Steady-state modulation of voltage-gated K+ channels in rat arterial smooth muscle by cyclic AMP-dependent protein kinase and protein phosphatase 2B

Voltage-gated potassium channels (Kv) are important regulators of membrane potential in vascular smooth muscle cells, which is integral to controlling intracellular Ca2+ concentration and regulating vascular tone. Previous work indicates that Kv channels can be modulated by receptor-driven alterations of cyclic AMP-dependent protein kinase (PKA) activity. Here, we demonstrate that Kv channel activity is maintained by tonic activity of PKA. Whole-cell recording was used to assess the effect of manipulating PKA signalling on Kv and ATP-dependent K+ channels of rat mesenteric artery smooth muscle cells. Application of PKA inhibitors, KT5720 or H89, caused a significant inhibition of Kv currents. Tonic PKA-mediated activation of Kv appears maximal as application of isoprenaline (a β-adrenoceptor agonist) or dibutyryl-cAMP failed to enhance Kv currents. We also show that this modulation of Kv by PKA can be reversed by protein phosphatase 2B/calcineurin (PP2B). PKA-dependent inhibition of Kv by KT5720 can be abrogated by pre-treatment with the PP2B inhibitor cyclosporin A, or inclusion of a PP2B auto-inhibitory peptide in the pipette solution. Finally, we demonstrate that tonic PKA-mediated modulation of Kv requires intact caveolae. Pre-treatment of the cells with methyl-β-cyclodextrin to deplete cellular cholesterol, or adding caveolin-scaffolding domain peptide to the pipette solution to disrupt caveolae-dependent signalling each attenuated PKA-mediated modulation of the Kv current. These findings highlight a novel, caveolae-dependent, tonic modulatory role of PKA on Kv channels providing new insight into mechanisms and the potential for pharmacological manipulation of vascular tone

Anti-müllerian hormone is not associated with cardiometabolic risk factors in adolescent females

<p>Objectives: Epidemiological evidence for associations of Anti-Müllerian hormone (AMH) with cardiometabolic risk factors is lacking. Existing evidence comes from small studies in select adult populations, and findings are conflicting. We aimed to assess whether AMH is associated with cardiometabolic risk factors in a general population of adolescent females.</p> <p>Methods: AMH, fasting insulin, glucose, HDLc, LDLc, triglycerides and C-reactive protein (CRP) were measured at a mean age 15.5 years in 1,308 female participants in the Avon Longitudinal Study of Parents and Children (ALSPAC). Multivariable linear regression was used to examine associations of AMH with these cardiometabolic outcomes.</p> <p>Results: AMH values ranged from 0.16–35.84 ng/ml and median AMH was 3.57 ng/ml (IQR: 2.41, 5.49). For females classified as post-pubertal (n = 848) at the time of assessment median (IQR) AMH was 3.81 ng/ml (2.55, 5.82) compared with 3.25 ng/ml (2.23, 5.05) in those classed as early pubertal (n = 460, P≤0.001). After adjusting for birth weight, gestational age, pubertal stage, age, ethnicity, socioeconomic position, adiposity and use of hormonal contraceptives, there were no associations with any of the cardiometabolic outcomes. For example fasting insulin changed by 0% per doubling of AMH (95%CI: −3%,+2%) p = 0.70, with identical results if HOMA-IR was used. Results were similar after additional adjustment for smoking, physical activity and age at menarche, after exclusion of 3% of females with the highest AMH values, after excluding those that had not started menarche and after excluding those using hormonal contraceptives.</p> <p>Conclusion: Our results suggest that in healthy adolescent females, AMH is not associated with cardiometabolic risk factors.</p&gt

"Open Innovation" and "Triple Helix" Models of Innovation: Can Synergy in Innovation Systems Be Measured?

The model of "Open Innovations" (OI) can be compared with the "Triple Helix of University-Industry-Government Relations" (TH) as attempts to find surplus value in bringing industrial innovation closer to public R&D. Whereas the firm is central in the model of OI, the TH adds multi-centeredness: in addition to firms, universities and (e.g., regional) governments can take leading roles in innovation eco-systems. In addition to the (transversal) technology transfer at each moment of time, one can focus on the dynamics in the feedback loops. Under specifiable conditions, feedback loops can be turned into feedforward ones that drive innovation eco-systems towards self-organization and the auto-catalytic generation of new options. The generation of options can be more important than historical realizations ("best practices") for the longer-term viability of knowledge-based innovation systems. A system without sufficient options, for example, is locked-in. The generation of redundancy -- the Triple Helix indicator -- can be used as a measure of unrealized but technologically feasible options given a historical configuration. Different coordination mechanisms (markets, policies, knowledge) provide different perspectives on the same information and thus generate redundancy. Increased redundancy not only stimulates innovation in an eco-system by reducing the prevailing uncertainty; it also enhances the synergy in and innovativeness of an innovation system.Comment: Journal of Open Innovations: Technology, Market and Complexity, 2(1) (2016) 1-12; doi:10.1186/s40852-016-0039-

Donor site morbidity in oral mucosa graft urethroplasty: implications of tobacco consumption

<p>Abstract</p> <p>Background</p> <p>The purpose of this prospective study was to evaluate the donor site morbidity in patients who have undergone oral mucosa graft urethroplasty for stricture of the urethra. The impact of smoking and oral consumption of tobacco and/or <it>paan masala </it>on the donor site was also assessed. This study is probably the first of its kind where the affect of smoking, <it>paan masala </it>and tobacco chewing on the donor site morbidity has been documented.</p> <p>Methods</p> <p>Forty-eight patients suffering from stricture of the urethra underwent oral mucosa graft urethroplasty between July 2005 and December 2007. The patients were divided into two groups (users or non-users) based on tobacco consumption and oral hygiene. The donor site was evaluated at frequent intervals for pain, swelling, numbness, bleeding, salivation and tightness of mouth.</p> <p>Results</p> <p>Donor site morbidity was more in users with poor oral hygiene. Pain scores were higher amongst the users and the morbidity persisted longer in the users compared to non-users with good oral hygiene.</p> <p>Conclusion</p> <p>Patients who consume tobacco and have poor oral hygiene should be warned regarding poorer outcomes after oral mucosa graft urethroplasty.</p

Huánuco : ¿cómo vamos en educación?

Reúne información estadística relevante de la situación actual de la educación de la región Huánuco, ha sido hecho para servir como una guía informativa amigable de todos aquellos actores de la región que puedan influir positivamente en su realidad educativa. Entre 2004 y 2013, la tasa media de crecimiento anual del gasto público educativo por alumno fue de 23.2% en inicial, 16.3% en primaria y 12.3% en secundaria. En los últimos tres años, el gasto por alumno en inicial ha crecido hasta llegar a ser mayor al gasto por alumno nacional, con 2,428 soles en el 2013, aunque el gasto por alumno en primaria y secundaria en Huánuco siguen siendo menores al gasto por alumno nacional. El porcentaje de escuelas con acceso a Internet de Huánuco en primaria pasó de 0.6% en 2005 a 12.4% en 2014. En secundaria el crecimiento fue mayor, pasando de 3.2% en 2005 a 29.6% en 2014. Sin embargo, estos valores siguen siendo menores a los valores nacionales de 27.9% en primaria y 52.0% en secundaria. El indicador en primaria muestra que en la mayoría de distritos el valor es menor a 20%. En secundaria hay mayor heterogeneidad, y son pocos los distritos con porcentajes altos de instituciones con acceso a internet. Entre 2004 y 2013, las tasas de asistencia de Huánuco para los tres niveles de educación básica regular han aumentado. En primaria, la tasa de asistencia pasó de 91.7% en 2004 a 96.3% en 2013, siendo este último valor mayor al nacional. El acceso a educación inicial aumentó en el período, pasando de 30.0% en 2004 a 71.7% en 2013. En el nivel secundaria, la tasa de asistencia pasó de 60.3% en 2004 a 72.2% en 2013. Huánuco presenta mayores tasas de desaprobación, atraso y retiro con respecto a todo el país, tanto en primaria como en secundaria. El porcentaje de desaprobados en primaria en las provincias de Huánuco fluctúa desde 4.8% en Lauricocha hasta 14.2% en Marañón, en tanto que en secundaria oscila entre 7.5% en Lauricocha y 14.9% en Huacaybamba. En ambos casos, la mayoría de distritos tiene menos del 10% de estudiantes con atraso escolar, aunque en secundaria la cantidad de distritos con más del 20% de atraso es mayor. En cuanto al porcentaje de retirados en 2013, la mayoría de distritos en primaria y secundaria se ubican debajo de 8.0%. Sin embargo, en secundaria hay una cantidad significativa de distritos con más de 8% de retiro

The Environment of the Malnourished Child

capítulo de libro -- Universidad de Costa Rica, 1976The study of the relation of man to his environment in developing countries emphasizes the inevitable need for societies to recognize the true causes of infection, malnutrition, and poverty. The need is for improvement in the quality of human life in less developed nations, a recommendation easy to prescribe but difficult to accomplish. Although our pool of knowledge is incomplete, it is adequate to suggest ways to diminish infection, increase food production, utilize food more efficiently, improve education, and provide systems of justice to protect the classes most in need. The physical environment in tropical and subtropical regions, and the socioeconomic characteristics of the population inhabiting such regions, favor maintenance and transmission of a variety of viruses, bacteria, and parasites that make agricultural progress and social development difficult, and that contribute to poor fetal growth, nutrient wastage, and deficient postnatal physical growth. accounting for most of the childhood morbidity and mortality. In this regard. infections contribute indirectly to the overall food problem in a similar fashion as pests do in terms of food losses and spoilage. The overall effect could be comparable or greater than that resulting from an inadequate capacity to produce or to purchase the food needed. Thus, my objective has been to stress, within the whole environment, the importance of infection and the need to diminish it. Waysto control and prevent infection are readily known. They have to do with education of the population to improve personal and environmental hygiene. Economic investment is necessary to improve housing and water supply sYstems, waste disposal, and such preventive measures as immunization programs. Although such measures may appear expensive when first implemented, they have long-lasting effects and many require minimal expenditure once they are established. Large segments of the population stand to benefit, and other development interventions can then be introduced. However, these measures should not be implemented singly. They should be accompanied by community development, family planning, social legislation-in other words, the holistic approach to health and welfare. To do otherwise may aggravate the problem by stimulating demographic growth, perpetuating malnutrition and infection, and maintaining underdevelopmentUniversidad de Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA

Prediction of MHC class II binding affinity using SMM-align, a novel stabilization matrix alignment method

<p>Abstract</p> <p>Background</p> <p>Antigen presenting cells (APCs) sample the extra cellular space and present peptides from here to T helper cells, which can be activated if the peptides are of foreign origin. The peptides are presented on the surface of the cells in complex with major histocompatibility class II (MHC II) molecules. Identification of peptides that bind MHC II molecules is thus a key step in rational vaccine design and developing methods for accurate prediction of the peptide:MHC interactions play a central role in epitope discovery. The MHC class II binding groove is open at both ends making the correct alignment of a peptide in the binding groove a crucial part of identifying the core of an MHC class II binding motif. Here, we present a novel stabilization matrix alignment method, SMM-align, that allows for direct prediction of peptide:MHC binding affinities. The predictive performance of the method is validated on a large MHC class II benchmark data set covering 14 HLA-DR (human MHC) and three mouse H2-IA alleles.</p> <p>Results</p> <p>The predictive performance of the SMM-align method was demonstrated to be superior to that of the Gibbs sampler, TEPITOPE, SVRMHC, and MHCpred methods. Cross validation between peptide data set obtained from different sources demonstrated that direct incorporation of peptide length potentially results in over-fitting of the binding prediction method. Focusing on amino terminal peptide flanking residues (PFR), we demonstrate a consistent gain in predictive performance by favoring binding registers with a minimum PFR length of two amino acids. Visualizing the binding motif as obtained by the SMM-align and TEPITOPE methods highlights a series of fundamental discrepancies between the two predicted motifs. For the DRB1*1302 allele for instance, the TEPITOPE method favors basic amino acids at most anchor positions, whereas the SMM-align method identifies a preference for hydrophobic or neutral amino acids at the anchors.</p> <p>Conclusion</p> <p>The SMM-align method was shown to outperform other state of the art MHC class II prediction methods. The method predicts quantitative peptide:MHC binding affinity values, making it ideally suited for rational epitope discovery. The method has been trained and evaluated on the, to our knowledge, largest benchmark data set publicly available and covers the nine HLA-DR supertypes suggested as well as three mouse H2-IA allele. Both the peptide benchmark data set, and SMM-align prediction method (<it>NetMHCII</it>) are made publicly available.</p

Antibodies against the Envelope Glycoprotein Promote Infectivity of Immature Dengue Virus Serotype 2

Cross-reactive dengue virus (DENV) antibodies directed against the envelope (E) and precursor membrane (prM) proteins are believed to contribute to the development of severe dengue disease by facilitating antibody-dependent enhancement of infection. We and others recently demonstrated that anti-prM antibodies render essentially non-infectious immature DENV infectious in Fcγ-receptor-expressing cells. Immature DENV particles are abundantly present in standard (st) virus preparations due to inefficient processing of prM to M during virus maturation. Structural analysis has revealed that the E protein is exposed in immature particles and this prompted us to investigate whether antibodies to E render immature particles infectious. To this end, we analyzed the enhancing properties of 27 anti-E antibodies directed against distinct structural domains. Of these, 23 bound to immature particles, and 15 enhanced infectivity of immature DENV in a furin-dependent manner. The significance of these findings was subsequently tested in vivo using the well-established West Nile virus (WNV) mouse model. Remarkably, mice injected with immature WNV opsonized with anti-E mAbs or immune serum produced a lethal infection in a dose-dependent manner, whereas in the absence of antibody immature WNV virions caused no morbidity or mortality. Furthermore, enhancement infection studies with standard (st) DENV preparations opsonized with anti-E mAbs in the presence or absence of furin inhibitor revealed that prM-containing particles present within st virus preparations contribute to antibody-dependent enhancement of infection. Taken together, our results support the notion that antibodies against the structural proteins prM and E both can promote pathogenesis by enhancing infectivity of prM-containing immature and partially mature flavivirus particles