Genotyping Array Design and Data Quality Control in the Million Veteran Program.
The Million Veteran Program (MVP), initiated by the Department of Veterans Affairs (VA), aims to collect biosamples with consent from at least one million veterans. Presently, blood samples have been collected from over 800,000 enrolled participants. The size and diversity of the MVP cohort, as well as the availability of extensive VA electronic health records, make it a promising resource for precision medicine. MVP is conducting array-based genotyping to provide a genome-wide scan of the entire cohort, in parallel with whole-genome sequencing, methylation, and other 'omics assays. Here, we present the design and performance of the MVP 1.0 custom Axiom array, which was designed and developed as a single assay to be used across the multi-ethnic MVP cohort. A unified genetic quality-control analysis was developed and conducted on an initial tranche of 485,856 individuals, leading to a high-quality dataset of 459,777 unique individuals. 668,418 genetic markers passed quality control and showed high-quality genotypes not only on common variants but also on rare variants. We confirmed that, with non-European individuals making up nearly 30%, MVP's substantial ancestral diversity surpasses that of other large biobanks. We also demonstrated the quality of the MVP dataset by replicating established genetic associations with height in European Americans and African Americans ancestries. This current dataset has been made available to approved MVP researchers for genome-wide association studies and other downstream analyses. Further data releases will be available for analysis as recruitment at the VA continues and the cohort expands both in size and diversity.


Multipoint Measurements of the Electron Jet of Symmetric Magnetic Reconnection with a Moderate Guide Field.
We report observations from the Magnetospheric Multiscale (MMS) satellites of the electron jet in a symmetric magnetic reconnection event with moderate guide field. All four spacecraft sampled the ion diffusion region and observed the electron exhaust. The observations suggest that the presence of the guide field leads to an asymmetric Hall field, which results in an electron jet skewed towards the separatrix with a nonzero component along the magnetic field. The jet appears in conjunction with a spatially and temporally persistent parallel electric field ranging from -3 to -5 mV/m, which led to dissipation on the order of 8 nW/m^{3}. The parallel electric field heats electrons that drift through it, and is associated with a streaming instability and electron phase space holes.


Critically examining diversity in end-of-life family caregiving: implications for equitable caregiver support and Canada's Compassionate Care Benefit.
Family (i.e., unpaid) caregiving has long been thought of as a 'woman's issue', which ultimately results not only in gendered, but also financial and health inequities. Because of this, gender-based analyses have been prioritized in caregiving research. However, trends in current feminist scholarship demonstrate that gender intersects with other axes of difference, such as culture, socio-economic status, and geography to create diverse experiences. In this analysis we examine how formal front-line palliative care providers understand the role of such diversities in shaping Canadian family caregivers' experiences of end-of-life care. In doing so we consider the implications of these findings for a social benefit program aimed at supporting family caregivers, namely the Compassionate Care Benefit (CCB). This analysis contributes to a utilization-focused evaluation of Canada's CCB, a social program that provides job security and limited income assistance to Canadian family caregivers who take a temporary leave from employment to provide care for a dying family member at end-of-life. Fifty semi-structured phone interviews with front-line palliative care providers from across Canada were conducted and thematic diversity analysis of the transcripts ensued. Findings reveal that experiences of caregiving are not homogenous and access to services and supports are not universal across Canada. Five axes of difference were commonly raised by front-line palliative care providers when discussing important differences in family caregivers' experiences: culture, gender, geography, lifecourse stage, and material resources. Our findings reveal inequities with regard to accessing needed caregiver services and resources, including the CCB, based on these axes of difference. We contend that without considering diversity, patterns in vulnerability and inequity are overlooked, and thus continually reinforced in health policy. Based on our findings, we demonstrate that re-framing categorizations of caregivers can expose specific vulnerabilities and inequities while identifying implications for the CCB program as it is currently administered. From a policy perspective, this analysis demonstrates why diversity needs to be acknowledged in policy circles, including in relation to the CCB, and seeks to counteract single dimensional approaches for understanding caregiver needs at end-of-life. Such findings illustrate how diversity analysis can dramatically enhance evaluative health policy research.


The challenges of genome-wide interaction studies: lessons to learn from the analysis of HDL blood levels.
Genome-wide association studies (GWAS) have revealed 74 single nucleotide polymorphisms (SNPs) associated with high-density lipoprotein cholesterol (HDL) blood levels. This study is, to our knowledge, the first genome-wide interaction study (GWIS) to identify SNP×SNP interactions associated with HDL levels. We performed a GWIS in the Rotterdam Study (RS) cohort I (RS-I) using the GLIDE tool which leverages the massively parallel computing power of Graphics Processing Units (GPUs) to perform linear regression on all genome-wide pairs of SNPs. By performing a meta-analysis together with Rotterdam Study cohorts II and III (RS-II and RS-III), we were able to filter 181 interaction terms with a p-value<1 · 10-8 that replicated in the two independent cohorts. We were not able to replicate any of these interaction term in the AGES, ARIC, CHS, ERF, FHS and NFBC-66 cohorts (Ntotal = 30,011) when adjusting for multiple testing. Our GWIS resulted in the consistent finding of a possible interaction between rs774801 in ARMC8 (ENSG00000114098) and rs12442098 in SPATA8 (ENSG00000185594) being associated with HDL levels. However, p-values do not reach the preset Bonferroni correction of the p-values. Our study suggest that even for highly genetically determined traits such as HDL the sample sizes needed to detect SNP×SNP interactions are large and the 2-step filtering approaches do not yield a solution. Here we present our analysis plan and our reservations concerning GWIS.


Localized sources of water vapour on the dwarf planet (1) Ceres.
The 'snowline' conventionally divides Solar System objects into dry bodies, ranging out to the main asteroid belt, and icy bodies beyond the belt. Models suggest that some of the icy bodies may have migrated into the asteroid belt. Recent observations indicate the presence of water ice on the surface of some asteroids, with sublimation a potential reason for the dust activity observed on others. Hydrated minerals have been found on the surface of the largest object in the asteroid belt, the dwarf planet (1) Ceres, which is thought to be differentiated into a silicate core with an icy mantle. The presence of water vapour around Ceres was suggested by a marginal detection of the photodissociation product of water, hydroxyl (ref. 12), but could not be confirmed by later, more sensitive observations. Here we report the detection of water vapour around Ceres, with at least 10(26) molecules being produced per second, originating from localized sources that seem to be linked to mid-latitude regions on the surface. The water evaporation could be due to comet-like sublimation or to cryo-volcanism, in which volcanoes erupt volatiles such as water instead of molten rocks.


Newly diagnosed gastroesophageal reflux disease increased the risk of acute exacerbation of chronic obstructive pulmonary disease during the first year following diagnosis--a nationwide population-based cohort study.
While prior studies have demonstrated that chronic obstructive pulmonary disease (COPD) is associated with gastroesophageal reflux disease (GERD), and that GERD is associated with acute exacerbations of COPD (AECOPD), no study to date has been able to establish temporality in this relationship. The purpose of this cohort study was to explore the impact of a new diagnosis of GERD on the risk of subsequent AECOPD. We used a retrospective population-based cohort design to analyse the data of 1976 COPD subjects with GERD as an exposure cohort and 3936 COPD subjects without GERD as a comparison group. We individually tracked each subject in this study for 12 months and identified those subjects who experienced an episode of AECOPD. Hazard ratios (HR) were calculated using Cox proportional hazards regression analysis. The incidence of AECOPD was 4.08 and 2.79 per 100 person-year in individuals with and without GERD, respectively (p = 0.012). Following adjustment for sex, age, ischaemic heart disease, heart failure, atrial fibrillation, hypertension, osteoporosis, anxiety, diabetes mellitus, angina, stroke, anaemia, dementia, occupational category, monthly insurance premium, number of OPD visits and COPD severity. The stepwise Cox regression analysis revealed that GERD was independently associated with an increased risk of AECOPD (HR = 1.48, 95% CI = 1.10-1.99). This study demonstrated that GERD is an independent risk factor for AECOPD. Caution should be exercised when assessing GERD symptoms in patients with COPD.


Effects of timing of pre-exercise ingestion of carbohydrate on subsequent metabolism and cycling performance.
The occurrence of rebound hypoglycaemia may depend on the timing of carbohydrate intake. The aim of the present study was to investigate the metabolic and performance responses to the ingestion of carbohydrate at differing times before exercise. Eight subjects [mean (SEM)] [28 (3) years, 74.5 (2.6) kg, maximal oxygen uptake 63.1 (3.1) ml.kg(-1).min(-1)] performed three experiments. They ingested 75 g glucose dissolved in 500 ml water, thereafter resting for either 15, 45 or 75 min (15-Pre, 45-Pre and 75-Pre) before exercising for 20 min at 65% maximal power output followed by a time trial [total work 685 (18) kJ]. There were no differences in performance between conditions [mean powers 268 (10), 269 (7) and 276 (12) W for 15-Pre, 45-Pre and 75-Pre, respectively]. There were significant differences in plasma glucose concentration between 15-Pre [6.6 (0.6) mmol.l(-1); P<0.05] and both 45-Pre [4.5 (0.2) mmol.l(-1)] and 75-Pre [3.7 (0.2) mmol.l(-1)] immediately before exercise. Insulin concentrations immediately before exercise were higher ( P<0.05) during 15-Pre [72.6 (10.4) microU.ml(-1)] than during 45-Pre [50.8 (9.9) microU.ml(-1)], which was higher ( P<0.05) than during 75-Pre [33.9 (5.5) microU.ml(-1)]. These differences disappeared within 10 min of exercise. Two subjects became hypoglycaemic (plasma glucose concentration of less than 3.5 mmol.l(-1)) in the 15-Pre while three and five subjects were transiently hypoglycaemic in the 45-Pre and 75-Pre, respectively. Performance and rating of perceived exertion did not seem to be related to hypoglycaemia. Altering the timing of the ingestion of carbohydrate before exercise resulted in differences in plasma glucose/insulin responses which disappeared within 10 min of exercise and which had no effect on performance. Hypoglycaemia was observed in some subjects during the first 10 min but this did not affect performance.


Comparison of outcomes after hospitalization for worsening heart failure, myocardial infarction, and stroke in patients with heart failure and reduced and preserved ejection fraction.
To investigate the prognostic significance of hospitalization for worsening heart failure (WHF), myocardial infarction (MI), and stroke in patients with chronic heart failure (HF). We studied 5011 patients with HF and reduced EF (HF-REF) in the CORONA trial and 4128 patients with HF and preserved EF (HF-PEF) in the I-Preserve trial. Adjusted hazard ratios (HRs) for death were estimated for 0-30 days and ≥31 days after first post-randomization WHF, MI, or stroke used as a time-dependent variable, compared with patients with none of these events. In CORONA, 1616 patients (32%) had post-randomization first events (1223 WHF, 216 MI, 177 stroke), and the adjusted HR for mortality ≤30 days after an event was: WHF 7.21 [95% confidence interval (CI) 2.05-25.40], MI 23.08 (95% CI 6.44-82.71), and stroke 32.15 (95% CI 8.93-115.83). The HR for mortality at >30 days was: WHF 3.62 (95% CI 3.11-4.21), MI 4.41 (95% CI 3.23-6.02), and stroke 3.19 (95% CI 2.21-4.61). In I-Preserve, 896 patients (22%) experienced a post-randomization event (638 WHF, 111 MI, 147 stroke). The HR for mortality ≤30 days was WHF 31.77 (95% CI 7.60-132.81), MI 154.77 (95% CI 34.21-700.17), and stroke 223.30 (95% CI 51.42-969.78); for >30 days it was WHF 3.36 (95% CI 2.79-4.05), MI 3.29 (95% CI 2.14-5.06), and stroke 5.13 (95% CI 3.61-7.29). In patients with both HF-REF and HF-PEF, hospitalization for WHF was associated with high early and late mortality. The early relative risk of death was not as great as following MI or stroke, but the longer term relative risk of death was similar following all three types of event. Numerically, more deaths occurred following WHF because it was a much more common event.


A general model for metabolic scaling in self-similar asymmetric networks.
How a particular attribute of an organism changes or scales with its body size is known as an allometry. Biological allometries, such as metabolic scaling, have been hypothesized to result from selection to maximize how vascular networks fill space yet minimize internal transport distances and resistances. The West, Brown, Enquist (WBE) model argues that these two principles (space-filling and energy minimization) are (i) general principles underlying the evolution of the diversity of biological networks across plants and animals and (ii) can be used to predict how the resulting geometry of biological networks then governs their allometric scaling. Perhaps the most central biological allometry is how metabolic rate scales with body size. A core assumption of the WBE model is that networks are symmetric with respect to their geometric properties. That is, any two given branches within the same generation in the network are assumed to have identical lengths and radii. However, biological networks are rarely if ever symmetric. An open question is: Does incorporating asymmetric branching change or influence the predictions of the WBE model? We derive a general network model that relaxes the symmetric assumption and define two classes of asymmetrically bifurcating networks. We show that asymmetric branching can be incorporated into the WBE model. This asymmetric version of the WBE model results in several theoretical predictions for the structure, physiology, and metabolism of organisms, specifically in the case for the cardiovascular system. We show how network asymmetry can now be incorporated in the many allometric scaling relationships via total network volume. Most importantly, we show that the 3/4 metabolic scaling exponent from Kleiber's Law can still be attained within many asymmetric networks.


A Novel MRI Tool for Evaluating Cortical Bone Thickness of the Proximal Femur.
Osteoporotic hip fractures heavily cost the health care system. Clinicians and patients can benefit from improved tools to assess bone health. Herein, we aim to develop a three-dimensional magnetic resonance imaging (MRI) method to assess cortical bone thickness and assess the ability of the method to detect regional changes in the proximal femur. Eighty-nine patients underwent hip magnetic resonance imaging. FireVoxel and 3DSlicer were used to generate three-dimensional proximal femur models. ParaView was used to define five regions: head, neck, greater trochanter, intertrochanteric region, and subtrochanteric region. Custom software was used to calculate the cortical bone thickness and generate a color map of the proximal femur. Mean cortical thickness values for each region were calculated. Statistical t-tests were performed to evaluate differences in cortical thickness based on proximal femur region. Measurement reliability was evaluated using coefficient of variation, intraclass correlation coefficients, and overlap metrics. Three-dimensional regional cortical thickness maps for all subjects were generated. The subtrochanteric region was found to have the thickest cortical bone and the femoral head had the thinnest cortical bone. There were statistically significant differences between regions (p < 0.01) for all possible comparisons. Cortical bone is an important contributor to bone strength, and its thinning results in increased hip fracture risk. We describe the development and measurement reproducibility of an MRI tool permitting assessment of proximal femur cortical thickness. This study represents an important step toward longitudinal clinical trials interested in monitoring the effectiveness of drug therapy on proximal femur cortical thickness.


Safety of acupuncture: results of a prospective observational study with 229,230 patients and introduction of a medical information and consent form.
To evaluate the safety of acupuncture in a large number of patients receiving conventional health care and, based on these results, to develop a new medical consent form for acupuncture. The prospective observational study included patients who received acupuncture treatment for chronic osteoarthritis pain of the knee or hip, low back pain, neck pain or headache, allergic rhinitis, asthma, or dysmenorrhoea. After treatment, all patients documented adverse events associated with acupuncture (defined as adverse effects). Patients who reported a need for treatment due to an adverse effect completed an additional standardised questionnaire on the most important adverse effect. Based on this data and considering ethical and legal aspects a new consent form was developed. A total of 229,230 patients received on average 10.2 +/- 3.0 acupuncture treatments. Altogether, 19,726 patients (8.6%) reported experiencing at least one adverse effect and 4,963 (2.2%) reported one which required treatment. Common adverse effects were bleedings or haematoma (6.1% of patients, 58% of all adverse effects), pain (1.7%) and vegetative symptoms (0.7%). Two patients experienced a pneumothorax (one needed hospital treatment, the other observation only). The longest duration of a side effect was 180 days (nerve lesion of the lower limb). The resulting medical consent form consists of five modules: Introduction to acupuncture and moxibustion, Risks of acupuncture treatment, Conditions which can increase the risk, Doctor's statement, and Consent. Acupuncture provided by physicians is a relatively safe treatment and the proposed consent form could support both patients and professionals in the process of obtaining informed consent.


Right, but not left, bundle branch block is associated with large anteroseptal scar.
This study sought to test the hypothesis that right bundle branch block (RBBB) patients have larger scar size than left bundle branch block (LBBB) patients do. A proximal septal perforating branch of the left anterior descending (LAD) coronary artery most commonly perfuses the right bundle branch and left anterior fascicle, but not the left posterior fascicle. Thus, proximal LAD occlusions should cause RBBB, not LBBB. We performed electrocardiograms and magnetic resonance imaging for scar quantification in 233 patients with left ventricular (LV) ejection fraction ≤35% who were receiving primary prevention implantable cardioverter-defibrillators (ICD cohort). Scar size and location were compared among patients with RBBB, LBBB, nonspecific LV conduction delay, and QRS <120 ms. A second cohort of 20 hypertrophic cardiomyopathy patients undergoing alcohol septal ablation was studied to determine whether controlled infarction in a proximal LAD septal perforator caused RBBB or LBBB. In the ICD cohort, LV ejection fraction was similar between RBBB and LBBB patients (24.9% vs. 25.0%; p = 0.98); however, RBBB patients had significantly larger scar size (24.0% vs. 6.5%; p < 0.0001). Patients with nonspecific LV conduction delay or QRS <120 ms had intermediate scar size (12.9% and 14.4%, respectively). Those with RBBB (compared with LBBB) were more likely to have ischemic heart disease (79% vs. 29%; p < 0.0001). In the alcohol septal ablation cohort, 15 of 20 patients (75%) developed RBBB, but no patients developed LBBB. In patients with LV ejection fraction ≤35%, RBBB is associated with significantly larger scar size than LBBB is, and occlusion of a proximal LAD septal perforator causes RBBB. In contrast, LBBB is most commonly caused by nonischemic pathologies.


Anesthesia for bariatric surgery in an achondroplastic dwarf with morbid obesity.
Achondroplasia is the most common form among the different types of osteochondrodysplasia that cause dwarfism. Dwarves develop obesity quite frequently and surgical treatment has shown greater efficacy, both for effective weight loss and long term maintenance. The objective of this report was to present the case of bariatric surgery with Y-en-Roux gastric bypass in an achondroplastic dwarf with morbid obesity. The different difficulties in the anesthetic management of this patient and the way they were dealt with were discussed in order to decrease intraoperative morbidity and mortality. This is a 29 years old female dwarf with achondroplasia and morbid obesity since childhood. She was 123 cm tall and weighed 144 kg at the time of admission to the Bariatric Surgery service. With a body mass index (BMI) of 95.18 kg.m2, she had several associated diseases especially of the respiratory system and osteoarticular system. After a long follow-up with diet, exercises, and psychological support, her clinical condition improved and she was referred for surgery: Y-en-Roux gastroplasty using the technique of Capella-Fobi. Intubation of the awake patient under direct laryngoscopy was difficult and a bronchofibroscope had to be used. Surgery was uneventful and the patient was maintained under total intravenous anesthesia with continuous infusion of remifentanil and propofol. She was extubated at the end of the surgery still in the operating room. The simultaneous comorbidities of achondroplasia and morbid obesity can hinder the anesthetic management, especially regarding the airways. A thorough pre-anesthetic evaluation is necessary to anticipate the conducts and minimize risks, therefore optimizing the evolution of anesthesia.


Functional Significance of Nuclear α Spectrin.
Nonerythroid alpha spectrin (αIISp) interacts in the nucleus with an array of different proteins indicating its involvement in a number of diverse functions. However, the significance of these interactions and their functional importance has been a relatively unexplored area. The best documented role of nuclear αIISp is in DNA repair where it is critical for repair of DNA interstrand cross-links (ICLs), acting as a scaffold recruiting proteins to sites of damage in genomic and telomeric DNA. A deficiency in αIISp can importantly impact DNA ICL repair as is seen in cells from patients with the genetic disorder, Fanconi anemia (FA), where loss of αIISp leads to not only defects in repair of both genomic and telomeric DNA but also to telomere dysfunction and chromosome instability. This previously unexplored link between αIISp and telomere function is important in developing an understanding of maintenance of genomic stability after ICL damage. In FA cells, these defects in chromosome instability after ICL damage can be corrected when levels of αIISp are returned to normal by knocking down μ-calpain, a protease which cleaves αIISp. These studies suggest a new direction for correcting a number of the phenotypic defects in FA and could serve as a basis for therapeutic intervention. More in depth, examination of the interactions of αIISp with other proteins in the nucleus is of major importance in development of insights into the interacting key elements involved in the diverse processes occurring in the nucleus and the consequences loss of αIISp has on them.


Changes in parameters of bone metabolism in postmenopausal women following a 12-month intervention period using dairy products enriched with calcium, vitamin D, and phylloquinone (vitamin K(1)) or menaquinone-7 (vitamin K (2)): the Postmenopausal Health Study II.
The objective of the present study was to examine the effect of dairy products enriched with calcium, vitamin D(3), and phylloquinone (vitamin K(1)) or menaquinone-7 (vitamin K(2)) on parameters of bone metabolism in postmenopausal women following a 12-month intervention. Postmenopausal women were divided into three intervention groups and a control group (CG). All three intervention groups attended biweekly sessions and received fortified dairy products providing daily 800 mg of calcium and 10 μg of vitamin D(3) (CaD). Furthermore, in two of the three intervention groups the dairy products were also enriched with vitamin K, providing daily 100 μg of either phylloquinone (CaDK1) or menaquinone-7 (CaDK2). The increase observed for serum 25(OH)D levels in all intervention groups and the increase observed for serum IGF-I levels in the CaDK2 group differed significantly compared to the changes observed in CG (P = 0.010 and P = 0.028, respectively). Furthermore, both the CaDK1 and CaDK2 groups had a significantly lower mean serum undercarboxylated osteocalcin to osteocalcin ratio and urine deoxypyridinoline levels at follow-up compared to the CaD and CG groups (P = 0.001 and P = 0.047, respectively). Significant increases in total-body BMD were observed in all intervention groups compared to CG (P < 0.05), while significant increases in lumbar spine BMD were observed only for CaDK1 and CaDK2 compared to CG (P < 0.05) after controlling for changes in serum 25(OH)D levels and dietary calcium intake. In conclusion, the present study revealed more favorable changes in bone metabolism and bone mass indices for the two vitamin K-supplemented groups, mainly reflected in the suppression of serum levels of bone remodeling indices and in the more positive changes in lumbar spine BMD for these two study groups.


Developing and using a structured, conversation-based intervention for clarifying values and preferences for end-of-life in the advance care planning-naïve Swedish context: Action research within the DöBra research program.
Sweden has no systematic advance care planning (ACP), nor legal recognition of end-of-life proxies. We describe our experiences and reflections from a participatory action research process, aiming at developing and initially using a conversation-based, structured ACP approach among community-dwelling, older adults in Sweden. Eco-mapping and DöBra cards were used with 65 people to catalyze discussions on preferences for the end-of-life. We found great individual variation in both Eco-map depictions of social networks and prioritization of the 37 DöBra card items. The DöBra cards were concluded to be a viable tool for stimulating person-centered conversations on preferences for future end-of-life care.


Dissimilar expression of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes.
The temporal expression of the Autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes in Spodoptera frugiperda cells was studied using virus recombinants in which either gene was replaced by the juvenile hormone esterase (JHE) gene of Heliothis virescens. The JHE served as a highly specific and sensitive reporter for gene expression. Activation of the p10 gene followed a pattern different to that of polyhedrin. The p10 gene was activated a few hours earlier than the polyhedrin gene, but its expression reached a lower maximum level. Northern blot analysis complemented and confirmed the results obtained from the JHE assays. Co-infection of sense recombinants and those containing an antisense copy of the JHE gene in place of the polyhedrin or p10 gene resulted in reduced levels of JHE gene expression. These experiments independently supported the hypothesis that the p10 gene promoter is more active at earlier times post-infection than that of the polyhedrin gene. The results also highlight the potential of the antisense strategy as an experimental approach for the study of baculovirus gene regulation and possibly insect metabolism.


PCTAIRE1 regulates p27 stability, apoptosis and tumor growth in malignant melanoma.
PCTAIRE1 is a cyclin-dependent kinase family protein that has been implicated in spermatogenesis. Although we recently revealed the function of PCTAIRE1 in tumorigenesis of epithelial carcinoma cells, its tumorigenic function in melanoma remains unclear. Interrogation of the Oncomine database revealed that malignant melanoma showed up-regulation of PCTAIRE1 mRNA compared to normal skin and benign melanocytic nevus tissues. In the melanoma cell lines A2058 and SK-MEL-28, PCTAIRE1 gene knockdown using siRNA or shRNA diminished melanoma cell proliferation as assessed by cellular ATP levels, cell counting and clonogenic assays. Moreover, FACS analyses of annexin V-PI staining and DNA content showed that PCTAIRE1 knockdown caused apoptosis in A2058 cells. In contrast, PCTAIRE1 does not appear to be involved in the proliferation of immortalized human keratinocyte HaCaT cells. Depletion of PCTAIRE1 by siRNA/shRNA led to p27 accumulation in melanoma cells but not HaCaT cells. In tumor xenografts of melanoma A2058 cells, conditional knockdown of PCTAIRE1 restored p27 protein expression and suppressed tumor growth. Our findings reveal a crucial role for PCTAIRE1 in regulating p27 protein levels and tumor growth in melanoma cells, suggesting that PCTAIRE1 could provide a target for melanoma treatment.


ImmunoMap: A Bioinformatics Tool for T-cell Repertoire Analysis.
Despite a dramatic increase in T-cell receptor (TCR) sequencing, few approaches biologically parse the data in a fashion that both helps yield new information about immune responses and may guide immunotherapeutic interventions. To address this issue, we developed a method, ImmunoMap, that utilizes a sequence analysis approach inspired by phylogenetics to examine TCR repertoire relatedness. ImmunoMap analysis of the CD8 T-cell response to self-antigen (Kb-TRP2) or to a model foreign antigen (Kb-SIY) in naïve and tumor-bearing B6 mice showed differences in the T-cell repertoire of self- versus foreign antigen-specific responses, potentially reflecting immune pressure by the tumor, and also detected lymphoid organ-specific differences in TCR repertoires. When ImmunoMap was used to analyze clinical trial data of tumor-infiltrating lymphocytes from patients being treated with anti-PD-1, ImmunoMap, but not standard TCR sequence analyses, revealed a clinically predicative signature in pre- and posttherapy samples. Cancer Immunol Res; 6(2); 151-62. ©2017 AACR.


caBIONet--A .NET wrapper to access and process genomic data stored at the National Cancer Institute's Center for Bioinformatics databases.
The National Cancer Institute's Center for Bioinformatics (NCICB) has developed a Java based data management and information system called caCORE. One component of this software suite is the object oriented API (caBIO) used to access the rich biological datasets collected at the NCI. This API can access the data using native Java classes, SOAP requests or HTTP calls. Non-Java based clients wanting to use this API have to use the SOAP or HTTP interfaces with the data being returned from the NCI servers as an XML data stream. Although the XML can be read and manipulated using DOM or SAX parsers, one loses the convenience and usability of an object oriented programming paradigm. caBIONet is a set of .NET wrapper classes (managers, genes, chromosomes, sequences, etc.) capable of serializing the XML data stream into local .NET objects. The software is able to search NCICB databases and provide local objects representing the data that can be manipulated and used by other .NET programs. The software was written in C# and compiled as a .NET DLL.


High dehydroepiandrosterone-sulfate predicts breast cancer progression during new aromatase inhibitor therapy and stimulates breast cancer cell growth in tissue culture: a renewed role for adrenalectomy.
Stage IV hormone-sensitive breast cancer is often treated with aromatase inhibitors (anastrozole, letrozole, exemestane), which block the conversion of dehydroepiandrosterone (DHEA) to estrone and estradiol. This is intended to obviate the need for steroid replacement and antiquate adrenalectomy. Patients who underwent oophorectomy and were being treated with new aromatase inhibitor therapy received serial measurements of serum estrone, estradiol, and DHEA-sulfate (DHEA-S). Steroid values during responsive and progressive phases of disease were compared. In vitro, human breast cancer cell lines T-47D (estrogen-receptor and progesterone-receptor positive) and HCC 1937 (estrogen-receptor and progesterone-receptor negative) were treated with DHEA-S. Proliferation rates were measured by colorimetric assay. Disease in 12 of the 19 patients progressed. DHEA-S was less than 89 microg/dL in patients during the responsive phase and more than or equal to 89 microg/dL during disease progression, with 1 exception (P < .0005). Estrone and estradiol remained suppressed. After disease progression, the condition of 9 patients stabilized with aminoglutethimide therapy (n = 8) or adrenalectomy (n = 1), and their DHEA-S levels were reduced to less than 89 microg/dL. In vitro, elevated DHEA-S induced cell proliferation in T-47D cells. DHEA-S levels more than or equal to 89 microg/dL predicted disease progression in states of low estrogen. Tissue culture results supported the role of DHEA-S as an estrogenic agent. Oophorectomies with either aminoglutethimide therapy or adrenalectomy were effective remedies for breast cancer progression due to high DHEA-S.


ARFI shear-wave elastography with simulation of acute urinary tract obstruction in an ex vivo porcine kidney model.
A total of 20 heparinized pig kidneys were investigated at 10 intrapelvic hydrostatic pressure steps (0-90 mmHg). SWE (ARFI; Virtual TouchTM IQ, Siemens) measurements were taken at three different measuring regions and in two measuring sequences using a linear ultrasonography probe (9L4, Siemens). Median values of 10 shear-wave speed (SWS) measurements were calculated for each pressure step. Logarithmic transformed median SWS values were analyzed in a linear mixed model. SWS increased significantly with increasing intrapelvic pressure. Median SWS for all kidneys in both measuring sequences and all measuring regions was 1.47 m/s (interquartile range [IQR], 0.38 m/s) at 0 mmHg, 1.94 m/s (IQR, 0.42 m/s) at 30 mmHg, 2.07 m/s (IQR, 0.43 m/s) at 60 mmHg, 2.24 m/s (IQR, 0.49 m/s) at 90 mmHg. The correlation between pelvic pressure increase and median SWS values for the central parenchyma was significantly higher compared with the peripheral parenchyma. Acutely increased renal pelvic pressure correlates with increasing SWS values in ARFI elastography in an ex vivo porcine kidney model.


Passive sweat collection and colorimetric analysis of biomarkers relevant to kidney disorders using a soft microfluidic system.
The rich range of biomarkers in sweat and the ability to collect sweat in a non-invasive manner create interest in the use of this biofluid for assessments of health and physiological status, with potential applications that range from sports and fitness to clinical medicine. This paper introduces two important advances in recently reported classes of soft, skin-interfaced microfluidic systems for sweat capture and analysis: (1) a simple, broadly applicable means for collection of sweat that bypasses requirements for physical/mental exertion or pharmacological stimulation and (2) a set of enzymatic chemistries and colorimetric readout approaches for determining the concentrations of creatinine and urea in sweat, throughout ranges that are physiologically relevant. The results allow for routine, non-pharmacological capture of sweat for patient populations, such as infants and the elderly, that cannot be expected to sweat through exercise, and they create potential opportunities in the use of sweat for kidney disease screening/monitoring. Studies on human subjects demonstrate these essential capabilities, with quantitative comparisons to standard methods. The results expand the range of options available in microfluidic sampling and sensing of sweat for disease diagnostics and health monitoring.


Transient impaired glucose tolerance in Pima Indians: is it important?
As part of a continuing epidemiological study of non-insulin dependent diabetes among Pima Indians 154 subjects who had had a transient impairment of glucose tolerance were followed up for 1.2-16.9 (median 5.8) years after their glucose tolerance had returned to normal. Of these, 49 subsequently developed diabetes; 26 subsequently developed impaired glucose tolerance; and 79 had normal glucose tolerance at the last examination. The cumulative incidence of diabetes was 16% and 48% at five and 10 years of follow up respectively, compared with 3% and 8% for a control group of 1245 members of the same population. After adjustment for age, sex, body mass index, and plasma glucose concentration two hours after glucose loading the incidence of diabetes among the subjects who had had transient impaired glucose tolerance was 3.0 times that among the controls (95% confidence interval 2.1 to 4.3). Proportional hazards function analysis indicated that obesity was the most important predictor of subsequent development of diabetes. The results suggest that transient impairment of glucose tolerance indicates, at least in some subjects, a predisposition to diabetes and should not be considered clinically unimportant.


Comparison of CT head interpretation between emergency physicians and neuroradiologists.
Cranial computed tomography (CT) of the head is widely used in the emergency department 24 h a day. We compared the accuracy of CT head interpretation between staff emergency physicians (EPs) and neuroradiologists. We conducted a health records review of patients who required head CT in the emergency department. Two independent reviewers rated disagreement as clinically normal, significant, or clinically insignificant findings using published definition criteria. We calculated concordance and prepared descriptive and kappa statistics with 95% confidence intervals using SAS 9.1 software. We included 442 for this study. CT heads were classified as: normal or nonacute 81.5% (360 cases), insignificant 3.8% (17 cases), and significant 14.7% (65 cases). The weighted kappa for agreement was 0.83 (95% confidence interval 0.76-0.90). None of these patients had adverse outcomes related to EP misinterpretation of the CT head. In conclusion, clinically important findings on CT head are not commonly missed by our EPs and patients rarely have inappropriate disposition.


Quality of life in Swedish children with congenital ichthyosis.
Congenital ichthyosis encompasses a large group of keratinizing disorders with widespread scaling and a variable degree of erythema. Little is known about the quality of life in children with congenital ichthyosis and the impact of the disease on their family. Fifteen children aged 5-16 years with lamellar ichthyosis, Netherton's syndrome, epidermolytic hyperkeratosis or Harlequin ichthyosis, were investigated concerning the effect of their ichthyosis on their quality of life. This was measured with the established Children's Dermatology Life Quality Index (CDLQI), and the Dermatitis Family Impact questionnaire (DFI) modified by substituting the word ichthyosis for eczema. The questionnaires covered the preceding seven days and each had a maximum score of 30: the higher the score, the greater the Quality of Life impairment. The median score was 9.0 (range 2-19) for the CDLQI and 9.0 (range 3-21) for DFI. There was a significant correlation between the DFI and the CDLQI scores. The item in the CDLQI questionnaire that showed the highest score was "itchy, scratchy, sore or painful skin" and the most highly scored item in the DFI questionnaire was effect on "housework, e.g. washing, cleaning"; both items related to the children's symptoms. The results of the study clearly establish that congenital ichthyosis impairs the quality of life of the affected children and their families.


The role of extreme events in the impacts of selective tropical forestry on erosion during harvesting and recovery phases at Danum Valley, Sabah.
Ten years' hydrological investigations at Danum have provided strong evidence of the effects of extremes of drought, as in the April 1992 El Niño southern oscillation event, and flood, as in January 1996. The 1.5 km2 undisturbed forest control catchment experienced a complete drying out of the stream for the whole 1.5 km of defined channel above the gauging station in 1992, but concentrated surface flow along every declivity from within a few metres of the catchment divide after the exceptional rains of 19 January 1996. Under these natural conditions, erosion is episodic. Sediment is discharged in pulses caused by storm events, collapse of debris dams and occasional landslips. Disturbance by logging accentuates this irregular regime. In the first few months following disturbance, a wave of sediment is moved by each storm, but over subsequent years, rare events scour sediment from bare areas, gullies and channel deposits. The spatial distribution of sediment sources changes with time after logging, as bare areas on slopes are revegetated and small gullies are filled with debris. Extreme storm events, as in January 1996, cause logging roads to collapse, with landslides leading to surges of sediment into channels, reactivating the pulsed sediment delivery by every storm that happened immediately after logging. These effects are not dampened out with increasing catchment scale. Even the 721 km2 Sungai Segama has a sediment yield regime dominated by extreme events, the sediment yield in that single day on 19 January 1996 exceeding the annual sediment load in several previous years. In a large disturbed catchment, such road failures and logging-activity-induced mass movements increase the mud and silt in floodwaters affecting settlements downstream. Management systems require long-term sediment reduction strategies. This implies careful road design and good water movement regulation and erosion control throughout the logging process.


De novo sequencing and analysis of the American ginseng root transcriptome using a GS FLX Titanium platform to discover putative genes involved in ginsenoside biosynthesis.
American ginseng (Panax quinquefolius L.) is one of the most widely used herbal remedies in the world. Its major bioactive constituents are the triterpene saponins known as ginsenosides. However, little is known about ginsenoside biosynthesis in American ginseng, especially the late steps of the pathway. In this study, a one-quarter 454 sequencing run produced 209,747 high-quality reads with an average sequence length of 427 bases. De novo assembly generated 31,088 unique sequences containing 16,592 contigs and 14,496 singletons. About 93.1% of the high-quality reads were assembled into contigs with an average 8-fold coverage. A total of 21,684 (69.8%) unique sequences were annotated by a BLAST similarity search against four public sequence databases, and 4,097 of the unique sequences were assigned to specific metabolic pathways by the Kyoto Encyclopedia of Genes and Genomes. Based on the bioinformatic analysis described above, we found all of the known enzymes involved in ginsenoside backbone synthesis, starting from acetyl-CoA via the isoprenoid pathway. Additionally, a total of 150 cytochrome P450 (CYP450) and 235 glycosyltransferase unique sequences were found in the 454 cDNA library, some of which encode enzymes responsible for the conversion of the ginsenoside backbone into the various ginsenosides. Finally, one CYP450 and four UDP-glycosyltransferases were selected as the candidates most likely to be involved in ginsenoside biosynthesis through a methyl jasmonate (MeJA) inducibility experiment and tissue-specific expression pattern analysis based on a real-time PCR assay. We demonstrated, with the assistance of the MeJA inducibility experiment and tissue-specific expression pattern analysis, that transcriptome analysis based on 454 pyrosequencing is a powerful tool for determining the genes encoding enzymes responsible for the biosynthesis of secondary metabolites in non-model plants. Additionally, the expressed sequence tags (ESTs) and unique sequences from this study provide an important resource for the scientific community that is interested in the molecular genetics and functional genomics of American ginseng.


Parens Patriae, Punishment, and Pandemics: The State's Responsibility for Incarcerated Persons During a Public Health Emergency.
This article looks at the nation's response to the COVID-19 pandemic since March 2020 and explores the commonalities and differences of states' actions to protect their citizens, especially the most vulnerable populations. The article discusses the government's obligations to jailees and prisoners during the COVID-19 pandemic and how incarcerated persons have been consistently failed by the institutions that are required to protect them. The article examines possible remedies for these governmental and institutional failings under the Eighth Amendment and §1983 civil rights claims. Ultimately the article proposes that monetary damages would provide relief to incarcerated individuals and their families where other remedies have often failed. Additionally, monetary damages will send the message that those under the protection of the state need tangible justice and that the government needs rigorous accountability during a public health emergency.


The Deubiquitinase USP47 Stabilizes MAPK by Counteracting the Function of the N-end Rule ligase POE/UBR4 in Drosophila.
RAS-induced MAPK signaling is a central driver of the cell proliferation apparatus. Disruption of this pathway is widely observed in cancer and other pathologies. Consequently, considerable effort has been devoted to understanding the mechanistic aspects of RAS-MAPK signal transmission and regulation. While much information has been garnered on the steps leading up to the activation and inactivation of core pathway components, comparatively little is known on the mechanisms controlling their expression and turnover. We recently identified several factors that dictate Drosophila MAPK levels. Here, we describe the function of one of these, the deubiquitinase (DUB) USP47. We found that USP47 acts post-translationally to counteract a proteasome-mediated event that reduces MAPK half-life and thereby dampens signaling output. Using an RNAi-based genetic interaction screening strategy, we identified UBC6, POE/UBR4, and UFD4, respectively, as E2 and E3 enzymes that oppose USP47 activity. Further characterization of POE-associated factors uncovered KCMF1 as another key component modulating MAPK levels. Together, these results identify a novel protein degradation module that governs MAPK levels. Given the role of UBR4 as an N-recognin ubiquitin ligase, our findings suggest that RAS-MAPK signaling in Drosophila is controlled by the N-end rule pathway and that USP47 counteracts its activity.


Reduction of death receptor 5 expression and apoptosis of CD4+ T cells from HIV controllers.
TNF-related apoptosis ligand (TRAIL) induces apoptosis of HIV-1-exposed CD4 T cells expressing the death receptor 5 (DR5) in vitro and has been associated with reduced CD4 T cell number in viremic HIV-1-infected patients. Alterations of the TRAIL/DR5 apoptotic pathway could be involved in the absence of massive CD4 T cell depletion in HIV-1-infected controllers (HIC). We studied here apoptosis of CD4 T cells from HIV-infected progressors and controllers. Reduced apoptosis of CD4 T cells from HIC was observed upon HIV stimulation. This lower apoptosis correlated with a deficiency of DR5 cell surface expression by CD4 T cells upon HIV-1 stimulation. The significant lower apoptosis observed in CD4 T cells after HIV exposure, associated with lower expression of membrane DR5 could explain the better survival of HIV-specific CD4 T cells from HIV controllers. The levels of DR5 cell surface expression on CD4 T cells could represent a new prognostic marker.


Fracture incidence and changes in quality of life in women with an inadequate clinical outcome from osteoporosis therapy: the Observational Study of Severe Osteoporosis (OSSO).
In this observational study of women with an inadequate clinical outcome to osteoporosis therapy, those with a fracture at baseline were more likely to sustain an incident fracture and have a worse health-related quality of life than those without prior fracture. The Observational Study of Severe Osteoporosis (OSSO) was designed to assess the fracture incidence and health-related quality of life (HRQoL) in women with an inadequate clinical outcome to osteoporosis therapy. Post-menopausal women (N=1,885) with established osteoporosis and an inadequate clinical response to osteoporosis drug therapy defined as: a) a fragility fracture despite therapy for one year (index fracture, N=988), or b) discontinued drug therapy due to adverse effects and/or non-compliance (N=897), were assessed during one year for HRQoL using the EQ-5D and the QUALEFFO questionnaires. One hundred and sixty-six (8.8%) women had a total of 209 incident fractures (1,139 fractures/10,000 women-years). Women with an index fracture were more likely to sustain an incident fracture than those without prior fractures (hazard ratio 1.91; 95% CI: 1.37-2.66; p<0.001). Co-morbidities or antidepressant use at baseline also increased the risk of incident fracture. Median total EQ-5D Health State Values and QUALEFFO scores were worse in women with an incident fracture regardless of index fracture status. The worst scores were reported in the EQ-5D sub-domains of self-care, usual activities and pain/discomfort. Women with an inadequate response to osteoporosis therapy had a high rate of incident fracture which had an adverse impact on HRQoL.


Expression of five cathepsins in murine melanomas of varying metastatic potential and normal tissues.
The relative levels of mRNAs for cathepsins B, D, H, L, and S in eight normal murine tissues and three murine melanoma variants, B16-F1, B16-F10, and B16a, have been analyzed by RNA dot blot and densitometry. A direct correlation was observed between the levels of cathepsin B mRNA and the metastatic potentials of these three melanoma variants. The relative amount of cathepsin B mRNA in B16a, which is the melanoma variant with the highest metastatic potential, was at least 3 times greater than that found in any of the normal murine tissues surveyed. Similar results were obtained in analyses of either solid tumors or of cultures of tumor cells, confirming that the tumor cells themselves were the source for the elevated expression of cathepsin B mRNA. Northern blot analysis revealed the presence of three cathepsin B transcripts of 5.0, 4.0, and 2.2 kilobases in the melanoma variants, while only the 2.2-kilobase transcript was seen in the normal murine tissues. Concurrently with the mRNA analysis, enzyme assays for cathepsin B activity were also performed using synthetic peptide substrates. The assays revealed increased cathepsin B activities in the melanoma variants, corresponding well with the increased cathepsin B mRNA levels, and in addition demonstrated that all three of the melanoma variants secreted a latent form of cathepsin B into conditioned medium, which could be activated by limited proteolysis with pepsin. The levels of the latent enzyme released by the murine melanoma variants correlated well with the levels of cathepsin B mRNA and with the metastatic potentials as determined by spontaneous metastasis form a s.c. site.


A national survey of practical airway training in UK anaesthetic departments. Time for a national policy?
The Fourth National Audit Project (NAP4) recommended airway training for trainee and trained anaesthetists. As the skills required for management of airway emergencies differ from routine skills and these events are rare, practical training is likely to require training workshops. In 2013, we surveyed all UK National Health Service hospitals to examine the current practices regarding airway training workshops. We received responses from 206 hospitals (62%) covering all regions. Regarding airway workshops, 16% provide none and 51% only for trainees. Of those providing workshops, more than half are run less than annually. Workshop content varies widely, with several Difficult Airway Society (DAS) guideline techniques not taught or only infrequently. Reported barriers to training include lack of time and departmental or individual interest. Workshop-based airway training is variable in provision, frequency and content, and is often not prioritised by departments or individual trainers. It could be useful if guidance on workshop organisation, frequency and content was considered nationally.


'They say we are money minded' exploring experiences of formal private for-profit health providers towards contribution to pro-poor access in post conflict Northern Uganda.
Background: The perception within literature and populace is that the private for-profit sector is for the rich only, and this characteristic results in behaviours that hinder advancement of Universal health coverage (UHC) goals. The context of Northern Uganda presents an opportunity for understanding how the private sector continues to thrive in settings with high poverty levels and history of conflict.Objective: The study aimed at understanding access mechanisms employed by the formal private for-profit providers (FPFPs) to enable pro-poor access to health services in post conflict Northern Uganda.Methods: Data collection was conducted in Gulu municipality in 2015 using Organisational survey of 45 registered formal private for-profit providers (FPFPs),10 life histories, and 13 key informant interviews. Descriptive statistics were generated for the quantitative findings whereas qualitative findings were analysed thematically.Results: FPFPs pragmatically employed various access mechanisms and these included fee exemptions and provision of free services, fee reductions, use of loan books, breaking down doses and partial payments. Most mechanisms were preceded by managers' subjective identification of the poor, while operationalisation heavily depended on the managers' availability and trust between the provider and the customer. For a few FPFPs, partnerships with Non-governmental organisations (NGOs) and government enabled provision of free, albeit mainly preventive services, including immunisation, consultations, screening for blood pressure and family planning. Challenges such as quality issues, information asymmetry and standardisation of charges arose during implementation of the mechanisms.Conclusion: The identification of the poor by the FPFPs was subjective and unsystematic. FPFPs implemented various innovations to ensure pro-poor access to health services. However, they face a continuous dilemma of balancing the profit maximization and altruism objectives. Implementation of some pro-poor mechanisms raises concerns included those related to quality and standardisation of pricing.


In vivo evaluation of drug dialyzability in a rat model of hemodialysis.
It is important to calculate the drug removal by hemodialysis (HD) for drug dosing regimens in HD patients. However, there are limited and inconsistent information about the dialyzability of drugs by HD. Therefore, the aim of our study is to evaluate drug removal by utilizing a rat model of HD (HD rat) and to extrapolate this result to the drug removal rate in HD patients. HD rats received bilateral nephrectomy and HD for 2 h. The dialysis removal of 6 drugs was evaluated in HD rats. Dialysis efficiency, plasma protein binding rate (PBR) and distribution volume (Vd) of drugs were also measured. Furthermore, we examined the correlation between the dialyzability of drug in HD rats and humans and constructed the prediction formula of the drug dialyzability in HD patients. The clearance of urea and creatinine and normalized dialysis dose in HD rats were 0.83 ± 0.07 mL/min, 0.70 ± 0.08 mL/min, and 0.13 ± 0.06, respectively. The drug dialyzability in HD rats was similar to reported clinical data except for doripenem. A higher correlation was observed between drug dialyzability in reported clinical data and HD rats which were adjusted for PBR (r2 = 0.936; p < 0.001) compared to unadjusted (r2 = 0.812; p = 0.009). Therefore, we constructed the prediction formula of the drug dialyzability in HD patients by utilizing the HD rat model and PBR. This study is useful for evaluating the dialyzability of high-risk drugs in a clinical setting and might provide appropriate preclinical dialyzability data for new drug.


Urinary TIMP-2 and IGFBP7 as early biomarkers of acute kidney injury and renal recovery following cardiac surgery.
Difficulties in prediction and early identification of (acute kidney injury) AKI have hindered the ability to develop preventive and therapeutic measures for this syndrome. We tested the hypothesis that a urine test measuring insulin-like growth factor-binding protein 7 (IGFBP7) and tissue inhibitor of metalloproteinases-2 (TIMP-2), both inducers of G1 cell cycle arrest, a key mechanism implicated in acute kidney injury (AKI), could predict AKI in cardiac surgery patients. We studied 50 patients at high risk for AKI undergoing cardiac surgery with cardiopulmonary bypass (CPB). Serial urine samples were analyzed for [TIMP-2]*[IGFBP7] concentrations. The primary outcome measure was AKI as defined by international consensus criteria following surgery. Furthermore, we investigated whether urine [TIMP-2]*[IGFBP7] could predict renal recovery from AKI prior to hospital discharge. 26 patients (52%) developed AKI. Diagnosis based on serum creatinine and/or oliguria did not occur until 1-3 days after CPB. In contrast, urine concentration of [TIMP-2]*[IGFBP7] rose from a mean of 0.49 (SE 0.24) at baseline to 1.51 (SE 0.57) 4 h after CPB in patients who developed AKI. The maximum urinary [TIMP-2]*[IGFBP7] concentration achieved in the first 24 hours following surgery (composite time point) demonstrated an area under the receiver-operating characteristic curve of 0.84. Sensitivity was 0.92, and specificity was 0.81 for a cutoff value of 0.50. The decline in urinary [TIMP-2]*[IGFBP7] values was the strongest predictor for renal recovery. Urinary [TIMP-2]*[IGFBP7] serves as a sensitive and specific biomarker to predict AKI early after cardiac surgery and to predict renal recovery. www.germanctr.de/, DRKS-ID: DRKS00005062.


High-performance light-emitting diodes based on carbene-metal-amides.
Organic light-emitting diodes (OLEDs) promise highly efficient lighting and display technologies. We introduce a new class of linear donor-bridge-acceptor light-emitting molecules, which enable solution-processed OLEDs with near-100% internal quantum efficiency at high brightness. Key to this performance is their rapid and efficient utilization of triplet states. Using time-resolved spectroscopy, we establish that luminescence via triplets occurs within 350 nanoseconds at ambient temperature, after reverse intersystem crossing to singlets. We find that molecular geometries exist at which the singlet-triplet energy gap (exchange energy) is close to zero, so that rapid interconversion is possible. Calculations indicate that exchange energy is tuned by relative rotation of the donor and acceptor moieties about the bridge. Unlike other systems with low exchange energy, substantial oscillator strength is sustained at the singlet-triplet degeneracy point.


The complete plastome sequence of Durian, Durio zibethinus L. (Malvaceae).
The complete plastome sequence of Durio zibethinus L. (Malvaceae) is determined in this study (NCBI acc. no. MG138151). D. zibethinus is an important fruit crop in Southeastern Asia and known as the 'king of fruit'. Our D. zibethinus plastome is the first reported sequences from the subfamily Helicteroideae of Malvaceae. The plastome sequence of D. zibethinus is 163,974 bp in length and it is composed of a pair of 23,679 bp inverted repeat regions separated by large and small single-copy regions of 95,704 bp and 20,912 bp, respectively. The gene order and structure of the D. zibethinus are similar to those of the typical plastome of land plants. The plastome encodes 113 genes, of which 79 are protein-coding genes, 30 are tRNA genes, and four are rRNA genes. Fifteen genes contain single intron and two genes have two introns. A total of 144 simple sequence repeats (SSR) were identified in the genome. Phylogenetic analysis show that D. zibethinus (Helicteroideae) is sister group of Tilia (Tilioideae) clade with 100% bootstrap support.


Tapered N-helical metamaterials with three-fold rotational symmetry as improved circular polarizers.
Chiral helix-based metamaterials can potentially serve as compact and broadband circular polarizers. We have recently shown that the physics of structures composed of multiple intertwined helices, so called N-helices with N being an integer multiple of 4, is distinct from that of structures made of single circular helices (N = 1). In particular, undesired circular polarization conversion is strictly eliminated for N = 4 helices arranged on a square lattice. However, the fabrication of such structures for infrared/visible operation wavelengths still poses very significant challenges. Thus, we here revisit the possibility of reducing N from 4 to 3, which would ease micro-fabrication considerably. We show analytically that N = 3 helices arranged on a hexagonal lattice exhibit strictly vanishing circular polarization conversion. N = 3 is the smallest option as N = 2 obviously leads to linear birefringence. To additionally improve the circular-polarizer operation bandwidth and the extinction ratio while maintaining high transmission for the wanted polarization and zero conversion, we also investigate by numerical calculations N = 3 helices with tapered diameter along the helix axis. We find operation bandwidths as large as 2.4 octaves.


Role of NADPH oxidase/ROS in pro-inflammatory mediators-induced airway and pulmonary diseases.
Reactive oxygen species (ROS) are products of normal cellular metabolism and are known to act as second messengers. Under physiological conditions, ROS participate in maintenance of cellular 'redox homeostasis' in order to protect cells against oxidative stress. In addition, regulation of redox state is important for cell activation, viability, proliferation, and organ function. However, overproduction of ROS, most frequently due to excessive stimulation of either reduced nicotinamide adenine dinucleotide phosphate (NADPH) by pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) or the mitochondrial electron transport chain and xanthine oxidase, results in oxidative stress. Oxidative stress is a deleterious process that leads to airway and lung damage and consequently to several respiratory inflammatory diseases/injuries, including acute respiratory distress syndrome (ARDS), asthma, cystic fibrosis (CF), and chronic obstructive pulmonary disease (COPD). Many of the known inflammatory target proteins, such as matrix metalloproteinase-9 (MMP-9), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), cyclooxygenase-2 (COX-2), and cytosolic phospholipase A(2) (cPLA(2)), are associated with NADPH oxidase activation and ROS overproduction in response to pro-inflammatory mediators. Thus, oxidative stress regulates both key inflammatory signal transduction pathways and target proteins involved in airway and lung inflammation. In this review, we discuss mechanisms of NADPH oxidase/ROS in the expression of inflammatory target proteins involved in airway and lung diseases. Knowledge of the mechanisms of ROS regulation could lead to the pharmacological manipulation of antioxidants in airway and lung inflammation and injury.


Cell death mechanism and protective effect of erythropoietin after focal ischemia in the whisker-barrel cortex of neonatal rats.
Cell death induced by the combined insult of hypoxia-ischemia in neonatal rodents has been extensively investigated. Ischemia-only-induced cell death, however, has been much less characterized. Based on the notion that 1) ischemic stroke is a relatively common disorder in human neonates, and 2) developing cells are more susceptible to apoptosis, the present study examined whether typical apoptosis was induced by cerebral ischemia in a new neonatal rat model. Erythropoietin (EPO; Epoetin) was tested for its protective effect against ischemia-induced cell death. Postnatal day 7 rats were subjected to permanent occlusion of the middle cerebral artery branch supplying the right whisker-barrel cortex. Terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeled-positive cells in the ischemic region were detectable 4 h after ischemia and reached a peak level 16 h later. The cell death was preceded by caspase activation and cytochrome c release. Cell body shrinkage was evident among damaged cells. Agarose gel electrophoresis showed DNA damage with a smear pattern as well as DNA laddering. Electron microscopy demonstrated apoptotic features such as cell shrinkage, chromatin condensation, and fragmentation; meanwhile, necrotic alterations coexisted in the cytoplasm. EPO treatment increased signal transducers and activators of transcription-5 and Bcl-2 levels, markedly attenuated apoptotic cell death, and reduced ischemic infarct in the cortex. It is suggested that focal ischemia in the developing brain causes cell death with prominent apoptotic features coexisting with some characteristics of necrosis. This is consistent with the concept of hybrid death described previously in cultures and adult or developing brain. EPO may be explored as a potential therapy for neonatal ischemic stroke.


Targeted disruption of hesr2 results in atrioventricular valve anomalies that lead to heart dysfunction.
Genes involved in the Notch signaling pathway have been shown to be critical regulators of cardiovascular development. In vitro studies have revealed that the Notch signaling pathway directly regulates transcription of hairy and enhancer of split-related (hesr) genes, encoding basic helix-loop-helix transcription factors. To assess the functional role of hesr genes in cardiovascular development, we generated mice with a targeted disruption of the hesr2 gene and used echocardiography to analyze heart function of the mutant mice. In the early postnatal period, a majority of hesr2 homozygous mice die as a result of congestive heart failure accompanied by pronounced heart enlargement. Transthoracic echocardiography on 5-day-old homozygous mice revealed tricuspid and mitral valve regurgitation and a dilated left ventricular chamber with markedly diminished fractional shortening of the left ventricle. The hemodynamic anomalies were accompanied by morphological changes, such as dysplastic atrioventricular (AV) valves, a perimembranous ventricular septal defect, and a secundum atrial septal defect. AV valve regurgitations attributable to dysplasia of the AV valves were most likely responsible for the heart dysfunction in hesr2 homozygous mice. These observations indicate that the Notch signaling target hesr2 plays an important role in the formation and function of the AV valves. In addition, hesr2 activity may be important for proper development of cardiomyocytes, thereby assuring normal left ventricular contractility. Because of the unique spectrum of cardiac anomalies expressed by hesr2-null mice, they represent a useful model system for elucidating the genetic basis of heart dysfunction.


Tuberculous spondylitis diagnosed through Xpert MTB/RIF assay in urine: a case report.
Extrapulmonary tuberculosis (EPTB) is associated with high rates of morbidity and mortality. Diagnosis of EPTB is challenging in resource-limited settings due to difficulties in obtaining samples, as well as the paucibacillarity of the specimens. Skeletal tuberculosis accounts for 10-35 % of EPTB cases, with vertebral osteomyelitis (Pott's disease) representing 50 % of the cases. We present two cases of suspected Pott's disease, diagnosed through GeneXpert MTB/RIF assay in urine at a rural Tanzanian hospital. Case I A 49-year old male, HIV-1 positive, on co-formulated tenofovir disoproxil fumarate/lamivudine/efavirenz since 2009 and CD4 counts of 205 cells/μL (13 %). He presented with lower back pain and progressive lower limb weakness for two weeks prior to admission. The physical examination revealed bilateral flaccid paraplegia with reduced reflexes, but otherwise unremarkable findings. A lateral lumbar X-ray showed noticeable reduction of intervertebral space between L4 and L5, and a small calcification in the anterior longitudinal ligament between L4 and L5, being compatible with focal spondylosis deformans but inconclusive with regard to tuberculous spondylitis. An abdominal ultrasound showed normal kidneys, bladder and prostate gland. The urinalysis and complete blood counts (CBC) were normal. M. Tuberculosis was detected through GeneXpert MTB/RIF in centrifuged urine, with no resistance to rifampicin. Case II A 76-year old female, HIV-1 negative, presented with lower back pain and progressive weakness and numbness of the lower limbs for two months prior to admission. The physical examination revealed paraplegia, but otherwise unremarkable findings. The lumbosacral X-ray findings were compatible with spondylosis deformans of the lumbar spine and possible tuberculous spondylitis in L3-L4. The abdominal and renal ultrasound showed normal kidneys and bladder. The urinalysis and CBC were normal. M. Tuberculosis was detected through GeneXpert MTB/RIF in centrifuged urine, with no resistance to rifampicin. We report two cases of suspected tuberculous spondylitis diagnosed through Xpert MTB/RIF in urine samples from a rural Tanzanian hospital. Urine testing using Xpert MTB/RIF reflects disseminated disease and renal involvement, and may offer a feasible additional diagnostic approach for Pott's disease in rural Africa.


Pharmacotherapy for hypertension in Sub-Saharan Africa: a systematic review and network meta-analysis.
The highest burden of hypertension is found in Sub-Saharan Africa (SSA) with a threefold greater mortality from stroke and other associated diseases. Ethnicity is known to influence the response to antihypertensives, especially in black populations living in North America and Europe. We sought to outline the impact of all commonly used pharmacological agents on both blood pressure reduction and cardiovascular morbidity and mortality in SSA. We used similar criteria to previous large meta-analyses of blood pressure agents but restricted results to populations in SSA. Quality of evidence was assessed using a risk of bias tool. Network meta-analysis with random effects was used to compare the effects across interventions and meta-regression to explore participant heterogeneity. Thirty-two studies of 2860 participants were identified. Most were small studies from single, urban centres. Compared with placebo, any pharmacotherapy lowered SBP/DBP by 8.51/8.04 mmHg, and calcium channel blockers (CCBs) were the most efficacious first-line agent with 18.46/11.6 mmHg reduction. Fewer studies assessing combination therapy were available, but there was a trend towards superiority for CCBs plus ACE inhibitors or diuretics compared to other combinations. No studies examined the effect of antihypertensive therapy on morbidity or mortality outcomes. Evidence broadly supports current guidelines and provides a clear rationale for promoting CCBs as first-line agents and early initiation of combination therapy. However, there is a clear requirement for more evidence to provide a nuanced understanding of stroke and other cardiovascular disease prevention amongst diverse populations on the continent. PROSPERO, CRD42019122490. This review was registered in January 2019.


Specificity of the human IgE response to inhaled acid anhydrides.
Patients with work-related respiratory symptoms caused by inhaled acid anhydrides (trimellitic (TMA), phthalic (PA), tetrachlorophthalic (TCPA), and maleic anhydrides) have specific IgE antibody. The antibody is specific for a conjugate of the sensitizing anhydride (the hapten) and human serum albumin (HSA). We have investigated the specificity of the reaction to determine whether the antibody is directed against (1) the anhydride, (2) new antigenic determinants formed by conjugation of albumin with the anhydride, or (3) the complete anhydride-HSA conjugate. For the patients sensitized to TCPA and TMA, RAST inhibition studies demonstrate the anhydride-HSA conjugate to be a more effective inhibitor of RAST than the sodium salt of the anhydride or an anhydride-bovine serum albumin conjugate, whereas for those sensitized to PA, the free hapten is almost as an effective inhibitor as the conjugate. With each sera HSA conjugates of anhydrides to which the patient is not sensitized are weaker inhibitors than the sensitizing anhydride-albumin conjugate. These results provide strong evidence that for the patients sensitized to TCPA and TMA, the antibody combines with the anhydride and the spatially adjacent portion of the HSA molecule, whereas in the patients sensitized to PA, the antibody is specific for the hapten.


Comparative evaluation of benzodiazepines for control of soman-induced seizures.
This study evaluated the ability of six benzodiazepines to stop seizures produced by exposure to the nerve agent soman. Guinea pigs, previously prepared with electrodes to record electroencephalographic (EEG) activity, were pretreated with pyridostigmine (0.026 mg/kg, i.m.) 30 min before challenge with soman (56 microg/kg, s.c.) and then treated 1 min after soman exposure with atropine (2.0 mg/kg, i.m.) and pralidoxime chloride (2-PAM Cl; 25 mg/kg, i.m.). All animals developed seizures following this treatment. Benzodiazepines (avizafone, clonazepam, diazepam, loprazolam, lorazepam, and midazolam) were given i.m. 5 or 40 min after seizure onset. All benzodiazepines were effective in stopping soman-induced seizures, but there were marked differences between drugs in the rapidity of seizure control. The 50% effective dose (ED50) values and latencies for anticonvulsant effect for a given benzodiazepine were the same at the two times of treatment delay. Midazolam was the most potent and rapidly acting compound at both treatment times. Since rapid seizure control minimizes the chance of brain damage, use of midazolam as an anticonvulsant may lead to improved clinical outcome in the treatment of nerve agent seizures.


Liposuction of Postmastectomy Arm Lymphedema Decreases the Incidence of Erysipelas.
The objective of this study was to assess erysipelas incidence before and after liposuction treatment for patients suffering from post-mastectomy lymphedema. A prospective cohort study of 130 patients at Skåne University Hospital in Malmö, Sweden with postmastectomy arm lymphedema, who had poor outcomes from prior conservative treatment and clinical signs of subcutaneous adipose tissue hypertrophy, underwent liposuction between 1993-2012. Pre- and postoperative incident data on erysipelas were available for all of them. Mean duration of lymphedema prior to liposuction was 8.8 years (range1-38, standard deviation (SD) 7.0 years). Mean age at liposuction was 63 years (range 39-89, SD 10 years). Total pre-liposuction observation years were 1147, and total post-liposuction observation years were 983. Erysipelas incidence dropped significantly (p<0.001) from 0.47 attacks/year (range 0-5.0, SD 0.8 attacks/year) to 0.06 attacks/year (range 0-3.0, SD 0.3 attacks/year) after liposuction, a reduction of 87%. Also, compared to 76 patients who experienced at least 1 erysipelas episode preoperatively, only 13 patients experienced erysipelas postoperatively. Of the 54 patients who did not have erysipelas preoperatively, 6 patients had erysipelas postoperatively. The total number of erysipelas attacks observed decreased from 534 to 60 bouts after liposuction. The excess arm volume of 1607 ml (range 570-3950, SD 707) was reduced to -43 ml (range -945 to 1390, SD 379) after 6 months and was maintained during the postoperative follow-up period of, at most, 18 years. Our data suggest that liposuction can significantly reduce incidence of erysipelas in patients with post mastectomy arm lymphedema who prior to the intervention suffered one or more attacks.


Sense of coherence and quality of life in older in-hospital patients without cognitive impairment--a 12 month follow-up study.
The relation between sense of coherence (SOC) and quality of life (QoL) among older persons has been found in some, but not all, studies and mostly in studies with cross-sectional design. We wanted to study if SOC was associated with domains of QoL at hospitalization and one year later among persons 65 years and above without cognitive impairment. At hospitalization (T1) and 12 month follow-up (T2) QoL and cognitive status were assessed using the WHOQOL-BREF and the Mini-Mental State Examination. At baseline, the 13-item version of the SOC scale was used to assess coping, the Hospital Anxiety and Depression Scale (HADS) was used to assess depressive and anxiety symptoms. Level of functioning was rated using Lawton and Brody's scales for physical self-maintenance and instrumental activities of daily living (personal and instrumental ADL). In total, 165 (80 men) persons with a mean age of 77.7 (SD 6.9) years were included. The proportion of people rating their overall QoL as high had decreased from T1 to T2. The mean score on QoL- physical domain had increased, while the mean score of QoL-environmental domain had decreased. In adjusted regression analyses at T1, a high level of SOC was positively associated with QoL in three of four domains, i.e. physical, psychological and environmental, but level of SOC assessed at T1 was not associated with any domain of QoL at T2. Personal ADL was associated with some domains of QOL at T1 and T2. The SOC level was associated with older adult's QoL during hospitalization but not their QoL one year after the hospital stay.


THE FINE STRUCTURE OF THE PIGMENT EPITHELIUM IN THE ALBINO RAT.
In this report, particular attention is paid to the inclusion bodies found in the apical cytoplasm of the pigment epithelial cell. These bodies are of variable size and form. The smallest (0.4 micro diameter) consist of a granular matrix enclosed by a single membrane, and are similar to the lysosomes of hepatic cells. Larger inclusion bodies contain areas of lamellated material in addition to granular matrix. The largest particles seen (2 micro diameter) are almost entirely lamellar. These different forms seem closely related, for it is possible to find all transitional stages between the smallest and largest particles. The relationship between the lamellar inclusion bodies and the rod outer segments is discussed.


Connected Audiological Rehabilitation: 21st Century Innovations.
Tele-audiology provides a means to offer audiologic rehabilitation (AR) in a cost-, resource-, and time-effective manner. If designed appropriately, it also has the capability of personalizing rehabilitation to the user in terms of content, depth of detail, etc., thus permitting selection of the best content for a particular individual. Synchronous/real-time data collection, store and forward telehealth, remote monitoring and mobile health using smartphone applications have each been applied to components of audiologic rehabilitation intervention (sensory management, instruction in the use of technology and control of the listening environment, perceptual and communication strategies training, and counseling). In this article, the current state of tele-audiological rehabilitation interventions are described and discussed. The provision of AR via tele-audiology potentially provides a cost-effective mechanism for addressing barriers to the routine provision of AR beyond provisions of hearing technology. Furthermore, if designed appropriately, it has the capability of personalizing rehabilitation to the user in terms of content, depth of detail, etc., thus permitting selection of the best content for a particular individual. However, effective widespread implementation of tele-audiology will be dependent on good education of patients and clinician alike, and researchers must continue to examine the effectiveness of these new approaches to AR in order to ensure clinicians provide effective evidence-based rehabilitation to their patients. While several barriers to the widespread use of tele-audiology for audiologic rehabilitation currently exist, it is concluded that through education of patients and clinicians alike, it will gain greater support from practitioners and patients over time and will become successfully and widely implemented.


Effect of human chorionic gonadotropin on uterine blood flow in intact and ovariectomized gilts.
This experiment was designed to test the hypothesis that human chorionic gonadotropin (hCG) can directly influence uterine blood flow in pigs. In the study, 18 crossbred gilts (95-100 kg) which had exhibited at least one estrous cycle were used: 8 cyclic gilts in the luteal phase of the estrous cycle and 10 gilts ovariectomized (OVX) three weeks before the experiment began. The OVX animals received i.m. injections of 1 mg estradiol benzoate and 50 mg progesterone daily for five consecutive days before transducer implantation. Precalibrated electromagnetic blood flow transducers were surgically implanted around the uterine artery under general anaesthesia. Beginning at 120 min (0 h) following the placement of probes, i.e. after recording had stabilized, blood flow values were recorded at 10 min intervals for the next 360 min. At 0 h, cyclic (n = 4) and OVX (n = 5) gilts received a bolus injection (5 ml) of 1000 IU hCG intravenously through an indwelling catheter and then the same dose of hCG continuously during 4 hours in 40 ml of saline. The remaining cyclic (n = 4) and OVX (n = 5) gilts were infused with saline only and served as control. The mean (SEM) uterine blood flow at 0 h in cyclic (n = 8) and OVX (n = 10) gilts was 104.0 +/- 25.7 and 58.6 +/- 6.0 ml/min, respectively. Because of the variability of initial blood flow, especially in the cyclic gilts, hCG induced changes were expressed as a percent change from the initial flow and were compared to pretreatment period 0 (-30 - 0 h). hCG produced a significant increase in the uterine blood flow in both treated groups (p < 0.05) reaching at 70 min 110.2 +/- 3.4 vs 80.0 +/- 10.5 and 122.4 +/- 12.1 vs 90.6 +/- 3.8, and then at 230 min 116.7 +/- 9.3 vs 62.0 +/- 10.0 and 121.8 +/- 13.2 vs 85.4 +/- 2.3 percent of initial blood flow in cyclic and OVX gilts, respectively. A comparison of nine 30-min periods (periods 1-9) to period 0 showed a significant decrease of uterine blood flow in both (cyclic and ovariectomized) saline control groups in periods 2-9 (50-360 min). The comparison of the same periods in cyclic and ovariectomized hCG treated gilts revealed a significant increase during periods 7-9 (250-360 min) and in period 6, 9 (210-240 and 330-360 min), respectively. Serum estradiol levels increased significantly from 7.4 +/- 1.5 to 351.9 +/- 50.2 pg/ml in cyclic, and from 43.3 0 +/- 7.0 to 224.8 +/- 24.1 in OVX hCG treated gilts at 180 and 120 min, respectively, and returned to the levels of the control groups 240-300 min after the beginning of hCG infusion. Those results indicate that the early response in uterine blood flow is produced directly by hCG, which overcomes the inhibitory pharmacological effect of pentobarbital on uterine blood flow. The second phase of response to hCG could be caused by estradiol which significantly increased an hour earlier before uterine blood flow elevation. We have also shown a novel source of estradiol in castrated pigs which is released by hCG.


Does Targeted Temperature Management Improve Neurological Outcome in Extracorporeal Cardiopulmonary Resuscitation (ECPR)?
Targeted temperature management (TTM) is a standard of care in patients after cardiac arrest for neuroprotection. Currently, the effectiveness and efficacy of TTM after extracorporeal cardiopulmonary resuscitation (ECPR) is unknown. We aimed to compare neurological and survival outcomes between TTM vs non-TTM in patients undergoing ECPR for refractory cardiac arrest. We searched PubMed and 5 other databases for randomized controlled trials and observational studies reporting neurological outcomes or survival in adult patients undergoing ECPR with or without TTM. Good neurological outcome was defined as cerebral performance category <3. Two independent reviewers extracted the data. Random-effects meta-analyses were used to pool data. We included 35 studies (n = 2,643) with the median age of 56 years (interquartile range [IQR]: 52-59). The median time from collapse to ECMO cannulation was 58 minutes (IQR: 49-82) and the median ECMO duration was 3 days (IQR: 2.0-4.1). Of 2,643, 1,329 (50.3%) patients received TTM and 1,314 (49.7%) did not. There was no difference in the frequency of good neurological outcome at any time between TTM (29%, 95% confidence interval [CI]: 23%-36%) vs. without TTM (19%, 95% CI: 9%-31%) in patients with ECPR (P = 0.09). Similarly, there was no difference in overall survival between patients with TTM (30%, 95% CI: 22%-39%) vs. without TTM (24%, 95% CI: 14%-34%) (P = 0.31). A cumulative meta-analysis by publication year showed improved neurological and survival outcomes over time. Among ECPR patients, survival and neurological outcome were not different between those with TTM vs. without TTM. Our study suggests that neurological and survival outcome are improving over time as ECPR therapy is more widely used. Our results were limited by the heterogeneity of included studies and further research with granular temperature data is necessary to assess the benefit and risk of TTM in ECPR population.


Fluorescence amplified fragment length polymorphism compared to pulsed field gel electrophoresis for Listeria monocytogenes subtyping.
Listeriosis is a severe infection which mainly affects pregnant women, neonates and immuno-compromised adults. ANSES's Laboratory for Food safety has been the European Union Reference Laboratory (EURL) for L. monocytogenes in the food chain since 2006. Pulsed Field Gel Electrophoresis (PFGE) is routinely used in the EURL for the surveillance of L. monocytogenes isolated from foods, animals and the environment. One of the main EURL activities is to evaluate alternative molecular subtyping methods to PFGE, and integrate their use within the National Reference Laboratories (NRL) network. Since 2008, the United Kingdom (UK)-NRL for L. monocytogenes at the Health Protection Agency (HPA), London, has used fluorescent Amplified Fragment Length Polymorphism (fAFLP) for the routine surveillance of L. monocytogenes isolated from human clinical cases, food and food processing environments in the UK. This study compares fAFLP with PFGE for subtyping L. monocytogenes. A panel of 109 L. monocytogenes isolates from either human cases of listeriosis, foods, food processing environments and animals were used for the comparative evaluation. Among these, 2 strains were tested from duplicate culture by both methods. The panel also included field isolates, isolates associated with outbreaks or sporadic cases and reference strains. The two strains tested in duplicate displayed the same fAFLP and PFGE types. Strains known to be epidemiologically associated with one another were found to have unique PFGE and fAFLP types. FAFLP and PFGE divided the strains into 76 and 82 distinct profiles, or types, respectively. The discriminatory index calculated was 0.993 and 0.996 for fAFLP and PFGE, respectively. The discriminatory ability of fAFLP was similar to that of PFGE for the subtyping of L. monocytogenes isolates. As a less labour intensive technique fAFLP may be a better method to use than PFGE in investigating outbreaks of human listeriosis and tracking the source of contamination in food processing facilities in real time.


The OpenPicoAmp: an open-source planar lipid bilayer amplifier for hands-on learning of neuroscience.
Understanding the electrical biophysical properties of the cell membrane can be difficult for neuroscience students as it relies solely on lectures of theoretical models without practical hands on experiments. To address this issue, we developed an open-source lipid bilayer amplifier, the OpenPicoAmp, which is appropriate for use in introductory courses in biophysics or neurosciences at the undergraduate level, dealing with the electrical properties of the cell membrane. The amplifier is designed using the common lithographic printed circuit board fabrication process and off-the-shelf electronic components. In addition, we propose a specific design for experimental chambers allowing the insertion of a commercially available polytetrafluoroethylene film. We provide a complete documentation allowing to build the amplifier and the experimental chamber. The students hand-out giving step-by step instructions to perform a recording is also included. Our experimental setup can be used in basic experiments in which students monitor the bilayer formation by capacitance measurement and record unitary currents produced by ionic channels like gramicidin A dimers. Used in combination with a low-cost data acquisition board this system provides a complete solution for hands-on lessons, therefore improving the effectiveness in teaching basic neurosciences or biophysics.


Climate sensitive size-dependent survival in tropical trees.
Survival rates of large trees determine forest biomass dynamics. Survival rates of small trees have been linked to mechanisms that maintain biodiversity across tropical forests. How species survival rates change with size offers insight into the links between biodiversity and ecosystem function across tropical forests. We tested patterns of size-dependent tree survival across the tropics using data from 1,781 species and over 2 million individuals to assess whether tropical forests can be characterized by size-dependent life-history survival strategies. We found that species were classifiable into four 'survival modes' that explain life-history variation that shapes carbon cycling and the relative abundance within forests. Frequently collected functional traits, such as wood density, leaf mass per area and seed mass, were not generally predictive of the survival modes of species. Mean annual temperature and cumulative water deficit predicted the proportion of biomass of survival modes, indicating important links between evolutionary strategies, climate and carbon cycling. The application of survival modes in demographic simulations predicted biomass change across forest sites. Our results reveal globally identifiable size-dependent survival strategies that differ across diverse systems in a consistent way. The abundance of survival modes and interaction with climate ultimately determine forest structure, carbon storage in biomass and future forest trajectories.


Adaptive responses to low-dose/low-dose-rate gamma rays in normal human fibroblasts: the role of growth architecture and oxidative metabolism.
To investigate low-dose/low-dose-rate effects of low-linear energy transfer (LET) ionizing radiation, we used gamma-irradiated cells adapted to grow in a three-dimensional architecture that mimics cell growth in vivo. We determined the cellular, molecular and biochemical changes in these cells. Quiescent normal human fibroblasts were irradiated with single acute or chronic doses (1-10 cGy) of (137)Cs gamma rays. Whereas exposure to an acute dose of 10 cGy increased micronucleus formation, protraction of the dose over 48 h reduced micronucleus frequency to a level similar to or lower than what occurs spontaneously. The protracted treatment also up-regulated the cellular content of the antioxidant glutathione. These changes correlated with modulation of phospho-TP53 (serine 15), a stress marker that was regulated by doses as low as 1 cGy. The DNA damage that occurred after exposure to an acute dose of 10 cGy was protected against in two ways: (1) up-regulation of cellular antioxidant enzyme activity by ectopic overexpression of MnSOD, catalase or glutathione peroxidase, and (2) inhibition of superoxide anion generation by flavin-containing oxidases. These results support a significant role for oxidative metabolism in mediating low-dose radiation effects and demonstrate that cell culture in three dimensions is ideal to investigate radiation-induced adaptive responses. Expression of connexin 43, a constitutive protein of gap junctions, and the G(1) checkpoint were more sensitive to regulation by gamma rays in cells maintained in a three-dimensional than in a two-dimensional configuration.


Curcumin protects against cerebral ischemia-reperfusion injury by activating JAK2/STAT3 signaling pathway in rats.
Curcumin (Cur) is the active component in Curcuma longa, and it has been reported to exhibit a variety of biological effects such as anti-inflammation and anti-oxidation. This study aims to investigate the effect of Cur on cerebral ischemia-reperfusion injury and whether the Janus kinase 2 and signal transducer and activator 3 of transcription (JAK2/STAT3) signaling pathway is involved in the neuroprotective effects of Cur. Rats were subjected to 2 h transient middle cerebral artery occlusion (MCAO), followed by 24 h reperfusion. Rats were randomly assigned into sham group, MCAO group, MCAO + Cur group. AG490, a JAK2 inhibitor, was utilized to throw light upon the underlying mechanism. The results suggested that compared to MCAO, Cur attenuated neurological deficits, reduced cerebral infarction area and lowered brain water content. In addition, Cur reduced the activity of IL-1β and IL-8. The results of western blot indicated that Cur enhanced the expression of p-JAK2 and p-STAT3, which was abolished by AG490 administration. Our results suggested that Cur protects effects against cerebral I/R injury through the activation of JAK2/STAT3 signaling pathway.


The effect of noise exposure on the vestibular systems of dental technicians.
Noise exposure is the primary cause of acquired hearing loss in several occupational settings, including dental laboratories and clinics. However, the impact of noise exposure on the vestibular system is not as well researched. To investigate the nature of vestibular damage caused by working in dental laboratories and clinics with high levels of noise exposure due to loud dental equipment. A descriptive, case study design was used to evaluate the vestibular function of dental technicians. Out of 30 dental technicians, 5 males who had been working for several years in dental settings were selected based on their reports of severe symptoms of imbalance. Audiologic evaluations were conducted in the vestibular unit of the Doctor Tarek Khrais Center in Amman, Jordan, for one year. Each subject underwent several hearing tests, which included otoscopic examination, pure tone audiometry (PTA), impedance measurements, and speech testing. Assessment of vestibular function was then conducted using a diagnostic test battery which included electrocochleography, ocular vestibular evoked myogenic potentials (oVEMP), cervical vestibular evoked myogenic potentials (cVEMP), positional testing using the Thomas Richard-Vitton (TRV) chair, and standing stability testing. All test subjects experienced some form of vestibular impairment, including benign paroxysmal positional vertigo (BPPV), endolymphatic hydrops (Meniere disease), or a combination of both. Three out of five cases displayed little or no hearing loss, indicating that vestibular function is more at risk than hearing acuity to continuous noise exposure in dental settings. Exposure to loud noise in dental laboratories severely impacts the functioning of the vestibular system of the inner ear more than the cochlea. The main clinical implication of this study is that regular vestibular assessments are a necessity for dental technicians.


First Report of CD4 Lymphopenia and Defective Neutrophil Functions in a Patient with Amebiasis Associated with CMV Reactivation and Severe Bacterial and Fungal Infections.
We report the case of a patient with acute necrotizing colitis due to invasive amebiasis associated with CD4 lymphopenia and impaired neutrophil responses. The course of the disease was characterized by CMV reactivation and severe and recurrent bacterial and fungal infections, which might be related to the decreased CD4 T cell count and the impaired functional capacities of neutrophils, respectively. The clinical outcome was positive with normalization of both CD4 cell count and neutrophil functions.


Saccadic momentum and attentive control in V4 neurons during visual search.
Saccadic momentum refers to the increased probability of making a saccade in a forward direction relative to the previous saccade. During visual search and free viewing conditions saccadic probability falls in a gradient from forward to backward directions. It has been considered to reflect an oculomotor bias for a continuing motor plan. Here we report that a saccadic momentum gradient is observed in nonhuman primate behavior and in the visual responses of cortical area V4 neurons during a conjunction style visual search task. This result suggests that saccadic momentum arises in part from a biased spatial distribution of visual responses to stimuli. The effect is independent of feature-based selective attention and overridden by directed spatial attention. The implications of saccadic momentum for search guidance are much broader and robust than the inhibition-of-return's presumed role in preventing refixation of recent locations.