Preservation of the glomerular capillary ultrafiltration coefficient during rat nephrotoxic serum nephritis by a specific leukotriene D4 receptor antagonist.
Leukotriene D4, a potent biologically active lipoxygenase derivative of arachidonic acid in activated leukocytes, depresses the glomerular capillary ultrafiltration coefficient (Kf) and contracts mesangial cells in culture. We therefore investigated its potential role in mediating the reduction in nephron filtration rate seen after induction of experimental nephrotoxic serum (NTS)-induced glomerulonephritis in the rat. Micropuncture measurements were performed in euvolemic Munich-Wistar rats 2 h after i.v. administration of 0.8 ml of rabbit serum (group 1, n = 6), 0.8 ml of rabbit anti-rat glomerular basement membrane antibody in the absence (group 2, n = 8), or presence (group 3, n = 7) of the new highly specific LTD4 receptor antagonist SK&F 104353. Quantitation of antibody binding and neutrophil infiltration revealed no differences between groups 2 and 3. Antagonism of endogenous LTD4 actions, however, was associated with prevention of the NTS-induced fall in SNGFR because of the abrogation of the fall in Kf which characterizes this form of experimental glomerulonephritis. Antagonism of endogenous LTD4 had no effect on the NTS-induced increases in pre- and postglomerular arteriolar resistances, and did not affect nephron plasma flow rate or net transcapillary hydraulic pressure difference. The observed highly localized protective action of the LTD4 antagonist on the glomerular capillary points to a possibly major functional role for intraglomerularly released LTD4, likely originating from infiltrating leukocytes, in the pathophysiology of this form of glomerulonephritis.


Design and control of an IPMC wormlike robot.
This paper presents an innovative wormlike robot controlled by cellular neural networks (CNNs) and made of an ionic polymer-metal composite (IPMC) self-actuated skeleton. The IPMC actuators, from which it is made of, are new materials that behave similarly to biological muscles. The idea that inspired the work is the possibility of using IPMCs to design autonomous moving structures. CNNs have already demonstrated their powerfulness as new structures for bio-inspired locomotion generation and control. The control scheme for the proposed IPMC moving structure is based on CNNs. The wormlike robot is totally made of IPMCs, and each actuator has to carry its own weight. All the actuators are connected together without using any other additional part, thereby constituting the robot structure itself. Worm locomotion is performed by bending the actuators sequentially from "tail" to "head," imitating the traveling wave observed in real-world undulatory locomotion. The activation signals are generated by a CNN. In the authors' opinion, the proposed strategy represents a promising solution in the field of autonomous and light structures that are capable of reconfiguring and moving in line with spatial-temporal dynamics generated by CNNs.


Association of oral health literacy with oral health behavior and oral health outcomes among adult dental patients.
To evaluate the association of oral health literacy (OHL) with oral health behavior and oral health outcomes among dental patients in Hyderabad City. Cross-sectional study among dental patients. A convenience sample of 605 adults >18 years of age visiting the out-patient Department of Public Health Dentistry of Government Dental College and Hospital, Hyderabad, were recruited. The five-item scale developed by Ishikawa was used to measure communication and critical OHL. Socioeconomic status was assessed using Modified Kuppuswamy's scale and questions for oral health behavior included frequency of toothbrushing, frequency of dental visit, and reason for visit. Dentition status, periodontal status, and loss of attachment were recorded according to World Health Organisation Survey methods. Frequency distribution was done and association between the variables and predictors (oral health behavior and oral health status) of OHL was calculated using odds ratio. The mean age of the study population was 31.5 + 11.2 years. None of the individuals' questions of OHL questionnaire had 50% response of strongly agree or agree. The oral health parameters of decayed and filled teeth emerged as a significant predictor for model 1 (adjusted by sex and age) and model 2 (adjusted by sex, age, and social class). Likewise, toothbrushing frequency was significantly associated with low OHL. This study shows a high prevalence of low OHL in the study population, with decayed teeth and filled teeth and oral health behavior like toothbrushing only once as a significant predictor for low OHL.


Vegetarian diets: what are the advantages?
A growing body of scientific evidence indicates that wholesome vegetarian diets offer distinct advantages compared to diets containing meat and other foods of animal origin. The benefits arise from lower intakes of saturated fat, cholesterol and animal protein as well as higher intakes of complex carbohydrates, dietary fiber, magnesium, folic acid, vitamin C and E, carotenoids and other phytochemicals. Since vegetarians consume widely divergent diets, a differentiation between various types of vegetarian diets is necessary. Indeed, many contradictions and misunderstandings concerning vegetarianism are due to scientific data from studies without this differentiation. In the past, vegetarian diets have been described as being deficient in several nutrients including protein, iron, zinc, calcium, vitamin B12 and A, n-3 fatty acids and iodine. Numerous studies have demonstrated that the observed deficiencies are usually due to poor meal planning. Well-balanced vegetarian diets are appropriate for all stages of the life cycle, including children, adolescents, pregnant and lactating women, the elderly and competitive athletes. In most cases, vegetarian diets are beneficial in the prevention and treatment of certain diseases, such as cardiovascular disease, hypertension, diabetes, cancer, osteoporosis, renal disease and dementia, as well as diverticular disease, gallstones and rheumatoid arthritis. The reasons for choosing a vegetarian diet often go beyond health and well-being and include among others economical, ecological and social concerns. The influences of these aspects of vegetarian diets are the subject of the new field of nutritional ecology that is concerned with sustainable life styles and human development.


Cytokine gene therapy in experimental allergic encephalomyelitis by injection of plasmid DNA-cationic liposome complex into the central nervous system.
Experimental allergic encephalomyelitis (EAE) is an autoimmune disease of the central nervous system with many similarities to multiple sclerosis. The main effector cells involved are CD4+ T cells, recognizing encephalitogenic epitopes within the central nervous system, and macrophages, both of which secrete proinflammatory cytokines, such as IFN-gamma and TNF. Studies have shown that immunomodulation of this inflammatory response by anti-inflammatory cytokines (IL-4, IL-10, IFN-beta, and TGF-beta) can reduce clinical severity in EAE. The importance of TNF in EAE has been demonstrated by using soluble TNF-receptor molecules to inhibit EAE. However, the limitation of this type of therapy is the necessity for frequent administration of cytokine proteins due to their short biologic half-life. This study demonstrates that EAE can be inhibited by a single injection of therapeutic cytokine (IL-4, IFN-beta, and TGF-beta) DNA-cationic liposome complex directly into the central nervous system. DNA coding for a novel, dimeric form of human p75 TNF receptor also ameliorated clinical EAE. Local administration of DNA-cationic liposome complex has identified gene targets that may be more efficiently exploited using vectors producing more stable expression for effective treatment of neuroimmunologic disease.


Cloning of cDNAs encoding two isoforms of 68-kDa type I phosphatidylinositol-4-phosphate 5-kinase.
Accumulating evidence suggests that phosphatidylinositol metabolism is essential for membrane traffic in the cell. Of particular importance, phosphatidylinositol transfer protein and the type I phosphatidylinositol- 4-phosphate 5-kinase (PI4P5K) have been identified as cytosolic components required for ATP-dependent, Ca2+-activated secretion. In order to identify PI4P5K isoforms that may play important roles in regulated insulin secretion from pancreatic beta-cells, we employed the polymerase chain reaction with degenerate primers and screening of a cDNA library of the murine pancreatic beta-cell line MIN6. Two novel cDNAs, designated PI4P5K-Ialpha and PI4P5K-Ibeta, were identified, which contained complete coding sequences encoding 539- or 546-amino acid proteins, respectively. These cDNAs were expressed in mammalian cells with an adenoviral expression vector. Proteins of both isoforms migrated at 68 kDa on SDS-polyacrylamide gel electrophoresis and exhibited phosphatidylinositol-4-phosphate 5-kinase activity, which was activated by phosphatidic acid, indicating that these proteins were type I isoforms. While these isoforms share a marked amino acid sequence homology in their central portion, the amino- and carboxyl-terminal regions differ significantly. Northern blot analysis depicted that tissue distributions differed between the two isoforms. Molecular identification of type I PI4P5K isoforms in insulin-secreting cells should provide insights into the role of phosphatidylinositol metabolism in regulated exocytosis of insulin-containing large dense core vesicles.


Improved evidence-based genome-scale metabolic models for maize leaf, embryo, and endosperm.
There is a growing demand for genome-scale metabolic reconstructions for plants, fueled by the need to understand the metabolic basis of crop yield and by progress in genome and transcriptome sequencing. Methods are also required to enable the interpretation of plant transcriptome data to study how cellular metabolic activity varies under different growth conditions or even within different organs, tissues, and developmental stages. Such methods depend extensively on the accuracy with which genes have been mapped to the biochemical reactions in the plant metabolic pathways. Errors in these mappings lead to metabolic reconstructions with an inflated number of reactions and possible generation of unreliable metabolic phenotype predictions. Here we introduce a new evidence-based genome-scale metabolic reconstruction of maize, with significant improvements in the quality of the gene-reaction associations included within our model. We also present a new approach for applying our model to predict active metabolic genes based on transcriptome data. This method includes a minimal set of reactions associated with low expression genes to enable activity of a maximum number of reactions associated with high expression genes. We apply this method to construct an organ-specific model for the maize leaf, and tissue specific models for maize embryo and endosperm cells. We validate our models using fluxomics data for the endosperm and embryo, demonstrating an improved capacity of our models to fit the available fluxomics data. All models are publicly available via the DOE Systems Biology Knowledgebase and PlantSEED, and our new method is generally applicable for analysis transcript profiles from any plant, paving the way for further in silico studies with a wide variety of plant genomes.


Handling Metadata in a Neurophysiology Laboratory.
To date, non-reproducibility of neurophysiological research is a matter of intense discussion in the scientific community. A crucial component to enhance reproducibility is to comprehensively collect and store metadata, that is, all information about the experiment, the data, and the applied preprocessing steps on the data, such that they can be accessed and shared in a consistent and simple manner. However, the complexity of experiments, the highly specialized analysis workflows and a lack of knowledge on how to make use of supporting software tools often overburden researchers to perform such a detailed documentation. For this reason, the collected metadata are often incomplete, incomprehensible for outsiders or ambiguous. Based on our research experience in dealing with diverse datasets, we here provide conceptual and technical guidance to overcome the challenges associated with the collection, organization, and storage of metadata in a neurophysiology laboratory. Through the concrete example of managing the metadata of a complex experiment that yields multi-channel recordings from monkeys performing a behavioral motor task, we practically demonstrate the implementation of these approaches and solutions with the intention that they may be generalized to other projects. Moreover, we detail five use cases that demonstrate the resulting benefits of constructing a well-organized metadata collection when processing or analyzing the recorded data, in particular when these are shared between laboratories in a modern scientific collaboration. Finally, we suggest an adaptable workflow to accumulate, structure and store metadata from different sources using, by way of example, the odML metadata framework.


SMRT sequencing only de novo assembly of the sugar beet (Beta vulgaris) chloroplast genome.
Third generation sequencing methods, like SMRT (Single Molecule, Real-Time) sequencing developed by Pacific Biosciences, offer much longer read length in comparison to Next Generation Sequencing (NGS) methods. Hence, they are well suited for de novo- or re-sequencing projects. Sequences generated for these purposes will not only contain reads originating from the nuclear genome, but also a significant amount of reads originating from the organelles of the target organism. These reads are usually discarded but they can also be used for an assembly of organellar replicons. The long read length supports resolution of repetitive regions and repeats within the organelles genome which might be problematic when just using short read data. Additionally, SMRT sequencing is less influenced by GC rich areas and by long stretches of the same base. We describe a workflow for a de novo assembly of the sugar beet (Beta vulgaris ssp. vulgaris) chloroplast genome sequence only based on data originating from a SMRT sequencing dataset targeted on its nuclear genome. We show that the data obtained from such an experiment are sufficient to create a high quality assembly with a higher reliability than assemblies derived from e.g. Illumina reads only. The chloroplast genome is especially challenging for de novo assembling as it contains two large inverted repeat (IR) regions. We also describe some limitations that still apply even though long reads are used for the assembly. SMRT sequencing reads extracted from a dataset created for nuclear genome (re)sequencing can be used to obtain a high quality de novo assembly of the chloroplast of the sequenced organism. Even with a relatively small overall coverage for the nuclear genome it is possible to collect more than enough reads to generate a high quality assembly that outperforms short read based assemblies. However, even with long reads it is not always possible to clarify the order of elements of a chloroplast genome sequence reliantly which we could demonstrate with Fosmid End Sequences (FES) generated with Sanger technology. Nevertheless, this limitation also applies to short read sequencing data but is reached in this case at a much earlier stage during finishing.


Is there a human health risk associated with indirect exposure to perfluoroalkyl carboxylates (PFCAs)?
The production and widespread use of poly- and perfluoroalkyl substances (PFAS) has led to their presence in the environment, wildlife, and humans. Particularly, the perfluoroalkyl carboxylates (PFCAs) are pervasive throughout the world and have been found at ng/mL concentrations in human blood. PFCAs, especially those having longer carbon chain lengths (≥C6), are associated with developmental and hormonal effects, immunotoxicity, and promote tumor growth in rodents through their role as PPARα agonists. Humans are directly exposed to PFCAs primarily through contaminated food, drinking water, and house dust. However, indirect exposure to PFCAs through the biotransformation of fluorotelomer-based substances may also be a significant, yet relatively underappreciated pathway. We are exposed to fluorotelomer-based substances through use of consumer products, ingestion of food, or from inhalation of dust particles, but the risk of this exposure has been largely uncharacterized. Here, we summarize the work that has been done to characterize toxicity of the classes of fluorotelomer-based substances shown to biotransform to PFCAs: the polyfluoroalkyl phosphate esters (PAPs), fluorotelomer alcohols (FTOHs), fluorotelomer iodides (FTIs), and fluorotelomer acrylate monomers (FTAcs). These fluorotelomer-based substances biotranform to yield PFCAs, yet also form bioactive intermediate metabolites, which have been observed to be more toxic than their corresponding PFCAs. We address what is known regarding the toxicity of the fluorotelomer-based substances and their metabolites, with focus on covalent binding to biological nucleophiles, such as glutathione, proteins, and DNA, as a possible mechanism of toxicity that may influence the risk of indirect exposure to PFCAs.


In-Hospital Management and Follow-Up Treatment of Venous Thromboembolism: Focus on New and Emerging Treatments.
Venous thromboembolism (VTE), encompassing pulmonary embolism (PE) and deep venous thrombosis (DVT), is a major cause of morbidity and mortality of particular relevance for intensivists and hospitalists. Acute VTE is usually managed with parenteral unfractionated heparin or low-molecular-weight heparin, followed by an oral vitamin K antagonist. Data are lacking for optimal treatment of less common occurrences, such as upper extremity DVT, and for approaches such as thrombolysis for PE associated with early signs of hemodynamic compromise or inferior vena cava filters when anticoagulation is contraindicated. Direct oral anticoagulants (DOACs) including apixaban, dabigatran, edoxaban, and rivaroxaban are now added to the armamentarium of agents available for acute management of VTE and/or reducing the risk of recurrence. This review outlines an algorithmic approach to acute VTE treatment: from aggressive therapies when anticoagulation may be inadequate, to alternative choices when anticoagulation is contraindicated, to anticoagulant options in the majority of patients in whom anticoagulation is appropriate. Evidence-based guidelines and the most recent DOAC clinical trial data are discussed in the context of the standard of care. Situations and treatment approaches for which data are unavailable or insufficient are identified. VTE therapy in care transitions is discussed, as are choices for secondary prevention.


Solution structure of the third TB domain from LTBP1 provides insight into assembly of the large latent complex that sequesters latent TGF-beta.
Almost all TGF-beta is secreted as part of a large latent complex. This complex is formed from three molecules, a latent transforming growth factor-beta binding protein (LTBP), which plays roles in targeting and activation, a latency associated peptide (LAP), which regulates latency, and the TGF-beta cytokine. LAP is the TGF-beta pro-peptide that is cleaved intracellularly prior to secretion, and TGF-beta binds non-covalently to LAP. Formation of the large latent complex is important for the efficient secretion of TGF-beta. Previous studies have revealed that the LTBP-LAP interaction is mediated by intracellular exchange of a single disulphide bond within the third, and only the third, TB domain (TB3) with LAP. We have previously reported the structure of a homologous TB domain from fibrillin-1. However, TB3 contains a two amino acid insertion, not found in fibrillin-1 TB domains, which is not amenable to molecular modelling. In order to clarify the basis of TB domain function, we have determined the solution NMR structure of TB3(LTBP1). Comparison with the fibrillin-1 TB domain reveals that the two-residue insertion is associated with a significant increase in solvent accessibility of one of the disulphide bonds (linking the second and sixth cysteine residues). Site-directed mutagenesis and NMR studies indicate that this is the only disulphide bond that can be removed without perturbing the TB domain fold. Furthermore, a ring of negatively charged residues has been identified that surrounds this disulphide bond. Homology modelling suggests that the surface properties of TB3 domains from different LTBP isoforms correlate with binding activities. This research provides testable hypotheses regarding the molecular basis of complex formation between LTBPs and LAPs.


The Role of Wnt/β-Catenin Signaling and K-Cadherin in the Regulation of Intraocular Pressure.
Wnt/β-catenin signaling in the trabecular meshwork (TM) is required for maintaining normal intraocular pressure (IOP), although the mechanism(s) behind this are unknown. We hypothesize that Wnt/β-catenin signaling regulates IOP via β-catenin's effects on cadherin junctions. Nonglaucomatous primary human TM (NTM) cells were treated with or without 100 ng/ml Wnt3a, 1 μg/ml sFRP1, or both for 4 to 48 hours. Cells were immunostained for β-catenin, total cadherins, or cadherin isoforms. Membrane proteins or whole-cell lysates were isolated for Western immunoblotting and probed for cadherin isoforms. RNA was extracted for cDNA synthesis and qPCR analysis of cadherin expression. Some NTM cells were cultured on electric plates for cell impedance assays. Ad5.CMV recombinant adenoviruses encoding K-cadherin, and/or sFRP1 were injected into eyes of 4- to 6-month-old female BALB/cJ mice (n = 8-10). Conscious IOPs were assessed for 35 days. Upon Wnt3a treatment, total cadherin expression increased and β-catenin accumulated at the TM cell membrane and on processes formed between TM cells. qPCR showed that Wnt3a significantly increased K-cadherin expression in NTM cells (P < 0.01, n = 3), and Western immunoblotting showed that Wnt3a increased K-cadherin in NTM cells, which was inhibited by the addition of sFRP1. Cell impedance assays showed that Wnt3a treatment increased transcellular resistance and anti-K-cadherin siRNA decreased transcellular resistance (P < 0.001, n = 4-6). Our in vivo study showed that K-cadherin significantly decreased sFRP1-induced ocular hypertension (P < 0.05, n = 6). Western immunoblotting also showed that K-cadherin alleviated sFRP1-induced β-catenin decrease in mouse anterior segments. Our results suggest that cadherins play important roles in the regulation of TM homeostasis and IOP via the Wnt/β-catenin pathway.


Immunoplatelet counting: potential for reducing the use of platelet transfusions through more accurate platelet counting.
Research is required to determine the optimal approach for prophylactic platelet transfusions in patients with haematological malignant disorders. It has been suggested that thresholds for prophylactic platelet transfusions of platelet counts below 10 x 109/l should be investigated, as these may be equivalent in clinical effectiveness and associated with lower costs and fewer complications. An important concern in such investigation is the accurate estimation of platelet counts below 10 x 109/l. This study aimed to further examine the potential reduction in platelet usage that could be made if a lowered platelet transfusion threshold of 5 x 109/l was used in conjunction with an immunoplatelet counting method. Clinical and laboratory data from 130 haematology patients were used. Standard platelet counting was performed using Bayer H3 and ABX Argos analysers. Immunoplatelet counting was performed by flow cytometry using anti-CD61. The potential for reducing platelet transfusions included consideration of clinical criteria that influence prophylactic platelet transfusion use. The results indicated that the use of an immunoplatelet count with a 5 x 109/l platelet transfusion threshold would potentially reduce the number of transfusions by 10.4% in comparison with a 10 x 109/l threshold and standard automated platelet counting with the ABX Argos analyser, and increase the number of transfusions by 5.4% in comparison with the same threshold using the Bayer H3 analyser. The immunoplatelet count may aid the clinical decision to transfuse platelets, but would not necessarily lead to a reduced use of platelet transfusions.


The Impact of First-Episode Psychosis on the Sibling Relationship.
Research and clinical practice in first-episode psychosis (FEP) has predominantly focused on parents and partners and has neglected siblings needs. This study aims to characterize the sibling relationship in FEP and to examine the illness-related variables that influence this relationship. Survey methodology explored the experience of 157 siblings during the first 18 months of their brother or sister's treatment. The Adult Sibling Relationship Questionnaire was used to measure the warmth, conflict, and rivalry within the relationship. A series of multivariate regression analyses were conducted to determine the relationship between illness characteristics and sibling relationship. When the young person experiencing FEP had a period of untreated psychosis longer than six months, required more than one hospital admission, had persisting psychotic symptoms, continued to use substances, and/or had a history of physical violence, warmth within the sibling relationship deteriorated. Regression analysis revealed that a history of violence was a significant predictor of the warmth, conflict, and rivalry within the sibling relationship. Suicide attempts were a significant predictor of conflict. This study has established associations between the sibling relationship and illness-related variables. This study promotes consideration of the importance of including siblings in early intervention. Given the powerful role this relationship can have as a protective factor, this study could inform future interventions involving siblings.


Pluripotent cell models of fanconi anemia identify the early pathological defect in human hemoangiogenic progenitors.
Fanconi anemia (FA) is a disorder of genomic instability characterized by progressive bone marrow failure (BMF), developmental abnormalities, and an increased susceptibility to cancer. Although various consequences in hematopoietic stem/progenitor cells have been attributed to FA-BMF, the quest to identify the initial pathological event is still ongoing. To address this issue, we established induced pluripotent stem cells (iPSCs) from fibroblasts of six patients with FA and FANCA mutations. An improved reprogramming method yielded iPSC-like colonies from all patients, and iPSC clones were propagated from two patients. Quantitative evaluation of the differentiation ability demonstrated that the differentiation propensity toward the hematopoietic and endothelial lineages is already defective in early hemoangiogenic progenitors. The expression levels of critical transcription factors were significantly downregulated in these progenitors. These data indicate that the hematopoietic consequences in FA patients originate from the early hematopoietic stage and highlight the potential usefulness of iPSC technology for elucidating the pathogenesis of FA-BMF.


Sexual contexts and the process of risk reduction.
Understanding the dynamics of sexual risks for HIV among men who have sex with men has been one of the ongoing challenges of HIV prevention. While the majority of HIV-prevention interventions target individual behaviour and decision making, multiple studies point to the importance of social context in shaping risk behaviour. Analysis of qualitative data from a study of men who have sex with men, drug use and sex found that sexual encounters were made up multiple contextual and interpersonal elements, which interacted to shape sexual practices and risk reduction strategies. Semi-structured interviews were conducted with 60 racially diverse men who have sex with men in NYC, recruited from multiple venues. The majority of respondents were gay-identified and half were 40 or older. Respondents described risk assessment and risk-reduction processes that develop throughout a sexual encounter, embedded in ongoing negotiations of sexual practices. Strategies of risk assessment and reduction draw on probability-based approaches to HIV prevention, presenting a challenge to health education.


Pharmacological interventions for sleepiness and sleep disturbances caused by shift work.
Shift work results in sleep-wake disturbances, which cause sleepiness during night shifts and reduce sleep length and quality in daytime sleep after the night shift. In its serious form it is also called shift work sleep disorder. Various pharmacological products are used to ameliorate symptoms of sleepiness or poor sleep length and quality. To evaluate the effects of pharmacological interventions to reduce sleepiness or to improve alertness at work and decrease sleep disturbances whilst off work, or both, in workers undertaking shift work in their present job and to assess their cost-effectiveness. We searched CENTRAL, MEDLINE, EMBASE, PubMed and PsycINFO up to 20 September 2013 and ClinicalTrials.gov up to July 2013. We also screened reference lists of included trials and relevant reviews. We included all eligible randomised controlled trials (RCTs), including cross-over RCTs, of pharmacological products among workers who were engaged in shift work (including night shifts) in their present jobs and who may or may not have had sleep problems. Primary outcomes were sleep length and sleep quality while off work, alertness and sleepiness, or fatigue at work. Two authors independently selected studies, extracted data and assessed risk of bias in included trials. We performed meta-analyses where appropriate. We included 15 randomised placebo-controlled trials with 718 participants. Nine trials evaluated the effect of melatonin and two the effect of hypnotics for improving sleep problems. One trial assessed the effect of modafinil, two of armodafinil and one examined caffeine plus naps to decrease sleepiness or to increase alertness.Melatonin (1 to 10 mg) after the night shift may increase sleep length during daytime sleep (mean difference (MD) 24 minutes, 95% confidence interval (CI) 9.8 to 38.9; seven trials, 263 participants, low quality evidence) and night-time sleep (MD 17 minutes, 95% CI 3.71 to 30.22; three trials, 234 participants, low quality evidence) compared to placebo. We did not find a dose-response effect. Melatonin may lead to similar sleep latency times as placebo (MD 0.37minutes, 95% CI - 1.55 to 2.29; five trials, 74 participants, low quality evidence).Hypnotic medication, zopiclone, did not result in significantly longer daytime sleep length compared to placebo in one low quality trial and we could not use the data from the study on lormetazepam.Armodafinil taken before the night shift probably reduces sleepiness by one point on the Karolinska Sleepiness Scale (KSS) (MD -0.99, 95% CI -1.32 to -0.67; range 1 to 10; two trials, 572 participants, moderate quality evidence) and increases alertness by 50 ms in a simple reaction time test (MD -50.0, 95% CI -85.5 to -15.5) at three months' follow-up in shift work sleep disorder patients. Modafinil probably has similar effects on sleepiness (KSS) (MD -0.90, 95% CI -1.45 to -0.35; one trial, 183 participants, moderate quality evidence) and alertness in the psychomotor vigilance test in the same patient group. Post-marketing, severe skin reactions have been reported. Adverse effects reported by trial participants were headache, nausea and a rise in blood pressure. There were no trials in non-patient shift workers.Based on one trial, caffeine plus pre-shift naps taken before the night shift decreased sleepiness (KSS) (MD -0.63, 95% CI -1.09 to -0.17).We judged most trials to have a low risk of bias even though the randomisation method and allocation concealment were often not described. There is low quality evidence that melatonin improves sleep length after a night shift but not other sleep quality parameters. Both modafinil and armodafinil increase alertness and reduce sleepiness to some extent in employees who suffer from shift work sleep disorder but they are associated with adverse events. Caffeine plus naps reduces sleepiness during the night shift, but the quality of evidence is low. Based on one low quality trial, hypnotics did not improve sleep length and quality after a night shift.We need more and better quality trials on the beneficial and adverse effects and costs of all pharmacological agents that induce sleep or promote alertness in shift workers both with and without a diagnosis of shift work sleep disorder. We also need systematic reviews of their adverse effects.


Relationship between fat distribution and cardiometabolic risk in Hispanic girls.
In adults, certain body fat depots have greater impact on cardiometabolic risk than total adiposity. Whether similar relationships exist in children is uncertain. The aim of this study was to examine the relationships among dual x-ray absorptiometry (DXA) measures of body fat distribution and total body adiposity with cardiometabolic risk factors in Hispanic girls. Measures of total percent body fat, percent of total fat within the android, gynoid, leg, and trunk regions, and cardiometabolic biomarkers (insulin, glucose, homeostatic model assessment of insulin resistance (HOMA-IR), triglycerides (TG), low and high lipoprotein cholesterol (LDL-C, HDL-C)) were obtained from 232 Hispanic girls (age 10.7 ±1.1 years). Regression models for each metabolic parameter were run against adiposity measures. Partial correlations of the adiposity measures were used to compare associations between adiposity measures and the cardiometabolic risk factors, controlling for somatic maturation. Total and regional adiposity were significantly related with cardiometabolic risk factors (P < 0.05) except fasting glucose. The partial correlations of total and regional adiposity measures with each cardiometabolic biomarker were similar. More variance was explained for insulin and the HOMA-IR (33%-43%) than other risk factors. Partial correlations for the percentage of total fat in the gynoid and leg regions with insulin, HOMA-IR, TG, and LDL-C were negative, and positive with HDL-C. Measures of total and regional fat perform similarly in predicting cardiometabolic risk in Hispanic girls. A higher proportion of fat distributed in the gynoid or leg region is associated with lower cardiometabolic risk.


Identification of Myocardial Disarray in Patients With Hypertrophic Cardiomyopathy and Ventricular Arrhythmias.
Myocardial disarray is a likely focus for fatal arrhythmia in hypertrophic cardiomyopathy (HCM). This microstructural abnormality can be inferred by mapping the preferential diffusion of water along cardiac muscle fibers using diffusion tensor cardiac magnetic resonance (DT-CMR) imaging. Fractional anisotropy (FA) quantifies directionality of diffusion in 3 dimensions. The authors hypothesized that FA would be reduced in HCM due to disarray and fibrosis that may represent the anatomic substrate for ventricular arrhythmia. This study sought to assess FA as a noninvasive in vivo biomarker of HCM myoarchitecture and its association with ventricular arrhythmia. A total of 50 HCM patients (47 ± 15 years of age, 77% male) and 30 healthy control subjects (46 ± 16 years of age, 70% male) underwent DT-CMR in diastole, cine, late gadolinium enhancement (LGE), and extracellular volume (ECV) imaging at 3-T. Diastolic FA was reduced in HCM compared with control subjects (0.49 ± 0.05 vs. 0.52 ± 0.03; p = 0.0005). Control subjects had a mid-wall ring of high FA. In HCM, this ring was disrupted by reduced FA, consistent with published histology demonstrating that disarray and fibrosis invade circumferentially aligned mid-wall myocytes. LGE and ECV were significant predictors of FA, in line with fibrosis contributing to low FA. Yet FA adjusted for LGE and ECV remained reduced in HCM (p = 0.028). FA in the hypertrophied segment was reduced in HCM patients with ventricular arrhythmia compared to patients without (n = 15; 0.41 ± 0.03 vs. 0.46 ± 0.06; p = 0.007). A decrease in FA of 0.05 increased odds of ventricular arrhythmia by 2.5 (95% confidence interval: 1.2 to 5.3; p = 0.015) in HCM and remained significant even after correcting for LGE, ECV, and wall thickness (p = 0.036). DT-CMR assessment of left ventricular myoarchitecture matched patterns reported previously on histology. Low diastolic FA in HCM was associated with ventricular arrhythmia and is likely to represent disarray after accounting for fibrosis. The authors propose that diastolic FA could be the first in vivo marker of disarray in HCM and a potential independent risk factor.


Arthroplasty for Unreconstructable Acute Fractures and Failed Fracture Fixation About the Hip and Knee in the Active Elderly: A New Paradigm.
The techniques, materials, and designs for total joint arthroplasty underwent major improvements in the past 30 years. During this time, trauma surgeons classified the severity of fractures as well as identified certain articular fractures that do not have good outcomes and have a high rate of failure after internal fixation. Advanced improvements in arthroplasty have increased its reliability and longevity. Total joint arthroplasty is becoming a standard of care for some acute articular fractures, particularly displaced femoral neck fractures in the active elderly. Total joint arthroplasty also has become the standard of care after failed internal fixation in patients who have very complicated fractures about the knee, hip, and shoulder. As the population ages, fractures worldwide continue to rapidly increase. Elderly patients have a high risk for fractures that result from falls because of their poor bone quality. The current active elderly population participates in higher risk activities than previous elderly populations, which places them at risk for more injuries. This has become both a worldwide healthcare problem and an economic problem. Surgeons need to manage fractures in the active elderly with the latest advancements in technology and patient selection to ensure rapid recovery and the reduction of complications.


Study of on-line monitoring of lactate based on optical fibre sensor and in-channel mixing mechanism.
A PDMS-based microfluidic device with integrated optical fibres was developed for online monitoring of lactate. The detection is based on the optical adsorption of the colour solution formed by in-channel mixing of sample and reagent solutions. Computational Fluid Dynamics (CFD) simulation and kinetic study of the colour development were conducted to determine the minimum channel length and optimum residential time for homogenous mixing and maximum optical adsorption respectively. The system was proved to be able to detect lactate with good linearity (R(2)=0.98), response time of about 130 s and estimated limitation of detection (LOD) of 0.52 mM (about 47 mg/L), which is sufficient for a general online lactate detection. Due to the miniaturization of sensing system and micro-scale mixing, higher detection sensitivity (0.15 V/mM) can be realized than the mixing in lab-scale equipments (0.08 V/mM).


Validity of the Generalized Anxiety Disorder-7 scale in an acute psychiatric sample.
Generalized anxiety disorder (GAD) is one of the most prevalent psychiatric presentations; however, GAD has the lowest diagnostic reliability of the anxiety disorders and is poorly recognized in clinical practice. A more reliable assessment of GAD could lead to earlier detection and treatment of the disorder, which has an otherwise debilitating course and significant associated impairment. The 7-item GAD Scale (GAD-7) has shown promise as a measure with good clinical utility and strong psychometric properties in primary care and community settings but has yet to be assessed in acute psychiatric populations. This study examined the validity of the GAD-7 in a sample of 232 patients enrolled in a partial hospital programme. Patients completed a diagnostic interview and a battery of self-report measures before and after treatment. Findings suggest that the GAD-7 has good internal consistency and good convergent validity with worry, anxiety, depression and stress, and the measure was sensitive to change over the course of a short intensive cognitive-behavioural therapy partial hospital programme. However, the confirmatory analysis failed to support the hypothesized unidimensional factor structure; and although the GAD-7 demonstrated good sensitivity (.83), specificity was poor (.46) in identifying patients with GAD. Overall, the GAD-7 appears to be a valid measure of generalized anxiety symptoms in this sample, on the basis of good internal consistency, convergent validity and sensitivity to change, but does not perform well as a screener for GAD. The GAD-7 Scale is an easy-to-score, self-report measure of core generalized anxiety disorder symptoms. The GAD-7 Scale has good internal consistency and convergent validity with depression, anxiety, stress and worry, and is sensitive to change. The GAD-7 Scale appears to be a good measure of generalized anxiety symptoms in an acute psychiatric sample. The GAD-7 Scale does not perform well as a screener for GAD and should not be used to identify cases of GAD in acute psychiatric samples.


Chronotype and environmental light exposure in a student population.
In humans and most other species, changes in the intensity and duration of light provide a critical set of signals for the synchronisation of the circadian system to the astronomical day. The timing of activity within the 24 h day defines an individual's chronotype, i.e. morning, intermediate or evening type. The aim of this study was to investigate the associations between environmental light exposure, due to geographical location, on the chronotype of university students. Over 6 000 university students from cities in the Northern Hemisphere (Oxford, Munich and Groningen) and Southern Hemisphere (Perth, Melbourne and Auckland) completed the Munich ChronoType Questionnaire. In parallel, light measures (daily irradiance, timing of sunrise and sunset) were compiled from satellite or ground stations at each of these locations. Our data shows that later mid-sleep point on free days (corrected for oversleep on weekends MFSsc) is associated with (i) residing further from the equator, (ii) a later sunset, (iii) spending more time outside and (iv) waking from sleep significantly after sunrise. However, surprisingly, MSFsc did not correlate with daily light intensity at the different geographical locations. Although these findings appear to contradict earlier studies suggesting that in the wider population increased light exposure is associated with an earlier chronotype, our findings are derived exclusively from a student population aged between 17 and 26 years. We therefore suggest that the age and occupation of our population increase the likelihood that these individuals will experience relatively little light exposure in the morning whilst encountering more light exposure later in the day, when light has a delaying effect upon the circadian system.


Variability in rates of arthrodesis procedures for patients with cervical spine injuries with and without associated spinal cord injury.
Cervical spine injury, with or without spinal cord injury, is an important cause of morbidity and mortality in the United States. While substantial regional variation has been shown in per capita rates of elective cervical spine surgery, similar data regarding arthrodesis rates for traumatic cervical injury have not been reported, to our knowledge. We assessed the rates of cervical spinal arthrodesis for patients who had a cervical spine injury with or without an associated spinal cord injury. The data for the present study came from the Healthcare Cost and Utilization Project Nationwide Inpatient Sample from 1998 to 2002. International Classification of Diseases, Ninth Revision, Clinical Modification codes were used to identify patients with a cervical vertebral fracture or dislocation with or without an associated spinal cord injury. Hospitals were grouped according to their teaching status, location (urban or rural), and volume of cervical spine injury patients. The rates of spinal arthrodesis and halo/tong placement were compared for patients within each diagnostic category. Twenty-eight thousand, five hundred and eighteen patients with a cervical spine injury were analyzed. Spinal arthrodesis was performed for 16.5% of patients who had a cervical fracture without an associated spinal cord injury, for 50.4% of patients who had a cervical spine fracture with an associated spinal cord injury, and for 44.1% of patients who had a cervical dislocation. With the numbers available, the rates of arthrodesis for patients who had a fracture without a spinal cord injury and for patients who had a cervical dislocation were not significantly different between high and low-volume centers, although the rate for patients who had a cervical fracture with a spinal cord injury was significantly higher at high-volume hospitals. The rates of arthrodesis did not vary significantly between urban teaching and nonteaching hospitals, with the numbers available, for patients in any of the three diagnostic categories. Individual hospitals had a threefold to fivefold variation in the arthrodesis rate for patients with a cervical spine injury, depending on the diagnostic category. The present study demonstrated substantial differences in the rate of arthrodesis for patients with cervical spine trauma, depending on the diagnostic category. The variations in the rates of arthrodesis within diagnostic categories appear to be lower than the previously reported variation in the rates of elective cervical spine procedures.


Dynamic regulatory features of the protein tyrosine kinases.
The SRC, Abelson murine leukemia viral oncogene homolog 1, TEC and C-terminal SRC Kinase families of non-receptor tyrosine kinases (collectively the Src module kinases) mediate an array of cellular signaling processes and are therapeutic targets in many disease states. Crystal structures of Src modules kinases provide valuable insights into the regulatory mechanisms that control activation and generate a framework from which drug discovery can advance. The conformational ensembles visited by these multidomain kinases in solution are also key features of the regulatory machinery controlling catalytic activity. Measurement of dynamic motions within kinases substantially augments information derived from crystal structures. In this review, we focus on a body of work that has transformed our understanding of non-receptor tyrosine kinase regulation from a static view to one that incorporates how fluctuations in conformational ensembles and dynamic motions influence activation status. Regulatory dynamic networks are often shared across and between kinase families while specific dynamic behavior distinguishes unique regulatory mechanisms for select kinases. Moreover, intrinsically dynamic regions of kinases likely play important regulatory roles that have only been partially explored. Since there is clear precedence that kinase inhibitors can exploit specific dynamic features, continued efforts to define conformational ensembles and dynamic allostery will be key to combating drug resistance and devising alternate treatments for kinase-associated diseases.


Trying to understand the extreme: school children's narratives of the mass killings in Norway July 22, 2011.
School-aged children have limited resources for coping with exposure to high-intensity media coverage of terrorist events. This study explores pupils' meaning-making process of their indirect, media-communicated encounters with a specific terrorist event in Norway. Qualitative in-depth interviews about the July 22, 2011 terror attacks were conducted with 54 pupils aged 6-8 years. Seven months after the attacks, the majority had unanswered questions based on more or less accurate knowledge of the events, and they still experienced fear. The media and peers appeared to be their major source of information and not parents or teachers. These children's narratives, characterized by some detailed facts, limited understanding, and a high degree of fiction, were inadequate for restoring calm and feelings of safety. Examples indicate how teacher-facilitated collaborative activities among pupils dealing with crisis can provide a way to construct meaning-making by stimulating conversations and reflections, and developing the narrative through a process of metacognition can provide for further learning and new insights. Implications for a proactive teacher role are indicated.


Coordinated defects in hepatic long chain fatty acid metabolism and triglyceride accumulation contribute to insulin resistance in non-human primates.
Non-alcoholic fatty liver disease (NAFLD) is characterized by accumulation of triglycerides (TG) in hepatocytes, which may also trigger cirrhosis. The mechanisms of NAFLD are not fully understood, but insulin resistance has been proposed as a key determinant. To determine the TG content and long chain fatty acyl CoA composition profile in liver from obese non-diabetic insulin resistant (IR) and lean insulin sensitive (IS) baboons in relation with hepatic and peripheral insulin sensitivity. Twenty baboons with varying grades of adiposity were studied. Hepatic (liver) and peripheral (mainly muscle) insulin sensitivity was measured with a euglycemic clamp and QUICKI. Liver biopsies were performed at baseline for TG content and LCFA profile by mass spectrometry, and histological analysis. Findings were correlated with clinical and biochemical markers of adiposity and insulin resistance. Obese IR baboons had elevated liver TG content compared to IS. Furthermore, the concentration of unsaturated (LC-UFA) was greater than saturated (LC-SFA) fatty acyl CoA in the liver. Interestingly, LC-FA UFA and SFA correlated with waist, BMI, insulin, NEFA, TG, QUICKI, but not M/I. Histological findings of NAFLD ranging from focal to diffuse hepatic steatosis were found in obese IR baboons. Liver TG content is closely related with both hepatic and peripheral IR, whereas liver LC-UFA and LC-SFA are closely related only with hepatic IR in non-human primates. Mechanisms leading to the accumulation of TG, LC-UFA and an altered UFA: LC-SFA ratio may play an important role in the pathophysiology of fatty liver disease in humans.


Effects of subchronic haloperidol and sulpiride on regional brain dopamine metabolism in the rat.
The DOPAC/dopamine and HVA/dopamine ratios in extracted whole tissue were determined to obtain an index of dopaminergic activity in various rat brain regions 24 h following 10 days' treatment with haloperidol (0.5 and 2 mg/kg) or sulpiride (10 and 100 mg/kg). Both neuroleptics caused a reduction in the metabolite/amine ratio in nucl. accumbens but not frontal cortex or substantia nigra. Haloperidol, but not sulpiride significantly reduced the HVA/dopamine ratio in striatum. A region-specific action of neuroleptics on brain dopamine neurons is discussed.


Effect of sphingosine-1-phosphate on L-type calcium current and Ca(2+) transient in rat ventricular myocytes.
Modulation of Ca(2+) homoeostasis in cardiac myocytes plays a major role in beat-to-beat regulation of heart function. Previous studies suggest that sphingosine-1-phosphate (S1P), a biologically active sphingomyelin metabolite, regulates Ca(2+) handling in cardiac myocytes, but the underlying mechanism is unclear. In the present study, we tested the hypothesis that S1P-induced functional alteration of intracellular Ca(2+) handling includes the L-type calcium channel current (ICa,L) via a signalling pathway involving P21-activated kinase 1 (Pak1). Our results show that, in rat ventricular myocytes, S1P (100 nM) does not affect the basal activity of ICa,L but is able to partially reverse the effect of the β-adrenergic agonist Isoproterenol (ISO, 100 nM) on ICa,L. S1P (25 nM) also significantly prevents ISO (5 nM)-induced Ca(2+) waves and diastolic Ca(2+) release in these cells. Our further molecular characterisation demonstrates that Pak1 activity is increased in myocytes treated with S1P (25 nM) compared with those myocytes without treatment of S1P. By immunoprecipitation we demonstrate that Pak1 and protein phosphatase 2A (PP2A) are associated in ventricular tissue indicating their functional interaction. Thus the results indicate that S1P attenuates β-adrenergic stress-induced alteration of intracellular Ca(2+) release and L-type Ca(2+) channel current at least in part via Pak1-PP2A-mediated signalling.


Radiomics feature stability of open-source software evaluated on apparent diffusion coefficient maps in head and neck cancer.
Radiomics is a promising technique for discovering image based biomarkers of therapy response in cancer. Reproducibility of radiomics features is a known issue that is addressed by the image biomarker standardisation initiative (IBSI), but it remains challenging to interpret previously published radiomics signatures. This study investigates the reproducibility of radiomics features calculated with two widely used radiomics software packages (IBEX, MaZda) in comparison to an IBSI compliant software package (PyRadiomics). Intensity histogram, shape and textural features were extracted from 334 diffusion weighted magnetic resonance images of 59 head and neck cancer (HNC) patients from the PREDICT-HN observational radiotherapy study. Based on name and linear correlation, PyRadiomics shares 83 features with IBEX and 49 features with MaZda, a sub-set of well correlated features are considered reproducible (IBEX: 15 features, MaZda: 18 features). We explore the impact of including non-reproducible radiomics features in a HNC radiotherapy response model. It is possible to classify equivalent patient groups using radiomic features from either software, but only when restricting the model to reliable features using a correlation threshold method. This is relevant for clinical biomarker validation trials as it provides a framework to assess the reproducibility of reported radiomic signatures from existing trials.


Surgical treatment for nontuberculous mycobacterial (NTM) cervicofacial lymphadenitis in children.
To compare surgical excision with surgical curettage in the treatment of nontuberculous mycobacterial (NMT) cervicofacial lymphadenitis in children. Fifty children, 22 boys and 28 girls, with a PCR- or cultured-confirmed diagnosis of cervicofacial NTM infection were included in the study. Twenty-five children were randomized to surgical excision of the involved lymph nodes, and 25 children to surgical curettage. The median age of the children was 36 months (range, 14-120 months). All children had a red, fluctuating lymphadenitis, and there were no marked differences between the treatment groups with respect to mean duration of lymph node swelling before presentation, location, and the size of the lymph node swelling. Most (84%) of the involved nodes were located in the submandibular region and 6% were located in the preauricular region. Multiple locations (both preauricular and submandibular) were observed in the remaining 10%. Mycobacterium avium (74%) and Mycobacterium haemophilum (22%) were the predominant NTM species. Mean wound healing time for the excision group was 3.6 ± 1.2 weeks versus 11.4 ± 5.1 weeks for the curettage group (P ≤ .05). Postoperative transient marginal mandibular nerve weakness of the facial nerve was seen in 4 patients (16%) of the excision group. In all these patients the function of the nerve returned to normal within 12 weeks. No facial nerve problems were observed in the curettage group. Postoperative infections were not observed. Surgical excision leads to a quick resolution of NTM cervicofacial lymphadenitis. Curettage leads to delayed healing but might be considered as an alternative if excision of the necrotized lymph nodes is technically difficult in cases of adherence of the facial nerve branche.


DNA compaction by the higher-order assembly of PRH/Hex homeodomain protein oligomers.
Protein self-organization is essential for the establishment and maintenance of nuclear architecture and for the regulation of gene expression. We have shown previously that the Proline-Rich Homeodomain protein (PRH/Hex) self-assembles to form oligomeric complexes that bind to arrays of PRH binding sites with high affinity and specificity. We have also shown that many PRH target genes contain suitably spaced arrays of PRH sites that allow this protein to bind and regulate transcription. Here, we use analytical ultracentrifugation and electron microscopy to further characterize PRH oligomers. We use the same techniques to show that PRH oligomers bound to long DNA fragments self-associate to form highly ordered assemblies. Electron microscopy and linear dichroism reveal that PRH oligomers can form protein-DNA fibres and that PRH is able to compact DNA in the absence of other proteins. Finally, we show that DNA compaction is not sufficient for the repression of PRH target genes in cells. We conclude that DNA compaction is a consequence of the binding of large PRH oligomers to arrays of binding sites and that PRH is functionally and structurally related to the Lrp/AsnC family of proteins from bacteria and archaea, a group of proteins formerly thought to be without eukaryotic equivalents.


Hand Rehabilitation Using a 3D Environment and Leap Motion Device.
The paper presents a 3D healthcare informatics support for Hand Rehabilitation after injury. As a novelty, the application uses the Leap Motion sensor for patient's gestures recognition, and videos to illustrate to the users the hand exercise to perform. The implemented application provides feedback to users regarding the correctness of the performed recovery gestures/exercises. The data related to hand rehabilitation is saved in a database and offers to the Medical Rehabilitation Experts the possibility to monitor the patients in a more consistent manner. To assess the efficiency and accuracy of the application the application will be tested following a usability plan.


Surveillance and modelling of HIV, STI, and risk behaviours in concentrated HIV epidemics.
HIV epidemics in most countries are highly concentrated among population subgroups such as female sex workers, injecting drug users, men who have sex with men, mobile populations, and their sexual partners. The perception that they are important only when they cause epidemic expansion to general populations has obscured a critical lack of coverage of preventive interventions in these groups, as well as appropriate methods for monitoring epidemic and behavioural risk trends. The difficulties in accessing such groups have likewise often cast doubt on the representativeness of observed disease and behavioural risk estimates and their validity and reliability, particularly those related to sampling and the measurement of risk behaviours. To review methodological obstacles in conducting surveillance with population subgroups in concentrated HIV epidemics, elaborate on recent advancements that partially overcome these obstacles, and illustrate the importance of modelling integrated HIV, STI, and behavioural surveillance data. Review of published HIV, STI, and behavioural surveillance data, research on epidemic dynamics, and case studies from selected countries. The population subgroups that merit regular and systematic surveillance in concentrated epidemics are best determined through extensive assessment and careful definition. Adherence to recently refined chain referral and time location sampling methods can help to ensure more representative samples. Finally, because of the inherent limitations of cross sectional surveys in understanding associations between complex sexual behaviours and HIV and STI transmission, mathematical models using multiple year data offer opportunities for integrated analysis of behavioural change and HIV/STI trends.


End-Effector Contact and Force Detection for Miniature Autonomous Robots Performing Lunar and Expeditionary Surgery.
The U.S. Space Force was stood up on December 20, 2019 as an independent branch under the Air Force consisting of about 16,000 active duty and civilian personnel focused singularly on space. In addition to the Space Force, the plans by NASA and private industry for exploration-class long-duration missions to the moon, near-earth asteroids, and Mars makes semi-independent medical capability in space a priority. Current practice for space-based medicine is limited and relies on a "life-raft" scenario for emergencies. Discussions by working groups on military space-based medicine include placing a Role III equivalent facility in a lunar surface station. Surgical capability is a key requirement for that facility. To prepare for the eventuality of surgery in space, it is necessary to develop low-mass, low power, mini-surgical robots, which could serve as a celestial replacement for existing terrestrial robots. The current study focused on developing semi-autonomous capability in surgical robotics, specifically related to task automation. Two categories for end-effector tissue interaction were developed: Visual feedback from the robot to detect tissue contact, and motor current waveform measurements to detect contact force. Using a pixel-to-pixel deep neural network to train, we were able to achieve an accuracy of nearly 90% for contact/no-contact detection. Large torques were predicted well by a trained long short-term memory recursive network, but the technique did not predict small torques well. Surgical capability on long-duration missions will require human/machine teaming with semi-autonomous surgical robots. Our existing small, lightweight, low-power miniature robots perform multiple essential tasks in one design including hemostasis, fluid management, suturing for traumatic wounds, and are fully insertable for internal surgical procedures. To prepare for the inevitable eventuality of an emergency surgery in space, it is essential that automated surgical robot capabilities be developed.


An amperometric cholesterol biosensor based on epoxy resin membrane bound cholesterol oxidase.
The use of epoxy resin membrane as a support for immobilization of enzyme has resulted into improved sensitivity and stability of biosensors for uric acid, ascorbic acid and polyphenols. The present work was aimed to prepare an improved amperometric biosensor for determination of serum cholesterol required in the diagnostics and management of certain pathological conditions. Epoxy resin membrane with immobilized cholesterol oxidase was mounted on the cleaned platinum (Pt) electrode with a parafilm to construct a working electrode. This working electrode along with Ag/AgCl as reference and Ag wire as an auxiliary electrode were connected through a three terminal electrometer to construct a cholesterol biosensor. The sensor showed optimum response within 25 sec at pH 7.0 and 45°C. The linear working range of biosensor was 1.0 to 8.0 mM cholesterol. K m and I max for cholesterol were 5.0 mM and 9.09 μA, respectively. The biosensor measured serum cholesterol. The minimum detection limit of the sensor was 1.0 mM. The mean analytical recoveries of added cholesterol in serum (2.84 and 4.13 mM) were 91.4 ± 2.8 and 92.3 ± 3.1 per cent (n=6), respectively. Within and between assay coefficient of variation (CV) were <2 and <4 per cent, respectively. Biosensor had a storage life of 6 months at 4 o C. The use of epoxy resin membrane as a support for immobilization of cholesterol oxidase has resulted into an improved amperometric cholesterol biosensor. The present biosensor had an advantage over the existing biosensors as it worked at comparatively lower potential.


SU-E-J-185: 4DCT Geometrical Eigenmode Model for Inter-Fraction Evaluation of Tumor Regression and Breathing Pattern Changes.
Evaluate the feasibility of using intra-fractional respiratory dominant eigenmodes of lung deformation as a method to track inter-fractional tumor regression and other physiological changes. Geometrical evaluation by dominant eigenmodes during free-breathing may be implemented as an IGART tool for assessing inter-fractional non-respiratory anatomical variation. Intra-fractional deformable image registration (DIR) is performed on 4DCT scans of lung cancer patients. Principal component analysis is conducted on a set of 10 deformation vector fields for each patient both with and without a lung mask. Leave-one-out cross-validation (LOOCV) is used to assess intra-fractional modeling error among a subset of respiratory phases. The first two dominant eigenmodes of PCA are compared across patients in axial, sagittal, and coronal slices. Eigenvalues between the first and second dominant respiratory modes decay by a factor of ten for all patients. Intra-fractional LOOCV maximum error is on the order of 1mm inside the lung and 4-6mm for the whole thorax for all patients using the first two dominant eigenmodes. Eigenvalue decay is fastest for patients where diaphragm motion is relatively larger in magnitude. The first two eigenmodes generally demonstrate a strong but smooth superior-inferior motion component with a lesser lateral and anterior-posterior component, while subsequent less dominant eigenmodes produce more random directional eigenvector fields. The two first principal components are consistent amongst patients. Inter-fractional eigenmode comparison may provide a means to track tumor regression and breathing pattern or physiological changes. We expect that patients with more dominant diaphragmatic respiration will allow for more delineated eigenmode classification. Clear dominant eigenmode delineation may allow subsequent less dominant eigenmodes to be inter-fractionally compared as a means to evaluate and compensate for tumor regression.


Transient Receptor Potential Melastatin-8 Activation Induces Relaxation of Pulmonary Artery by Inhibition of Store-Operated Calcium Entry in Normoxic and Chronic Hypoxic Pulmonary Hypertensive Rats.
Pulmonary hypertension (PH) is characterized by enhanced vasoconstriction and vascular remodeling, which are attributable to the alteration of Ca2+ homeostasis in pulmonary arterial smooth muscle cells (PASMCs). It is well established that store-operated Ca2+ entry (SOCE) is augmented in PASMCs during PH and that it plays a crucial role in PH development. Our previous studies showed that the melastatin-related transient receptor potential 8 (TRPM8) is down-regulated in PASMCs of PH animal models, and activation of TRPM8 causes relaxation of pulmonary arteries (PAs). However, the mechanism of TRPM8-induced PA relaxation is unclear. Here we examined the interaction of TRPM8 and SOCE in PAs and PASMCs of normoxic and chronic hypoxic pulmonary hypertensive (CHPH) rats, a model of human group 3 PH. We found that TRPM8 was down-regulated and TRPM8-mediated cation entry was reduced in CHPH-PASMCs. Activation of TRPM8 with icilin caused concentration-dependent relaxation of cyclopiazonic acid (CPA) and endothelin-1 contracted endothelium-denuded PAs, and the effect was abolished by the SOCE antagonist Gd3+ Application of icilin to PASMCs suppressed CPA-induced Mn2+ quenching and Ca2+ entry, which was reversed by the TRPM8 antagonist N-(3-aminopropyl)-2-([(3-methylphenyl)methyl])-oxy-N-(2-thienylmethyl)benzamide hydrochloride salt (AMTB). Moreover, the inhibitory effects of icilin on SOCE in PA and PASMCs of CHPH rats were significantly augmented due to enhanced SOCE activity in PH. Our results, therefore, demonstrated a novel mechanism of TRPM8-mediated inhibition of SOCE in pulmonary vasculature. Because SOCE is important for vascular remodeling and enhanced vasoconstriction, down-regulation of TRPM8 in PASMCs of CHPH rats may minimize its inhibitory influence to allow unimpeded SOCE activity for PH development.


Pediatric kidney recipients with small capacity, defunctionalized urinary bladders receiving adult-sized kidney without prior bladder augmentation.
Children with small capacity, defunctionalized urinary bladders present unique operative challenges. Thus, traditional practice has included pretransplant bladder augmentation, but this has several adverse consequences. A single-institutional, retrospective review from January 1, 2004 to December 31, 2008 was conducted. Twelve pediatric patients, whom had not undergone pretransplant bladder augmentation, did not have neurogenic bladders or require preoperative catheterization, and a small capacity defunctionalized bladders were included. All were managed by the same surgeon with a previously described ureteral implantation, and a 7F ureteral stent attached to a large diameter suprapubic catheter was removed in a joint manner without cystoscopy at 2 weeks. Data were collected on patient and graft survival, rejection episodes, urinary tract infection (UTI) requiring antibiotics, grade of vesicoureteral reflux, and posttransplant bladder capacity. One-year patient and graft survival rates were 100%. One patient experienced a clinical rejection episode, which was successfully treated. Five patients (41.7%) had a UTI requiring abx treatment within the first postoperative year, but at 1 year, all patients had sterile urinary tracts. After removal of suprapubic catheters and ureteral stents, all patients were able to void spontaneously. Seven patients had no posttransplant ureteral reflux, three had grade 1 reflux, and two had grade 3 reflux (both successfully treated). The average age estimated pretransplant bladder and 1 year posttransplant bladder capacity was 14.5% and 84% of expected, respectively. In pediatric end-stage renal disease patients with a small capacity defunctionalized bladder, it is reasonable to proceed with kidney transplantation without pretransplant bladder augmentation, thus avoiding an unnecessary surgery.


Chronic intermittent hypoxia sensitizes acute hypothalamic-pituitary-adrenal stress reactivity and Fos induction in the rat locus coeruleus in response to subsequent immobilization stress.
Obstructive sleep apnea (OSA) is associated with several pathophysiological conditions, including hypertension, obesity, insulin resistance, hypothalamic-pituitary-adrenal (HPA) dysregulation, and other endocrine and metabolic disturbances comprising the "metabolic syndrome." Repeated episodes of hypoxia in OSA may represent a chronic intermittent stress, leading to HPA dysregulation. Alterations in HPA reactivity could then contribute to or exacerbate other pathophysiological processes. We showed previously that another metabolic stressor, chronic intermittent cold stress, enhanced noradrenergic facilitation of acute HPA stress reactivity. In this study, we investigated whether chronic intermittent hypoxia (CIH), a rat model for the arterial hypoxemia that accompanies OSA, similarly sensitizes the HPA response to novel acute stress. Rats were exposed to CIH (alternating cycles of normoxia [3 min at 21% O(2)] and hypoxia [3 min at 10% O(2)], repeated continuously for 8 h/day during the light portion of the cycle for 7 days). On the day after the final CIH exposure, there were no differences in baseline plasma adrenocorticotropic hormone (ACTH), but the peak ACTH response to 30 min acute immobilization stress was greater in CIH-stressed rats than in controls. Induction of Fos expression by acute immobilization stress was comparable following CIH in several HPA-modulatory brain regions, including the paraventricular nucleus, bed nucleus of the stria terminalis, and amygdala. Fos induction was attenuated in lateral hypothalamus, an HPA-inhibitory region. By contrast, acute Fos induction was enhanced in noradrenergic neurons in the locus coeruleus following CIH exposure. Thus, similar to chronic cold stress, CIH sensitized acute HPA and noradrenergic stress reactivity. Plasticity in the acute stress response is important for long-term adaptation, but may also contribute to pathophysiological conditions associated with states of chronic or repeated stress, such as OSA. Determining the neural mechanisms underlying these adaptations may help us better understand the etiology of such disorders, and inform the development of more effective treatments.


Local deep tissue penetration of compounds after dermal application: structure-tissue penetration relationships.
Topically applied compounds can penetrate directly into deeper underlying tissues. In this report, an attempt has been made to establish structure-deep tissue penetration relationships for a variety of solutes with diverse physicochemical properties. Stepwise multiple linear regression analysis was performed with the concentration in the immediately overlying tissue, the solute molecular size and the octanol/water partition coefficient as independent variables. During the initial period of direct penetration, the concentration of any solute in a given tissue was dependent on the concentration in the preceding tissue. The presence of molecular weight and lipophilicity terms as independent variables improved the regressions for some tissues. The solute concentration in the deeper tissues of sacrificed animals was higher for the smaller solutes. Due to the dominance of solute clearance by blood perfusing the tissues, the dependence of solute concentrations in anesthetized animal tissues on size was less than observed for sacrificed animals. The predictions from these regression analyses yielded predictions similar to those based on a physiological pharmacokinetic model. However, only about 50% of the data was explained by both models. Based on the preliminary qualitative and quantitative analysis, deep tissue penetration of solutes after application, as aqueous solutions, to the epidermis is greater for smaller solutes with adequate lipophilicity.


Prediction of postpartum depression by sociodemographic, obstetric and psychological factors: a prospective study.
Many studies have documented serious effects of postpartum depression. This prospective study sought to determine predictive factors for postpartum depression. Pregnant women (n = 239) were enrolled before 24 weeks in their pregnancy. At 6 weeks postpartum, 30 women who had postpartum depression and 30 non-depressed mothers were selected. The Edinburgh Postnatal Depression Scale (EPDS), the Beck Depression Inventory (BDI), the Beck Anxiety Inventory (BAI), the Rosenberg Self-Esteem Scale (RSES) Marital Satisfaction Scale (MSS), and the Childcare Stress Inventory (CSI) were administered to all 60 mothers at 24 weeks pregnancy, 1 week postpartum, and 6 weeks postpartum. The differences in most of the diverse sociodemographic and obstetric factors assessed were not statistically significant. There were significant differences in MSS scores at 24 weeks pregnancy (P = 0.003), and EPDS (P < 0.001; P = 0.002), BDI (P = 0.001; P = 0.031), and BAI (P < 0.001; P < 0.001) at both 24 weeks pregnant and 1 week postpartum, while there was no significant difference in the RSES scores at 24 weeks pregnant (P = 0.065). A logistic regression analysis was performed on the following factors: 'depressive symptoms immediately after delivery' (EPDS and BDI at 1 week postpartum), 'anxiety' (BAI prepartum), 'stress factors from relationships' (MSS prepartum and CSI at 1 week postpartum) or 'self-esteem' (RSES prepartum). When these four factors were added individually to a model of the prepartum depressive symptoms (EPDS and BDI prepartum), no additional effect was found. The optimum psychological predictor is prepartum depression, and other psychological measures appear to bring no significant additional predictive power.


Microfluidics for electrophysiology, imaging, and behavioral analysis of Hydra.
The nervous system of the cnidarian Hydra vulgaris exhibits remarkable regenerative abilities. When cut in two, the bisected tissue reorganizes into fully behaving animals in approximately 48 hours. Furthermore, new animals can reform from aggregates of dissociated cells. Understanding how behaviors are coordinated by this highly plastic nervous system could reveal basic principles of neural circuit dynamics underlying behaviors. However, Hydra's deformable and contractile body makes it difficult to manipulate the local environment while recording neural activity. Here, we present the first microfluidic technologies capable of simultaneous electrical, chemical, and optical interrogation of these soft, deformable organisms. Specifically, we demonstrate devices that can immobilize Hydra for hours-long simultaneous electrical and optical recording, and chemical stimulation of behaviors revealing neural activity during muscle contraction. We further demonstrate quantitative locomotive and behavioral tracking made possible by confining the animal to quasi-two-dimensional micro-arenas. Together, these proof-of-concept devices show that microfluidics provide a platform for scalable, quantitative cnidarian neurobiology. The experiments enabled by this technology may help reveal how highly plastic networks of neurons provide robust control of animal behavior.


Clinical importance of elevated CK-MB and troponin I levels in congestive heart failure.
Myocyte necrosis has been considered to play a fundamental role in the pathophysiology of congestive heart failure (CHF), which has usually evolved as a consequence of depletion of compensatory mechanisms and contractile reserve of myocardium. Elevated levels of creatine kinase MB (CK-MB) and troponin I (Tn-I) have been regarded as biochemical markers of myocyte necrosis. This study was planned to investigate the specificity and sensitivity of Tn-I and CK-MB in CHF and to examine the correlation of these markers with disease severity. A total of 104 patients (38 female, 66 male; mean age, 66 y [range, 36-89]) with symptoms and signs of heart failure on admission and with a reduced left ventricular ejection fraction (EF; by transthoracic echocardiography) were labeled "the patient group," and 58 patients (40 female,18 male; mean age, 61 y [range, 34-77]) with no signs or symptoms of CHF and with a normal EF detected by transthoracic echocardiography were included in the study as "the control group." Left ventricular EFs, end-diastolic diameters, and end-systolic diameters of patients in both groups were measured. Blood samples were drawn from all patients in both groups on admission, so that levels of CK-MB and Tn-I could be measured. All patients in both groups also underwent coronary angiography. Conditions leading to elevation of CK-MB or Tn-I were considered exclusion criteria. The 2 groups failed to show any significant differences in terms of mean age and the presence of coronary artery disease, hypertension, or diabetes mellitus (P>.05). Mean EF in the patient group was lower than that in the control group (P<.05). Mean CK-MB and Tn-I in the patient group were significantly higher than in the control group (P<.05). In the patient group, hypertensive patients were found to have significantly higher mean values of CK-MB than were seen in normotensive patients in the same group (P<.05). In the patient group, 52 cases were considered to be class I-II (New York Heart Association [NYHA]) (group 1), and 52 were considered to be class III-IV (group 2). Group 1, group 2, and the control group did not differ significantly from one another with regard to the presence of coronary artery disease, hypertension, and diabetes mellitus (P>.05). The mean EF in group 2 was significantly lower than that in group 1 and in the control group (P<.05); the mean EF in group 1 was significantly lower than that in the control group (P<.05). Group 1 values did not differ significantly from those of group 2 or the control group in terms of enzymatic markers (P>.05), but group 2 had significantly higher mean values of CK-MB and Tn-I than were noted in the control group (P<.05). The uphill course of CK-MB and Tn-I values from the control group to group 2 (NYHA class III-IV) was statistically significant (P<.05). Serum concentrations of CK-MB and Tn-I may become elevated in severely symptomatic patients with CHF (particularly NYHA class III-IV), demonstrating a relationship between clinical severity of the disease and elevation of myocardial enzymes (CK-MB and Tn-I).


Tuning the isoelectric point of graphene by electrochemical functionalization.
The ability to control the charge-potential landscape at solid-liquid interfaces is pivotal to engineer novel devices for applications in sensing, catalysis and energy conversion. The isoelectric point (pI)/point of zero charge (pzc) of graphene plays a key role in a number of physico-chemical phenomena occurring at the graphene-liquid interface. Supported by theory, we present here a methodology to identify the pI/pzc of (functionalized) graphene, which also allows for estimating the nature and extent of ion adsorption. The pI of bare graphene (as-prepared, chemical vapor deposition (CVD)-grown) is found to be less than 3.3, which we can continuously modify up to 7.5 by non-covalent electrochemical attachment of aromatic amino groups, preserving the favorable electronic properties of graphene throughout. Modelling all the observed results with detailed theory, we also show that specific adsorption of ions and the substrate play only an ancillary role in our capability to tune the pI of graphene.


A Multifaceted Quality Improvement Programme to Improve Acute Kidney Injury Care and Outcomes in a Large Teaching Hospital.
Acute kidney injury (AKI) is now widely recognised as a serious health care issue, occurring in up to 25% of hospital in-patients, often with worsening of outcomes. There have been several reports of substandard care in AKI. This quality improvement (QI) programme aimed to improve AKI care and outcomes in a large teaching hospital. Areas of documented poor AKI care were identified and specific improvement activities implemented through sequential Plan-Do-Study-Act (PDSA) cycles. An electronic alert system (e-alert) for AKI was developed, a Priority Care Checklist (PCC) was tested with the aid of specialist nurses whilst targeted education activities were carried out and data on care processes and outcomes monitored. The e-alert had a sensitivity of 99% for the detection of new cases of AKI. Key aspects of the PCC saw significant improvements in their attainment: Detection of AKI within 24 hours from 53% to 100%, fluid assessment from 42% to 90%, drug review 48% to 95% and adherence to nine key aspects of care from 40% to 90%. There was a significant reduction in variability of delivered AKI care. AKI incidence reduced from 9% of all hospitalisations at baseline to 6.5% (28% reduction), AKI related length of stay reduced from 22.1 days to 17 days (23% reduction) and time to recovery (AKI days) 15.5 to 9.8 days (36% reduction). AKI related deaths also showed a trend towards reduction, from an average of 38 deaths to 34 (10.5%). The number of cases of hospital acquired AKI were reduced by 28% from 120 to 86 per month. This study demonstrates significant improvements related to a QI programme combining e-alerts, a checklist implemented by a nurse and education in improving key processes of care. This resulted in sustained improvement in key patient outcomes.


Japanese Clinical Guidelines for Endoscopic Treatment of Pancreatolithiasis.
In addition to surgery, procedures for patients with pancreatolithiasis are developing; therefore, establishing practical guidelines for the management of pancreatolithiasis is required. Three committees (the professional committee for asking clinical questions (CQs) and statements by Japanese endoscopists, the expert panel committee for rating statements by the modified Delphi method, and the evaluating committee by moderators) were organized. Eight endoscopists and a surgeon for pancreatolithiasis made the CQs and statements from a total of 694 reports of published literature by PubMed search (from 1983 to 2012). The expert panelists individually rated these clinical statements using a modified Delphi approach, in which a clinical statement receiving a median score greater than 7 on a 9-point scale from the panel was regarded as valid. The professional committee made 3, 7, and 10 CQs and statements for the concept and pathogenesis, diagnosis, and treatment, respectively. The expert panelists regarded them as valid after a 2-round modified Delphi approach. After evaluation by the moderators, the Japanese clinical guidelines for pancreatolithiasis were established. Further discussions and studies for international guidelines are needed.


Data on assessing fluoride risk in bottled waters in Iran.
The general goal of this data was to determine the concentration of fluoride and assess its risk in waters bottled in Iran. Seventy-one samples of different brands of bottled water were collected. Then, the fluoride concentration was measured through standard method for water and wastewater experiments. The non-carcinogenicity risk of fluoride for different groups of infants, children, teenagers, and adults was calculated by proper formula. The data presented here indicated that the mean concentration of fluoride in bottled waters was 0.272 mg/L, which is lower than the minimum world health organization (WHO) guideline. Further, the mean hazard quotient (HQ) values for fluoride across the groups of infants, children, teenagers, and adults with respective values of 0.0363, 0.2568, 0.1813, and 0.1452 were observed in 0, 1, 1, and 0 cases of HQ>1. Generally, in most of the tested brands, HQ value was less than 1, and this value was above 1 in only one brand of bottled water.


The transcriptional coactivator TAZ regulates reciprocal differentiation of TH17 cells and Treg cells.
An imbalance in the lineages of immunosuppressive regulatory T cells (Treg cells) and the inflammatory TH17 subset of helper T cells leads to the development of autoimmune and/or inflammatory disease. Here we found that TAZ, a coactivator of TEAD transcription factors of Hippo signaling, was expressed under TH17 cell-inducing conditions and was required for TH17 differentiation and TH17 cell-mediated inflammatory diseases. TAZ was a critical co-activator of the TH17-defining transcription factor RORγt. In addition, TAZ attenuated Treg cell development by decreasing acetylation of the Treg cell master regulator Foxp3 mediated by the histone acetyltransferase Tip60, which targeted Foxp3 for proteasomal degradation. In contrast, under Treg cell-skewing conditions, TEAD1 expression and sequestration of TAZ from the transcription factors RORγt and Foxp3 promoted Treg cell differentiation. Furthermore, deficiency in TAZ or overexpression of TEAD1 induced Treg cell differentiation, whereas expression of a transgene encoding TAZ or activation of TAZ directed TH17 cell differentiation. Our results demonstrate a pivotal role for TAZ in regulating the differentiation of Treg cells and TH17 cells.


Maternal morbidity and risk of death at delivery hospitalization.
To examine the effect of underlying maternal morbidities on the odds of maternal death during delivery hospitalization. We used data that linked birth certificates to hospital discharge diagnoses from singleton live births at 22 weeks of gestation or later during 1995-2003 in New York City. Maternal morbidities examined included prepregnancy weight more than 114 kilograms (250 pounds), chronic hypertension, pregestational or gestational diabetes mellitus, chronic cardiovascular disease, pulmonary hypertension, chronic lung disease, human immunodeficiency virus (HIV), and preeclampsia or eclampsia. Associations with maternal mortality were estimated using multivariate logistic regression. During the specified time period, 1,084,862 live singleton births and 132 maternal deaths occurred. Patients with increasing maternal age, non-Hispanic black ethnicity, self-pay or Medicaid, primary cesarean delivery, and premature delivery had higher rates of maternal mortality during delivery hospitalization. From the entire study population, 4.1% had preeclampsia or eclampsia (n=44,004), 1.8% had chronic hypertension (n=19,647), 1.1% of patients were classified as obese (n=11,936), 0.7% had pregestational diabetes (n=7,474), 0.4% had HIV (n=4,665), and 0.01% had pulmonary hypertension (n=166). Preeclampsia or eclampsia (adjusted odds ratio [OR], 8.1; 95% confidence interval [CI], 5.5-12.1), chronic hypertension (adjusted OR, 7.7; 95% CI 4.7-12.5), underlying maternal obesity (adjusted OR, 2.9; 95% CI 1.1-8.1), pregestational diabetes (adjusted OR, 3.3; 95% CI 1.3-8.1), HIV (adjusted OR, 7.7; 95% CI 3.4-17.8), and pulmonary hypertension (adjusted OR, 65.1; 95% CI 15.8-269.3) were associated with an increased risk of death during the delivery hospitalization. The presence of maternal disease significantly increases the odds of maternal mortality at the time of delivery hospitalization. II.


Combination therapy of Nigella sativa and human parathyroid hormone on bone mass, biomechanical behavior and structure in streptozotocin-induced diabetic rats.
Extracts of the seeds of Nigella sativa (NS), an annual herbaceous plant of the Ranunculaceae family, have been used for many years for therapeutic purposes, including their potential anti-diabetic properties. The aim of the present study was to test the hypothesis that combined treatment with NS and human parathyroid hormone (hPTH) is more effective than treatment with NS or hPTH alone in improving bone mass, connectivity, biomechanical behaviour and strength in insulin-dependent diabetic rats. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ) at a single dose of 50mg/kg. The diabetic rats received NS (2ml/kg/day, i.p.), hPTH (6microg/kg/day, i.p.) or NS and hPTH combined for 4 weeks, starting 8 weeks after STZ injection. The beta-cells of the pancreatic islets of Langerhans were examined by immunohistochemical methods. In addition, bone sections of femora were processed for histomorphometry and biomechanical analysis. In diabetic rats, the beta-cells were essentially negative for insulin-immunoreactivity. NS treatment (alone or in combination with hPTH) significantly increased the area of insulin immunoreactive beta-cells in diabetic rats; however, hPTH treatment alone only led to a slightly increase in the insulin-immunoreactivity. These results suggest that NS might be used in a similar manner to insulin as a safe and effective therapy for diabetes and might be useful in the treatment of diabetic osteopenia.


Impact of AFM-induced nano-pits in a-Si:H films on silicon crystal growth.
Conductive tips in atomic force microscopy (AFM) can be used to localize field-enhanced metal-induced solid-phase crystallization (FE-MISPC) of amorphous silicon (a-Si:H) at room temperature down to nanoscale dimensions. In this article, the authors show that such local modifications can be used to selectively induce further localized growth of silicon nanocrystals. First, a-Si:H films by plasma-enhanced chemical vapor deposition on nickel/glass substrates are prepared. After the FE-MISPC process, yielding both conductive and non-conductive nano-pits in the films, the second silicon layer at the boundary condition of amorphous and microcrystalline growth is deposited. Comparing AFM morphology and current-sensing AFM data on the first and second layers, it is observed that the second deposition changes the morphology and increases the local conductivity of FE-MISPC-induced pits by up to an order of magnitude irrespective of their prior conductivity. This is attributed to the silicon nanocrystals (<100 nm) that tend to nucleate and grow inside the pits. This is also supported by micro-Raman spectroscopy.


Widespread occurrence of Mycobacterium tuberculosis DNA from 18th-19th century Hungarians.
A large number (265) of burials from 1731-1838 were discovered in sealed crypts of the Dominican Church, Vác, Hungary in 1994. Many bodies were naturally mummified, so that both soft tissues and bones were available. Contemporary archives enabled the determination of age at death, and the identification of family groups. In some cases, symptoms before death were described and, occasionally, occupation. Initial radiological examination of a small number of individuals had indicated calcified lung lesions and demonstrable acid-fast bacteria suggestive of tuberculosis infection. Tuberculosis was endemic in 18th-19th century Europe, so human remains should contain detectable Mycobacterium tuberculosis complex (MTB) DNA, enabling comparisons with modern isolates. Therefore, a comprehensive examination of 168 individuals for the presence of MTB DNA was undertaken. Specific DNA amplification methods for MTB showed that 55% of individuals were positive and that the incidence varied according to age at death and sampling site in the body. Radiographs were obtained from 27 individuals and revealed an association between gross pathology and the presence of MTB DNA. There was an inverse relationship between PCR positivity and MTB target sequence size. In some cases, the preservation of MTB DNA was excellent, and several target gene sequences could be detected from the same sample. This information, combined with MTB DNA sequencing data and molecular typing techniques, will enable us to study the past epidemiology of TB infection, and extends the timeframe for studying changes in molecular fingerprints.


Outgrowth-regulating actions of glutamate in isolated hippocampal pyramidal neurons.
The present study examined the effects of glutamate on the outgrowth of dendrites and axons in isolated hippocampal pyramidal-like neurons in cell culture. During the first day of culture the survival and outgrowth of these neurons was unaffected by high concentrations (up to 1 nM) of glutamate, quisqualic acid (QA), kainic acid (KA), and N-methyl-D-aspartic acid. Beginning on day 2 of culture high levels of glutamate, KA and QA were toxic to the majority of pyramidal neurons, while subtoxic levels of these agents caused a well-defined, dose-dependent, sequence of effects on dendritic outgrowth. At increasing concentrations of glutamate, QA, and KA, the following events were observed: (1) dendritic outgrowth rates were reduced, while axonal elongation rates were unaffected; (2) dendritic length was reduced, while axons continued to grow; (3) dendrites regressed dramatically, and axonal outgrowth rate was reduced. These dendrite-specific effects of glutamate were apparently mediated at the growth cones since focal application of glutamate to individual dendritic growth cones resulted in suppression of growth cone activity and a regression of the dendrite; axons were unaffected by focal glutamate application. Pharmacological tests using glutamate receptor agonists and antagonists demonstrated that receptors of the KA/QA type mediated the glutamate effects on outgrowth and survival. The calcium channel blocker Co2+ prevented both glutamate neurotoxicity and glutamate-induced dendritic regression. Ionophore A23187 and elevations in extracellular K+ levels each caused a dose-dependent series of outgrowth and survival responses similar to those caused by glutamate. Taken together, these results indicate that activation of glutamate receptors leads to the opening of voltage-dependent calcium channels; the resulting increases in calcium influx lead to the observed alterations in dendritic outgrowth and neuronal survival.


Revisiting the hetero-fertilization phenomenon in maize.
Development of a seed DNA-based genotyping system for marker-assisted selection (MAS) has provided a novel opportunity for understanding aberrant reproductive phenomena such as hetero-fertilization (HF) by observing the mismatch of endosperm and leaf genotypes in monocot species. In contrast to conventional approaches using specific morphological markers, this approach can be used for any population derived from diverse parental genotypes. A large-scale experiment was implemented using seven F(2) populations and four three-way cross populations, each with 534 to 1024 individuals. The frequency of HF within these populations ranged from 0.14% to 3.12%, with an average of 1.46%. The highest frequency of HF in both types of population was contributed by the pollen gametes. Using three-way crosses allowed, for the first time, detection of the HF contributed by maternal gametes, albeit at very low frequency (0.14%-0.65%). Four HF events identified from each of two F(2) populations were tested and confirmed using 1032 single nucleotide polymorphic markers. This analysis indicated that only 50% of polymorphic markers can detect a known HF event, and thus the real HF frequency can be inferred by doubling the estimate obtained from using only one polymorphic marker. As expected, 99% of the HF events can be detected by using seven independent markers in combination. Although seed DNA-based analysis may wrongly predict plant genotypes due to the mismatch of endosperm and leaf DNA caused by HF, the relatively low HF frequencies revealed with diverse germplasm in this study indicates that the effect on the accuracy of MAS is limited. In addition, comparative endosperm and leaf DNA analysis of specific genetic stocks could be useful for revealing the relationships among various aberrant fertilization phenomena including haploidy and apomixis.