2-Pyrrolidinone and Succinimide as Clinical Screening Biomarkers for GABA-Transaminase Deficiency: Anti-seizure Medications Impact Accurate Diagnosis.
Broad-scale untargeted biochemical phenotyping is a technology that supplements widely accepted assays, such as organic acid, amino acid, and acylcarnitine analyses typically utilized for the diagnosis of inborn errors of metabolism. In this study, we investigate the analyte changes associated with 4-aminobutyrate aminotransferase (ABAT, GABA transaminase) deficiency and treatments that affect GABA metabolism. GABA-transaminase deficiency is a rare neurodevelopmental and neurometabolic disorder caused by mutations in ABAT and resulting in accumulation of GABA in the cerebrospinal fluid (CSF). For that reason, measurement of GABA in CSF is currently the primary approach to diagnosis. GABA-transaminase deficiency results in severe developmental delay with intellectual disability, seizures, and movement disorder, and is often associated with death in childhood. Using an untargeted metabolomics platform, we analyzed EDTA plasma, urine, and CSF specimens from four individuals with GABA-transaminase deficiency to identify biomarkers by comparing the biochemical profile of individual patient samples to a pediatric-centric population cohort. Metabolomic analyses of over 1,000 clinical plasma samples revealed a rich source of biochemical information. Three out of four patients showed significantly elevated levels of the molecule 2-pyrrolidinone (Z-score ≥2) in plasma, and whole exome sequencing revealed variants of uncertain significance in ABAT. Additionally, these same patients also had elevated levels of succinimide in plasma, urine, and CSF and/or homocarnosine in urine and CSF. In the analysis of clinical EDTA plasma samples, the levels of succinimide and 2-pyrrolidinone showed a high level of correlation (R = 0.73), indicating impairment in GABA metabolism and further supporting the association with GABA-transaminase deficiency and the pathogenicity of the ABAT variants. Further analysis of metabolomic data across our patient population revealed the association of elevated levels of 2-pyrrolidinone with administration of vigabatrin, a commonly used anti-seizure medication and a known inhibitor of GABA-transaminase. These data indicate that anti-seizure medications may alter the biochemical and metabolomic data, potentially impacting the interpretation and diagnosis for the patient. Further, these data demonstrate the power of combining broad scale genotyping and phenotyping technologies to diagnose inherited neurometabolic disorders and support the use of metabolic phenotyping of plasma to screen for GABA-transaminase deficiency.


Thermal and Cardiovascular Strain Mitigate the Potential Benefit of Carbohydrate Mouth Rinse During Self-Paced Exercise in the Heat.
To determine whether a carbohydrate mouth rinse can alter self-paced exercise performance independently of a high degree of thermal and cardiovascular strain. Eight endurance-trained males performed two 40-km cycling time trials in 35°C, 60% RH while swilling a 20-ml bolus of 6.5% maltodextrin (CHO) or a color- and taste-matched placebo (PLA) every 5 km. Heart rate, power output, rectal temperature (Tre), and mean skin temperature (Tsk) were recorded continuously; cardiac output, oxygen uptake (VO2), mean arterial pressure (MAP), and perceived exertion (RPE) were measured every 10 min. Performance time and mean power output were similar between treatments, averaging 63.9 ± 3.2 and 64.3 ± 2.8 min, and 251 ± 23 and 242 ± 18 W in CHO and PLA, respectively. Power output, stroke volume, cardiac output, MAP, and VO2 decreased during both trials, increasing slightly or remaining stable during a final 2-km end-spurt. Tre, Tsk, heart rate, and RPE increased throughout exercise similarly with both treatments. Changes in RPE correlated with those in Tre (P < 0.005) and heart rate (P < 0.001). These findings suggest that carbohydrate mouth rinsing does not improve ~1-h time trial performance in hot-humid conditions, possibly due to a failure in down-regulating RPE, which may be influenced more by severe thermal and cardiovascular strain.


Evaluation of a manualised speech and language therapy programme for children with social communication disorder: the SCIP feasibility study.
Children with Social (Pragmatic) Communication Disorder (SPCD) have long-term needs in using and processing social language and have a high risk of later mental health difficulties. A manualised speech and language therapy programme, the Social Communication Intervention Programme (SCIP) provides therapy content for SPCD. A feasibility study is required to derive more precise estimates of key parameters for a future trial of SCIP. To assess the feasibility of conducting a substantive randomized controlled trial of SCIP for children with SPCD. A questionnaire was distributed to paediatric speech and language therapists in England. Survey questions addressed number of eligible children, routine intervention provision and trial recruitment factors. In the second phase, a single-arm intervention feasibility study was completed. Fifteen speech and language practitioners identified 24 children aged 5-11 years with SPCD. Practitioners received training/supervision to deliver 20 SCIP therapy sessions to each child. At time 1, parents of participating children provided three communication goals; expected steps in each goal were defined. After intervention, parents and practitioners independently rated each goal compared to baseline ability. Two research practitioners compared parent post-intervention commentaries with outcome scores to derive guidance about clinical significance. All practitioners recorded audio commentaries on therapy experiences. Post-intervention interviews were conducted with 6 practitioners and 6 parents. An expert panel completed a Delphi consultation on trial design. Routine practice for SPCD varies widely. Children tend to be embedded in autism provision. Participation in a future trial was well supported provided resources are available to services. Outcomes analysis indicated all children except one made some progress on parent ratings; all children made progress on practitioner ratings. A power analysis for a future trial was carried out using current outcome measure as putative primary endpoint. Practitioners' audio-diaries provided suggestions for training and adaption in a future trial. Outcomes and therapy methods were acceptable to practitioners and parents. The feasibility study evaluated a novel outcome measure of social communication skills in SPCD. A power calculation indicated a feasible framework for a trial within a realistic period of time. Recommendations for recruitment methods, adaptation of manual and training were supported by practitioners and an expert panel. Title: Speech-language therapy for child social communication disorderTrial ID: ISRCTN48030419. Date registered: January 1, 2017. Registered retrospectively.


Palsies of the fifth cervical nerve root after cervical decompression: prevention using continuous intraoperative electromyography monitoring.
A desire to prevent complications resulting from spinal surgery led to the development of intraoperative monitoring. Intraoperative electromyography (EMG) provides useful diagnostic information regarding nerve root function during spinal and peripheral nerve surgeries. The C-5 nerve root is considered particularly vulnerable to injury during cervical surgery. Despite advances in techniques, the incidence of postoperative C-5 palsy has not changed. The authors reviewed prospectively collected data obtained in 161 patients who underwent 171 cervical procedures. In 116 procedures, operative monitoring was modified to include continuous C-5 EMG from the deltoid muscle. In cases in which spontaneous C-5 activity occurred, an appropriate change in operative manipulation was made. A historical control group consisted of a retrospective review of 55 procedures that were monitored using conventional techniques. In the retrospective cohort, four (7.3%) of 55 patients presented after undergoing surgery for C-5 nerve root palsy. In each patient conventional monitoring revealed unremarkable findings. In the prospective cohort, intraoperative spontaneous EMG activity necessitated a change in either positioning or operative technique in three cases. Only one patient (0.9%) experienced postoperative C-5 palsy. Postoperative C-5 palsy occurred in no patient in whom there was no intra-operative evidence of root irritation (p < 0.03, chi-square test). The incidence of postoperative C-5 palsies was reduced from 7.3% to 0.9% due to intraoperative continuous EMG monitoring. No patient suffered a postoperative C-5 palsy when intraoperative evidence of root irritation was absent.


Electron microscopic evaluation of adhesion of Helicobacter pylori to the gastric epithelial cells in chronic gastritis.
The adhesion of H. pylori to the gastric epithelial cells may be an essential step for the pathophysiology of various H. pylori-induced gastrointestinal diseases. The purpose of this study was to investigate the ultrastructural relation of H. pylori and gastric epithelial cells in their adhesion. Endoscopic biopsy of gastric antrum and body was performed from 15 patients (9 men, 6 women) with chronic gastritis and H. pylori infection. The specimens were processed for electron microscopy and observed with a transmission electron microscope (Hitachi H-600). On the basis of morphological appearances, the adhesions of H. pylori to the gastric epithelial cells were categorized into three types; filamentous connection, adhesion pedestals and membrane fusion. Coccoid and undetermined forms adhered mainly by the filamentous connection, whereas the bacillary forms adhered primarily by the adhesion pedestals and membrane fusion. Various types of adhesion were associated with H. pylori and gastric epithelium. Further studies are needed to evaluate the influence of different types of adhesion to the pathophysiology of H. pylori.


Comparison of efficacy of bupivacaine and fentanyl with bupivacaine and sufentanil for epidural labor analgesia.
A study to compare the efficacy between fentanyl and sufentanil combined with low concentration (0.0625%) of bupivacaine for epidural labor analgesia in laboring women Fifty full term parturients received an initial bolus dose of a 10 ml solution containing 0.125% bupivacaine. The patients were randomly divided into two: group F received 0.0625% bupivacaine with 2.5 mcg/ml fentanyl and group S received 0.0625% bupivacaine with 0.25 mcg/ml sufentanil. Verbal analogue pain scores, need of supplementary/rescue boluses dose of bupivacaine consumed, mode of delivery, maternal satisfaction, and neonatal Apgar scores were recorded. No significant difference was observed between both groups. Both the groups provided equivalent labor analgesia and maternal satisfaction. The chances of cesarean delivery were also not increased in any group. No difference in the cephalad extent of sensory analgesia, motor block or neonatal Apgar score were observed. Although mean pain scores throughout the labor and delivery were similar in both groups, more patients in fentanyl group required supplementary boluses though not statistically significant. We conclude that both 0.0625% bupivacaine-fentanyl (2.5 μg/ml) and 0.0625% bupivacaine-sufentanil (0.25 μg/ml) were equally effective by continuous epidural infusion in providing labor analgesia with hemodynamic stability achieving equivalent maternal satisfaction without serious maternal or fetal side effects. We found that sufentanil was 10 times more potent than fentanyl as an analgesic for continuous epidural labor analgesia.


Enhancement of Adriamycin cytotoxicity in a multidrug resistant Chinese hamster ovary (CHO) subline, CHO-Adrr, by toremifene and its modulation by alpha 1 acid glycoprotein.
The effects of a new antioestrogen, toremifene, on multidrug resistance have been studied in a Chinese hamster ovary parental line, CHO-K1, and in a multidrug resistance subline, CHO-Adrr. Toremifene at subinhibitory concentrations increased the cytotoxic effectiveness of Adriamycin in both cell lines. The degree of potentiation was greater in the CHO-Adrr lines for any given concentration of toremifene. Toremifene is 99.7% bound to human serum proteins (Sipila et al. Pharmacol Toxicol 1988, 63, 62-64), which includes binding to an acute phase plasma protein, alpha 1 acid glycoprotein (AAG). Since AAG is normally absent from tissue culture media, we have assessed the effect of AAG on toremifene mediated potentiation of Adriamycin cytotoxicity. In the presence of increasing concentrations of AAG, there was a dose-related reversal of the effect of toremifene on Adriamycin cytotoxicity in both cell lines. These results show that toremifene is effective in enhancement of Adriamycin cytotoxicity in CHO-K1 and CHO-Adrr cell lines, and this modulation can be altered by AAG. The clinical implication is that patients should be selected for such therapy by measurement of AAG levels.


Man, You Might Look Like a Woman-If a Child Is Next to You.
Gender categorization seems prone to a pervasive bias: Persons about whom null or ambiguous gender information is available are more often considered male than female. Our study assessed whether such a male-bias is present in non-binary choice tasks and whether it can be altered by social contextual information. Participants were asked to report their perception of an adult figure's gender in three context conditions: (1) alone, (2) passively besides a child, or (3) actively helping a child (n = 10 pictures each). The response options male, female and I don't know were provided. As a result, participants attributed male gender to most figures and rarely used the I don't know option in all conditions, but were more likely to attribute female gender to the same adult figure if it was shown with a child. If such social contextual information was provided in the first rather than the second block of the experiment, subsequent female gender attributions increased for adult figures shown alone. Additionally, female gender attributions for actively helping relative to passive adults were made more often. Thus, we provide strong evidence that gender categorization can be altered by social context even if the subject of gender categorization remains identical.


Surgical Resection of Deep-Seated Arteriovenous Malformations Through Stereotactically Guided Tubular Retractor Systems: A Case Series.
Arteriovenous malformations (AVMs) in the subcortical and/or periventricular regions can cause significant intraventricular and intracranial hemorrhage. These AVMs can pose a unique surgical challenge because traditional, open approaches to the periventricular region require significant cortical/white matter retraction to establish sufficient operative corridors, which may result in risk of neurological injury. Minimally invasive tubular retractor systems represent a novel, feasible surgical option for treating deep-seated AVMs. To explore 5 cases of NICO BrainPath-assisted resection of subcortical/periventricular AVMs. Five patients from a single institution were operated on for deep-seated AVMs using tubular retractor systems. Collected data included demographics, AVM specifications, preoperative neurological status, postoperative neurological status, and postoperative/intraoperative angiogram results. Five patients, ranging from age 10 to 45 years, underwent mini-craniotomy for stereotactically guided tubular retractor-assisted AVM resection using neuronavigation for selecting a safe operative corridor. No preoperative embolization was necessary. Mean maximum AVM nidal diameter was 8.2 mm. All deep-seated AVMs were completely resected without complications. All AVMs demonstrated complete obliteration on intraoperative angiogram and on 6-month follow-up angiogram. Minimally invasive tubular retractors are safe and present a promising surgical option for well-selected deep-seated AVMs. Furthermore, study may elucidate whether tubular retractors improve outcomes after microsurgical AVM resection secondary to mitigation of iatrogenic retraction injury risk.


Classifiers for Accelerometer-Measured Behaviors in Older Women.
Machine learning methods could better improve the detection of specific types of physical activities and sedentary behaviors from accelerometer data. No studies in older populations have developed and tested algorithms for walking and sedentary time in free-living daily life. Our goal was to rectify this gap by leveraging access to data from two studies in older women. In study 1, algorithms were developed and tested in a sample of older women (N = 39, age range = 55-96 yr) in the field. Women wore accelerometers and SenseCam (ground truth annotation) devices for 7 d, yielding 3191 h and 320 d of data. Images were annotated and time matched to accelerometer data, and random forest classifiers labeled behaviors (sitting, riding in a vehicle, standing still, standing moving, and walking/running). In study 2, we examined the concurrent validity of the algorithms using accelerometer data from an observed 400-m walk test (2983 min of data available) and 6 d of wearing both accelerometers and global positioning systems devices in a sample of 222 women (age range = 67-100; 313,290 min of data available). Analyses included sensitivity, specificity balanced accuracy, and precision, as appropriate, averaged over each test participant at the minute level for each behavior. In study 1, the algorithms had 82.2% balanced accuracy. In study 2, the classifier had 87.9% accuracy for predicting walking. Overall machine learning classifiers and global positioning systems had 88.6% agreement. Free-living algorithms for walking and sedentary time yielded high levels of accuracy and concurrent validity and can be applied to existing accelerometer data from older women.


Depressive symptoms, anxiety and academic motivation in youth: Do schools and families make a difference?
This longitudinal study aimed to examine the association between depressive and anxiety symptoms and academic motivation by gender, and whether positive school and family factors would be associated with academic motivation, in spite of the presence of such symptoms. Study participants were predominantly economically disadvantaged youths aged 13-15 years in a Northeastern US urban public school system. The Social and Health Assessment (SAHA) served as the basis for a survey undertaken in 2003 and 2004 with information being used from students who participated at both time points (N = 643). Multiple linear regression analyses showed that depressive symptoms were negatively associated with academic motivation, while anxiety was positively related to academic motivation in both genders. Teacher support, school attachment and parental control were positively related to academic motivation even in the presence of internalizing problems. The negative association of depressive symptoms with academic motivation may be potentially decreased by attachment to school.


Lifestyle intervention for prevention of diabetes: determinants of success for future implementation.
Lifestyle interventions are reported to reduce the risk of type 2 diabetes in high-risk individuals after mid- and long-term follow-up. Information on determinants of intervention outcome and adherence and the mechanisms underlying diabetes progression are valuable for a more targeted implementation. Weight loss seems a major determinant of diabetes risk reduction, whereas physical activity and dietary composition may contribute independently. Body composition and genetic variation may also affect the response to intervention. Lifestyle interventions are cost-effective and should be optimized to increase adherence and compliance, especially for individuals in the high-risk group with a low socioeconomic status, so that public health policy can introduce targeted implementation programs nationwide. The aims of this review are to summarize the mid- and long-term effects of lifestyle interventions on impaired glucose tolerance and type 2 diabetes mellitus and to provide determinants of intervention outcome and adherence, which can be used for future implementation of lifestyle interventions.


Overcoming Language Barriers in Paramedic Care With an App Designed to Improve Communication With Foreign-Language Patients: Nonrandomized Controlled Pilot Study.
Communication across language barriers is a particular challenge for health care providers. In emergency medical services, interpreters are mostly not available on rescue scenes, which jeopardizes safe and high-quality medical care. In a cocreative process together with paramedics and software designers, we developed a fixed-phrase translation app with 600 phrases and 18 supported languages that supports paramedics when providing care to foreign-language patients. This paper reports on the results of a pilot study to evaluate the app's effect on paramedic-patient communication. This study aims to gain insights into the efficacy and feasibility of a multilingual app that helps paramedics to communicate with patients who are not proficient in the local language. A 3-armed nonrandomized interventional pilot study was conducted in 4 rescue stations in the German Federal State of Lower Saxony: 3 rural areas and 1 in urban environment. The intervention group comprised rescue missions with patients with limited German language proficiency (LGP) with whom the app was used; control group 1 comprised LGP patients without app usage; and control group 2 consisted of rescue missions with German-speaking patients. For each rescue operation with LGP patients, paramedics filled out questionnaires about the communications with patients. From standardized Rescue Service Case Protocols, we extracted information on patient demographics (age and sex), clinical aspects (preliminary diagnosis and Glasgow Coma Scale), and rescue operation characteristics (time spent on emergency scene and additional dispatch of emergency physicians). The primary outcome was the paramedics' perceived quality of communication with LGP patients. The secondary outcome was the ability to obtain necessary information from patients and the ability to provide important information to patients. A linear regression model was applied to assess the impact of the app on perceived communication, controlling demographic factors, and severity of illness. A total of 22 LGP patients were recruited into the intervention group and 122 into control group 1. The control group 2 included 27,212 German-speaking patients. LGP patients were more than 2 decades younger than German-speaking patients. App usage among LGP patients was associated with higher perceived overall quality of communication (0.7 points on a 5-point Likert scale, P=.03). Applying a linear regression model controlling for age, sex, and Glasgow Coma Scale, the quality of communication was associated with an increase of 0.9 points (95% CI 0.2-1.6, P=.01). Compared to either German-speaking patients or LGP patients, paramedics spent 6-7 minutes longer on an emergency scene when the app was used (P=.24). The use of the app suggests a relevant improvement in communication with patients with limited proficiency in the locally spoken language in paramedic care. The small sample size and the lack of randomization reduce the generalizability of the findings. German Clinical Trials Register DRKS00016719; https://drks.de/search/de/trial/DRKS00016719.


Role of antioxidants in generalised anxiety disorder and depression.
Anxiety and depression form commonest stress-induced psychiatric disorders. To combat the biochemical changes which occur as a result of stress, there is antioxidant defence in the biological system. Secondary defence is by the nonenzymatic antioxidants like vitamins E (alphatocopherol), C (ascorbic acid), and β-carotene. Therefore, the authors interest was aroused to examine the status of these antioxidants in the biological system of patients suffering from stress-induced psychiatric disorders. This study was carried out to find out whether patients with generalized anxiety disorder (GAD) and depression have any difference in blood serum levels of vitamins A (β-carotene), C, and E in comparison to the normal healthy control group and whether supplementation of adequate doses of vitamins A (β-carotene), C, and E leads to improvement in anxiety and depression and reduction in scores of the patients. Eighty subjects in the age group of 20-60 years, who attended a psychiatric clinic of a private hospital and who met inclusion and exclusion criteria of the study and consented for psychological evaluation and blood screening to find out the serum levels of vitamins A, C, and E, were included in the study. Approval was sought from the institutional ethics committee for collecting the blood sample of these subjects before and after vitamins A, C, and E supplements given for a period of 6 weeks. It was observed that patients with GAD and depression had significantly lower levels of vitamins A, C, and E in comparison to healthy controls. After dietary supplementation of these vitamins for a period of 6 weeks, a significant reduction in anxiety and depression scores of patients was observed (P<0.001). A significant increase in the blood levels of antioxidants was observed in patients (P<0.05) except that of vitamin E in the group of depressed patients. The findings suggest that antioxidant supplement therapy as an adjuvant therapy is useful in patients with stress-induced psychiatric disorders and the results have been discussed.


[3H]tiagabine binding to GABA uptake sites in human brain.
The binding of [3H]tiagabine ((RS-1-(4,4-(3-methyl-2-thienyl)-3-butenyl)-3-piperidine carboxylic acid) to homogenates of frozen post-mortem human brain has been characterized. Inhibition experiments with gamma-aminobutyric acid (GABA), GABA uptake inhibitors, ligands active at postsynaptic GABA receptors and receptors for other neurotransmitters, suggest that [3H]tiagabine binds with high affinity to GABA uptake sites. Inhibition and kinetic experiments suggests that 70%-80% of the binding is to a high affinity site. Saturation experiments showed that the binding was saturable. Bmax was 3.4 pmol/mg protein and Kd 16 nM in frontal cortex. The dissociation constants (Kd) measured in kinetic and equilibrium experiments were in the same range (16-56 nM). The regional distribution was studied in nine brain regions and the binding was heterogenous, with the highest binding in frontal cortex and parietal cortex and the lowest binding in nucleus caudatus and putamen. This is, to our knowledge, the first study on [3H]tiagabine binding in human tissue. It is concluded that [3H]tiagabine binding can be used as a specific marker for the GABA transporter GAT-1 in homogenates of human brain.


Neuroserpin, a neuroprotective factor in focal ischemic stroke.
Because recent studies have indicated that tissue plasminogen activator (tPA) aggravates neurodegenerative processes in many neural pathologies, we studied whether the endogenous tPA antagonist neuroserpin has a neuroprotective effect in an animal model of focal ischemic stroke. After induction of a focal ischemic stroke in the mouse by occlusion of the middle cerebral artery, we found that microglial cells accumulated in the marginal zone of the infarct are the most important source for both plasminogen activators, tPA and uPA. To investigate the effect of neuroserpin on the size and the histology of the infarct we produced transgenic mice overexpressing neuroserpin approximately sixfold in the nervous system. In the brain of these mice the total tPA activity in the uninjured tissue was strongly reduced. After induction of a focal ischemic stroke in the transgenic mice by a permanent occlusion of the middle cerebral artery (MCA), the infarcts were 30% smaller than in the wild-type mice. Immunohistochemical analyses and in situ hybridization revealed an attenuation of the microglial activation in the reactive zone. Concomitantly, the microglial production of tPA and uPA, as well as the PA-activity in the infarct region was markedly reduced. Thus, our results indicate that neuroserpin reduces microglial activation and, therefore, the PA activity and has a neuroprotective role after focal ischemic stroke.


Intestinal carcinogenesis: IKK can go all the way.
Chronic inflammation has long been suspected to support tumorigenesis in a variety of cancers. The IκB kinase (IKK)/NF-κB pathway is the critical signal transduction pathway regulating inflammation, and loss-of-function studies have demonstrated its involvement in tumorigenesis. In this issue of the JCI, Vlantis et al. present evidence that persistent genetic activation of IKK/NF-κB signaling in intestinal epithelial cells not only accelerates tumorigenesis in models of both carcinogen- and mutation-induced colorectal cancer, but also is sufficient to induce intestinal tumors.


Grip strength at four years in relation to birth weight.
Consistent positive relationships have been found between birth weight and grip strength in adults but evidence in children is limited. In a prospective general population birth cohort (Southampton Women's Survey) grip strength and anthropometry (height and weight) were measured in 968 children at age 4 years. Mean (standard deviation (S.D.)) birth weight was 3.48 (0.52) kg. Birth weight, adjusted for sex and gestational age, was positively associated with grip strength (β = 0.22 kg/S.D. increase in adjusted birth weight; 95% CI 0.11, 0.34). The relationship was attenuated after adjustment for current height and weight such that it became non-significant (β = 0.03 kg/S.D. increase in adjusted birth weight; 95% CI -0.08, 0.14), suggesting that body size may be on the causal pathway. Early influences on muscle development appear to impact on grip strength in children as well as adults.


Structural Insights into Thioether Bond Formation in the Biosynthesis of Sactipeptides.
Sactipeptides are ribosomally synthesized peptides that contain a characteristic thioether bridge (sactionine bond) that is installed posttranslationally and is absolutely required for their antibiotic activity. Sactipeptide biosynthesis requires a unique family of radical SAM enzymes, which contain multiple [4Fe-4S] clusters, to form the requisite thioether bridge between a cysteine and the α-carbon of an opposing amino acid through radical-based chemistry. Here we present the structure of the sactionine bond-forming enzyme CteB, from Clostridium thermocellum ATCC 27405, with both SAM and an N-terminal fragment of its peptidyl-substrate at 2.04 Å resolution. CteB has the (β/α)6-TIM barrel fold that is characteristic of radical SAM enzymes, as well as a C-terminal SPASM domain that contains two auxiliary [4Fe-4S] clusters. Importantly, one [4Fe-4S] cluster in the SPASM domain exhibits an open coordination site in absence of peptide substrate, which is coordinated by a peptidyl-cysteine residue in the bound state. The crystal structure of CteB also reveals an accessory N-terminal domain that has high structural similarity to a recently discovered motif present in several enzymes that act on ribosomally synthesized and post-translationally modified peptides (RiPPs), known as a RiPP precursor peptide recognition element (RRE). This crystal structure is the first of a sactionine bond forming enzyme and sheds light on structures and mechanisms of other members of this class such as AlbA or ThnB.


Pandemic influenza and health system resource gaps in Bali: an analysis through a resource transmission dynamics model.
The failure to contain pandemic influenza A(H1N1) 2009 in Mexico has shifted global attention from containment to mitigation. Limited surveillance and reporting have, however, prevented detailed assessment of mitigation during the pandemic, particularly in low- and middle-income countries. To assess pandemic influenza case management capabilities in a resource-limited setting, the authors used a health system questionnaire and density-dependent, deterministic transmission model for Bali, Indonesia, determining resource gaps. The majority of health resources were focused in and around the provincial capital, Denpasar; however, gaps are found in every district for nursing staff, surgical masks, and N95 masks. A relatively low pathogenicity pandemic influenza virus would see an overall surplus for physicians, antivirals, and antimicrobials; however, a more pathogenic virus would lead to gaps in every resource except antimicrobials. Resources could be allocated more evenly across Bali. These, however, are in short supply universally and therefore redistribution would not fill resource gaps.


[Markers of coagulation and fibrinolysis do not detect or predict the presence of left atrial appendage thrombus in patients with atrial fibrillation].
This study was designed to evaluate the role of hemostatic variables in arterial blood serum in left atrial thrombosis and to define any hemostatic variables, such as serum biomarkers, that could potentially reduce the need for transesophageal echocardiography. This study included patients with non-valvular asymptomatic atrial fibrillation (AF), either paroxysmal, persistent, or chronic. The presence of an left atrial appendix (LAA) thrombus was used to form 2 groups: thrombus (+) and thrombus (-). The serum levels of the thrombotic/fibrinolytic markers including beta-thromboglobulin, prothrombin fragment 1+2, thrombin/antithrombin complex, human plasminogen activator inhibitor-1/tissue plasminogen activator complex, and D-dimer were compared between 2 groups. The mean age of the study population was 65.6±12.2 years (range: 30-96 years), and 33 (61.1%) patients were male. Fourteen (25.9%) patients had an LAA thrombus and 40 patients did not. Two groups did not differ significantly with regard to any of the coagulation/fibrinolysis markers. The LAA thrombus (+) group had significantly higher rates of heart failure, peripheral artery disease, coronary artery disease, and chronic obstructive pulmonary disease (<0.05). Neither the serum levels of the study markers nor demographic and clinical parameters were predictive of an LAA thrombus in binary logistic regression analysis. The arterial blood serum markers did not differ significantly between groups with and without an LAA thrombus and did not predict an LAA thrombus in patients presenting with AF.


Neonatal sepsis bacterial isolates and antibiotic susceptibility patterns at a NICU in a tertiary care hospital in western Nepal: a retrospective analysis.
Neonatal sepsis is one of the commonest causes of neonatal mortality in the developing world. The neonatal intensive care units (NICUs) today face one common problem of tackling sepsis and neonatologists remain constantly baffled by the changing patterns of microbial flora and their sensitivity patterns. With the neonatal services coming of age in Nepal this retrospective analysis spread over a period of six years has become very pertinent. We conducted a retrospective study over a period of six years to study the prevalence of different organisms causing septicaemia in the community and at our hospital and the antibiotic susceptibility pattern. In all 265 cases of suspected sepsis were screened using a panel consisting of CRP, ANC and I/T ratio and subsequently confirmed by cultures. The cases were early onset (n=44), late onset (n=56) and nosocomial groups (n=40). The data for the intramural (n=32) and extramural (n=68) cases was analysed separately. One hundred nineteen cultures out of the 131 positives were obtained from blood (44.92%) and the remaining were isolated from urine (6.11 %) and CSF (4.58 %). The most common organism to be isolated was staphylococcus aureus (42.75%) followed by klebsiella pneumoniae (18.32%)and escherechia coli (12.21%). Staphylococcus was isolated from 36.84%, 45.16% and 31.81% of the cultures obtained from neonates in the in-born, out-born and the nosocomial groups respectively while klebsiella pneumoniae [18.32 %] was seen in 21.05 %, 17.39 % and 36.36 % in each of the three groups. Pseudomonas aeruginosa [6.11 %] was isolated from 13.64 % of the nosocomial cultures compared to 8.7 % of the out-borns while it was not seen in the in-borns. Other organisms isolated were much less in number, included - pathogenic streptococci, acinetobacter and enterobacter species. Coagulase negative staphylococci (CoNS) was seen in 4.39 % [n=4] and 9.09 % [n=4] of the same groups respectively. The gram positive organisms displayed a high degree of resistance to most penicillins and cephalosporins but glycopeptides and monobactams were effective in them. There was a high incidence of resistance noted with most third generation cephalosporins and aminoglycosides amongst most gram negative organisms where-in cefepime and imepenem were effective in most cases. Staphylococcal sepsis is not only common in community acquired infections but also in nosocomial sepsis. There is an emerging resistance to cephalosposrins probably attributable to extended spectrum betalactamases. Further large-scale multicentre studies are required to generalise the data for the whole country.


No association of IL-12p40 pro1.1 polymorphism with juvenile idiopathic arthritis.
IL-12p40 plays an important role in the activation of the T-cell lines like Th17 and Th1-cells. Theses cells are crucial in the pathogenesis of juvenile idiopathic arthritis. A polymorphism in its promoter region and the genotype IL12p40 pro1.1 leads to a higher production of IL-12p40. We studied whether there is a difference in the distribution of the genotype in patients with JIA and the healthy population. In 883 patients and 321 healthy controls the IL-12p40 promoter genotype was identified by ARMS-PCR. There is no association of IL-12p40 pro polymorphism neither in patients with JIA compared to controls nor in subtypes of JIA compared to oligoarthritis. We found a non-significant tendency of a higher prevalence of the genotype pro1.1 in systemic arthritis (32.4%) and in rheumatoid factor negative polyarthritis (30.5%) and a lower pro1.1 genotype in persistent oligoarthritis (20.7%) and in enthesitis-related arthritis (17%). Likelihood of the occurrence of genotype IL12-p40 pro1.1 in patients with systemic arthritis (OR 1.722, CI 95% 1.344-2.615, p 0.0129) and RF-negative polyarthritis (OR 1.576, CI 95% 1.046-2.376, p 0.0367) compared to persistent oligoarthritis was significantly higher. This was also true for comparison of their homozygous genotypes IL-12p40 pro 1.1 and 2.2 in systemic arthritis (OR 1.779, CI 95 % 1.045-3.029, p 0.0338). However, in Bonferroni correction for multiple hypothesis this was not significant. A tendency of a higher prevalence of the genotype IL-12p40 pro1.1 in systemic arthritis and in rheumatoid factor negative polyarthritis was observed but not significant. Further investigations should be done to clarify the role IL-12p40 in the different subtypes of JIA.


Genes Translocated into the Plastid Inverted Repeat Show Decelerated Substitution Rates and Elevated GC Content.
Plant chloroplast genomes (plastomes) are characterized by an inverted repeat (IR) region and two larger single copy (SC) regions. Patterns of molecular evolution in the IR and SC regions differ, most notably by a reduced rate of nucleotide substitution in the IR compared to the SC region. In addition, the organization and structure of plastomes is fluid, and rearrangements through time have repeatedly shuffled genes into and out of the IR, providing recurrent natural experiments on how chloroplast genome structure can impact rates and patterns of molecular evolution. Here we examine four loci (psbA, ycf2, rps7, and rps12 exon 2-3) that were translocated from the SC into the IR during fern evolution. We use a model-based method, within a phylogenetic context, to test for substitution rate shifts. All four loci show a significant, 2- to 3-fold deceleration in their substitution rate following translocation into the IR, a phenomenon not observed in any other, nontranslocated plastid genes. Also, we show that after translocation, the GC content of the third codon position and of the noncoding regions is significantly increased, implying that gene conversion within the IR is GC-biased. Taken together, our results suggest that the IR region not only reduces substitution rates, but also impacts nucleotide composition. This finding highlights a potential vulnerability of correlating substitution rate heterogeneity with organismal life history traits without knowledge of the underlying genome structure.


Evaluations of and reasoning about normative and deviant ingroup and outgroup members: development of the black sheep effect.
Research with adults has demonstrated a "black sheep effect" (BSE) whereby, relative to evaluations of normative group members, ingroup deviants are derogated more than outgroup deviants. The developmental subjective group dynamics (DSGD) model holds that the BSE should develop during middle childhood when children apply wider social norms. Three hundred and thirty-eight children who were between 5 and 12 years old judged a normative (socially desirable) and a deviant (socially undesirable) member from an ingroup or an outgroup school. Results confirmed a developmental increase in the BSE, the first time this has been demonstrated. Children's own evaluations of group members were mediated by their expectations about ingroup peers' evaluations. In line with DSGD and social domain theories, with age, children's explanations of peer evaluations for ingroup deviance focused relatively more on loyalty. Practical and theoretical implications for peer inclusion and exclusion are discussed.


IFN regulatory factor 8 restricts the size of the marginal zone and follicular B cell pools.
Transcriptional control of marginal zone (MZ) and follicular (FO) B cell development remains incompletely understood. The transcription factor, IFN regulatory factor (IRF)8, is known to play important roles in the differentiation of early B cells. In this article, we demonstrate that IRF8 is also required for normal development of MZ and FO B cells. Mice with a conventional knockout of Irf8 (IRF8(-/-)) or a point mutation in the IRF association domain of IRF8 had increased numbers of MZ B cells. To determine the B cell-intrinsic effects of IRF8 deficiency, we generated mice with a conditional allele of Irf8 crossed with CD19-Cre mice (designated IRF8-conditional knockout [CKO]). These mice had enlarged MZ and increased numbers of MZ and FO B cells compared with controls. The FO B cells of CKO mice exhibited reduced expression of CD23 and moderately increased expression of CD21. Gene-expression profiling showed that increased B cell production in IRF8-CKO mice was associated with changes in expression of genes involved in regulation of transcription, signaling, and inflammation. Functional studies showed that IRF8-CKO mice generated normal Ab responses to T-independent and T-dependent Ags. Thus, IRF8 controls the expansion and maturation of MZ and FO B cells but has little effect on B cell function.


Prosthetic hip replacement for pathologic or impending pathologic fractures in myeloma.
The authors retrospectively reviewed 53 hip replacements performed in 50 patients (mean age, 65 years) for plasmacytoma (four lesions) or multiple myeloma (49 lesions) of the hip region. Perioperatively, 49 patients received chemotherapy. Twenty-four hip lesions had radiation therapy. Within a mean followup period of 32.6 months, 15 (28%) complications occurred. Late deep infection, dislocation, and aseptic loosening with medial migration of the acetabular component occurred in one patient each. Three patients died during the first postoperatively month. All of the remaining patients postoperatively regained their previous ambulatory status; hip pain relief was achieved in 98% of patients. At the time of the latest followup evaluation, 84% of the patients had died (median survival time, 18 months) and 16% were still alive (median survival time, 76 months). Patients with plasmacytoma had longer survival than did patients with multiple myeloma (median survival, 6.3 years versus 18 months, respectively). No significant association was found between patient survival and the stage of disease at diagnosis, age, gender, type or location of the myeloma lesion, or the type of implant used. Because of the relatively long survival time, prosthetic replacement is indicated for extensive hip lesions in patients with plasmacytoma or multiple myeloma.


Reliability of a schedule for rating personality disorders.
The inter-situational, inter-rater and temporal reliability of a schedule for rating personality disorders is described. In an initial study with a simplified form of the schedule in patients from different wards of a psychiatric hospital inter-situational reliability between raters was higher for patients with personality disorders than with no personality disorder. Using the full schedule, inter-rater reliability, using audiotaped and separate interviews, and temporal reliability at interviews conducted a mean of 12.5 months apart all reached a satisfactory level, suggesting that the schedule may be a useful instrument for measuring deviant personality traits. The interview may be used with a subject or an informant but agreement between ratings made with informants and psychiatric patients during illness was low, and the schedule is not recommended for use with patients alone during acute episodes of illness.


Management of Coronary Artery Calcium and Coronary CTA Findings.
Coronary artery calcium (CAC) testing and coronary computed tomography angiography (CTA) have significant data supporting their ability to identify coronary artery disease (CAD) and classify patient risk for atherosclerotic cardiovascular disease (ASCVD). Evidence regarding CAC use for screening has established an excellent prognosis in patients with no detectable CAC, and the ability to risk re-classify the majority of asymptomatic patients considered intermediate risk by traditional risk scores. While data regarding the ideal management of CAC findings are limited, evidence supports statin consideration in patients with CAC > 0 and individualized aspirin therapy accounting for CAD risk factors, CAC severity, and factors which increase a patient's risk of bleeding. In patients with stable or acute symptoms undergoing coronary CTA, a normal CTA predicts excellent prognosis, allowing reassurance and disposition without further testing. When CTA identifies nonobstructive CAD (<50 % stenosis), observational data support consideration of statin use/intensification in patients with extensive plaque (at least four coronary segments involved) and patients with high-risk plaque features. In patients with both nonobstructive and obstructive CAD, multiple studies have now demonstrated an ability of CTA to guide management and improve CAD risk factor control. Still, significant under-treatment of cardiovascular risk factors and high-risk image findings remain, among concerns that CTA may increase invasive angiography and revascularization. To fully realize the impact of atherosclerosis imaging for ASCVD prevention, patient engagement in lifestyle changes and the modification of ASCVD risk factors remain the foundation of care. This review provides an overview of available data and recommendations in the management of CAC and CTA findings.


The directed growth of retinal axons towards surgically transposed tecta in Xenopus; an examination of homing behaviour by retinal ganglion cell axons.
The growth of optic axons towards experimentally rotated tecta has been studied. In stage 24/25 embryos, a piece of the dorsal neural tube, containing the dorsal midbrain rudiment, was rotated through 180 degrees. At later stages of development, the pathways of growing optic axons were investigated by labelling with either horseradish peroxidase or fluorescent dye. It is shown that retinal ganglion cell axons followed well-defined pathways, in spite of the abnormal structure of the brain, and were able to locate displaced tecta. This directed outgrowth of retinal axons in the optic tracts appears to be related either to the tectum or to some other component included in the graft operations. In tadpoles in which the midbrain rudiment was removed, optic axons still followed the normal course of the optic tract. This observation argues against long-range target attraction as being essential in guiding growing retinal axons towards the tectum. An alternative axon guidance mechanism, selective fasciculation, is discussed as a possible alternative to explain the directed axon outgrowth which occurs in both the normal and in these experimentally manipulated tadpoles.


Development of a rice-based peptide vaccine for Japanese cedar and cypress pollen allergies.
Peptide immunotherapy using dominant T-cell epitopes is a safe treatment alternative to conventional subcutaneous injection of natural crude allergen extract, which is sometimes accompanied by anaphylactic shock. For Japanese cedar pollinosis (JCP), hybrid peptides composed of six to seven major T-cell epitopes (7Crp peptide) from the causative allergens Cry j 1 and Cry j 2 have been developed on the basis of different human leukemia antigen class II restrictions, because of the diversity of patients' genetic backgrounds. However, other dominant T-cell epitopes that are produced in some patients are not covered by these peptides. To develop a more universal peptide vaccine for JCP, we generated transgenic rice seeds containing seven new T-cell epitopes (Crp3) in addition to the T-cell epitopes used in the 7Crp peptide. Next, we co-expressed unique T-cell epitopes (6Chao) from the Japanese cypress pollen allergens Cha o 1 and Cha o 2 in transgenic rice seeds, with 7Crp and Crp3. These transgenic rice seeds, containing many highly homologous T-cell epitopes derived from cedar and cypress allergens, are expected to be applicable to a wide range of patients suffering from these pollen allergies.


Visualizing facial shape regression upon 2nd to 4th digit ratio and testosterone.
Sex steroids are supposed to moderate the differences between male and female facial characteristics. Studies on women's preferences for male faces reported increased preferences for facial architecture developed under the influence of testosterone as this may indicate masculinity, dominance and social status. Recent research demonstrates that facial sexual dimorphism does not only develop at puberty but may be organized much earlier in ontogeny. However, the actual cause and timing of variation in facial shape due to sex-steroids remains speculative. This study uses data from Neave and colleagues who measured digit ratio (2D:4D) as a proxy to prenatal testosterone and also salivary testosterone samples in order to study differential effects of androgens on perceived male facial shape. Male facial shape was regressed upon 2D:4D ratio and circulating levels of testosterone by means of geometric morphometric methods. We found some evidence for opposite effects of early androgen action (via 2D:4D ratio) on the upper and the lower face respectively (i.e. low 2D:4D ratio results in a relatively robust and prominent lower face), whereas circulating testosterone seems to cause a rather uniform elongation of the face. Local deformations primarily show pronounced and medially tailed eyebrows for the shapes associated with increasing salivary testosterone. These preliminary results suggest that prenatal and pubertal testosterone have differential effects on male facial shape that should be considered in future studies on women's preferences towards male facial appearance.


Trajectory Inspection: A Method for Iterative Clinician-Driven Design of Reinforcement Learning Studies.
Reinforcement learning (RL) has the potential to significantly improve clinical decision making. However, treatment policies learned via RL from observational data are sensitive to subtle choices in study design. We highlight a simple approach, trajectory inspection, to bring clinicians into an iterative design process for model-based RL studies. We identify where the model recommends unexpectedly aggressive treatments or expects surprisingly positive outcomes from its recommendations. Then, we examine clinical trajectories simulated with the learned model and policy alongside the actual hospital course. Applying this approach to recent work on RL for sepsis management, we uncover a model bias towards discharge, a preference for high vasopressor doses that may be linked to small sample sizes, and clinically implausible expectations of discharge without weaning off vasopressors. We hope that iterations of detecting and addressing the issues unearthed by our method will result in RL policies that inspire more confidence in deployment.


Role of MicroRNA-34a in Anti-Apoptotic Effects of Granulocyte-Colony Stimulating Factor in Diabetic Cardiomyopathy.
Recent studies have shown that microRNAs (miRNAs) are involved in the process of cardiomyocyte apoptosis. We have previously reported that granulocyte-colony stimulating factor (G-CSF) ameliorated diastolic dysfunction and attenuated cardiomyocyte apoptosis in a rat model of diabetic cardiomyopathy. In this study, we hypothesized a regulatory role of cardiac miRNAs in the mechanism of the anti-apoptotic effect of G-CSF in a diabetic cardiomyopathy rat model. Rats were given a high-fat diet and low-dose streptozotocin injection and then randomly allocated to receive treatment with either G-CSF or saline. H9c2 rat cardiomyocytes were cultured under a high glucose (HG) condition to induce diabetic cardiomyopathy in vitro. We examined the extent of apoptosis, miRNA expression, and miRNA target genes in the myocardium and H9c2 cells. G-CSF treatment significantly decreased apoptosis and reduced miR-34a expression in diabetic myocardium and H9c2 cells under the HG condition. G-CSF treatment also significantly increased B-cell lymphoma 2 (Bcl-2) protein expression as a target for miR-34a. In addition, transfection with an miR-34a mimic significantly increased apoptosis and decreased Bcl-2 luciferase activity in H9c2 cells. Our results indicate that G-CSF might have an anti-apoptotic effect through down-regulation of miR-34a in a diabetic cardiomyopathy rat model.


Variation within and among species in gene expression: raw material for evolution.
Heritable variation in regulatory or coding regions is the raw material for evolutionary processes. The advent of microarrays has recently promoted examination of the extent of variation in gene expression within and among taxa and examination of the evolutionary processes affecting variation. This review examines these issues. We find: (i) microarray-based measures of gene expression are precise given appropriate experimental design; (ii) there is large inter-individual variation, which is composed of a minor nongenetic component and a large heritable component; (iii) variation among populations and species appears to be affected primarily by neutral drift and stabilizing selection, and to a lesser degree by directional selection; and (iv) neutral evolutionary divergence in gene expression becomes nonlinear with greater divergence times due to functional constraint. Evolutionary analyses of gene expression reviewed here provide unique insights into partitioning of regulatory variation in nature. However, common limitations of these studies include the tendency to assume a linear relationship between expression divergence and species divergence, and failure to test explicit hypotheses that involve the ecological context of evolutionary divergence.


Enhanced treatment of petroleum refinery wastewater by short-term applied voltage in single chamber microbial fuel cell.
Electrochemically active anodic biofilm that has adapted under mild applied potentials in the range 100-500 mV was evaluated for its improved bioelectrogenesis and bioelectrochemical treatment of petroleum refinery wastewater (PRW) in a single chamber air cathode microbial fuel cell (MFC). MFC operation with 500 mV as supplemental voltage has exhibited a maximum power density of 132 mW/m2, which was three times higher than control MFC (45 mW/m2). Similarly, highest substrate removal efficiency (48%) was also obtained with the MFC of 500 mV, followed by 300 mV (37%), 100 mV (32%) and control (27%). Adaptation under applied potential conditions also exhibited enhanced degradation efficiency of diesel range organics (DROs)/straight chain-alkanes. The strategy efficiently reduced DROs with the maximum efficiency of 89% (500 mV), which is almost 50% higher than that of the control system (59%), demonstrating the effectiveness of using supplemented voltage in treating PRW.


Hispanic Male's Perspectives of Health Behaviors Related to Weight Management.
Hispanic males have the highest prevalence of overweight and obesity among men in the United States; yet are significantly underrepresented in weight loss research. The purpose of the current study was to examine Hispanic male's perspectives of health behaviors related to weight management to refine the methodologies to deliver a gender-sensitive and culturally sensitive weight loss intervention. From October 2014 to April 2015, semistructured interviews were conducted with 14 overweight Hispanic men of ages 18 to 64 years. The interviews lasted approximately 60 minutes. Participants also completed a brief questionnaire and body weight/height were measured. Grounded in a deductive process, a preliminary codebook was developed based on the topics included in the interview guides. A thematic analysis facilitated the identification of inductive themes and the finalization of the codebook used for transcript analysis. Four overarching themes were identified: (a) general health beliefs of how diet and physical activity behaviors affect health outcomes, (b) barriers to healthy eating and physical activity, (c) motivators for change, and (d) viable recruitment and intervention approaches. Future research should examine feasible and appropriate recruitment and intervention strategies identified by Hispanic males to improve weight management in this vulnerable group.


The Influence of Lifestyle Behaviors on the Incidence of Frailty.
Frailty is a clinical state defined as an increase in an individual's vulnerability to developing adverse health-related outcomes. We propose that healthy behaviors could lower the incidence of frailty. The aim is to describe the association between healthy behaviors (physical activity, vaccination, tobacco use, and cancer screening) and the incidence of frailty. This is a secondary longitudinal analysis of the Mexican Health and Aging Study (MHAS) cohort. MHAS is a population-based cohort, of community-dwelling Mexican older adults. With five assessments currently available, for purposes of this work, 2012 and 2015 waves were used. A total of 6,087 individuals 50-year or older were included. Frailty was defined using a 39-item frailty index. Healthy behaviors were assessed with questions available in MHAS. Individuals without frailty in 2012 were followed-up three years in order to determine their frailty incidence, and its association with healthy behaviors. Multivariate logistic regression models were used to assess the odds of frailty occurring according to the four health-related behaviors mentioned above. At baseline (2012), 55.2% of the subjects were male, the mean age was 62.2 (SD ± 8.5) years old. The overall incidence (2015) of frailty was 37.8%. Older adults physically active had a lower incidence of frailty (48.9% vs. 42.2%, p< 0.0001). Of the activities assessed in the adjusted multivariate models, physical activity was the only variable that was independently associated with a lower risk of frailty (odds ratio: 0.79, 95% confidence interval 0.71-0.88, p< 0.001). Physically active older adults had a lower 3-year incidence of frailty even after adjusting for confounding variables. Increasing physical activity could therefore represent a strategy for reducing the incidence of frailty. Other so-called healthy behaviors were not associated with incident frailty, however there is still uncertainty on the interpretation of those results.


Can perineural invasion detected in prostate needle biopsy specimens predict surgical margin positivity in D'Amico low risk patients?
In this study, our aim was to estimate the value of perineural invasion (PNI) in prostate needle biopsy (PNB) specimens in the prediction of surgical margin positivity (SMP) and its prognostic significance (upgrade Gleason Score) in patients who had undergone radical retropubic prostatectomy (RRP) with low risk prostate cancer according to D'Amico risk assessment. We retrospectively analyzed the data of 65 patients who were diagnosed as clinical stage T1c prostate cancer (PC) and underwent RRP between January 2010 and June 2013. Pathological specimens of PNB and RRP were separately examined for the parameters of PNI, vascular invasion (VI), Gleason Score (GS) and SMP. The patients' mean age was 63.65 ± 4.93 (range 47- 75) years. PNI in PNB specimens were identified in 12 of 65 patients and 11 of 12 patients showed SMP on RRP specimens. While 53 of 65 patients had not PNI on PNB, only 11 of them demonstrated SMP on RRP specimens. SMP was 30.64-fold more frequently encountered in PNB specimens obtained from PNI-positive patients relative to PNI-negative patients. In our study, PNI detected in PNB specimens was statistically significantly associated with SMP on RRP specimens (P = 0.0001). It is well known that higher PSA values and GS were independent predictors of SMP in clinically localized prostate cancer (CLPC). We think that PNI in PNB specimens may be a useful prognostic factor for predicting SMP in cases with CLPC.


Anatomy and muscle activity of the dorsal fins in bamboo sharks and spiny dogfish during turning maneuvers.
Stability and procured instability characterize two opposing types of swimming, steady and maneuvering, respectively. Fins can be used to manipulate flow to adjust stability during swimming maneuvers either actively using muscle control or passively by structural control. The function of the dorsal fins during turning maneuvering in two shark species with different swimming modes is investigated here using musculoskeletal anatomy and muscle function. White-spotted bamboo sharks are a benthic species that inhabits complex reef habitats and thus have high requirements for maneuverability. Spiny dogfish occupy a variety of coastal and continental shelf habitats and spend relatively more time cruising in open water. These species differ in dorsal fin morphology and fin position along the body. Bamboo sharks have a larger second dorsal fin area and proportionally more muscle insertion into both dorsal fins. The basal and radial pterygiophores are plate-like structures in spiny dogfish and are nearly indistinguishable from one another. In contrast, bamboo sharks lack basal pterygiophores, while the radial pterygiophores form two rows of elongated rectangular elements that articulate with one another. The dorsal fin muscles are composed of a large muscle mass that extends over the ceratotrichia overlying the radials in spiny dogfish. However, in bamboo sharks, the muscle mass is divided into multiple distinct muscles that insert onto the ceratotrichia. During turning maneuvers, the dorsal fin muscles are active in both species with no differences in onset between fin sides. Spiny dogfish have longer burst durations on the outer fin side, which is consistent with opposing resistance to the medium. In bamboo sharks, bilateral activation of the dorsal in muscles could also be stiffening the fin throughout the turn. Thus, dogfish sharks passively stiffen the dorsal fin structurally and functionally, while bamboo sharks have more flexible dorsal fins, which result from a steady swimming trade off.


Chronic periodontitis with multiple risk factor syndrome: a case report.
Multiple risk factor syndrome is a clustering of cardiovascular risk factors, such as diabetes, dyslipidemia, hypertension, and obesity associated epidemiologically with insulin resistance. This report describes the clinical course of a patient suffering from severe periodontitis with multiple risk factor syndrome, and discusses the association between periodontal infection and systemic health. The patient had a history of type 2 diabetes, dyslipidemia, and hypertension for over 10 years. At baseline, her hemoglobin A1 c was 8.1%. However, she had no diabetic complications except periodontitis. The IgG antibody titers against Porphyromonas gingivalis FDC 381 and SU63 were elevated above the mean of healthy subjects +2 standard deviations. Intensive periodontal treatment, including periodontal surgery, was performed to reduce periodontal infection and bacteremia. Her systemic and periodontal conditions were evaluated longitudinally for 10 years. Following periodontal treatment, antibody titers against Porphyromonas gingivalis and hemoglobin A1c values were significantly improved. The other clinical data and medication for her systemic condition also remained stable during supportive periodontal therapy. However, she developed myocardial infarction, and showed continuous deterioration of hemoglobin A1 c level and periodontitis. The long-term clustering of risk factors, such as diabetes, dyslipidemia, hypertension, and periodontitis, are associated with the development of myocardial infarction. Treatment of systemic conditions in combination with comprehensive periodontal treatment is important in management of patients with multiple risk factor syndrome.


Architecture of the DNA polymerase B-proliferating cell nuclear antigen (PCNA)-DNA ternary complex.
DNA replication in archaea and eukaryotes is executed by family B DNA polymerases, which exhibit full activity when complexed with the DNA clamp, proliferating cell nuclear antigen (PCNA). This replication enzyme consists of the polymerase and exonuclease moieties responsible for DNA synthesis and editing (proofreading), respectively. Because of the editing activity, this enzyme ensures the high fidelity of DNA replication. However, it remains unclear how the PCNA-complexed enzyme temporally switches between the polymerizing and editing modes. Here, we present the three-dimensional structure of the Pyrococcus furiosus DNA polymerase B-PCNA-DNA ternary complex, which is the core component of the replisome, determined by single particle electron microscopy of negatively stained samples. This structural view, representing the complex in the editing mode, revealed the whole domain configuration of the trimeric PCNA ring and the DNA polymerase, including protein-protein and protein-DNA contacts. Notably, besides the authentic DNA polymerase-PCNA interaction through a PCNA-interacting protein (PIP) box, a novel contact was found between DNA polymerase and the PCNA subunit adjacent to that with the PIP contact. This contact appears to be responsible for the configuration of the complex specific for the editing mode. The DNA was located almost at the center of PCNA and exhibited a substantial and particular tilt angle against the PCNA ring plane. The obtained molecular architecture of the complex, including the new contact found in this work, provides clearer insights into the switching mechanism between the two distinct modes, thus highlighting the functional significance of PCNA in the replication process.


Influence on effectiveness of early treatment with anti-TNF therapy in rheumatoid arthritis.
To evaluate the association between starting early treatment with anti-TNF and effectiveness as well as the possibility of applying therapeutic spacing in daily practice in patients with rheumatoid arthritis (RA). Observational, retrospective study conducted in two universitary hospitals in Spain. RA patients who received the first anti-TNF (adalimumab: ADA, etanercept: ETN or infliximab: IFX) during the study period (October 2006-2010) were included. Demographic data, time since diagnosis, disease activity (DAS28-ESR) and anti-TNF dosage were analyzed. Therapeutic objective was defined as DAS28 DAS28 < 2.6. Also the response related to criteria of the European League Against Rheumatism (EULAR) was evaluated. Therapeutic spacing was defined as the use of a lower dose or a higher interval according to label doses. The main endpoint was to assess the association between the effectiveness and the moment when the anti-TNF therapy begins. The secondary target was to evaluate the association between RA activity at the beginning of treatment with anti-TNF and dose used. Results. 82 patients were included. The prescription profile was: ADA (48.8%), ETN (31.7%) and IFX (19.5%). 71.4% of patients treated with anti-TNF during the first year since diagnosis, 57.1% of those who started after 1-5 years and 30.6% of patients who started after 5 years were in remission when the study ended. De-escalation strategy was performed in 25.6% of patients: ETN (38.5%), ADA (20.0%) and IFX (18.8%). The patients treated with a higher dose according to label doses were: IFX (81%), ADA, (12.5%) and ETN (7.7%). Results suggest that early treatment with anti-TNF can achieve a higher percentage of remissions. Therapeutic spacing is established as a strategy that improves the efficiency in those patients in remission, being the ETN the anti-TNF most susceptible for spacing, although a relation between the early beginning with anti-TNF and the used dose was not found.


Time to death and the forecasting of macro-level health care expenditures: some further considerations.
Although the effect of time to death (TTD) on health care expenditures (HCE) has been investigated using individual level data, the most profound implications of TTD have been for the forecasting of macro-level HCE. Here we estimate the TTD model using macro-level data from the Netherlands consisting of mortality rates and age- and gender-specific per capita health expenditures for the years 1981-2007. Forecasts for the years 2008-2020 of this macro-level TTD model were compared to forecasts that excluded TTD. Results revealed that the effect of TTD on HCE in our macro model was similar to those found in micro-econometric studies. As the inclusion of TTD pushed growth rate estimates from unidentified causes upwards, however, the two models' forecasts of HCE for the 2008-2020 were similar. We argue that including TTD, if modeled correctly, does not lower forecasts of HCE.


Perioperative cardiac evaluation, monitoring, and risk reduction strategies in noncardiac surgery patients.
Cardiac complications after noncardiac surgery cause significant morbidity and mortality. This review will discuss recent developments in risk stratification, monitoring, and risk reduction strategies. The addition of biomarkers for ischemia, left ventricular function, and atherosclerosis to classic cardiac risk factors improves the prediction of both short-term and long-term outcome after noncardiac surgery. Intraoperative monitoring, using continuous 12-lead ECG assessment and transesophageal echocardiography, may timely identify treatable myocardial ischemia and arrhythmias. A prudent perioperative beta-blocker and statin regimen can reduce cardiac complications and mortality without increasing the risk of stroke in intermediate to high-risk patients. The use of circulatory assist devices might improve outcomes after major surgery in patients with severely reduced left ventricular function. Systematic preoperative assessment can identify patients at high risk of cardiac complications and guide the application of appropriate risk reduction strategies.


Interleukin 15 upregulates the expression of PD-1 and TIM-3 on CD4+ and CD8+ T cells.
Virus-specific T cell-mediated immunity is severely impaired in chronic hepatitis B virus (HBV) patients. HBV-specific T cells in chronic HBV patients show a low ability to produce cytokines and to exert their cytotoxic activity. A prominent characteristic of these exhausted T cells is overexpression of inhibitory receptor molecules which negatively regulate T cell function. In this study, we examined in vitro regulation of two inhibitory receptor expressions, programmed death 1 (PD-1) and T cell immunoglobulin mucin domain-containing molecule 3 (TIM-3). Peripheral blood mononuclear cells (PBMCs) obtained from healthy individuals were in vitro stimulated with a panel of cytokines. PD-1 and TIM-3 expression levels on CD4+ and CD8+ T cells were examined at days 2 and 7 post stimulation. We demonstrated that PD-1 and TIM-3 were induced via polyclonal (anti-CD3) and cytokine (interleukin 15 [IL-15]) stimulations. Noteworthy, there was a significantly increased induction of TIM-3 on CD8+ T cells as compared to CD4+ T cells. Our study thus contributes to further understanding the regulation of T cell exhaustion markers PD-1 and TIM-3.


Effect of a pressor infusion of angiotensin II on sympathetic activity and heart rate in normal humans.
We tested the hypothesis that pressor infusions of angiotensin II (AII) could stimulate the sympathetic nervous system as reflected by norepinephrine (NE) spillover in humans. AII was infused at 5 ng/kg/min in six healthy volunteers, with vehicle and phenylephrine infusions as controls, on 3 separate days. Heart rate, mean arterial pressure, plasma NE, NE clearance, and NE spillover were assessed before and after 30-minute infusions of AII, vehicle, or phenylephrine in the supine position and then after 15 minutes of head-up and 15 minutes of head-down tilt. Both AII and phenylephrine raised mean arterial pressure (88 +/- 9.6 to 103 +/- 14 mm Hg, p less than 0.001, and 91 +/- 7.6 to 104 +/- 9.2 mm Hg, p less than 0.001, respectively), whereas heart rate fell only with phenylephrine (60 +/- 6 to 51 +/- 6.3 beats/min, p less than 0.001). Neither plasma NE nor NE spillover was affected by either infusion, and NE clearance declined slightly with both. No changes occurred in any variable during vehicle infusions in the supine position. During upright tilt, NE spillover increases were attenuated by both AII and phenylephrine while NE clearance changes were slightly greater, leaving plasma NE increases similar on each day. During head-down tilt, NE and NE spillover declined comparably on each study day.(ABSTRACT TRUNCATED AT 250 WORDS)


Phasic alertness enhances processing of face and non-face stimuli in congenital prosopagnosia.
Congenital prosopagnosia (CP) is a severe face processing impairment that occurs in the absence of any obvious brain damage and has often been associated with a more general deficit in deriving holistic relations between facial features or even between non-face shape dimensions. Here we further characterized this deficit and examined a potential way to ameliorate it. To this end we manipulated phasic alertness using alerting cues previously shown to modulate attention and enhance global processing of visual stimuli in normal observers. Specifically, we first examined whether individuals with CP, similarly to controls, would show greater global processing when exposed to an alerting cue in the context of a non-facial task (Navon global/local task). We then explored the effect of an alerting cue on face processing (upright/inverted face discrimination). Confirming previous findings, in the absence of alerting cues, controls showed a typical global bias in the Navon task and an inversion effect indexing holistic processing in the upright/inverted task, while CP failed to show these effects. Critically, when alerting cues preceded the experimental trials, both groups showed enhanced global interference and a larger inversion effect. These results suggest that phasic alertness may modulate visual processing and consequently, affect global/holistic perception. Hence, these findings further reinforce the notion that global/holistic processing may serve as a possible mechanism underlying the face processing deficit in CP. Moreover, they imply a possible route for enhancing face processing in individuals with CP and thus shed new light on potential amelioration of this disorder.


Engineering GST M2-2 for high activity with indene 1,2-oxide and indication of an H-site residue sustaining catalytic promiscuity.
The substrate-binding H-site of human glutathione transferase (GST) M2-2 was subjected to iterative saturation mutagenesis in order to obtain an efficient enzyme with the novel epoxide substrate indene 1,2-oxide. Residues 10, 116, and 210 were targeted, and the activities with the alternative substrates, benzyl isothiocyanate and the prodrug azathioprine, undergoing divergent chemical reactions were monitored for comparison. In general, increased activities were found when the smaller residues Gly, Ser, and Ala replaced the original Thr210. The most active mutant T210G was further mutated at position 116, but no mutant showed enhanced catalytic activity. However, saturation mutagenesis of position 10 identified one double mutant T210G/I10C with 100-fold higher specific activity with indene 1,2-oxide than wild-type GST M2-2. This enhanced epoxide activity of 50 μmol min(-1) mg(-1) resulted primarily from an increased k(cat) value (70 s(-1)). The specific activity is 24-fold higher than that of wild-type GST M1-1, which is otherwise the most proficient GST enzyme with epoxide substrates. A second double mutant T210G/I10W displayed 30-fold increased activity with azathioprine, 0.56 μmol min(-1) mg(-1). In both double mutants, the replacement of Ile10 led to narrowed acceptance of alternative substrates. Ile10 is evolutionarily conserved in related class Mu GSTs. Conservation usually indicates preservation of a particular function, and in the Mu class, it would appear that the conserved Ile10 is not necessary to maintain catalytic functions but to prevent loss of broad substrate acceptance. In summary, our data underscore the facile transition between alternative substrate selectivity profiles in GSTs by a few mutations.


Particle backscatter, extinction, and lidar ratio profiling with Raman lidar in south and north China.
Aerosol Raman lidar observations of profiles of the particle extinction and backscatter coefficients and the respective extinction-to-backscatter ratio (lidar ratio) were performed under highly polluted conditions in the Pearl River Delta (PRD) in southern China in October 2004 and at Beijing during a clear period with moderately polluted to background aerosol conditions in January 2005. The anthropogenic haze in the PRD is characterized by volume light-extinction coefficients of particles ranging from approximately 200 to 800 Mm(-1) and lidar ratios mostly between 40 and 55 sr (average of 47+/-6 sr). Almost clean air masses were observed throughout the measurements of the Beijing campaign. These air masses originated from arid desert-steppe-like regions (greater Gobi area). Extinction values usually varied between 100 and 300 Mm(-1), and the lidar ratios were considerably lower (compared with PRD values) with values mostly from 30 to 45 sr (average of 38+/-7 sr). Gobi dust partly influenced the observations. Unexpectedly low lidar ratios of approximately 25 sr were found for a case of background aerosol with a low optical depth of 0.05. The low lidar ratios are consistent with Mie-scattering calculations applied to ground-based observations of particle size distributions.


Inactivation of surface viruses by gaseous ozone.
Environmental surfaces may be contaminated with viruses and contribute to their transmission. Concerns have arisen in trying to control viruses because of an increasing incidence of viral infections. Ozone is considered to be a promising method to inactivate viruses on surfaces. In this investigation, the effects of ozone concentration, contact time, different capsid architecture of viruses, and relative humidity (RH) on inactivating viruses by ozone were evaluated. The authors observed that the survival fraction of viruses on surfaces decreased exponentially with increasing ozone dose. Viruses required ozone doses of 20-112 min(mg/m3) (contact time [min] multiplied by ozone concentration [mg/m3) for 90% inactivation and 47-223 min(mg/m3) for 99% inactivation. The ozone dose for 99% inactivation was two times higher than for 90% inactivation. The required ozone concentration at 85% RH was lower than at 55% RH. In summary, ozone should be an effective method for reducing the viral number between 1 and 3 logs on surfaces.


Modulators of Acetylcholinesterase Activity: From Alzheimer's Disease to Anti-Cancer Drugs.
Acetylcholinesterase (AChE) is involved in the termination of impulse transmission by rapid hydrolysis of the neurotransmitter acetylcholine in numerous cholinergic pathways in the central and peripheral nervous systems. The enzyme inactivation leads to acetylcholine accumulation, hyperstimulation of nicotinic and muscarinic receptors, and disrupted neurotransmission. Hence, acetylcholinesterase inhibitors, interacting with the enzyme as their primary target, are applied as relevant drugs for different neurodegenerative diseases (such as Alzheimer's and Parkinson's) as well as toxins. At the same time, there are increasing evidence that in non-neuronal context, AChE is involved in the regulation of cell proliferation, differentiation, apoptosis and cell-cell interaction. An irregular expression of AChE has been found in different types of tumors, suggesting the involvement of AChE in the regulation of tumor development. Having all this in mind, there is a possibility that some AChE inhibitors could be used as anti-cancer agents. This contribution will discuss a broad range of possible application of different AChE inhibitors as drugs, from well-known anti-Alzheimer's disease drugs to their use in cancer treatment in future. Emphasis will be put on various known AChE inhibitors classes, whose application as drugs could be controversy, as well as on newly investigated natural products, which can also modulate AChE activity. It is not clear a patient treated for neurodegenerative condition prone to increased risk for some types of cancer and vice versa. This is necessary to keep in mind during rational drug design process for all therapies, which are based on AChE as a target molecule.


Chronic treatment with the peroxisome proliferator-activated receptor alpha agonist Wy-14,643 attenuates myocardial respiratory capacity and contractile function.
We investigated whether chronic in vivo treatment with the peroxisome proliferator-activated receptor alpha agonist Wy-14,643 attenuates cardiac contractile function by impairing mitochondrial respiration. Wy-14,643 (25 mg kg(-1) day(-1)) was administered to Wistar rats by oral gavage for 14 consecutive days, after which ex vivo heart function, myocardial mitochondrial respiratory capacity, and metabolic gene expression were determined. Body and heart weights were not significantly altered following 14 days of Wy-14,643 administration. Heart perfusion studies showed significantly reduced systolic and developed pressures, while the rate pressure product declined by 36 +/- 2.6% (P < 0.01 vs. vehicle) after 14 days of Wy-14,643 treatment. State 3 mitochondrial respiration was lower in the Wy-14,643 group (P = 0.06 vs. vehicle). State 4 respiration and oligomycin-insensitive proton leak were significantly increased compared with matched controls. The rate of ADP phosphorylation was also decreased by 44.9 +/- 1.9% (P < 0.05 vs. vehicle). Pyruvate dehydrogenase kinase 4 (PDK4) and uncoupling protein 3 (UCP3) transcript levels were upregulated, while cytochrome oxidase II (COXII) expression was decreased following Wy-14,643 treatment. This study demonstrates that chronic in vivo Wy-14,643 administration impaired cardiac contractile function in parallel with decreased mitochondrial respiratory function and increased uncoupling.


Divergent Evolutionary Trajectories of Two Young, Homomorphic, and Closely Related Sex Chromosome Systems.
There exists extraordinary variation among species in the degree and nature of sex chromosome divergence. However, much of our knowledge about sex chromosomes is based on comparisons between deeply diverged species with different ancestral sex chromosomes, making it difficult to establish how fast and why sex chromosomes acquire variable levels of divergence. To address this problem, we studied sex chromosome evolution in two species of African clawed frog (Xenopus), both of whom acquired novel systems for sex determination from a recent common ancestor, and both of whom have female (ZW/ZZ) heterogamy. Derived sex chromosomes of one species, X. laevis, have a small region of suppressed recombination that surrounds the sex determining locus, and have remained this way for millions of years. In the other species, X. borealis, a younger sex chromosome system exists on a different pair of chromosomes, but the region of suppressed recombination surrounding an unidentified sex determining gene is vast, spanning almost half of the sex chromosomes. Differences between these sex chromosome systems are also apparent in the extent of nucleotide divergence between the sex chromosomes carried by females. Our analyses also indicate that in autosomes of both of these species, recombination during oogenesis occurs more frequently and in different genomic locations than during spermatogenesis. These results demonstrate that new sex chromosomes can assume radically different evolutionary trajectories, with far-reaching genomic consequences. They also suggest that in some instances the origin of new triggers for sex determination may be coupled with rapid evolution sex chromosomes, including recombination suppression of large genomic regions.


Profiling individual clinical responses by high-frequency serum neurofilament assessment in MS.
To evaluate individual neurofilament light chain (NfL) variation over the time of disease course and the potential of NfL measurement to predict treatment response in patients with MS. We investigated 15 patients with MS after immune reconstitution treatment with alemtuzumab (ATZ). Monthly serum NfL (sNFL) measurements were correlated with Expanded Disability Status Scale (EDSS), MRI, and relapse activity over an observational period of up to 102 months. Before ATZ, sNfL was significantly increased in correlation with previous relapse/MRI activity. After ATZ, sNfL decreased quickly within the first 6 months. In patients classified as NEDA-3, sNfL declined and persisted at an individual low steady-state level of <8 pg/mL. During follow-up, 34 sNfL peaks with a >20 fold increase could be detected, which were associated with clinical or MRI disease activity. Even patient-reported relapse-suspicious symptoms, which have not been confirmed because relapses were accompanied by sNfL, increase, proposing sNfL assessment as a marker for relapse activity. sNfL started to increase earliest 5 months before, peaked at clinical onset, and recovered within 4-5 months. sNfL presented at higher levels in active patients requiring ATZ retreatment compared with responder patients. During 2 documented pregnancies, sNfL was at a low level, whereas a postpartum transient sNfL increase was seen without any signs of activity. This study applied a long-term high-frequency sNfL assessment in an ATZ-treated cohort, allowing a holistic profiling on the individual level and highlighted that sNfL can eminently complement the individual clinical and MRI monitoring in clinical practice.


Electrofluidics fabricated by space-selective metallization in glass microfluidic structures using femtosecond laser direct writing.
Space-selective metallization of the inside of glass microfluidic structures using femtosecond laser direct-write ablation followed by electroless plating is demonstrated. Femtosecond laser direct writing followed by thermal treatment and successive chemical etching allows us to fabricate three-dimensional microfluidic structures inside photosensitive glass. Then, femtosecond laser ablation followed by electroless metal plating enables flexible deposition of patterned metal films on desired locations of not only the top and bottom walls but also the sidewalls of fabricated microfluidic structures. A volume writing scheme for femtosecond laser irradiation inducing homogeneous ablation on the sidewalls of microfluidic structures is proposed for sidewall metallization. The developed technique is used to fabricate electrofluidics in which microelectric components are integrated into glass microchannels. The fabricated electrofluidics are applied to control the temperature of liquid samples in the microchannels for the enhancement of chemical reactions and to manipulate the movement of biological samples in the microscale space.


Associations Between Hepatitis B Virus Infection and Risk of All Cancer Types.
Hepatitis B virus (HBV) has been identified as a major risk factor for hepatocellular carcinoma. However, the associations between HBV infection and other cancer types are not well understood. To assess the associations between chronic HBV infection and risk of all cancer types. This population-based study involved 3 cohorts in China. The China Kadoorie Biobank (CKB) prospective cohort study, conducted between June 2004 and July 2008, used a dipstick assay for detection of serum hepatitis B surface antigen (HBsAg) among 496 732 participants to determine the association between HBV infection and risk of all cancer types. Two cohort studies were used to validate the associations by applying more precise serum HBsAg detection assays: the Qidong cohort (37 336 participants enrolled from November 2007 to April 2011) and the Changzhou nested case-control study (17 723 participants enrolled from June 2004 to September 2005). A total of 97 samples of stomach cancer tissues, 10 samples of pancreatic cancer tissues, and 9 samples of lung cancer tissues were included to assess the presence of HBV replication and expression. Statistical analysis was performed from December 2016 to October 2018. Serum HBsAg status in the population-based stage and HBV DNA status, the expression of hepatitis B X protein, and hepatitis B core antibody (anti-HBc) in the tissue-based stage. Incidence of all cancer types during follow-up. In the CKB cohort, the mean (SD) age of the 496 732 participants was 51.5 (10.7) years; 59.0% of the participants were women. After 4.4 million person-years of follow-up, participants who were HBsAg seropositive (n = 15 355) had a higher risk of hepatocellular carcinoma (hazard ratio [HR], 15.77; 95% CI, 14.15-17.57), stomach cancer (HR, 1.41; 95% CI, 1.11-1.80), colorectal cancer (HR, 1.42; 95% CI, 1.12-1.81), oral cancer (HR, 1.58; 95% CI, 1.01-2.49), pancreatic cancer (HR, 1.65; 95% CI, 1.03-2.65), and lymphoma (HR, 2.10; 95% CI, 1.34-3.31) when compared with participants who were HBsAg seronegative (n = 481 377). Because of the limitation of sample size, only associations of HBV infection with hepatocellular carcinoma and stomach cancer were validated in the Qidong cohort (hepatocellular carcinoma: HR, 17.51; 95% CI, 13.86-22.11; stomach cancer: HR, 2.02; 95% CI, 1.24-3.29); the Changzhou nested case-control study validated only an association between HBV infection and stomach cancer (odds ratio, 1.76; 95% CI, 1.04-2.98). Moreover, among 22 participants with stomach cancer from the Qidong cohort who were anti-HBc seropositive, 12 samples (54.5%) of cancer tissues were HBV DNA positive, while among 25 participants with stomach cancer who were anti-HBc seronegative, no HBV DNA was detected. The same negative and positive rate was observed in the validation set from Zhejiang Tumor Hospital (19 of 35 samples [54.3%] were HBV DNA positive). Moreover, among the 8 patients with stomach cancer from the Qidong cohort who were anti-HBc seropositive, anti-HBc and hepatitis B X protein were expressed in all of their stomach cancer tissue samples. The same phenomenon was observed in the patients with pancreatic cancer but not in the patients with lung cancer, which was consistent with the population-based results of the CKB cohort. This study found that HBV infection was also associated with the risk of nonliver cancer, especially digestive system cancers among adults in China.


The inflammatory gene pathway is not a major contributor to polycystic ovary snydrome.
Although inflammation is clearly associated with obesity, diabetes, and insulin resistance, the role of chronic inflammation in the etiology of polycystic ovary syndrome (PCOS) is unclear. To determine whether chronic inflammation plays a causal role in the etiology of PCOS, we tested for an association between PCOS and genetic markers mapping to 80 members of the inflammatory pathway. This was a case-control association study. The setting was an academic medical center. A total of 905 index case patients with PCOS and 955 control women (108 intensively phenotyped subjects with normal androgen levels and regular menses and 847 minimally phenotyped subjects with regular menses and no history of PCOS). Subjects were genotyped at single nucleotide polymorphisms mapping to 80 inflammatory genes. Logistic regression was used to test for an association between 822 single nucleotide polymorphisms and PCOS after adjustment for population stratification, body mass index, and/or age. In the index patients, we also tested for association with 11 quantitative traits (body mass index and testosterone, fasting insulin, fasting glucose, 2-hour postchallenge glucose, LH, FSH, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglyceride levels). The evidence for an association with PCOS and with 11 quantitative traits was investigated. Nominally significant evidence for an association was observed with MAP3K7, IKBKG, TNFRS11A, AKT2, IL6R, and IRF1, but no results remained statistically significant after adjustment for multiple testing. Genetic variation in the inflammatory pathway is not a major contributor to the etiology of PCOS or related quantitative traits in women with PCOS.


Highly tunable interfacial adhesion of glass fiber by hybrid multilayers of graphene oxide and aramid nanofiber.
The performance of fiber-reinforced composites is governed not only by the nature of each individual component comprising the composite but also by the interfacial properties between the fiber and the matrix. We present a novel layer-by-layer (LbL) assembly for the surface modification of a glass fiber to enhance the interfacial properties between the glass fiber and the epoxy matrix. Solution-processable graphene oxide (GO) and an aramid nanofiber (ANF) were employed as active components for the LbL assembly onto the glass fiber, owing to their abundant functional groups and mechanical properties. We found that the interfacial properties of the glass fibers uniformly coated with GO and ANF multilayers, such as surface free energy and interfacial shear strength, were improved by 23.6% and 39.2%, respectively, compared with those of the bare glass fiber. In addition, the interfacial adhesion interactions between the glass fiber and the epoxy matrix were highly tunable simply by changing the composition and the architecture of layers, taking advantage of the versatility of the LbL assembly.


Effect of creatine supplementation and resistance-exercise training on muscle insulin-like growth factor in young adults.
The purpose of this study was to compare changes in muscle insulin-like growth factor-I (IGF-I) content resulting from resistance-exercise training (RET) and creatine supplementation (CR). Male (n=24) and female (n=18) participants with minimal resistance-exercise-training experience (=1 year) who were participating in at least 30 min of structured physical activity (i.e., walking, jogging, cycling) 3-5 x/wk volunteered for the study. Participants were randomly assigned in blocks (gender) to supplement with creatine (CR: 0.25 g/kg lean-tissue mass for 7 days; 0.06 g/kg lean-tissue mass for 49 days; n=22, 12 males, 10 female) or isocaloric placebo (PL: n=20, 12 male, 8 female) and engage in a whole-body RET program for 8 wk. Eighteen participants were classified as vegetarian (lacto-ovo or vegan; CR: 5 male, 5 female; PL: 3 male, 5 female). Muscle biopsies (vastus lateralis) were taken before and after the intervention and analyzed for IGF-I using standard immunohistochemical procedures. Stained muscle cross-sections were examined microscopically and IGF-I content quantified using image-analysis software. Results showed that RET increased intramuscular IGF-I content by 67%, with greater accumulation from CR (+78%) than PL (+54%; p=.06). There were no differences in IGF-I between vegetarians and nonvegetarians. These findings indicate that creatine supplementation during resistance-exercise training increases intramuscular IGF-I concentration in healthy men and women, independent of habitual dietary routine.


Light-Driven Expansion of Spiropyran Hydrogels.
The incorporation of molecular switches in organic structures is of great interest in the chemical design of stimuli-responsive materials that mimic the complex functions of living systems. Merocyanine dyes that convert to spiropyran moieties upon exposure to visible light have been extensively studied as they can be incorporated in hydrated covalent networks that will expel water when this conversion occurs and induce a volumetric shrinkage. We report here on a sulfonate-based water-soluble photoswitch that, in contrast to the well-known systems, triggers a volumetric expansion in hydrogels upon exposure to photons. Contraction is in turn observed under dark conditions in a highly reversible manner. The novel behavior of the photoswitch incorporated in the covalent network was predicted by coarse-grained simulations of the system's chemical structure. Using pH control and polymeric structures that differ in lower critical solution temperature, we were able to develop hydrogels with highly tunable volumetric expansion. The novel molecular function of the systems developed here led to materials with the negative phototaxis observed in plants and could expand the potential use of hydrogels as sensors, soft robots, and actuators.