Bone health of immigrant Chinese women living in New York City.
Osteoporosis is a serious national and global public health problem, but data on bone health are limited for Asian women living in the U.S., the majority of whom are Chinese. For this study, we measured bone mineral density (BMD) by dual energy X-ray densitometry (DXA) at the lumbar spine and hip region in 300 immigrant Chinese women, ages 40-90 y, living in New York City. We also collected demographic and health data, information about knowledge and care for osteoporosis, and anthropometric measures, and estimated calcium intake from the women. In our sample, 55% had osteoporosis and 38% had low bone mass (osteopenia). Older age, lower body mass index (BMI), and shorter height were associated with lower BMD at all sites. Years lived in the U.S. and number of children were also associated with lower BMD of the lumbar spine. Chinese women who emigrated from Mainland China had lower BMD at the lumbar spine and femoral neck than Chinese women who emigrated from Hong Kong, after adjusting for potential confounders. Both groups of immigrant women had lower BMD at all sites than a national sample of U.S. Caucasian women. Although the women in our study had generally poor knowledge about osteoporosis, most could identify at least one food rich in calcium. The large number of immigrant Chinese women in New York City with osteoporosis calls for major efforts to increase awareness, prevention, diagnosis, and treatment of this condition in this susceptible population.


The differential impact of two anesthetic techniques on cortisol levels in Nigerian surgical patients.
Surgical procedures are associated with a complexity of stress response characterized by neurohumoral, immunologic, and metabolic alterations. The aim was to compare the effects on the stress response by isoflurane-based intratracheal general anesthesia (ITGA) and bupivacaine-based epidural anesthesia (EA), using cortisol as a biochemical marker. Following the approval of the Hospital Ethical Board, informed written consent from patients recruited into this study was obtained. One group received general anesthesia with relaxant technique (group A) while the other group had bupicaine epidural anesthesia with catheter placement for top-ups (group B) for their surgeries. Both groups were assessed for plasma cortisol levels - baseline, 30 minutes after skin the start of surgery and at skin closure. There was no statistically significant difference in the baseline mean heart rate, mean arterial pressure (mean MAP) and the mean duration of surgery between the two groups; the baseline mean plasma cortisol level was 88.70 ± 3.85 ng/ml for group A and 85.55 ± 2.29 ng/ml for group B, P=0.148. At 30 minutes after the start of surgery the plasma cortisol level in the GA group was 361.60 ± 31.27 ng/ml while it was 147.45 ± 22.36 ng/ml in the EA group, showing a significant difference, P=0.001. At skin closure the mean plasma cortisol value of 384.65 ± 48.04 ng/ml recorded in the GA group was found to be significantly higher than the value of 140.20 ± 10.74 ng/ml in the GA group, P<0.002. Using plasma cortisol as a measure, bupivacaine-based epidural anesthesia significantly reduces the stress response to surgical stimuli when compared with isoflurane-based tracheal general anesthesia.


An aeration control strategy for oxidation ditch processes based on online oxygen requirement estimation.
A feedforward-feedback aeration control strategy based on online oxygen requirements (OR) estimation is proposed for oxidation ditch (OD) processes, and it is further developed for intermittent aeration OD processes, which are the most popular type in Japan. For calculating OR, concentrations of influent biochemical oxygen demand (BOD) and total Kjeldahl nitrogen (TKN) are estimated online by the measurement of suspended solids (SS) and sometimes TKN is estimated by NH4-N. Mixed liquor suspended solids (MLSS) and temperature are used to estimate the required oxygen for endogenous respiration. A straightforward parameter named aeration coefficient, Ka, is introduced as the only parameter that can be tuned automatically by feedback control or manually by the operators. Simulation with an activated sludge model was performed in comparison to fixed-interval aeration and satisfying result of OR control strategy was obtained. The OR control strategy has been implemented at seven full-scale OD plants and improvements in nitrogen removal are obtained in all these plants. Among them, the results obtained in Yumoto wastewater treatment plant were presented, in which continuous aeration was applied previously. After implementing intermittent OR control, the total nitrogen concentration was reduced from more than 5 mg/L to under 2 mg/L, and the electricity consumption was reduced by 61.2% for aeration or 21.5% for the whole plant.


Continuous subcutaneous infusion of pramipexole protects against lipopolysaccharide-induced dopaminergic cell death without affecting the inflammatory response.
The D2/D3 dopamine receptor agonist pramipexole, protects against toxin-induced dopaminergic neuronal destruction but its mechanism of action is unknown. Inflammation following glial cell activation contributes to cell death in Parkinson's disease and we now report on the effects of acute or chronic administration of pramipexole on lipopolysaccharide (LPS) induced inflammation and nigral dopaminergic cell death in the rat. At 48 h and 30 days following supranigral administration of LPS, approximately 70% of tyrosine hydroxylase (TH) immunoreactive (-ir) cells in substantia nigra had degenerated with a corresponding loss of TH-ir terminals in the striatum. In rats acutely treated with pramipexole (2x1 mg/kg; s.c.) 48 h following LPS application, there was no difference in the number of TH-ir cells or terminals compared to LPS-treated rats receiving vehicle. However, the continuous subcutaneous infusion of pramipexole for 7 days prior to LPS and 21 days subsequently, produced a marked preservation of both TH-ir cells and terminals. At 48 h or 30 days, LPS induced an up-regulation of ubiquitin-ir within the nigral TH-ir neurones, which was reduced by pramipexole treatment. Thirty days following supranigral LPS administration (9 days after the end of infusion), (+)-amphetamine (5 mg/kg, i.p.) caused robust ipsiversive rotation. In rats treated with LPS but receiving continuous subcutaneous administration of pramipexole, (+)-amphetamine-induced rotation was markedly reduced. LPS-induced increase in the levels of inflammatory markers, were not affected by either acute administration or continuous infusion of pramipexole. Continuous infusion of pramipexole protected dopaminergic neurones against inflammation induced degeneration but without modification of the inflammatory response.


Detection of carbon monoxide and water absorption lines in an exoplanet atmosphere.
Determining the atmospheric structure and chemical composition of an exoplanet remains a formidable goal. Fortunately, advancements in the study of exoplanets and their atmospheres have come in the form of direct imaging--spatially resolving the planet from its parent star--which enables high-resolution spectroscopy of self-luminous planets in jovian-like orbits. Here, we present a spectrum with numerous, well-resolved molecular lines from both water and carbon monoxide from a massive planet orbiting less than 40 astronomical units from the star HR 8799. These data reveal the planet's chemical composition, atmospheric structure, and surface gravity, confirming that it is indeed a young planet. The spectral lines suggest an atmospheric carbon-to-oxygen ratio that is greater than that of the host star, providing hints about the planet's formation.


Couch potatoes or french fries: are sedentary behaviors associated with body mass index, physical activity, and dietary behaviors among adolescents?
To describe the demographic characteristics of adolescent boys and girls who engage in three sedentary behaviors (television/video use, computer use, and reading/homework), and to explore how each sedentary activity is associated with body mass index (BMI), dietary behaviors, and leisure time physical activity. This study draws on data collected from Project EAT (Eating Among Teens), a school-based survey examining personal, behavioral, and socioenvironmental factors that are associated with nutritional intake among adolescents. The study sample consists of 4746 middle and high school students from 31 public schools in a metropolitan area of the upper Midwest. All students were invited to participate. The overall response rate for Project EAT was 81.5%. Data collection was completed during the 1998-1999 school year. Multivariate linear regression was used for examining associations between independent and dependent variables, controlling for age, race/ethnicity, and socioeconomic status. All differences were considered statistically significant at P<.05. Among boys, television/video use and time spent reading/doing homework were positively associated with BMI (P<.05), whereas for girls television/video and computer use were positively associated with BMI (P<.05). High television/video use among boys and girls was associated with more unhealthful dietary behaviors (eg, increased consumption of soft drinks, fried foods, and snacks) (P<.05). In contrast, time spent reading/doing homework was associated with more healthful dietary behaviors (eg, increased consumption of fruits and vegetables) (P<.05). Leisure time physical activity was not associated with television/video use among boys or girls, but was positively associated with computer use and time spent reading/doing homework (P<.05). Applications/Conclusions Messages and advice aimed at reducing time spent in sedentary activities should be targeted at television/video use instead of time spent reading, doing homework, or using a computer. Nutrition education should incorporate messages about the influence of the media and advertising on dietary behaviors.


Association between type of medication instruction and patients' knowledge, side effects, and compliance.
The authors measured knowledge about medication and its side effects, impact of side effects, and compliance in 30 chronic outpatients before and after they participated in two instruction sessions about their medication held one month apart. Instruction consisted of a verbal or a written and verbal presentation and minimum or maximum information about side effects. All patients' medication knowledge increased after both sessions. Those on high doses of neuroleptics given verbal and written information gained significantly more medication knowledge than those given only verbal information. After instruction, more patients knew about specific side effects, including tardive dyskinesia, and both patients given only verbal instruction and those given minimum information about side effects had fewer problems with side effects. Instruction did not affect compliance.


microRNA regulation of human pancreatic cancer stem cells.
microRNAs (miRNAs) are a group of small non-coding RNAs that function primarily in the post transcriptional regulation of gene expression in plants and animals. Deregulation of miRNA expression in cancer cells, including pancreatic cancer cells, is well documented, and the involvement of miRNAs in orchestrating tumor genesis and cancer progression has been recognized. This review focuses on recent reports demonstrating that miRNAs are involved in regulation of pancreatic cancer stem cells (CSCs). A number of miRNA species have been identified to be involved in regulating pancreatic CSCs, including miR-21, miR-34, miR-1246, miR-221, the miR-17-92 cluster, the miR-200 and let-7 families. Furthermore, the Notch-signaling pathway and epithelial-mesenchymal transition (EMT) process are associated with miRNA regulation of pancreatic CSCs. Given the significant contribution of CSCs to chemo-resistance and tumor progression, a better understanding of how miRNAs function in pancreatic CSCs could provide novel strategies for the development of therapeutics and diagnostics for this devastating disease.


Assessment of oxidative stress markers and hearing thresholds in patients with obstructive sleep apnea-hypopnoea treated with cysteine and superoxide dismutase therapy.
In OSAHS, the hypoxia and reoxygenation cicles, maintain a state of oxidative stress, which seems to cause a change in the oxidative balance. Our aim is to compare the markers of oxidative stress with audiological findings and OSAHS severity, in OSAHS patients untreated and also treated ones, with cysteine and superoxide dismutase. 65 patients (42 Men, 23 Women) with 30-65 years age range have been enrolled, with a mean age of 52.6 ± 13.3 years with moderate OSAHS. We have analyzed plasma and lymphocyte markers of oxidative stress (glutathione, thioredoxin and heat shock protein) and they were underwent tonal audiometry. Patients were divided in two groups: Group A (32 patients) included patients treated for 8 weeks with cysteine and superoxide dismutase; Group B (33 patients) included patients untreated. The research showed a significant increase in reduced glutathione levels (p<0.05) in OSAHS patients treated; conversely, it showed a decrease of oxidized glutathione level (p<0.05) in treated patients than OSAHS untreated ones. The thioredoxin values, in untreated OSAHS patients, appear to be reduced than in OSAHS patients treated (p<0.05), and that the heat shock protein values were more elevated in untreated OSAHS patients (p<0.05). Finally, it was found that a correlation exists between the severity of OSAHS and auditory dysfunction. The study of the oxidative stress markers has produced results which lead to support the idea that, in a personalized therapy context, the use of antioxidant therapy can cooperate effectively the first choice treatment.


Wavelet-based estimation of a semiparametric generalized linear model of fMRI time-series.
This paper addresses the problem of detecting significant changes in fMRI time series that are correlated to a stimulus time course. This paper provides a new approach to estimate the parameters of a semiparametric generalized linear model of fMRI time series. The fMRI signal is described as the sum of two effects: a smooth trend and the response to the stimulus. The trend belongs to a subspace spanned by large scale wavelets. The wavelet transform provides an approximation to the Karhunen-Loève transform for the long memory noise and we have developed a scale space regression that permits to carry out the regression in the wavelet domain while omitting the scales that are contaminated by the trend. In order to demonstrate that our approach outperforms the state-of-the art detrending technique, we evaluated our method against a smoothing spline approach. Experiments with simulated data and experimental fMRI data, demonstrate that our approach can infer and remove drifts that cannot be adequately represented with splines.


Human cancer cells retain modest levels of enzymatically active matriptase only in extracellular milieu following induction of zymogen activation.
The type 2 transmembrane serine protease matriptase is broadly expressed in human carcinomas and hematological cancers. The proteolytic activity of matriptase is a potential target of drugs and imaging probes. We assessed the fate of active matriptase following the induction of matriptase zymogen activation. Exposing eight human carcinoma cells to pH 6.0 buffer induced robust matriptase zymogen activation followed by rapid inhibition of the nascent active matriptase by hepatocyte growth factor activator inhibitor (HAI)-1. Consequently, no enzymatically active matriptase was detected in these cells. Some active matriptase is, however, rapidly shed to the extracellular milieu by these carcinoma cells. The lack of cell-associated active matriptase and the shedding of active matriptase were also observed in two hematological cancer lines. Matriptase shedding is correlated closely with the induction of matriptase activation, suggesting that matriptase activation and shedding are kinetically coupled. The coupling allows a proportion of active matriptase to survive HAI-1 inhibition by rapid shedding from cell surface. Our study suggests that cellular free, active matriptase is scarce and might not be an effective target for in vivo imaging and drug development.


The role of cardiovascular disease in the relationship between air pollution and incident dementia: a population-based cohort study.
Evidence suggests a link between air pollution and dementia. Cardiovascular disease (CVD) may be a potential determinant of dementia. This motivated us to quantify the contribution of CVD to the association between air pollution and dementia. A cohort of Canadian-born residents of Ontario, who participated in the 1996-2003 Canadian Community Health Surveys, was followed through 2013 or until dementia diagnosis. Exposure to nitrogen dioxide (NO2) and fine particulate matter (PM2.5) was estimated with a 3-year average and 5-year lag before dementia diagnosis. Incident CVD was evaluated as a mediator. We used multi-level Cox proportional and Aalen additive hazard regression models, adjusting for individual- and neighbourhood-level risk factors to estimate associations with NO2 and PM2.5. We estimated the total, direct and indirect effects of air pollution on dementia through cardiovascular disease. This study included 34 391 older adults. At baseline, the mean age of this cohort was 59 years. The risk of dementia was moderately higher among those more exposed to NO2 (hazard ratio (HR) 1.10, 95% confidence interval (CI) 0.99-1.19; and 100 additional cases per 100 000 [standard error (SE) <100x10-5]) and PM2.5 [(HR 1.29, 95% CI 0.99-1.64; 200 additional cases per 100 000] [SE 100x10-5]) after adjusting for covariates; however, these estimates are imprecise. A greater proportion of the relationship between PM2.5 and dementia was mediated through CVD than NO2 for both scales. These results suggest some of the association between air pollution and dementia is mediated through CVD, indicating that improving cardiovascular health may prevent dementia in areas with higher exposure to air pollution.


Long-Term Implications of Welfare Reform for the Development of Adolescents and Young Adults.
We draw upon the 3-wave longitudinal dataset called Welfare Children and Families: A Three-City Study to examine the long-term implications for adolescents and young adults (N=783) of mothers' welfare receipt and labor force participation from 1999 to 2005. In general, changes in mothers' work and welfare patterns were not associated with deterioration or improvement in youth development (ages 16 to 20 years at wave 3). The few significant associations suggested that youth whose mothers increased employment (net of welfare participation) were more likely to show declines in serious behavior problems and delinquency compared to youth whose mothers were unemployed or employed part-time during the study period. Welfare roll exits (controlling for employment experiences) were unrelated to adolescent and young adult outcomes. Mothers' employment transitions were linked to improvements in household income and mothers' self esteem in addition to reductions in financial strain and their own illegal activities. However, these associations did not explain the relation between maternal employment and youths' improved behavior. These results do not support the predictions of either the supporters or the opponents of welfare reform, an outcome we discuss.


Hippocampal pyramidal neurons switch from a multipolar migration mode to a novel "climbing" migration mode during development.
The hippocampus plays important roles in brain functions. Despite the importance of hippocampal functions, recent analyses of neuronal migration have mainly been performed on the cerebral neocortex, and the cellular mechanisms responsible for the formation of the hippocampus are not yet completely understood. Moreover, why a prolonged time is required for hippocampal neurons to complete their migration has been unexplainable for several decades. We analyzed the migratory profile of neurons in the developing mouse hippocampal CA1 region and found that the hippocampal pyramidal neurons generated near the ventricle became postmitotic multipolar cells and accumulated in the multipolar cell accumulation zone (MAZ) in the late stage of development. The hippocampal neurons passed through the pyramidal layer by a unique mode of migration. Their leading processes were highly branched and made contact with many radial fibers. Time-lapse imaging revealed that the migrating cells changed their scaffolds from the original radial fibers to other radial fibers, and as a result they proceed in a zigzag manner, with long intervals. The migrating cells in the hippocampus reminded us of "rock climbers" that instead of using their hands to pull up their bodies were using their leading processes to pull up their cell bodies. Because this mode of migration had never been described, we called it the "climbing" mode. The change from the "climbing" mode in the hippocampus to the "locomotion" mode in the neocortex may have contributed to the brain expansion during evolution.


The Antiresorptive Effect of GIP, But Not GLP-2, Is Preserved in Patients With Hypoparathyroidism-A Randomized Crossover Study.
Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-2 (GLP-2) are gut hormones secreted postprandially. In healthy humans, both hormones decrease bone resorption accompanied by a rapid reduction in parathyroid hormone (PTH). The aim of this study was to investigate whether the changes in bone turnover after meal intake and after GIP- and GLP-2 injections, respectively, are mediated via a reduction in PTH secretion. This was tested in female patients with hypoparathyroidism given a standardized liquid mixed-meal test (n = 7) followed by a peptide injection test (n = 4) using a randomized crossover design. We observed that the meal- and GIP- but not the GLP-2-induced changes in bone turnover markers were preserved in the patients with hypoparathyroidism. To understand the underlying mechanisms, we examined the expression of the GIP receptor (GIPR) and the GLP-2 receptor (GLP-2R) in human osteoblasts and osteoclasts as well as in parathyroid tissue. The GIPR was expressed in both human osteoclasts and osteoblasts, whereas the GLP-2R was absent or only weakly expressed in osteoclasts. Furthermore, both GIPR and GLP-2R were expressed in parathyroid tissue. Our findings suggest that the GIP-induced effect on bone turnover may be mediated directly via GIPR expressed in osteoblasts and osteoclasts and that this may occur independent of PTH. In contrast, the effect of GLP-2 on bone turnover seems to depend on changes in PTH and may be mediated through GLP-2R in the parathyroid gland. © 2021 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).


Study of central nervous system depressant and behavioral activity of an ethanol extract of Achyranthes aspera (Agadha) in different animal models.
Achyranthes aspera Linn., an indigenous herb, has been reported to have antifertility, antihyperlipidemic, antidiabetic, immunomodulatory, anticarcinogenic, diuretic, cardiotonic, analgesic anti-inflammatory, hypnotic, antifungal, antibacterial, and central antinociceptive activities. This study was designed to evaluate depressant effects on central nervous system (CNS) and behavioral effects of ethanol extract of A. aspera (EEAA) and to find the phytochemical responsible for these activities. The pharmacological assays used to study CNS depressant effect in albino mice were rota rod and actophotometer performance test. Effects on behavioral activity were studied using open field test. The extract was given intraperitoneally (i.p.) at a dose of 400 mg/kg. Diazepam (2 mg/kg body weight i.p.) was used as standard. Data were analyzed by using analysis of variance followed by Dunnett's test. P < 0.05 was considered significant. Phytochemical screening revealed presence of triterpenoids, saponins, alkaloids (betaine, achyranthine), and steroids as major constituents. The result of this study reflected that EEAA (400 mg/kg i.p.) decreased locomotor activity, produced muscle relaxation, and showed anxiolytic activity. EEAA exhibit CNS depressant and significant anxiolytic activity comparable to diazepam.


Complex master slave interferometry.
A general theoretical model is developed to improve the novel Spectral Domain Interferometry method denoted as Master/Slave (MS) Interferometry. In this model, two functions, g and h are introduced to describe the modulation chirp of the channeled spectrum signal due to nonlinearities in the decoding process from wavenumber to time and due to dispersion in the interferometer. The utilization of these two functions brings two major improvements to previous implementations of the MS method. A first improvement consists in reducing the number of channeled spectra necessary to be collected at Master stage. In previous MSI implementation, the number of channeled spectra at the Master stage equated the number of depths where information was selected from at the Slave stage. The paper demonstrates that two experimental channeled spectra only acquired at Master stage suffice to produce A-scans from any number of resolved depths at the Slave stage. A second improvement is the utilization of complex signal processing. Previous MSI implementations discarded the phase. Complex processing of the electrical signal determined by the channeled spectrum allows phase processing that opens several novel avenues. A first consequence of such signal processing is reduction in the random component of the phase without affecting the axial resolution. In previous MSI implementations, phase instabilities were reduced by an average over the wavenumber that led to reduction in the axial resolution.


Superiority of radiobinding assay over ELISA for detection of IAAs in newly diagnosed type I diabetic children.
Liquid- or solid-phase assays have been used for insulin autoantibody (IAA) determination, and the method of IAA measurement has not been standardized. IAAs were determined by radiobinding assay (RBA) and enzyme-linked immunosorbent assay (ELISA) in two large age-matched groups of nondiabetic and newly diagnosed insulin-dependent (type I) diabetic children. Positivity for IAA by RBA (greater than or equal to nondiabetic mean + 3SD) was 2 of 178 (1.1%) and 55 of 173 (32%) in nondiabetic and diabetic children, respectively. Prevalence of IAA by RBA was significantly higher in the youngest age-group (63% between 0-4 yr). Positivity for IAA by ELISA was 1 of 178 (0.6%) and 8 of 169 (4.7%) in nondiabetic and diabetic children, respectively. Concordance rates between both assays were 0 of 3 (0%) in control subjects and 5 of 58 (8.6%) in diabetic children. We conclude that RBA is more appropriate than ELISA for IAA detection at the onset of the disease. In addition, because available data suggest that IAAs detected by RBA only are high-affinity antibodies, it is tempting to speculate that IAAs reflect a mature immune reaction against endogenous insulin.


Method of targeted delivery of laser beam to isolated retinal rods by fiber optics.
A method of controllable light delivery to retinal rod cells using an optical fiber is described. Photo-induced current of the living rod cells was measured with the suction electrode technique. The approach was tested with measurements relating the spatial distribution of the light intensity to photo-induced current. In addition, the ion current responses of rod cells to polarized light at two different orientation geometries of the cells were studied.


Identification of a mutation in porcine ryanodine receptor associated with malignant hyperthermia.
Malignant hyperthermia (MH) causes neurological, liver, and kidney damage and death in humans and major economic losses in the swine industry. A single point mutation in the porcine gene for the skeletal muscle ryanodine receptor (ryr1) was found to be correlated with MH in five major breeds of lean, heavily muscled swine. Haplotyping suggests that the mutation in all five breeds has a common origin. Assuming that this is the causal mutation for MH, the development of a noninvasive diagnostic test will provide the basis for elimination of the MH gene or its controlled inclusion in swine breeding programs.


The EVITA framework for evidence-based mental health policy agenda setting in low- and middle-income countries.
The burden of mental illness is excessive, but many countries lack evidence-based policies to improve practice. Mental health research evidence translation into policymaking is a 'wicked problem', often failing despite a robust evidence base. In a recent systematic review, we identified a gap in frameworks on agenda setting and actionability, and pragmatic, effective tools to guide action to link research and policy are needed. Responding to this gap, we developed the new EVITA 1.1 (EVIdence To Agenda setting) conceptual framework for mental health research-policy interrelationships in low- and middle-income countries (LMICs). We (1) drafted a provisional framework (EVITA 1.0); (2) validated it for specific applicability to mental health; (3) conducted expert in-depth interviews to (a) validate components and mechanisms and (b) assess intelligibility, functionality, relevance, applicability and effectiveness. To guide interview validation, we developed a simple evaluation framework. (4) Using deductive framework analysis, we coded and identified themes and finalized the framework (EVITA 1.1). Theoretical agenda-setting elements were added, as targeting the policy agenda-setting stage was found to lead to greater policy traction. The framework was validated through expert in-depth interviews (n = 13) and revised. EVITA 1.1 consists of six core components [advocacy coalitions, (en)actors, evidence generators, external influences, intermediaries and political context] and four mechanisms (capacity, catalysts, communication/relationship/partnership building and framing). EVITA 1.1 is novel and unique because it very specifically addresses the mental health research-policy process in LMICs and includes policy agenda setting as a novel, effective mechanism. Based on a thorough methodology, and through its specific design and mechanisms, EVITA has the potential to improve the challenging process of research evidence translation into policy and practice in LMICs and to increase the engagement and capacity of mental health researchers, policy agencies/planners, think tanks, NGOs and others within the mental health research-policy interface. Next, EVITA 1.1 will be empirically tested in a case study.


Dutasteride in the long-term management of stuttering priapism.
The objective of this study is to review our 12-year experience with the 5-α reductase inhibitor dutasteride as a potential long-term treatment option for stuttering priapism. Dutasteride has a uniquely long half-life of 35 days which offers a theoretical advantage as a chronic therapy for management of stuttering priapism. We retrospectively reviewed patients with stuttering priapism in our database from 2006-2018 treated with dutasteride. Men with concurrent use of medications other than dutasteride to treat stuttering priapism were excluded. Patients were started on a dose of 0.5 mg daily and tapered to a more infrequent dosing schedule, ranging from 0.5 mg every other day to once weekly. The frequency of priapism episodes before and after initiation of dutasteride therapy was analyzed. Among 21 cases, 13 patients met our inclusion criteria (mean age 43 years). Median follow-up on daily dutasteride was 79 days, and median follow-up on tapered dutasteride was 607 days. A total of 11/13 (85%) men treated with dutasteride had some degree of improvement-5/13 (38%) had complete resolution of their symptoms and 6/13 (46%) had reduced frequency and/or severity of their episodes. Among 5/13 (38%) men who had >2 emergency room (ER) visits for ischemic priapism prior to therapy, most (3/5, 60%) did not require any ER visits while on dutasteride therapy. Among the five men who received chronic, tapered-dose therapy, all reported continued suppression of priapistic episodes. Among 4 patients with sickle cell disease (SCD), 3/4 (75%) ultimately chose more invasive therapy including androgen deprivation therapy (ADT) and penile prosthesis. Side effects were minimal and included gynecomastia (8%), decreased libido (8%), and fatigue (8%). In patients with stuttering priapism, daily dutasteride therapy is a promising treatment option to reduce the frequency and severity of priapistic episodes without significant side effects. Therapy can effectively be tapered to once weekly dosing without a reduction in efficacy.


Clinical and virological course of SARS-CoV 2 infected patients in a tertiary care hospital in Pakistan.
Acute respiratory illness caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) involved the whole globe within no time. Various studies published globally have shown variable severity of disease and mortality. The objective of our study was to describe clinical and epidemiological characteristics of the disease in our setup. in this descriptive case series, individuals with signs and symptoms of Coronavirus disease-19 (COVID-19) and asymptomatic patients with history of close contact to confirmed COVID-19 patients were considered for SARS-CoV-2 Polymerase chain reaction (PCR) assay. Epidemiological and clinical features of only PCR positive cases were recorded. Data regarding hospitalization status, exposure to known COVID-19 patients, clinical feature and clinical outcome of patients was collected and interpreted. A total of 266 patients were found to be SARS-CoV 2 PCR positive which were included in the study. Mean age of patients was 39.45±31.9 years and majority of the patients in our study were male, i.e., 238 (89.5%). Most common clinical features among COVID-19 symptomatic patients were fever and dry cough followed by myalgias and sore throat. Eighteen (7%) out of 266 died in our setup. Time duration of viral shedding after initial positive PCR varied between 11 days to up to more than 55 days. Coronavirus disease-19 (COVID-19) can present with wide range of clinical spectrum and disease can be life threatening. Severity of disease, requirement of ICU care and mortality were directly related to age of the patient and underlying comorbidities. Rigorous precautionary measures are of utmost importance particularly in this high-risk population.


Phase- and tilt-shift determinations by analysis of spectra sidebands for phase-shift interferometers.
A spatial frequency domain method is presented to deal with tilt-shift errors and random phase shift in temporal phase-shift interferometry. The proposed method determines tilt shift and phase shift by analyzing positions and phase variances of sidebands in spatial frequency domain. The method is computationally fast for it is noniterative and needs only one 2D Fourier transform for each spatial carrier interferogram. No initial estimations are required and no ambiguous results are generated with the proposed method. Simulations indicate that the proposed method could detect tilt shift and piston phase shift with high accuracy. Results of experiments conducted in the presence of vibration demonstrate that the proposed method could alleviate fluctuations in the retrieved phase map. The method could be applied to interferometers that are uncalibrated or with an unbalanced piezoelectric transducer, besides interferometers in unsteady conditions.


Isolation, nucleotide sequence analysis, and disruption of the MDH2 gene from Saccharomyces cerevisiae: evidence for three isozymes of yeast malate dehydrogenase.
The major nonmitochondrial isozyme of malate dehydrogenase (MDH2) in Saccharomyces cerevisiae cells grown with acetate as a carbon source was purified and shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to have a subunit molecular weight of approximately 42,000. Enzyme assays and an antiserum prepared against the purified protein were used to screen a collection of acetate-nonutilizing (acetate-) yeast mutants, resulting in identification of mutants in one complementation group that lack active or immunoreactive MDH2. Transformation and complementation of the acetate- growth phenotype was used to isolate a plasmid carrying the MDH2 gene from a yeast genomic DNA library. The amino acid sequence derived from complete nucleotide sequence analysis of the isolated gene was found to be extremely similar (49% residue identity) to that of yeast mitochondrial malate dehydrogenase (molecular weight, 33,500) despite the difference in sizes of the two proteins. Disruption of the MDH2 gene in a haploid yeast strain produced a mutant unable to grow on minimal medium with acetate or ethanol as a carbon source. Disruption of the MDH2 gene in a haploid strain also containing a disruption in the chromosomal MDH1 gene encoding the mitochondrial isozyme produced a strain unable to grow with acetate but capable of growth on rich medium with glycerol as a carbon source. The detection of residual malate dehydrogenase activity in the latter strain confirmed the existence of at least three isozymes in yeast cells.


Sexual Functioning among Married Iranian Women with Polycystic Ovary Syndrome.
This study aimed to assess sexual functioning among women with polycystic ovary syndrome (PCOS) in Iran. A cross-sectional study was conducted to ascertain factors re- lated to sexual functioning in 300 PCOS patients attending to the private practice centers in Kashan, Isfahan Province, Iran, from May to October 2012. The Female Sexual Function Index (FSFI) was used to measure sexual functioning. Moreover, the socio-demo-graphic details and clinical information of PCOS including obesity, hirsutism, acne, mestrual cycle disturbances, infertility and endocrine profile were recorded for each patient. Overall the prevalence of female sexual dysfunction (FSD) was 16.6%. In particular patients indicated poorer sexual functioning for the desire (48.3%) and the arousal (44.7%) subscales. Multiple logistic regression analysis suggested patients with lower educational level (OR: 2.94; 95% CI: 1.46-5.92) and irregular menstrual status (OR: 4.61; 95% CI: 1.93-11) were more likely to report sexual dysfunction. The findings suggest that desire and arousal were the most prevalent sexual disorders reported in this patient population. In addition, findings suggested that women with limited or no formal education and a history of menstrual irregularities were the most likely to report female sexual dysfunction. Further investigations are needed to examine female sexual functioning among women with PCOS, to educate their health care providers, and to develop therapeutic interventions.


Urbanization and health: The effects of the built environment on chronic disease risk factors among women in Tanzania.
Sub-Saharan Africa is experiencing rapid urban growth. Cities enable greater access to health services and improved water and sanitation infrastructure, leading to some improvements in health. However, urban settings may also be associated with more sedentary, stressful lifestyles and consumption of less nutritious food. C-reactive protein (CRP) is a measure of chronic inflammation predictive of cardiovascular disease, and high body mass index (BMI), a ratio of weight to height, indicates overweight or obesity and is associated with an increased risk of many chronic diseases. To explore the association between urbanicity and these two markers, we overlaid data from the 2010 Tanzania Demographic and Health Survey (DHS) with a satellite-derived measure of built environment. Linear regression models were constructed for the outcomes of BMI and CRP, by 1) administratively defined urban/rural categorization from the DHS, 2) satellite derived built environment, and 3) built environment stratified by urban/rural. A total of 2,212 women were included; 23% had elevated CRP, 21% were overweight or obese. A third (33%) lived in a highly built up area and 29% lived in an area classified as urban. A strong positive association between both CRP and BMI and built environment was detected; log CRP increased 0.43 in the highest built up areas compared to not built up (p<0.05); log BMI increased 0.02 in the most built up areas compared to not built up (p<0.05). However, comparing urban to rural category was only significant in unadjusted models. Models stratified by urban/rural category highlight that the variation in CRP and BMI by built environment is mainly driven by rural areas; within urban areas there is less variation. Our findings highlight the potential negative effects of urbanicity on chronic disease markers, with potentially more change detected for those transitioning from rural to urban lifestyles. Satellite-derived urbanicity measures are reproducible and provide more nuanced understanding of effects of built environment on health.


Solvent nanostructure, the solvophobic effect and amphiphile self-assembly in ionic liquids.
The ability of ionic liquids (ILs) to support amphiphile self-assembly into a range of mesophase structures has been established as a widespread phenomenon. From the ILs evaluated as self-assembly media, the vast majority have supported some lyotropic liquid crystal phase formation. Many neat ionic liquids have been shown to segregate into polar and non-polar domains to form a nanostructured liquid. A very strong correlation between the nanostructure of the ionic liquid and its characteristics as an amphiphile self-assembly solvent has been found. In this review we discuss ionic liquids as amphiphile self-assembly media, and identify trends that can be used to distinguish which ionic liquids are likely to have good promotion properties as self-assembly media. In particular these trends focus on the nanostructure of neat ionic liquids, their solvent cohesive energy density, and the related solvophobic effect. We forecast that many more ILs will be identified as amphiphile self-assembly solvents in the future.


The INTERPRET-DD study of diabetes and depression: a protocol.
People with diabetes are at an increased risk of developing depression and other psychological disorders. However, little is known about the prevalence, correlates or care pathways in countries other than the UK and the USA. A new study, the International Prevalence and Treatment of Diabetes and Depression Study (INTERPRET-DD) aims to address this dearth of knowledge and identify optimal pathways to care across the globe. INTERPRET-DD is a 2-year longitudinal study, taking place in 16 countries' diabetes outpatients' facilities, investigating the recognition and management of depressive disorders in people with Type 2 diabetes. Clinical interviews are used to diagnose depression, with clinical and other data obtained from medical records and through patient interviews. Pathways to care and the impact of treatment for previously unrecognized (undocumented) depression on clinical outcomes and emotional well-being are being investigated. Initial evidence indicates that a range of pathways to care exist, with few of them based on available recommendations for treatment. Pilot data indicates that the instruments we are using to measure both the symptoms and clinical diagnosis of depression are acceptable in our study population and easy to use. Our study will increase the understanding of the impact of comorbid diabetes and depression and identify the most appropriate (country-specific) pathways via which patients receive their care. It addresses an important public health problem and leads to recommendations for best practice relevant to the different participating centres with regard to the identification and treatment of people with comorbid diabetes and depression.


The murine Bapx1 homeobox gene plays a critical role in embryonic development of the axial skeleton and spleen.
Our previous studies in both mouse and human identified the Bapx1 homeobox gene, a member of the NK gene family, as one of the earliest markers for prechondrogenic cells that will subsequently undergo mesenchymal condensation, cartilage production and, finally, endochondral bone formation. In addition, Bapx1 is an early developmental marker for splanchnic mesoderm, consistent with a role in visceral mesoderm specification, a function performed by its homologue bagpipe, in Drosophila. The human homologue of Bapx1 has been identified and mapped to 4p16.1, a region containing loci for several skeletal diseases. Bapx1 null mice are affected by a perinatal lethal skeletal dysplasia and asplenia, with severe malformation or absence of specific bones of the vertebral column and cranial bones of mesodermal origin, with the most severely affected skeletal elements corresponding to ventral structures associated with the notochord. We provide evidence that the failure of the formation of skeletal elements in Bapx1 null embryos is a consequence of a failure of cartilage development, as demonstrated by downregulation of several molecular markers required for normal chondroblast differentiation (&agr; 1(II) collagen, Fgfr3, Osf2, Indian hedgehog, Sox9), as well as a chondrocyte-specific alpha1 (II) collagen-lacZ transgene. The cartilage defects are correlated with failed differentiation of the sclerotome at the time when these cells are normally initiating chondrogenesis. Loss of Bapx1 is accompanied by an increase in apoptotic cell death in affected tissues, although cell cycling rates are unaltered.


Expression of cytokines in gingival crevicular fluid associated with tooth movement induced by aligners: a pilot study.
The search for more aesthetic and comfortable orthodontic devices has led to an increase in the use of clear aligners. To increase knowledge on biological mechanisms of orthodontic tooth movement using Invisalign aligners. This study included 11 patients with a mean age of 23.6 ± 4.8 years. Cases planning included alignment and leveling of lower incisors using Invisalign aligners. Gingival crevicular fluid samples were collected from the lower incisors on the day of delivery of aligner number 1 (T0) and after 1 (T24h), 7 (T7d), and 21 (T21d) days. During the observation period of the study, the patients used only the aligner number 1. Levels of nine cytokines were quantified using Luminex's multi-analysis technology. Non-parametric tests were used for comparisons between cytokine expression levels over time. Cytokine expression levels remained constant after 21 days of orthodontic activation, except those of MIP-1β, which presented a statistical difference between T24h and T21d with a decrease in the concentration levels. IL-8, GM-CSF, IL-1β, MIP-1β, and TNF-α showed the highest concentrations over time. The different behavior in the levels of the investigated cytokines indicates a role of these biomarkers in the tissue remodeling induced by Invisalign.


The WHO maternal near-miss approach and the maternal severity index model (MSI): tools for assessing the management of severe maternal morbidity.
To validate the WHO maternal near-miss criteria and develop a benchmark tool for severe maternal morbidity assessments. In a multicenter cross-sectional study implemented in 27 referral maternity hospitals in Brazil, a one-year prospective surveillance on severe maternal morbidity and data collection was carried out. Diagnostic accuracy tests were used to assess the validity of the WHO maternal near-miss criteria. Binary logistic regression was used to model the death probability among women with severe maternal complications and benchmark the management of severe maternal morbidity. Of the 82,388 women having deliveries in the participating health facilities, 9,555 women presented pregnancy-related complications, including 140 maternal deaths and 770 maternal near misses. The WHO maternal near-miss criteria were found to be accurate and highly associated with maternal deaths (Positive likelihood ratio 106.8 (95% CI 99.56-114.6)). The maternal severity index (MSI) model was developed and found to able to describe the relationship between life-threatening conditions and mortality (Area under the ROC curve: 0.951 (95% CI 0.909-0.993)). The identification of maternal near-miss cases using the WHO list of pregnancy-related life-threatening conditions was validated. The MSI model can be used as a tool for benchmarking the performance of health services managing women with severe maternal complications and provide case-mix adjustment.


Cell-mediated lympholysis of trinitrophenyl-modified autologous lymphocytes. Effector cell specificity to modified cell surface components controlled by H-2K and H-2D serological regions of the murine major histocompatibility complex.
Splenic lymphocytes from four C57BL/10 congenic resistant mouse strains were sensitized in vitro with trinitrophenyl (TNP)-modified autologous spleen cellsmthe effector cells generated were incubated with 51-Cr-labeled unmodified or TNP-modified spleen or tumor target cells, and the percentage of specific lympholysis determined. The results obtained using syngeneic-, congenic-, recombinante, and allogeneic-modified target cells indicated that TNP modification of the target cells was a necessary but insufficient requirement for lympholysis. Intra-H-2 homology either between modified stimulating cells and modified target cells or between responding lymphocytes and modified target cells was also important in the specificity for lysis. Homology at the K serological region or at K plus I-A in the B10.A and B10BR strains, and at either the D serological region or at some other region (possibly K) in the B10.D2 and C57BL/10 strains were shown to be necessary in order to detect lympholysis. Experiments using (B10itimes C57BL/10)F1 responding lymphocytes sensitized and assayed with TNP-modified parental cells indicated that the homology required for lympholysis was between modified stimulating and modified target cellsmthe possibility is raised that histocompatibility antigens may serve in the autologous system as cell surface components which are modified by viruses or autoimmune complexes to form cell-bound modified-self antigens, which are particularly suited for cell-mediated immune reactions. Evidence is presented suggesting that H-2-linked Ir genes are expressed in the TNP-modified autologous cytotoxic system. These findings imply that the major histocompatibility complex can be functionally involved both in the response potential to and in the formation of new antigenic determinants involving modified-self components.


Effect of high-dose simvastatin on brain atrophy and disability in secondary progressive multiple sclerosis (MS-STAT): a randomised, placebo-controlled, phase 2 trial.
Secondary progressive multiple sclerosis, for which no satisfactory treatment presently exists, accounts for most of the disability in patients with multiple sclerosis. Simvastatin, which is widely used for treatment of vascular disease, with its excellent safety profile, has immunomodulatory and neuroprotective properties that could make it an appealing candidate drug for patients with secondary progressive multiple sclerosis. We undertook a double-blind, controlled trial between Jan 28, 2008, and Nov 4, 2011, at three neuroscience centres in the UK. Patients aged 18-65 years with secondary progressive multiple sclerosis were randomly assigned (1:1), by a centralised web-based service with a block size of eight, to receive either 80 mg of simvastatin or placebo. Patients, treating physicians, and outcome assessors were masked to treatment allocation. The primary outcome was the annualised rate of whole-brain atrophy measured from serial volumetric MRI. Analyses were by intention to treat and per protocol. This trial is registered with ClinicalTrials.gov, number NCT00647348. 140 participants were randomly assigned to receive either simvastatin (n=70) or placebo (n=70). The mean annualised atrophy rate was significantly lower in patients in the simvastatin group (0·288% per year [SD 0·521]) than in those in the placebo group (0·584% per year [0·498]). The adjusted difference in atrophy rate between groups was -0·254% per year (95% CI -0·422 to -0·087; p=0·003); a 43% reduction in annualised rate. Simvastatin was well tolerated, with no differences between the placebo and simvastatin groups in proportions of participants who had serious adverse events (14 [20%] vs nine [13%]). High-dose simvastatin reduced the annualised rate of whole-brain atrophy compared with placebo, and was well tolerated and safe. These results support the advancement of this treatment to phase 3 testing. The Moulton Foundation [charity number 1109891], Berkeley Foundation [268369], the Multiple Sclerosis Trials Collaboration [1113598], the Rosetrees Trust [298582] and a personal contribution from A Pidgley, UK National Institute of Health Research (NIHR) University College London Hospitals/UCL Biomedical Research Centres funding scheme.


Structural and stereochemical diversity in prenylated indole alkaloids containing the bicyclo[2.2.2]diazaoctane ring system from marine and terrestrial fungi.
Covering: up to February 2017 Various fungi of the genera Aspergillus, Penicillium, and Malbranchea produce prenylated indole alkaloids possessing a bicyclo[2.2.2]diazaoctane ring system. After the discovery of distinct enantiomers of the natural alkaloids stephacidin A and notoamide B, from A. protuberus MF297-2 and A. amoenus NRRL 35660, another fungi, A. taichungensis, was found to produce their diastereomers, 6-epi-stephacidin A and versicolamide B, as major metabolites. Distinct enantiomers of stephacidin A and 6-epi-stephacidin A may be derived from a common precursor, notoamide S, by enzymes that form a bicyclo[2.2.2]diazaoctane core via a putative intramolecular hetero-Diels-Alder cycloaddition. This review provides our current understanding of the structural and stereochemical homologies and disparities of these alkaloids. Through the deployment of biomimetic syntheses, whole-genome sequencing, and biochemical studies, a unified biogenesis of both the dioxopiperazine and the monooxopiperazine families of prenylated indole alkaloids constituted of bicyclo[2.2.2]diazaoctane ring systems is presented.


The Role of Culture and Evolution for Human Cognition.
Since the emergence of our species at least, natural selection based on genetic variation has been replaced by culture as the major driving force in human evolution. It has made us what we are today, by ratcheting up cultural innovations, promoting new cognitive skills, rewiring brain networks, and even shifting gene distributions. Adopting an evolutionary perspective can therefore be highly informative for cognitive science in several ways: It encourages us to ask grand questions about the origins and ramifications of our cognitive abilities; it equips us with the means to investigate, explain, and understand key dimensions of cognition; and it allows us to recognize the continued and ubiquitous workings of culture and evolution in everyday instances of cognitive behavior. Taking advantage of this reorientation presupposes a shift in focus, though, from human cognition as a general, homogenous phenomenon to the appreciation of cultural diversity in cognition as an invaluable source of data.


Potential Prognostic Value of Handgrip Strength in Older Hospitalized Patients.
Objective measures of physical function are useful prognostic tools also for hospitalized elders. Low handgrip strength is predictive of poor outcomes and it can be assessed also in a sitting position, representing a potential alternative measure in bedridden patients. We evaluated grip strength prognostic value in hospitalized older patients. Prospective cohort study. Geriatric, medical ward of an academic medical center in Ferrara, Italy. Patients aged 65 and older (N = 88) admitted to the hospital for an acute medical condition. Patients were evaluated for grip strength at hospital admission and were re-evaluated at discharge. After discharge, they were followed every 3 months for 1 year by telephone interviews to assess new hospitalizations and vital status. The mean age of the sample was 77.3 years, 47% were women. At admission, mean height standardized handgrip strength was 15.7±5 kg/m; men had greater strength (p<0.001). There was a direct relationship of admission grip strength with BMI (p<0.05), serum albumin (p=0.07), and Short Physical Performance Battery score (p<0.05), and an inverse relationship with age (gender-adjusted p value <0.01). In multiple regression analysis, after adjustment for possible confounders, patients in third tertile of grip strength had a shorter hospital stay compared to those in the first tertile (β -2.8; p<0.05). Patients with higher grip strength at discharge also had a lower risk of rehospitalization or death over the follow-up, although the result was not statistically significant (OR: 0.68; 95% CI: 0.30-1.52). In older hospitalized medical patients, grip strength assessment might provide useful prognostic information.


Clinical utility of a monoclonal direct fluorescent reagent specific for Legionella pneumophila: comparative study with other reagents.
Twenty-four lower respiratory tract samples taken from patients with culture-confirmed Legionella pneumophila infection were examined with three different direct immunofluorescent antisera to L. pneumophila, as were 29 samples from similar sources taken from patients without Legionnaires disease. The reagents studied were Genetic Systems Corp. (GS) monoclonal L. pneumophila conjugate, which reacts with all known serogroups of L. pneumophila, BioDx polyvalent L. pneumophila serogroups 1 through 6 conjugate, and Centers for Disease Control polyvalent pool A L. pneumophila serogroups 1 through 4 conjugate. The specimens had been frozen at -70 degrees C for 0.5 to 5 years. Randomization was used in coding the samples, which were stained and read by an independent observer. All three conjugates correctly identified all positive and negative samples. No difference was noted among the conjugates in the absolute numbers of fluorescent L. pneumophila bacteria per sample. The GS conjugate had a much cleaner background than did the other two reagents. Mean staining intensity scores were 3.4, 3.9, and 3.7 for the GS, BioDx, and Centers for Disease Control conjugates, respectively. This study demonstrates that the diagnostic efficiency of all three conjugates is equivalent. Since the GS conjugate is easier to read, does not cross-react with non-L. pneumophila bacteria, and reacts with serogroups 1 through 10 of L. pneumophila, it appears to be preferable for use in diagnostic testing on nonhistopathologically processed specimens.


Conventional and Molecular Breeding Tools for Accelerating Genetic Gain in Faba Bean (Vicia Faba L.).
Faba bean is a cool-season grain legume crop, which is grown worldwide for food and feed. Despite a decrease in area under faba bean in the past, the interest in growing faba bean is increasing globally due to its high seed protein content and its excellent ecological service. The crop is, however, exposed to diverse biotic and abiotic stresses causing unstable, low grain yield. Although, sources of resistance to main diseases, such as ascochyta blight (Ascochyta fabae Speg.), rust (Uromyces viciae-fabae (Pers.) Schroet.), chocolate spot (Botrytis fabae Sard.) and gall disease (Physioderma viciae), have been identified, their resistance is only partial and cannot prevent grain yield losses without agronomical practices. Tightly associated DNA markers for host plant resistance genes are needed to enhance the level of resistance. Less progress has been made for abiotic stresses. Different breeding methods are proposed, but until now line breeding, based on the pedigree method, is the dominant practice in breeding programs. Nonetheless, the low seed multiplication coefficient and the requirement for growing under insect-proof enclosures to avoid outcrossing hampers breeding, along with the lack of tools such as double haploid system and cytoplasmic male sterility. This reduces breeding population size and speed of breeding hence the chances of capturing rare combinations of favorable alleles. Availability and use of the DNA markers such as vicine-convicine (vc -) and herbicide tolerance in breeding programs have encouraged breeders and given confidence in marker assisted selection. Closely linked QTL for several biotic and abiotic stress tolerance are available and their verification and conversion in breeder friendly platform will enhance the selection process. Recently, genomic selection and speed breeding techniques together with genomics have come within reach to accelerate the genetic gains in faba bean. Advancements in genomic resources with other breeding tools, methods and platforms will enable to accelerate the breeding process for enhancing genetic gain in this species.


Growing a backbone - functional biomaterials and structures for intervertebral disc (IVD) repair and regeneration: challenges, innovations, and future directions.
Back pain and associated maladies can account for an immense amount of healthcare cost and loss of productivity in the workplace. In particular, spine related injuries in the US affect upwards of 5.7 million people each year. The degenerative disc disease treatment almost always arises due to a clinical presentation of pain and/or discomfort. Preferred conservative treatment modalities include the use of non-steroidal anti-inflammatory medications, physical therapy, massage, acupuncture, chiropractic work, and dietary supplements like glucosamine and chondroitin. Artificial disc replacement, also known as total disc replacement, is a treatment alternative to spinal fusion. The goal of artificial disc prostheses is to replicate the normal biomechanics of the spine segment, thereby preventing further damage to neighboring sections. Artificial functional disc replacement through permanent metal and polymer-based components continues to evolve, but is far from recapitulating native disc structure and function, and suffers from the risk of unsuccessful tissue integration and device failure. Tissue engineering and regenerative medicine strategies combine novel material structures, bioactive factors and stem cells alone or in combination to repair and regenerate the IVD. These efforts are at very early stages and a more in-depth understanding of IVD metabolism and cellular environment will also lead to a clearer understanding of the native environment which the tissue engineering scaffold should mimic. The current review focusses on the strategies for a successful regenerative scaffold for IVD regeneration and the need for defining new materials, environments, and factors that are so finely tuned in the healthy human intervertebral disc in hopes of treating such a prevalent degenerative process.


Glial scaffold required for cerebellar granule cell migration is dependent on dystroglycan function as a receptor for basement membrane proteins.
Cobblestone lissencephaly is a severe neuronal migration disorder associated with congenital muscular dystrophies (CMD) such as Walker-Warburg syndrome, muscle-eye-brain disease, and Fukuyama-type CMD. In these severe forms of dystroglycanopathy, the muscular dystrophy and other tissue pathology is caused by mutations in genes involved in O-linked glycosylation of alpha-dystroglycan. While cerebellar dysplasia is a common feature of dystroglycanopathy, its pathogenesis has not been thoroughly investigated. Here we evaluate the role of dystroglycan during cerebellar development. Brain-selective deletion of dystroglycan does not affect overall cerebellar growth, yet causes malformations associated with glia limitans disruptions and granule cell heterotopia that recapitulate phenotypes found in dystroglycanopathy patients. Cerebellar pathology in these mice is not evident until birth even though dystroglycan is lost during the second week of embryogenesis. The severity and spatial distribution of glia limitans disruption, Bergmann glia disorganization, and heterotopia exacerbate during postnatal development. Astrogliosis becomes prominent at these same sites by the time cerebellar development is complete. Interestingly, there is spatial heterogeneity in the glia limitans and granule neuron migration defects that spares the tips of lobules IV-V and VI. The full spectrum of developmental pathology is caused by loss of dystroglycan from Bergmann glia, as neither granule cell- nor Purkinje cell-specific deletion of dystroglycan results in similar pathology. These data illustrate the importance of dystroglycan function in radial/Bergmann glia, not neurons, for normal cerebellar histogenesis. The spatial heterogeneity of pathology suggests that the dependence on dystroglycan is not uniform.


Estimating leaf area index in Southeast Alaska: a comparison of two techniques.
The relationship between canopy structure and light transmission to the forest floor is of particular interest for studying the effects of succession, timber harvest, and silviculture prescriptions on understory plants and trees. Indirect measurements of leaf area index (LAI) estimated using gap fraction analysis with linear and hemispheric sensors have been commonly used to assess radiation interception by the canopy, although the two methods often yield inconsistent results. We compared simultaneously obtained measurements of LAI from a linear ceptometer and digital hemispheric photography in 21 forest stands on Prince of Wales Island, Alaska. We assessed the relationship between these estimates and allometric LAI based on tree diameter at breast height (LAIDBH). LAI values measured at 79 stations in thinned, un-thinned controls, old-growth and clearcut stands were highly correlated between the linear sensor (AccuPAR) and hemispheric photography, but the latter was more negatively biased compared to LAIDBH. In contrast, AccuPAR values were more similar to LAIDBH in all stands with basal area less than 30 m(2)ha(-1). Values produced by integrating hemispheric photographs over the zenith angles 0-75° (Ring 5) were highly correlated with those integrated over the zenith angles 0-60° (Ring 4), although the discrepancies between the two measures were significant. On average, the AccuPAR estimates were 53% higher than those derived from Ring 5, with most of the differences in closed canopy stands (unthinned controls and old-growth) and less so in clearcuts. Following typical patterns of canopy closure, AccuPAR LAI values were higher in dense control stands than in old-growth, whereas the opposite was derived from Ring 5 analyses. Based on our results we advocate the preferential use of linear sensors where canopy openness is low, canopies are tall, and leaf distributions are clumped and angles are variable, as is common in the conifer forests of coastal Alaska.


Career choices for obstetrics and gynaecology: recent updates from 40 years of national surveys of UK medical graduates.
To report doctors' early career choices for obstetrics and gynaecology, their eventual career destinations and factors influencing their career pathways. Multi-cohort multi-purpose national questionnaire surveys of medical graduates in selected graduation years between 1974 and 2015. UK. UK-trained medical graduates. Career specialty choices; certainty about specialty choice; factors which influenced doctors' career choices; career specialty destinations 10 years after graduation. Obstetrics and Gynaecology was the first choice of career for 5.7% of post-2002 graduates in year 1, 4.3% in year 3 and 3.8% in year 5. A much higher percentage of women than men specified Obstetrics and Gynaecology as their first choice: in year 1, 7.7% of women and 2.3% of men did so. The gender gap has widened since the 1970s and 1980s. In recent years, of those who specified Obstetrics and Gynaecology as their first choice in year 1 after graduation, 48% were working in Obstetrics and Gynaecology in year 10 (63% of men, 45% of women). Looking backwards from career destinations, 85% of doctors working in Obstetrics and Gynaecology in year 10 had specified Obstetrics and Gynaecology as a first, second or third choice of preferred career in year 1. Interest in Obstetrics and Gynaecology among UK graduates appears to be exceeding the demand for new specialists. Policy needs to address risks of over-production of trainees and ensure that some graduates interested in Obstetrics and Gynaecology consider alternative careers. The large gender imbalance should encourage consideration of the reasons for men choosing Obstetrics and Gynaecology in falling numbers.


Resuscitative hypothermia protects the neonatal rat brain from hypoxic-ischemic injury.
The effect of 24 h of hypothermic recovery on moderate hypoxic-ischemic brain damage in P7-rats was investigated for 42 d after the insult, using magnetic resonance and histopathology. Occlusion of right common carotid artery and 90 min exposure to 8% O2 at 37 degrees C body temperature produced cytotoxic edema of 51(+/-11)% brain volume (BV) and depression of brain energy metabolism (PCr/Pi) from 1.43(+/-0.21) to 0.14(+/-0.11). During recovery, the body temperature was reduced to 30 degrees C for 24 h in 36 animals, but was kept at 37 degrees C in 34 animals. The edema waned upon reoxygenation leaving only the core lesion at 2 h, but reappeared reaching a maximal extent of 11+/-8% BV under hypothermia compared to 45(+/-10)% under normothermia at around 24 h. PCr/Pi recovered transiently within 13 h and declined again to 1.07(+/-0.19) under hypothermia and to 0.48(+/-0.22) under normothermia at around 24 h. Hypothermia led to significant long term brain protection, leaving permanent tissue damage of 12(+/-6)% BV compared to 35(+/-12)% BV under normothermia. However, animals with severe initial injury developed large infarctions, despite hypothermic treatment. Even then, the time to develop infarction was significantly prolonged, leaving the opportunity for additional therapeutic intervention.


Expression and localization of laminin-5 subunits during mouse tooth development.
Tooth morphogenesis is regulated by epithelial-mesenchymal interactions mediated by the basement membrane (BM). Laminins are major glycoprotein components of the BMs, which are involved in several cellular activities. The expression and localization of the alpha3, beta3, and gamma2 laminin-5 subunits have been analyzed by in situ hybridization and immunohistochemistry during mouse molar development. Initially (E12), mRNAs of all subunits were detected in the entire dental epithelium and the corresponding proteins were located in the BM. During cap formation (E13-14), transcripts for the alpha3 and gamma2 subunits were localized in the outer dental epithelium (ODE), whereas the beta3 subunit mRNA was present in the inner dental epithelium (IDE). During the early bell stage (E16), immunoreactivity for all subunits disappeared from the BM along the IDE, although intense signals for beta3 mRNA were detectable in cells of the IDE. Subsequently, when the dentinal matrix was secreted by odontoblasts (E18-19.5), mRNAs of all three subunits were re-expressed by ameloblasts, and the corresponding proteins were detected in ameloblasts and in the enamel matrix. Tissue recombination experiments demonstrated that when E16 IDE or ODE was associated with E18 dental papilla mesenchyme, immunostaining for all laminin-5 subunits disappeared from the BM, whereas when cultured with non-dental limb bud mesenchyme, they remained positive after 48 hr of culture. These results suggest that the temporospatial expression of laminin-5 subunits in tooth development, which appears to be differentially controlled by the dental mesenchyme, might be related to the enamel organ histo-morphogenesis and the ameloblast differentiation.


Chryseobacterium indologenes pneumonia in a patient with non-Hodgkin's lymphoma.
A 42-year-old man diagnosed with gastric non-Hodgkin's lymphoma 2 years earlier, for which he had undergone treatment, presented with expectorative cough, exertional shortness of breath and left-sided chest pain of 3 months duration. Respiratory system examination was suggestive of left-sided pneumonia with pleural effusion. Routine haemogram showed leukocytosis with high erythrocyte sedimentation rate. Chest radiograph showed blunting of left-sided cardiophrenic angle. Sputum culture grew Chryseobacterium indologenes. Diagnostic thoracocentesis was suggestive of lymphomatous metastasis. Pleural fluid culture was sterile. Contrast-enhanced CT (CECT) of the thorax showed left lower lobe consolidation with bilateral pleural effusion. The patient was treated with antibiotics, following which his cough improved and total leukocyte count normalised. Sputum culture repeated after the antibiotic course showed no growth of C. indologenes. However, the pleural effusion soon aggravated along with features suggestive of multiple metastasis. The patient finally succumbed to his underlying advanced malignancy.


Involvement of the Ku autoantigen in the cellular response to DNA double-strand breaks.
The Ku autoantigen is a well-characterized heterodimer of 70 and 86 kDa that binds to DNA ends, but its cellular function has been obscure. An electrophoretic mobility-shift assay and Ku antisera were used to show that Ku or a closely related protein was deficient in three mutant hamster cell lines from x-ray-sensitive complementation group 5, which is characterized by defects in DNA double-strand break repair and V(D)J recombination. Furthermore, Ku protein expression was restored when the cells reverted to x-ray resistance. The Ku p86 gene maps to human chromosome 2q33-35, and group 5 cells are rescued by almost precisely the same region, 2q34-36. Thus, biochemical and genetic evidence suggests that Ku is involved in pathways for DNA recombination and repair. By its association with a DNA-dependent protein kinase activated by DNA ends, Ku may also initiate a signaling pathway induced by DNA damage, perhaps for cell cycle arrest.


Assessment of urinary angiotensinogen as a marker of podocyte injury in proteinuric nephropathies.
Urinary protein (UP) is widely used as a clinical marker for podocyte injury; however, not all proteinuric nephropathies fit this model. We previously described the elevation of urinary angiotensinogen (AGT) accompanied by AGT expression by injured podocytes in a nitric oxide inhibition rat model (Eriguchi M, Tsuruya K, Haruyama N, Yamada S, Tanaka S, Suehiro T, Noguchi H, Masutani K, Torisu K, Kitazono T. Kidney Int 87: 116-127, 2015). In this report, we performed the human and animal studies to examine the significance and origin of urinary AGT. In the human study, focal segmental glomerulosclerosis (FSGS) patients presented with higher levels of urinary AGT, corrected by UP, than minimal-change disease (MCD) patients. Furthermore, AGT was evident in podocin-negative glomerular segmental lesions. We also tested two different nephrotic models induced by puromycin aminonucleoside in Wistar rats. The urinary AGT/UP ratio and AGT protein and mRNA expression in sieved glomeruli from FSGS rats were significantly higher than in MCD rats. The presence of AGT at injured podocytes in FSGS rats was detected by immunohistochemistry and immunoelectron microscopy. Finally, we observed the renal tissue and urinary metabolism of exogenous injected human recombinant AGT (which is not cleaved by rodent renin) in FSGS and control rats. Significant amounts of human AGT were detected in the urine of FSGS rats, but not of control rats. Immunostaining for rat and human AGT identified that only rat AGT was detected in injured podocytes, and filtered human AGT was seen in superficial proximal tubules, but not in injured podocytes, suggesting AGT generation by injured podocytes. In conclusion, the urinary AGT/UP ratio represents a novel specific marker of podocyte injury.


Habitual Myofibrillar Protein Synthesis Is Normal in Patients with Upper GI Cancer Cachexia.
Skeletal muscle wasting and weight loss are characteristic features of cancer cachexia and contribute to impaired function, increased morbidity, and poor tolerance of chemotherapy. This study used a novel technique to measure habitual myofibrillar protein synthesis in patients with cancer compared with healthy controls. An oral heavy water (87.5 g deuterium oxide) tracer was administered as a single dose. Serum samples were taken over the subsequent week followed by a quadriceps muscle biopsy. Deuterium enrichment was measured in body water, serum alanine, and alanine in the myofibrillar component of muscle using gas chromatography-pyrolysis-isotope ratio mass spectrometry and the protein synthesis rate calculated from the rate of tracer incorporation. Net change in muscle mass over the preceding 3 months was calculated from serial CT scans and allowed estimation of protein breakdown. Seven healthy volunteers, 6 weight-stable, and 7 weight-losing (≥5% weight loss) patients undergoing surgery for upper gastrointestinal cancer were recruited. Serial CT scans were available in 10 patients, who lost skeletal muscle mass preoperatively at a rate of 5.6%/100 days. Myofibrillar protein fractional synthetic rate was 0.058%, 0.061%, and 0.073%/hour in controls, weight-stable, and weight-losing patients, respectively. Weight-losing patients had higher synthetic rates than controls (P = 0.03). Contrary to previous studies, there was no evidence of suppression of myofibrillar protein synthesis in patients with cancer cachexia. Our finding implies a small increase in muscle breakdown may account for muscle wasting.


[Urinary incontinence after radical prostatectomy: surgical technique historic evolution and present functional outcomes].
In patients with localized prostate cancer and life expectancy longer than 10 years, radical prostatectomy (RP) remains the Gold Standard. Radical surgery must achieve good oncological and functional outcomes with early continence and potency. In expert hands RP offers very low post-prostatectomy urinary incontinence rates. To analyze functional outcomes regarding urinary incontinence (UI) after RP in our centre in the last ten years. We have performed a retrospective study of the 137 patients who had a radical prostatectomy (open retropubic or laparoscopic) on the General University Hospital of Elche from 1998 to 2008. Two patient groups were analyzed, patients who had surgery from 1998 to 2003, and those who had surgery from 2004 to 2008. Stress UI rates after RP were determined in both groups at 1, 3, 6 and 12 months following surgery with patient pad day usage. Urethrovesical anastomosis strictures were also analyzed in each group and their resolution. 15 patients were excluded from the initial 137 because cT3 stage was detected, remaining 122 prostatectomies with <cT2c stage in the study. Average age was 67 years (50-74). Mean preoperative PSA was 8.1 ng/ml (4.5-25). Biopsy Gleason score was < 6 in 70% (85/122) and 7-8 in 30 % (37/122). In the period between 1998 and 2003, 49 patients underwent surgery and results were: initially continent 28.5% (14/49), continent after one month: 6.1% (3/49), at 3 months: 12.2% (6/49), six months 8.1% ( 4/49) and at a year: 22.4%(11/49). 20.4% (10/49) remain with mild incontinence and moderate/severe 2% (1/49). In this period 78% of the patients remain continent and 22% incontinent. In the period 2004-2008, 73 radical prostatectomies were performed with the following continence rates: initial continence 44% (32/73), after one month 0 patients, 3 months 6.8 % (5/73), 6 months 12.3 % (9/73), a year later 24.6 % (18/73). Mild incontinence was observed in 8.2% ( 6/73) and moderate-severe in 4% (3/73). 88% of the patients achieved continence and 12% remain incontinent. Urinary incontinence is a low frequency disorder in patients who undergo RP. Continence rates have been improving during the last years along with surgical technique and urologic expertise. Anti-incontinence devices after PR are rarely necessary and use is overrated.


Should the scope of addictive behaviors be broadened to include pathological gambling?
Pathological gambling is currently considered an impulse control disorder in the Diagnostic and Statistical Manual of Mental Disorders (DSM). This paper considers the prospect of reclassifying and broadening the substance use disorders to include non-pharmacological addictions such as pathological gambling. Literature reviews were conducted on areas relevant to pathological gambling and its classification and similarities to substance use disorders. The diagnostic criteria for pathological gambling are outlined across the three versions of the DSM since its initial introduction. The paper also describes instruments that have been used to assess the disorder, basic epidemiology and some biological and genetic data, especially in terms of potential overlap with substance use disorders. Similarities and differences with respect to treatments are reviewed as well. Both advantages and disadvantages should be considered with respect to expanding the classification system to include pathological gambling within the context of addictive disorders.


Serratia vespertilionis (García-Fraile et al. 2015) is a later heterotypic synonym of Serratia ficaria (Grimont et al. 1981).
A previous 16S rRNA gene sequence comparison had demonstrated that the type strains of Serratia vespertilionis and Serratia ficaria shared 99.5 % sequence similarity. Despite the 56.2 % homology by DNA-DNA hybridization previously found between these strains, the results of an in silico whole-genome sequence comparison and a new DNA-DNA hybridization study have clearly demonstrated that the genomes of the type strain of S. vespertilionis deposited in different Culture Collections (52T=CECT 8595T=DSM 28727T) and the type strain of S. ficaria (culture DSM 4569T), cannot support such a species differentiation. Tests for substrate utilization redone on the deposited cultures of these strains has also shown very few differences between the type strains of both species. Based on these results, and since the name S. ficaria was validly published earlier, S. vespertilionis should be considered as a later heterotypic synonym of S. ficaria, in application of the priority rule. The type strain of the species S. ficaria is strain 4024T=DSM 4569T=NCTC 12148T=ATCC 33105T=CIP 79.23T=ICPB 4050T.


Inhibitors of Bacillus subtilis DNA polymerase III. Structure-activity relationships of 6-(phenylhydrazino)uracils.
6-(Phenylhydrazino)uracils inhibit the replication-specific enzyme DNA polymerase III of Bacillus subtilis by forming a strong, reversible complex with template-primer DNA and enzyme. The phenyl ring interacts with a hydrophobic enzyme site which, on the basis of structure-activity relationships of substituted analogues, appears to possess the following characteristics: (1) planarity or near-planarity; (2) a finite capacity to accommodate bulky substituents; and (3) location near the domain of the enzyme active site. A mutant DNA polymerase III, derived from a mutant strain of B. subtilis selected for resistance to 6-(p-hydroxyphenylazo)pyrimidines, is resistant only to inhibitors bearing p-hydroxy or amino groups and is hypersensitive to inhibitors containing nonpolar substituents; these results suggest the existence of mutable, secondary regions of the binding site which interact with para substituents and, thus, influence the strength of the primary phenyl-enzyme interaction.


The SPECTRUM population model of the impact of screening and treatment on U.S. breast cancer trends from 1975 to 2000: principles and practice of the model methods.
This stochastic simulation model was developed to estimate the impact of screening and treatment diffusion on U.S. breast cancer mortality between 1975 and 2000. We use an event-driven continuous-time state transition model. Women who are destined to develop breast cancer may be screen detected, present with symptoms, or die of other causes before cancer is diagnosed. At presentation, the cancer has a stage assigned on the basis of mode of detection. Cancers are assumed to be estrogen receptor (ER) positive or negative. Data on screening and treatment diffusion are based on national datasets; other parameters are based on a synthesis of the evidence available in the literature. The model is calibrated to predict incidence and stage distribution (in situ, local, regional, and distant). Other than screening or treatment, background events that affect mortality are not explicitly modeled but are captured in the deviation between model projections of mortality trends and actual trends. We assume that: 1) tumors progress more slowly in older age groups, 2) screen- and clinically detected disease have the same survival conditional on age and stage, 3) women do not die of breast cancer within the "lead time" period, 4) screening benefits are captured by shifts in stage at diagnosis, 4) tamoxifen benefits only ER-positive women, and 5) preclinical sojourn time and dwell times in each of the clinical stages are stochastically independent. Dissemination of screening and therapeutic advances had a substantial impact on mortality trends. We estimate that, by the year 2000, diffusion of screening lowered mortality by 12.4% and treatment improvements and dissemination lowered mortality by 14.6%. Models such as this one can be useful to translate clinical trial findings to general populations. This model can also be used inform policy debates about how to best achieve targeted reductions in breast cancer morbidity and mortality.


Molecular dynamics simulations of methane hydrate using polarizable force fields.
Molecular dynamics simulations of methane hydrate have been carried out using the polarizable AMOEBA and COS/G2 force fields. Properties calculated include the temperature dependence of the lattice constant, the OC and OO radial distribution functions, and the vibrational spectra. Both the AMOEBA and COS/G2 force fields are found to successfully account for the available experimental data, with overall somewhat better agreement with experiment being found for the AMOEBA model. Comparison is made with previous results obtained using TIP4P and SPC/E effective two-body force fields and the polarizable TIP4P-FQ force field, which allows for in-plane polarization only. Significant differences are found between the properties calculated using the TIP4P-FQ model and those obtained using the other models, indicating an inadequacy of restricting explicit polarization to in-plane only.


Enhanced photosynthetic performance and growth as a consequence of decreasing mitochondrial malate dehydrogenase activity in transgenic tomato plants.
Transgenic tomato (Solanum lycopersicum) plants expressing a fragment of the mitochondrial malate dehydrogenase gene in the antisense orientation and exhibiting reduced activity of this isoform of malate dehydrogenase show enhanced photosynthetic activity and aerial growth under atmospheric conditions (360 ppm CO2). In comparison to wild-type plants, carbon dioxide assimilation rates and total plant dry matter were up to 11% and 19% enhanced in the transgenics, when assessed on a whole-plant basis. Accumulation of carbohydrates and redox-related compounds such as ascorbate was also markedly elevated in the transgenics. Also increased in the transgenic plants was the capacity to use L-galactono-lactone, the terminal precursor of ascorbate biosynthesis, as a respiratory substrate. Experiments in which ascorbate was fed to isolated leaf discs also resulted in increased rates of photosynthesis providing strong indication for an ascorbate-mediated link between the energy-generating processes of respiration and photosynthesis. This report thus shows that the repression of this mitochondrially localized enzyme improves both carbon assimilation and aerial growth in a crop species.


Local Climate Heterogeneity Shapes Population Genetic Structure of Two Undifferentiated Insular Scutellaria Species.
Spatial climate heterogeneity may not only affect adaptive gene frequencies but could also indirectly shape the genetic structure of neutral loci by impacting demographic dynamics. In this study, the effect of local climate on population genetic variation was tested in two phylogenetically close Scutellaria species in Taiwan. Scutellaria taipeiensis, which was originally assumed to be an endemic species of Taiwan Island, is shown to be part of the widespread species S. barbata based on the overlapping ranges of genetic variation and climatic niches as well as their morphological similarity. Rejection of the scenario of "early divergence with secondary contact" and the support for multiple origins of populations of S. taipeiensis from S. barbata provide strong evolutionary evidence for a taxonomic revision of the species combination. Further tests of a climatic effect on genetic variation were conducted. Regression analyses show nonlinear correlations among any pair of geographic, climatic, and genetic distances. However, significantly, the bioclimatic variables that represent the precipitation from late summer to early autumn explain roughly 13% of the genetic variation of our sampled populations. These results indicate that spatial differences of precipitation in the typhoon season may influence the regeneration rate and colonization rate of local populations. The periodic typhoon episodes explain the significant but nonlinear influence of climatic variables on population genetic differentiation. Although, the climatic difference does not lead to species divergence, the local climate variability indeed impacts the spatial genetic distribution at the population level.


Covalently linked dengue virus envelope glycoprotein dimers reduce exposure of the immunodominant fusion loop epitope.
A problem in the search for an efficient vaccine against dengue virus is the immunodominance of the fusion loop epitope (FLE), a segment of the envelope protein E that is buried at the interface of the E dimers coating mature viral particles. Anti-FLE antibodies are broadly cross-reactive but poorly neutralizing, displaying a strong infection enhancing potential. FLE exposure takes place via dynamic 'breathing' of E dimers at the virion surface. In contrast, antibodies targeting the E dimer epitope (EDE), readily exposed at the E dimer interface over the region of the conserved fusion loop, are very potent and broadly neutralizing. We here engineer E dimers locked by inter-subunit disulfide bonds, and show by X-ray crystallography and by binding to a panel of human antibodies that these engineered dimers do not expose the FLE, while retaining the EDE exposure. These locked dimers are strong immunogen candidates for a next-generation vaccine.


Primary structural documentation of the major urinary protein of the Indian commensal rat (Rattus rattus) using a proteomic platform.
A number of proteome studies have been performed recently to identify pheromone-related protein expression and their molecular function using genetically modified rodents' urine. However, no such studies have used Indian commensal rodents; interestingly, in a previous investigation, we confirmed the presence of volatile molecules in commensal rodents urine and these molecules seem to be actively involved in pheromonal communication. Therefore, the present study aims to identify the major urinary protein [MUP] present in commensal rat urine, which will help us to understand the protein's expression pattern and intrinsic properties among the rodents globally. Initially, the total urinary proteins were separated by 1-D and 2-D electrophoresis and then subsequently analyzed by Matrix Assisted Laser Desorption Ionization-Time of Flight and Mass Spectrometer (MALDI-TOF/MS). Furthermore, they were then fragmented with the aid of a Tandem Mass Spectrometer (TOF/TOF) and the identified sequences aligned and confirmed using similarity with the deduced primary structures of members of the lipocalin superfamily. The SDS-PAGE protein profiles showed distinct proteins with molecular masses of 15, 22.4, 25, 28, 42, 50, 55, 68, and 91 kDa. Of these proteins, the 22.4 kDa protein was considered to be target candidate. When 2D elctrophoresis was carried out, about approximately 50 spots were detected with different masses and various pI ranges. The 22.4 kDa protein was found to have a pI of about 4.9.This 22.4 kDa protein spot was digested and subjected to mass spectrometry; it was identified as rat MUP. The fragmented peptides from the rat MUP at 935, 1026, 1192, and 1303 m/z were further fragmented with the aid of MS/MS and generated de novo sequence and this confirmed this protein to be the MUP present in the urine of commensal rats. The present investigation confirms the presence of MUP with a molecular mass of 22.4 kDa in the urine of commensal rats. This protein may be involved in the binding of volatile molecules and opens up a discussion about how volatile and non-volatile molecules in the commensal rats' urine may contribute chemo-communication.


Genome-Based Genetic Tool Development for Bacillus methanolicus: Theta- and Rolling Circle-Replicating Plasmids for Inducible Gene Expression and Application to Methanol-Based Cadaverine Production.
Bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene. For independent complementary gene expression control, the heterologous xylose-inducible system from B. megaterium was employed and a two-plasmid gene expression system was developed. Four different replicons for expression vectors were compared with respect to their copy number and stability. As an application example, methanol-based production of cadaverine was shown to be improved from 11.3 to 17.5 g/L when a heterologous lysine decarboxylase gene cadA was expressed from a theta-replicating rather than a rolling-circle replicating vector. The current work on inducible promoter systems and compatible theta- or rolling circle-replicating vectors is an important extension of the poorly developed B. methanolicus genetic toolbox, valuable for genetic engineering and further exploration of this bacterium.


The role of the extracellular matrix in ovarian follicle development.
Regulation of ovarian follicle development depends on endocrine- and paracrine-acting hormones, the 3-dimensional architecture of the follicle, and the physical rigidity of the surrounding tissue. These 3 forces are integrated throughout the life cycle of the follicle to ensure appropriate hormone secretion, differentiation of the somatic cells, and maturation of the oocyte. The process of in-follicle maturation provides a new tool for understanding ovarian follicle development under the influence of these factors.


An Integrative Model for the Effectiveness of Biofeedback Interventions for Anxiety Regulation: Viewpoint.
Biofeedback has shown to be a promising tool for the treatment of anxiety; however, several theoretical as well as practical limitations have prevented widespread adaptation until now. With current technological advances and the increasing interest in the use of self-monitoring technology to improve mental health, we argue that this is an ideal time to launch a new wave of biofeedback training. In this viewpoint paper, we reflect on the current state of biofeedback training, including the more traditional techniques and mechanisms that have been thought to explain the effectiveness of biofeedback such as the integration of operant learning and meditation techniques, and the changes in interoceptive awareness and physiology. Subsequently, we propose an integrative model that includes a set of cognitive appraisals as potential determinants of adaptive trajectories within biofeedback training such as growth mindset, self-efficacy, locus of control, and threat-challenge appraisals. Finally, we present a set of detailed guidelines based on the integration of our model with the mechanics and mechanisms offered by emerging interactive technology to encourage a new phase of research and implementation using biofeedback. There is a great deal of promise for future biofeedback interventions that harness the power of wearables and video games, and that adopt a user-centered approach to help people regulate their anxiety in a way that feels engaging, personal, and meaningful.


Studies on Xenopus laevis intestine reveal biological pathways underlying vertebrate gut adaptation from embryo to adult.
To adapt to its changing dietary environment, the digestive tract is extensively remodeled from the embryo to the adult during vertebrate development. Xenopus laevis metamorphosis is an excellent model system for studying mammalian gastrointestinal development and is used to determine the genes and signaling programs essential for intestinal development and maturation. The metamorphosing intestine can be divided into four distinct developmental time points and these were analyzed with X. laevis microarrays. Due to the high level of conservation in developmental signaling programs and homology to mammalian genes, annotations and bioinformatics analysis were based on human orthologs. Clustering of the expression patterns revealed co-expressed genes involved in essential cell processes such as apoptosis and proliferation. The two largest clusters of genes have expression peaks and troughs at the climax of metamorphosis, respectively. Novel conserved gene ontology categories regulated during this period include transcriptional activity, signal transduction, and metabolic processes. Additionally, we identified larval/embryo- and adult-specific genes. Detailed analysis revealed 17 larval specific genes that may represent molecular markers for human colonic cancers, while many adult specific genes are associated with dietary enzymes. This global developmental expression study provides the first detailed molecular description of intestinal remodeling and maturation during postembryonic development, which should help improve our understanding of intestinal organogenesis and human diseases. This study significantly contributes towards our understanding of the dynamics of molecular regulation during development and tissue renewal, which is important for future basic and clinical research and for medicinal applications.