakage-fusion-bridge cycles and completed by simultaneously acquired, localized clusters of breakpoints. Aflatoxin B1 induced substitutions of guanines in a GpC context, as observed in aflatoxin-induced liver cancers. Mutational burden increased with impaired nucleotide excision repair. Cisplatin and mechlorethamine, DNA crosslinking agents, caused dose- and genotype-dependent signatures among indels, substitutions, and rearrangements. Strikingly, both agents induced clustered rearrangements resembling "chromoanasynthesis," a replication-based mutational signature seen in constitutional genomic disorders, suggesting that interstrand crosslinks may play a pathogenic role in such events. Cisplatin mutagenicity was most pronounced in xpf-1 mutants, suggesting that this gene critically protects cells against platinum chemotherapy. Thus, experimental model systems combined with genome sequencing can recapture and mechanistically explain mutational signatures associated with human disease.


Pol12, the B subunit of DNA polymerase alpha, functions in both telomere capping and length regulation.
The regulation of telomerase action, and its coordination with conventional DNA replication and chromosome end "capping," are still poorly understood. Here we describe a genetic screen in yeast for mutants with relaxed telomere length regulation, and the identification of Pol12, the B subunit of the DNA polymerase alpha (Pol1)-primase complex, as a new factor involved in this process. Unlike many POL1 and POL12 mutations, which also cause telomere elongation, the pol12-216 mutation described here does not lead to either reduced Pol1 function, increased telomeric single-stranded DNA, or a reduction in telomeric gene silencing. Instead, and again unlike mutations affecting POL1, pol12-216 is lethal in combination with a mutation in the telomere end-binding and capping protein Stn1. Significantly, Pol12 and Stn1 interact in both two-hybrid and biochemical assays, and their synthetic-lethal interaction appears to be caused, at least in part, by a loss of telomere capping. These data reveal a novel function for Pol12 and a new connection between DNA polymerase alpha and Stn1. We propose that Pol12, together with Stn1, plays a key role in linking telomerase action with the completion of lagging strand synthesis, and in a regulatory step required for telomere capping.


Articulatory Changes in Vowel Production following STN DBS and Levodopa Intake in Parkinson's Disease.
Purpose. To investigate the impact of deep brain stimulation of the subthalamic nucleus (STN DBS) and levodopa intake on vowel articulation in dysarthric speakers with Parkinson's disease (PD). Methods. Vowel articulation was assessed in seven Quebec French speakers diagnosed with idiopathic PD who underwent STN DBS. Assessments were conducted on- and off-medication, first prior to surgery and then 1 year later. All recordings were made on-stimulation. Vowel articulation was measured using acoustic vowel space and formant centralization ratio. Results. Compared to the period before surgery, vowel articulation was reduced after surgery when patients were off-medication, while it was better on-medication. The impact of levodopa intake on vowel articulation changed with STN DBS: before surgery, levodopa impaired articulation, while it no longer had a negative effect after surgery. Conclusions. These results indicate that while STN DBS could lead to a direct deterioration in articulation, it may indirectly improve it by reducing the levodopa dose required to manage motor symptoms. These findings suggest that, with respect to speech production, STN DBS and levodopa intake cannot be investigated separately because the two are intrinsically linked. Along with motor symptoms, speech production should be considered when optimizing therapeutic management of patients with PD.


Role of CT in the assessment of muscular venous branch invasion in patients with renal cell carcinoma.
The purpose of this article is to determine whether the relationship between a renal cell carcinoma and the renal sinus fat on contrast-enhanced CT could predict muscular venous branch invasion and the type of surgery needed. A total of 115 consecutive patients underwent pre-operative contrast-enhanced CT between August 2011 and December 2011. Without access to histopathologic information, on nephrographic phase contrast-enhanced CT images, two radiologists independently determined whether the renal tumor was in contact with the renal sinus fat or separated from the renal sinus fat. Interreader agreements and performance characteristics of imaging tests were calculated, and histopathologic analysis served as the standard of reference. Histopathologic analysis identified 115 renal tumors, 90% (103/115) of which were renal cell carcinomas. Thirty-nine percent (31/80) of renal cell carcinomas that abutted the renal sinus fat on CT displayed muscular venous branch invasion on histopathologic analysis. Patients with renal cell carcinomas separated from the renal sinus fat were more likely to undergo partial nephrectomies (96% [22/23]; p = 0.013). Sensitivity and specificity for the identification of muscular venous branch invasion on CT were 94% (95% CI, 80-99%) and 30% (20-42%), respectively. Interreader agreement of visual assessment was excellent (κ = 0.87; 95% CI, 0.81-0.92). If a renal cell carcinoma was separated from the renal sinus fat on CT, the likelihood of muscular venous branch invasion being identified by histopathologic analysis was significantly decreased, and the patient was more likely to undergo a partial nephrectomy.


Endoscopic selective neck dissection in a porcine model.
To investigate the feasibility of accomplishing a selective neck dissection (SND) endoscopically. Prospective, nonrandomized experimental investigation in a porcine model. Unilateral endoscopic SNDs were performed in Yorkshire pigs. A spacious operative pocket was developed using a combination of hernia balloon expansion followed by low-pressure (4 mm Hg) carbon dioxide insufflation. The sternomastoid muscle, thymus, submandibular gland, lymph nodes, and fibrofatty tissue were removed in a procedure approximating a human SND. Data (operative time, blood loss, arterial blood gas values, weight of the specimen, and complications) were prospectively recorded. The specimens were analyzed by a pathologist, and the number and size of lymph nodes were recorded. Fourteen endoscopic SNDs were successfully performed. No conversions to open surgery were necessary. The median operative time was 131 minutes (range, 95-235 minutes). The median estimated blood loss was 4 mL (range, 0-150 mL). The mean +/- SD specimen weight was 42.9 +/- 8.3 g; the mean number +/- SD of nodes retrieved from the neck specimen was 4.8 +/- 2.2, and the mean +/- SD maximal nodal dimension was 2.4 +/- 0.5 cm. The arterial PCO2 increased by an average of only 3.9 mm Hg from the beginning to the end of the surgery; correspondingly, the pH fell by only 0.02. There were no major complications, and no animals had to be euthanized prior to the completion of the procedure. Endoscopic neck dissection in a porcine model can be accomplished with a combination of strategies to overcome the dilemma of creating and maintaining an operative pocket. The merger of SND with endoscopic technology offers the promise of truly minimally invasive surgery for the node-negative neck.


Predicting Age From Optical Coherence Tomography Scans With Deep Learning.
To assess whether age can be predicted from deep learning analysis of peripapillary spectral-domain optical coherence tomography (SD-OCT) B-scans and to determine the importance of specific retinal areas on the predictions. Deep learning (DL) convolutional neural networks were developed to predict chronological age in healthy subjects using peripapillary SD-OCT B-scan images. Models were built using the whole B-scan, as well as using specific regions through image ablation. Cross-validation was used for training and testing the model. Mean absolute error (MAE) and correlations between predicted and observed age were used to evaluate model performance. A total of 7271 images from 542 eyes of 278 healthy subjects were included. DL predictions of age using the whole B-scan were strongly correlated with chronological age (MAE = 5.82 years; r = 0.860, P < 0.001). The model also accurately discriminated between the lowest and highest tertiles of age, with an area under the receiver operating characteristic curve of 0.962. In general, class activation maps tended to show a diffuse pattern of activation throughout the scan image. For specific structures of the B-scan, the layers with the strongest correlations with chronological age were the choroid and vitreous (both r = 0.736), whereas retinal nerve fiber layer had the lowest correlation (r = 0.492). A DL algorithm was able to accurately predict age from whole peripapillary SD-OCT B-scans. DL models applied to SD-OCT scans suggest that aging appears to affect several layers in the posterior eye segment.


ACONITASE 3 is part of theANAC017 transcription factor-dependent mitochondrial dysfunction response.
Mitochondria are tightly embedded within metabolic and regulatory networks that optimize plant performance in response to environmental challenges. The best-known mitochondrial retrograde signaling pathway involves stress-induced activation of the transcription factor NAC DOMAIN CONTAINING PROTEIN 17 (ANAC017), which initiates protective responses to stress-induced mitochondrial dysfunction in Arabidopsis (Arabidopsis thaliana). Posttranslational control of the elicited responses, however, remains poorly understood. Previous studies linked protein phosphatase 2A subunit PP2A-B'γ, a key negative regulator of stress responses, with reversible phosphorylation of ACONITASE 3 (ACO3). Here we report on ACO3 and its phosphorylation at Ser91 as key components of stress regulation that are induced by mitochondrial dysfunction. Targeted mass spectrometry-based proteomics revealed that the abundance and phosphorylation of ACO3 increased under stress, which required signaling through ANAC017. Phosphomimetic mutation at ACO3-Ser91 and accumulation of ACO3S91D-YFP promoted the expression of genes related to mitochondrial dysfunction. Furthermore, ACO3 contributed to plant tolerance against ultraviolet B (UV-B) or antimycin A-induced mitochondrial dysfunction. These findings demonstrate that ACO3 is both a target and mediator of mitochondrial dysfunction signaling, and critical for achieving stress tolerance in Arabidopsis leaves.


Spatial classification of youth physical activity patterns.
Physical activity is an essential element in reducing the prevalence of obesity, but much is unknown about the intensity and location of physical activity among youth-this is important because adolescent health behaviors are predictive of behaviors in adults. This study aims to identify the locations where youth moderate-to-vigorous physical activity (MVPA) occurs, and to examine how MVPA varies according to urbanicity (urban, suburban, rural). Participants included adolescent students (N=380, aged 12-16 years) from Halifax, Nova Scotia. Locations of MVPA were measured using accelerometers and GPS data loggers for up to 7 days. Specialized software was developed to integrate and process the data. Frequencies of MVPA by location were determined, and differences in MVPA were assessed for association with urbanicity. Active commuting accounted for the largest proportion of time in MVPA among urban and suburban students. Rural students achieved most MVPA at school. Other residential locations, shopping centers, and green spaces accounted for a majority of the remaining MVPA. Minutes in MVPA varied significantly overall (196.6 ± 163.8, 84.9 ± 103.2, 81.7 ± 98.2); at school (45.7 ± 45.2, 18.6 ± 28.0, 29.8 ± 39.7); while commuting (110.3 ± 107.1, 31.5 ± 55.2, 19.5 ± 39.7); and at other activity locations (19.7 ± 27.1, 14.8 ± 26.8, 12.0 ± 22.1) and by urbanicity. Findings reveal that the journeys between locations are as important as home and school settings in contributing to greater MVPA in adolescent youth. The relative importance of context as a contributor to MVPA varies with urbanicity. Combining actimetry and GPS data provides a precise link between physical activity measurements and contexts of the built environment.


Predictive and prognostic impact of TP53 mutations and MDM2 promoter genotype in primary breast cancer patients treated with epirubicin or paclitaxel.
TP53 mutations have been associated with resistance to anthracyclines but not to taxanes in breast cancer patients. The MDM2 promoter single nucleotide polymorphism (SNP) T309G increases MDM2 activity and may reduce wild-type p53 protein activity. Here, we explored the predictive and prognostic value of TP53 and CHEK2 mutation status together with MDM2 SNP309 genotype in stage III breast cancer patients receiving paclitaxel or epirubicin monotherapy. Each patient was randomly assigned to treatment with epirubicin 90 mg/m(2) (n = 109) or paclitaxel 200 mg/m(2) (n = 114) every 3rd week as monotherapy for 4-6 cycles. Patients obtaining a suboptimal response on first-line treatment requiring further chemotherapy received the opposite regimen. Time from last patient inclusion to follow-up censoring was 69 months. Each patient had snap-frozen tumor tissue specimens collected prior to commencing chemotherapy. While TP53 and CHEK2 mutations predicted resistance to epirubicin, MDM2 status did not. Neither TP53/CHEK2 mutations nor MDM2 status was associated with paclitaxel response. Remarkably, TP53 mutations (p = 0.007) but also MDM2 309TG/GG genotype status (p = 0.012) were associated with a poor disease-specific survival among patients having paclitaxel but not patients having epirubicin first-line. The effect of MDM2 status was observed among individuals harbouring wild-type TP53 (p = 0.039) but not among individuals with TP53 mutated tumors (p>0.5). TP53 and CHEK2 mutations were associated with lack of response to epirubicin monotherapy. In contrast, TP53 mutations and MDM2 309G allele status conferred poor disease-specific survival among patients treated with primary paclitaxel but not epirubicin monotherapy.


Nutrition intervention is beneficial in oncology outpatients receiving radiotherapy to the gastrointestinal or head and neck area.
Malnutrition occurs frequently in patients with cancer of the gastrointestinal (GI) or head and neck area and can lead to negative outcomes. The aim of this study is to determine the impact of early and intensive nutrition intervention (NI) on body weight, body composition, nutritional status, global quality of life (QoL) and physical function compared to usual practice in oncology outpatients receiving radiotherapy to the GI or head and neck area. Outpatients commencing at least 20 fractions of radiotherapy to the GI or head and neck area were randomised to receive intensive, individualised nutrition counselling by a dietitian using a standard protocol and oral supplements if required, or the usual practice of the centre (general advice and nutrition booklet). Outcome parameters were measured at baseline and 4, 8 and 12 weeks after commencing radiotherapy using valid and reliable tools. A total of 60 patients (51 M : 9 F; mean age 61.9+/-14.0 years) were randomised to receive either NI (n=29) or usual care (UC) (n=31). The NI group had statistically smaller deteriorations in weight (P<0.001), nutritional status (P=0.020) and global QoL (P=0.009) compared with those receiving UC. Clinically, but not statistically significant differences in fat-free mass were observed between the groups (P=0.195). Early and intensive NI appears beneficial in terms of minimising weight loss, deterioration in nutritional status, global QoL and physical function in oncology outpatients receiving radiotherapy to the GI or head and neck area. Weight maintenance in this population leads to beneficial outcomes and suggests that this, rather than weight gain, may be a more appropriate aim of NI.


Systematic review of the measurement properties of self-report physical activity questionnaires in healthy adult populations.
This systematic review evaluated the measurement properties of current self-report physical activity questionnaires (SRPAQs) completed within healthy adult populations. Two reviewers independently searched seven electronic databases and hand searched for articles investigating measurement properties of a SRPAQ evaluating physical activity over the previous 6 months. Articles published from 1 May 2001 to 4 December 2014 were systematically screened and eligible studies were not limited to English language sources. Articles investigating specific race, gender or socioeconomic populations were excluded. 10 studies investigating 10 SRPAQs were included. The methodological quality of the included studies was evaluated using COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) and ranged from 'poor' to 'good'. The Recent Physical Activity Questionnaire, International Physical Activity Questionnaires and Physical Activity Assessment Tool demonstrated good/excellent test-retest reliability (intra-class coefficient (ICC)=0.76, p<0.0001; r=0.627-0.91; r=0.618, p<0.001, respectively), but variable criterion validity (r=0.67, p<0.0001; r=-0.02-0.43; r=0.392, p<0.01, respectively). The single-item measure showed significant criterion validity against an accelerometer (for moderate to vigorous physical activity (MVPA) k=0.23, 95% CI 0.05 to 0.41; and physical activity ≥10 min bouts 0.39 (95% CI 0.14 to 0.64). Construct validity of the six-point scale and Human Activity Profile varied significantly with age, marital status and presence of comorbidities (p<0.05, <0.01, <0.000 and p<0.05, <0.05, <0.000, respectively). The 1 week Godlin-Shephard recall demonstrated 'moderate' validity with the gold standard measure of accelerometry (r=0.43). Inconclusive evidence exists. Further investigation of criterion validity of the short-form International Physical Activity Questionnaire is required, as it demonstrated excellent test-retest reliability. CRD42012002484.


Cause-specific mortality among physicians with differing life-styles.
We compare the mortality experience of medical school graduates from Loma Linda University (LLU [n = 4342]) and the University of Southern California (USC [n = 2832]) with each other and with that of contemporaneous, white, American men. When compared with US white men from the general population, both USC and LLU graduates had below expected deaths for all causes (USC, standardized mortality ratio [SMR] = 76; LLU, SMR = 56), although deaths due to cerebrovascular disease, airplane accidents, and suicides were elevated for USC (SMRs = 132, 360, and 218, respectively). The LLU graduates had a risk similar to that of the USC graduates for fatal cancer, with a mortality ratio (MR) of 0.92 (95% confidence interval, 0.67 to 1.26); but half the risk of fatal atherosclerotic disease, with MRs of 0.58 (0.46 to 0.73) and 0.66 (0.43 to 0.99) for coronary and cerebrovascular disease, respectively; and three times the risk of fatal airplane accidents. The overall mortality rate of LLU physicians was only 75% as high as that of the USC physicians and only 56% as high as that of the US male population at large. We attribute this reduced mortality mainly to the low cardiovascular mortality rates, which may be accounted for by the life-style of the substantial proportion of Seventh-day Adventists among LLU graduates.


Cellular and metabolic basis of Clara cell tolerance to multiple doses of cytochrome P450-activated cytotoxicants. I: Bronchiolar epithelial reorganization and expression of cytochrome P450 monooxygenases in mice exposed to multiple doses of naphthalene.
The objectives of this study were to quantitatively define alterations in the epithelial populations and expression of cytochrome P450 monooxygenases in distal airways which are associated with the tolerance resulting from repeated exposure of mice to the acute Clara cell toxicant, naphthalene. Bronchiolar epithelium of mice treated daily for 7 days with an acutely cytotoxic dose of naphthalene (200 mg/kg/day) quantitatively resembles the bronchiolar epithelium of control animals in terms of proportions of ciliated and nonciliated cells and nuclear and cytoplasmic volumes. Subsequent treatment of tolerant animals with higher doses (300 mg/kg) does not produce the same Clara cell injury observed in previously untreated mice after a single treatment with 300 mg/kg. After repeated exposures to naphthalene, cellular expression of immunoreactive cytochrome P450 monooxygenases 2B and 2F, P450 reductase and Clara cell secretory protein significantly decreased in bronchiolar epithelium. Although metabolism of naphthalene to the 1R,2S-naphthalene oxide is depressed in microsomes derived from whole-lung homogenates of tolerant animals, metabolism of naphthalene in distal airways isolated by microdissection is unchanged from the controls at saturating substrate concentrations. When substrate concentrations are less than the apparent K(m) for naphthalene (< 0.05 mM), the rate of naphthalene metabolite production is markedly lower in distal airways of tolerant mice compared with controls. Covalent binding of reactive naphthalene metabolites in lungs of tolerant mice is also unchanged from control. In conclusion, after repeated exposure to short-term cytotoxic doses of naphthalene distal bronchioles of tolerant mice 1) are lined by epithelium which resembles that of controls; 2) express lower levels of P450 proteins; 3) have reduced levels of naphthalene monooxygenase activity, but only at less than saturating concentrations and 4) have no decrease in covalent binding of reactive metabolites to proteins.


Visibility Graph Analysis of Intraspinal Pressure Signal Predicts Functional Outcome in Spinal Cord Injured Patients.
To guide management of patients with acute spinal cord injuries, we developed intraspinal pressure monitoring from the injury site. Here, we examine the complex fluctuations in the intraspinal pressure signal using network theory. We analyzed 7097 h of intraspinal pressure data from 58 patients with severe cord injuries. Intraspinal pressure signals were split into hourly windows. Each window was mapped into a visibility graph as follows. Vertical bars were drawn at 0.1 Hz representing signal amplitudes. Each bar produced a node, thus totalling 360 nodes per graph. Two nodes were linked with an edge if the straight line through the nodes did not intersect a bar. We computed several topological metrics for each graph including diameter, modularity, eccentricity, and small-worldness. Patients were followed up for 20 months on average. Our data show that the topological structure of intraspinal pressure visibility graphs is highly sensitive to pathological events at the injury site, including cord compression (high intraspinal pressure), ischemia (low spinal cord perfusion pressure), and deranged autoregulation (high spinal pressure reactivity index). These pathological changes correlate with long graph diameter, high modularity, high eccentricity and reduced small-worldness. In a multivariate logistic regression model, age, neurological status on admission, and average node eccentricity were independent predictors of neurological improvement. We conclude that analysis of intraspinal pressure fluctuations after spinal cord injury as graphs, rather than as time series, captures clinically important information. Our novel technique may be applied to other signals recorded from injured central nervous system (CNS); for example, intracranial pressure, tissue metabolite, and oxygen levels.


Effect of movement on SEPs generated by dorsal column nuclei.
To investigate the effect of the voluntary movement on the amplitude of the somatosensory evoked potentials (SEPs) recorded by an epidural electrode at level of the dorsal column nuclei (DCN). Five patients, suffering from chronic pain resistant to pharmacological treatment, underwent an epidural electrode implant at high cervical spinal cord level (C2) for neuromodulation. After tibial nerve stimulation, SEPs were recorded from the epidural electrode contacts, from a Cz lead, and from two electrodes placed over the 12th dorsal vertebra and 4th lumbar vertebra, respectively. SEPs were recorded at rest and during a voluntary flexo-extension movement of the stimulated foot. Beyond the low-frequency SEPs, also the high-frequency oscillations (HFOs), obtained by filtering the recorded traces by means of a 1000-2000 Hz bandpass offline, were analysed. The epidural electrode contacts recorded a triphasic potential (P1-N1-P2), whose negative peak showed a latency similar to that of the P30 far-field response obtained from the scalp. The epidural potential amplitude was significantly decreased by the voluntary movement, as compared to the rest (p<0.01). A main HFO peak, identifiable at around 1200 Hz, was significantly lower in amplitude during movement than at rest (p=0.008). Our findings suggest that the epidural C2 triphasic wave is a potential arising from DCN. The low-frequency epidural SEP component is subtended by a 1200 Hz HFO, probably generated by post-synaptic events. The amplitude reduction of the DCN response during movement is possibly due to decreased excitability of the DCN neurons receiving the somatosensory ascending input.


Advanced Steel Microstructural Classification by Deep Learning Methods.
The inner structure of a material is called microstructure. It stores the genesis of a material and determines all its physical and chemical properties. While microstructural characterization is widely spread and well known, the microstructural classification is mostly done manually by human experts, which gives rise to uncertainties due to subjectivity. Since the microstructure could be a combination of different phases or constituents with complex substructures its automatic classification is very challenging and only a few prior studies exist. Prior works focused on designed and engineered features by experts and classified microstructures separately from the feature extraction step. Recently, Deep Learning methods have shown strong performance in vision applications by learning the features from data together with the classification step. In this work, we propose a Deep Learning method for microstructural classification in the examples of certain microstructural constituents of low carbon steel. This novel method employs pixel-wise segmentation via Fully Convolutional Neural Network (FCNN) accompanied by a max-voting scheme. Our system achieves 93.94% classification accuracy, drastically outperforming the state-of-the-art method of 48.89% accuracy. Beyond the strong performance of our method, this line of research offers a more robust and first of all objective way for the difficult task of steel quality appreciation.


Characterising and predicting cyanobacterial blooms in an 8-year amplicon sequencing time course.
Cyanobacterial blooms occur in lakes worldwide, producing toxins that pose a serious public health threat. Eutrophication caused by human activities and warmer temperatures both contribute to blooms, but it is still difficult to predict precisely when and where blooms will occur. One reason that prediction is so difficult is that blooms can be caused by different species or genera of cyanobacteria, which may interact with other bacteria and respond to a variety of environmental cues. Here we used a deep 16S amplicon sequencing approach to profile the bacterial community in eutrophic Lake Champlain over time, to characterise the composition and repeatability of cyanobacterial blooms, and to determine the potential for blooms to be predicted based on time course sequence data. Our analysis, based on 135 samples between 2006 and 2013, spans multiple bloom events. We found that bloom events significantly alter the bacterial community without reducing overall diversity, suggesting that a distinct microbial community-including non-cyanobacteria-prospers during the bloom. We also observed that the community changes cyclically over the course of a year, with a repeatable pattern from year to year. This suggests that, in principle, bloom events are predictable. We used probabilistic assemblages of OTUs to characterise the bloom-associated community, and to classify samples into bloom or non-bloom categories, achieving up to 92% classification accuracy (86% after excluding cyanobacterial sequences). Finally, using symbolic regression, we were able to predict the start date of a bloom with 78-92% accuracy (depending on the data used for model training), and found that sequence data was a better predictor than environmental variables.


Implementation of a successful incentive-based ultrasound credentialing program for emergency physicians.
With the rapid expansion of emergency ultrasound, resident education in ultrasound has become more clearly developed and broadly integrated. However, there still exists a lack of guidance in the training of physicians already in practice to become competent in this valuable skill. We sought to employ a step-wise, goal-directed, incentive-based credentialing program to educate emergency physicians in the use of emergency ultrasound. Successful completion of this program was the primary outcome. The goal was for the physicians to gain competency in 8 basic ultrasound examinations types: aorta, focused assessment with sonography in trauma, cardiac, renal, biliary, transabdominal pelvic, transvaginal pelvic, and deep venous thrombosis. We separated the 2.5 year training program into 4 distinct blocks, with each block focusing on 2 of the ultrasound examination types. Each block consisted of didactic and hands-on sessions with the goal of the physician completing 25 technically-adequate studies of each examination type. There was a financial incentive associated with completion of these requirements. A total of 31 physicians participated in the training program. Only one physician, who retired prior to the end of the 2.5 year period, did not successfully complete the program. All have applied for and received hospital privileging in emergency ultrasound and incorporated it into their daily practice. We found that a step-wise, incentive-based ultrasound training program with a combination of didactics and ample hands-on teaching was successful in the training of physicians already in practice.


Longitudinal Imaging of Heart Development With Optical Coherence Tomography.
Optical coherence tomography (OCT) has great potential for deciphering the role of mechanics in normal and abnormal heart development. OCT images tissue microstructure and blood flow deep into the tissue (1-2mm) at high spatiotemporal resolutions allowing unprecedented images of the developing heart. Here, we review the advancement of OCT technology to image heart development and report some of our recent findings utilizing OCT imaging under environmental control for longitudinal imaging. Precise control of the environment is absolutely required in longitudinal studies that follow the growth of the embryo or studies comparing normal versus perturbed heart development to obtain meaningful in vivo results. These types of studies are essential to tease out the influence of cardiac dynamics on molecular expression and their role in the progression of congenital heart defects.


Spirituality as experienced by occupational therapists in practice.
Despite considerable literature describing the potential place of spirituality in occupational therapy, surveys repeatedly demonstrate that therapists are uncomfortable with this concept in practice. To gain a better understanding of how spirituality might inform practice, we interviewed eight occupational therapists who stated that they considered spirituality while working with patients. Participants defined spirituality as one's beliefs about the world and one's place in it and how one lives out these beliefs, through reflection and conscious actions. Four themes of consideration of spirituality in practice emerged. In the first, addressing religious concerns, therapists dealt with patients' religious questions and issues. In the second, addressing suffering, therapists assisted patients to deal with their feelings related to loss and pain, attempted to relieve patients' distress and helped patients move towards increased functioning. In the third, encouraging the self, therapists worked to assist patients to acknowledge their own worth and to use their unique gifts and interests. In the fourth, growing as a person, therapists themselves were transformed as a result of the therapeutic encounter. This study represents an early attempt to determine what occupational therapists who consider spirituality actually do in practice. Future research should go deeper into the experience of such therapists, to gain a richer understanding of the phenomenon.


Faster than fiber: over 100-Gb/s signal delivery in fiber wireless integration system.
We summarize several different approaches for the realization of large capacity (>100Gb/s) fiber wireless integration system, including optical polarization-division-multiplexing (PDM) combined with multiple-input multiple-output (MIMO) reception, advanced multi-level modulation, optical multi-carrier modulation, electrical multi-carrier modulation, antenna polarization multiplexing and multi-band multiplexing. These approaches can effectively reduce the signal baud rate as well as the required bandwidth for optical and electrical devices. We also investigate the problems, such as wireless multi-path effect due to different wireless transmission distance, existing in the large capacity fiber wireless integration system. We demonstrate these problems can be effectively solved based on advanced digital-signal-processing (DSP) algorithms including classic constant modulus algorithm (CMA). Moreover, based on the combination of these approaches as well as advanced DSP algorithms, we have successfully demonstrated a 400G fiber wireless integration system, which creates a capacity record of wireless delivery and ushers in a new era of ultra-high bit rate (>400Gb/s) optical wireless integration communications at mm-wave frequencies.


Nonlinear Elastic and Inelastic Properties of Cells.
Mechanical forces play an important role in various physiological processes, such as morphogenesis, cytokinesis, and migration. Thus, in order to illuminate mechanisms underlying these physiological processes, it is crucial to understand how cells deform and respond to external mechanical stimuli. During recent decades, the mechanical properties of cells have been studied extensively using diverse measurement techniques. A number of experimental studies have shown that cells are far from linear elastic materials. Cells exhibit a wide variety of nonlinear elastic and inelastic properties. Such complicated properties of cells are known to emerge from unique mechanical characteristics of cellular components. In this review, we introduce major cellular components that largely govern cell mechanical properties and provide brief explanations of several experimental techniques used for rheological measurements of cell mechanics. Then, we discuss the representative nonlinear elastic and inelastic properties of cells. Finally, continuum and discrete computational models of cell mechanics, which model both nonlinear elastic and inelastic properties of cells, will be described.


What's therapeutic about the therapeutic milieu?
While the milieu of an inpatient facility is considered a treatment modality, extant literature focuses on the staff's role in creating the milieu rather than the patient's perception of it. Not since Goffman's Asylums (1961) has there been an in-depth examination of the phenomenal world of the hospitalized psychiatric patient. In this study, eight inpatients (ages 23 to 58) on the acute psychiatric unit of a metropolitan general hospital participated in phenomenological interviews about their experience of the environment. The essential meaning of the hospital was refuge from self-destructiveness. Prominent aspects of patients' experience within the place of refuge were three interrelated themes (like me/not like me, possibilities/no possibilities, and connection/disconnection). Universally, patients perceived peer-administered "therapy" as the most beneficial aspect of their hospitalization. They expressed longing for a deeper connection with staff and more intensive insight-oriented therapies. Although their needs for safety, structure, and medication were met, patients were not gaining greater understanding of their dysfunctional patterns of behavior. Renewed emphasis must be placed on the nurse-patient relationship and the therapeutic alliance.


Abnormal synaptic protein expression and cell death in murine scrapie.
Reduced expression of synaptophysin p38, synaptic-associated protein of molecular weight 25,000 (SNAP-25), syntaxin-1, synapsin-1, and alpha- and beta-synuclein, matching the distribution of spongiform degeneration, was found in the neurological phase of scrapie-infected mice. In addition, synaptophysin and SNAP-25 were accumulated in isolated neurons, mainly in the thalamus, midbrain and pons, and granular deposits of alpha- and beta-synuclein were present in the neuropil of the same areas. No modifications in the steady state levels of Bcl-2, Bax, Fas and Fas ligand were observed following infection. Yet antibodies against the c-Jun N-terminal peptide, which cross-react with products emerging after caspase-mediate proteolysis, recognize coarse granular deposits in the cytoplasm of reactive microglia. In situ end-labeling of nuclear DNA fragmentation showed positive nuclei with extreme chromatin condensation in the thalamus, pons, hippocampus and, in particular, the granular layer of the cerebellum. More importantly, expression of cleaved caspase-3, a major executioner of apoptosis, was seen in a few cells in the same regions, thus indicating that cell death by apoptosis in scrapie-infected mice is associated with caspase-3 activation. The present findings support the concept that synaptic pathology is a major substrate of neurological impairment and that caspase-3 activation may play a pivotal role in apoptosis in experimental scrapie. However, there is no correlation between decreased synaptic protein expression and caspase-3-associated apoptosis, which suggests that in addition to abnormal prion protein deposition, there may be other factors that distinctively influence synaptic vulnerability and cell death in murine scrapie.


Evaluation of Earbud and Wristwatch Heart Rate Monitors during Aerobic and Resistance Training.
Assessment of biometrics during exercise is evolving to create devices that are "all-inclusive", in an effort to decrease the number of devices required during exercise while providing comprehensive and accurate biometric measures. The purpose of this study was to determine the accuracy of two optical heart rate monitors, the Jabra earbud and the Mio Alpha wristwatch, during aerobic and anaerobic exercise. Twenty-two recreationally active participants (25.4 ± 6.9 years, 171 ± 11 cm, 73.9 ± 3.1 kg, and 25.2 ± 9.2% body fat) completed this study. Participants completed 30 minutes of treadmill activity, 25 minutes of high-intensity interval exercise (HIT), and 40 minutes of continuous outdoor activity of their choice, walking or running. Three heart rate (HR) monitors, (Polar chest strap, Mio Alpha, Jabra earbud) were worn during all exercises, with the Polar chest strap serving as the benchmark. HR was assessed in one-second intervals. Analyses included mean bias, mean absolute percent error (MAPE), and Lin's concordance coefficient. Overall, the Mio Alpha had a MAPE of 5.73 ± 10.19% and a moderate correlation with the benchmark, r(c) = 0.771, performing better in the treadmill and outdoor conditions. The Jabra earbud had a MAPE of 3.14 ± 6.13%, and a high correlation with the benchmark, r(c) = 0.939, performing well in all three conditions. Placing a HR monitor in an earbud is a viable option for obtaining an accurate HR assessment during different types of exercise. The accuracy of the Mio Alpha was likely affected by wrist movements during the HIT training.


Association of muscle strength, power, and optimal shortening velocity with functional abilities of women with chronic osteoarthritis participating in a multi-modal exercise program.
We assessed the relative association of quadriceps muscle strength and power as well as optimal shortening velocity (υ(opt)) to physical functioning in 28 women aged 50-87 years with chronic osteoarthritis participating in a three-week multimodal exercise program. Quadriceps muscle strength, power, υ(opt), and functional performance using the Activities of Daily Living (ADL) scale, Timed Up & Go (TUG) test, Tinetti test, and 6-Minute Walking Test (6-MWT) were assessed pre- and postrehabilitation. With rehabilitation, patients improved the values of strength, power, and the results of all functional tests. Both at baseline and postrehabilitation, functional status was more strongly related to power and υ(opt) than to strength. Functional gains obtained with rehabilitation were not related to changes in power or υ(opt), and only very modestly related to changes in strength. Future studies should assess the benefits and feasibility of power- and velocity-oriented training in patients with osteoarthritis.


Impact of antibiotic treatment duration on outcomes in older men with suspected urinary tract infection: Retrospective cohort study.
Clinical guidelines recommend at least 7 days of antibiotic treatment for older men with urinary tract infection (UTI). There may be potential benefits for patients, health services, and antimicrobial stewardship if shorter antibiotic treatment resulted in similar outcomes. We aimed to determine if treatment duration could be reduced by estimating risk of adverse outcomes according to different prescription durations. This retrospective cohort study included men aged greater than or equal to 65 years old with a suspected UTI. We compared outcomes in men prescribed 3, 5, 7, and 8 to 14 days of antibiotic treatment in a multivariable logistic regression analysis and 3 versus 7 days in a propensity-score matched analysis. Our outcomes were reconsultation and represcription (proxy for treatment failure), hospitalisation for UTI, sepsis, or acute kidney injury (AKI), and death. Of 360 640 men aged greater than or equal to 65 years, 33 745 (9.4%) had a UTI. Compared with 7 days, men prescribed 3-day treatment had greater odds of reconsultation and represcription (adjusted OR 1.48; 95% CI, 1.25-1.74) but lower odds of AKI hospitalisation (adjusted OR 0.66; 95% CI, 0.45-0.97). We estimated that treating 150 older men with 3 days instead of 7 days of antibiotics could result in four extra reconsultation and represcriptions and one less AKI hospitalisation. We estimated annual prescription cost savings at around £2.2 million. Antibiotic treatment for older men with suspected UTI could be reduced to 3 days, albeit with a small increase in risk of treatment failure. A definitive randomised trial is urgently needed.


Psychosocial, Physical, and Neurophysiological Risk Factors for Chronic Neck Pain: A Prospective Inception Cohort Study.
The purpose of this investigation was to identify modifiable risk factors for the development of first-onset chronic neck pain among an inception cohort of healthy individuals working in a high-risk occupation. Candidate risk factors identified from previous studies were categorized into psychosocial, physical, and neurophysiological domains, which were assessed concurrently in a baseline evaluation of 171 office workers within the first 3 months of hire. Participants completed monthly online surveys over the subsequent year to identify the presence of chronic interfering neck pain, defined as a Neck Disability Index score ≥5 points for 3 or more months. Data were analyzed using backward logistic regression to identify significant predictors within each domain, which were then entered into a multivariate regression model adjusted for age, sex, and body mass index. Development of chronic interfering neck pain was predicted by depressed mood (odds ratio [OR] = 3.36, 95% confidence interval [CI] = 1.10-10.31, P = .03), cervical extensor endurance (OR = .92, 95% CI, .87-.97, P = .001), and diffuse noxious inhibitory control (OR = .90, 95% CI, .83-.98, P = .02) at baseline. These findings provide the first evidence that individuals with preexisting impairments in mood and descending pain modulation may be at greater risk for developing chronic neck pain when exposed to peripheral nociceptive stimuli such as that produced during muscle fatigue. Depressed mood, poor muscle endurance, and impaired endogenous pain inhibition are predisposing factors for the development of new-onset chronic neck pain of nonspecific origin in office workers. These findings may assist with primary prevention by allowing clinicians to screen for individuals at risk of developing chronic neck pain.


Effect of cervical spinal adjustments on lumbar paraspinal muscle tone: evidence for facilitation of intersegmental tonic neck reflexes.
To determine whether spinal adjustments, delivered to the upper vs. lower cervical spine, might result in tonic neck reflex-induced alterations in the activity of the lumbar paraspinal musculature. Randomized, double-blind, before/after treatment comparisons. Cervical Ergonomics Laboratory, Palmer College of Chiropractic-West, Sunnyvale, CA. Healthy, nonsymptomatic chiropractic college students, about evenly divided with respect to gender and ranging from 23-38 yr of age. Modified "diversified" spinal adjustments, delivered bilaterally to either the upper (C2) or lower (C7) cervical region. Tissue compliance measures using a tissue compliance meter, obtained from each subject at sites 2 cm on either side of the spinous processes of L1, L2, L3, L4 and L5 both prior to and within 15 min following treatments. Upper cervical adjustments produced changes in lumbar tissue compliance which were only slight (p < .05) and not significantly different from that which occurred following upper cervical sham manipulation (p > .1). However, lower cervical adjustments induced increases in tissue compliance (decreases in tone) which were highly significant (p < .001) and relatively robust compared to those found following upper cervical adjustments (p < .01). Furthermore, the greatest effects were observed on either side of the L4 and L5 spinous processes, suggesting influences on the gluteal musculature in particular. Results indicate that cervical spinal manipulation can have significant effects on the tone of the lumbopelvic musculature, presumably by facilitating tonic neck reflexes involving intersegmental spinal pathways.


A Modification of the Fibular Osteotomy for Total Ankle Replacement Through the Lateral Transfibular Approach.
One disadvantage of lateral transfibular total ankle arthroplasty is the rate of symptoms related to the implant and wound issues requiring implant removal in association with the traditional fibular osteotomy. In the present study, lateral total ankle arthroplasty involving the traditional short oblique fibular osteotomy was compared with arthroplasty involving a long oblique osteotomy (the Foot & Ankle Reconstruction Group osteotomy). We retrospectively reviewed all primary lateral total ankle arthroplasties that had been performed by a single surgeon from May 2013 to October 2016 and had a minimum of 2 years of follow-up. Clinical assessment included patient demographics, wound complications, the need for implant removal, the pain score on a visual analog scale, the American Orthopaedic Foot & Ankle Society score, and the Short Form-12 Mental and Physical Component Summary scores. Weight-bearing radiographs were used to assess tibiotalar alignment, implant alignment, and fibular osteotomy healing. One hundred and fifty-nine total ankle arthroplasties were identified. The traditional short fibular osteotomy was used in 50 ankles, and the long oblique osteotomy was used in 109. The rate of survival of the tibial and talar components of the ankle replacements was 100%, and there were no osteotomy nonunions. There was improvement in all clinical parameters, with no significant difference between the 2 groups. Radiographs showed excellent arthroplasty alignment at all time points, with no significant difference between the groups. There was a significant difference in the rates of wound dehiscence (p = 0.011) and fibular implant removal (p < 0.0001), with the long oblique osteotomy having lower rates of both (2.8% and 2.8%, respectively) compared with the short oblique osteotomy (14% and 28%, respectively). In total ankle arthroplasty, modification of the traditional short oblique fibular osteotomy to a long oblique configuration provided excellent 2-year survival rates with good clinical and radiographic outcomes and decreased rates of wound complications and symptoms related to the fibular implant. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


Sulfated oligosaccharides (heparin and fucoidan) binding and dimerization of stromal cell-derived factor-1 (SDF-1/CXCL 12) are coupled as evidenced by affinity CE-MS analysis.
Chemokine stromal cell-derived factor-1 (SDF-1) is a potent chemoattractant involved in leukocyte trafficking and metastasis. Heparan sulfate on the cell surface binds SDF-1 and may modulate its function as a coreceptor of this chemokine. A major effect of the glycosaminoglycan binding may be on the quaternary structure of SDF-1, which has been controversially reported as a monomer or a dimer. We have investigated the effect of sulfated oligosaccharides on the oligomerization of SDF-1 and of its mutated form SDF-1 (3/6), using affinity capillary electrophoresis (ACE) hyphenated to mass spectrometry (MS). Coupled to MS, ACE allowed the study for the first time of the effect of size-defined oligosaccharides on the quaternary organization of SDF-1 in muM range concentrations, i.e., lower values than the mM values previously reported in NMR, light scattering, and ultracentrifugation experiments. Our results showed that in the absence of sulfated oligosaccharides, SDF-1 is mostly monomeric in solution. However, dimer formation was observed upon interaction with heparin-sulfated oligosaccharides despite the mM Kd values for dimerization. A SDF-1/oligosaccharide 2/1 complex was detected, indicating that oligosaccharide binding promoted the dimerization of SDF-1. Heparin tetrasaccharide but not disaccharide promoted dimer formation, suggesting that the dimer required to be stabilized by a long enough bound oligosaccharide. The SDF-1/oligosaccharide 1/1 complex was only observed with heparin disaccharide and fucoidan pentasaccharide, pointing out the role of specific structural determinants in promoting dimer formation. These results underline the importance of dimerization induced by glycosaminoglycans for chemokine functionality.


Adaptive variation for growth and resistance to a novel pathogen along climatic gradients in a foundation tree.
Natural ecosystems are under pressure from increasing abiotic and biotic stressors, including climate change and novel pathogens, which are putting species at risk of local extinction, and altering community structure, composition and function. Here, we aim to assess adaptive variation in growth and fungal disease resistance within a foundation tree, Corymbia calophylla to determine local adaptation, trait heritability and genetic constraints in adapting to future environments. Two experimental planting sites were established in regions of contrasting rainfall with seed families from 18 populations capturing a wide range of climate origins (~4,000 individuals at each site). Every individual was measured in 2015 and 2016 for growth (height, basal diameter) and disease resistance to a recently introduced leaf blight pathogen (Quambalaria pitereka). Narrow-sense heritability was estimated along with trait covariation. Trait variation was regressed against climate-of-origin, and multivariate models were used to develop predictive maps of growth and disease resistance. Growth and blight resistance traits differed significantly among populations, and these differences were consistent between experimental sites and sampling years. Growth and blight resistance were heritable, and comparisons between trait differentiation (Q ST) and genetic differentiation (F ST) revealed that population differences in height and blight resistance traits are due to divergent natural selection. Traits were significantly correlated with climate-of-origin, with cool and wet populations showing the highest levels of growth and blight resistance. These results provide evidence that plants have adaptive growth strategies and pathogen defence strategies. Indeed, the presence of standing genetic variation and trait heritability of growth and blight resistance provide capacity to respond to novel, external pressures. The integration of genetic variation into adaptive management strategies, such as assisted gene migration and seed sourcing, may be used to provide greater resilience for natural ecosystems to both biotic and abiotic stressors.


REM-related bradyarrhythmia syndrome.
Cardiac arrhythmias during sleep are relatively common and include a diverse etiology, from benign sinus bradycardia to potentially fatal ventricular arrhythmias. Predisposing factors include obstructive sleep apnea and cardiac disease. Rapid eye movement (REM)-related bradyarrhythmia syndrome (including sinus arrest and complete atrioventricular block with ventricular asystole) in the absence of an underlying cardiac or physiologic sleep disorder was first described in the early 1980s. Although uncertain, the underlying pathophysiology likely reflects abnormal autonomic neural-cardiac inputs during REM sleep. The autonomic nervous system (ANS) is a known key modulator of heart rate fluctuations and rhythm during sleep and nocturnal heart rate reflects a balance between the sympathetic-parasympathetic systems. Whether the primary trigger for REM-related bradyarrhythmias reflects abnormal centrally mediated control of the ANS during REM sleep or anomalous baroreflex parasympathetic influences is unknown. This review focuses on the salient features of the REM-related bradyarrhythmia syndrome and explores potential mechanisms with a particular assessment of the relationship between the ANS and nocturnal heart rate fluctuations.


Steroid hormone determination in water using an environmentally friendly membrane based extraction technique.
In this study, a method was developed for determination of steroid hormones (17beta-estradiol, estrone, 17alpha-ethynylestradiol) in tap and sewage water samples from Sweden. Sample preparation and analysis were performed by a hollow-fibre microporous membrane liquid-liquid extraction (HF-MMLLE) set-up combined with gas chromatography-mass spectrometry (GC-MS). In this approach, only the organic liquid in the lumen (10microL) of the hollow-fibre membrane was utilised for depleting extraction. Several parameters were studied, including: type of organic solvent, sample pH, salt and humic acid content. The optimised method allowed the determination of the analyte at the low ngL(-1) level in tap and sewage water. A linear plot gave correlation coefficients better than 0.995 and resulted in a method limit of detection of 1.6, 3 and 10ngL(-1) for 17beta-estradiol, estrone, and 17alpha-ethynylestradiol, respectively, in sewage water. Enrichment factors were over 1400 after derivatisation. The repeatabilities at 50 and 600ngL(-1) were better than 10% and 6%, respectively.


Osteoporosis and jawbones in women.
Osteoporosis is a major health problem affecting one in three women over the age of 50 and may not be detected until fractures occur. Since osteoporotic fractures are a health burden worldwide, identifying subjects with a high risk of osteoporosis and preventing osteoporosis-related mortality and morbidity are a very important health strategy. Women show an estrogen-related bone loss starting at menopause, predominantly occurring in trabecular bone. Diagnosis of osteoporosis is usually based on the bone mineral density measurement, but this is not a practical and economical technique for early detection. Therefore, investigators are interested in the possibility of detecting osteoporosis from the panoramic radiographs. Mandibular cortical bone undergoes resorptive activity in osteoporotic patients, leading to a decreased thickness and more porous inferior border. Therefore, studies have demonstrated the usefulness of cortical width and shape, determined from panoramic radiographs, in identifying elderly individuals with undetected osteoporosis, especially postmenopausal women. In conclusion, postmenopausal women with C3 category, Mental Index (MI) <3 mm, and panoramic mandibular index (PMI) <0.3 may be considered for further osteoporosis investigation.