Effects of contractile protein phosphorylation on force development in permeabilized rat cardiac myocytes

The phosphorylation status of myofibrillar proteins influences the Ca2+ responsiveness of the myofilaments,but the contribution of and the interaction between the individual components is poorly characterized. Therefore, in Langendorff perfused rat hearts (n=30), the phosphorylation levels of cardiac myosin binding protein-C (cMyBP-C), troponin I and T (cTnI, cTnT) and myosin light chain 1 and 2 (MLC-1, MLC-2) were determined by 1- and 2-dimensional gel electrophoresis. Isometric force development, its Ca2+-sensitivity, the rate of tension redevelopment (ktr) and passive force (Fpas) were studied at optimal sarcomere length (2.2 μm) in mechanically isolated,permeabilized cardiomyocytes at 15 °C. Protein phosphorylation was varied by: 1) blocking spontaneous cardiac activity by lidocaine (0.35 mM; Quiescence); 2) electrical stimulation of the hearts at 5 Hz (Contraction) and 3. treatment of contracting hearts with Isoprenaline (1 μM). MLC-2 phosphorylation was increased in the Contraction group almost 2-fold, relative to the Quiescence group, whereas cMyBP-C and cTnI phosphorylation remained the same. Isoprenaline resulted in 3.7-fold increases in both cMyBP-C and cTnI phosphorylation, but did not result in a further increase in MLC-2 phosphorylation.No significant differences were found in maximum force and ktr between groups, both before and after protein kinase A (PKA) treatment. Ca2+-sensitivity in the Contraction and Isoprenaline groups was significantly reduced in comparison to the Quiescence group. These differences were largely abolished by PKA and Fpas was reduced. These results highlight the impact of PKA-dependent phosphorylation on Ca2+-sensitivity and provide evidence for an interaction between the effects of TnI and MLC-2 phosphorylation

Analysis and design of solid-state circuits utilizing the NASA analysis computer program Annual report

Network Analysis for Systems Application Program /NASAP/ applicable in analysis and design of solid state circuit

Interrogating technology-led experiments in sustainability governance

Solutions to global sustainability challenges are increasingly technology‐intensive. Yet, technologies are neither developed nor applied to governance problems in a socio‐political vacuum. Despite aspirations to provide novel solutions to current sustainability governance challenges, many technology‐centred projects, pilots and plans remain implicated in longer‐standing global governance trends shaping the possibilities for success in often under‐recognized ways. This article identifies three overlapping contexts within which technology‐led efforts to address sustainability challenges are evolving, highlighting the growing roles of: (1) private actors; (2) experimentalism; and (3) informality. The confluence of these interconnected trends illuminates an important yet often under‐recognized paradox: that the use of technology in multi‐stakeholder initiatives tends to reduce rather than expand the set of actors, enhancing instead of reducing challenges to participation and transparency, and reinforcing rather than transforming existing forms of power relations. Without recognizing and attempting to address these limits, technology‐led multi‐stakeholder initiatives will remain less effective in addressing the complexity and uncertainty surrounding global sustainability governance. We provide pathways for interrogating the ways that novel technologies are being harnessed to address long‐standing global sustainability issues in manners that foreground key ethical, social and political considerations and the contexts in which they are evolving

Computerised cognitive behaviour therapy for depression in adolescents: Study protocol for a feasibility randomised controlled trial

\ua9 2014, BMJ Publishing Group. All rights reserved. Introduction: The 1 year prevalence of depression in adolescents is about 2%. Treatment with antidepressant medication is not recommended for initial treatment in young people due to concerns over high side effects, poor efficacy and addictive potential. Evidence suggests that cognitive behaviour therapy (CBT) is an effective treatment for depression and is currently one of the main treatment options recommended in adolescents. Given the affinity young people have with information technology they may be treated effectively, more widely and earlier in their illness evolution using computeradministered CBT (CCBT). Currently little is known about the clinical and resource implications of implementing CCBT within the National Health Service for adolescents with low mood/depression. We aim to establish the feasibility of running a fully powered randomised controlled trial (RCT). Methods and analysis: Adolescents aged 12-18 with low mood/depression, (scoring ≥20 on the Mood and Feelings Questionnaire (MFQ)), will be approached to participate. Consenting participants will be randomised to either a CCBT programme (Stressbusters) or accessing selected websites providing information about low mood/depression. The primary outcome measure will be the Beck Depression Inventory (BDI). Participants will also complete generic health measures (EQ5D-Y, HUI2) and resource use questionnaires to examine the feasibility of cost-effectiveness analysis. Questionnaires will be completed at baseline, 4 and 12-month follow-ups. Progress and risk will be monitored via the MFQ administered at each treatment session. The acceptability of a CCBT programme to adolescents; and the willingness of clinicians to recruit participants and of participants to be randomised, recruitment rates, attrition rates and questionnaire completion rates will be collected for feasibility analysis. We will estimate \u27numbers needed\u27 to plan a fully powered RCT of clinical and cost-effectiveness. Ethics and dissemination: The current trial protocol received a favourable ethical opinion from Leeds (West) Research and Ethics Committee. (Reference: 10/H1307/137). Trial registration number: ISRCTN31219579

The TRAF1/C5 region is a risk factor for polyarthritis in juvenile idiopathic arthritis

Juvenile idiopathic arthritis (JIA) is a chronic disorder in which both genetic and environmental factors are involved. Recently, we identified the TRAF1/C5 region (located on chromosome 9q33-34) as a risk factor for rheumatoid arthritis (RA) (p(combined) = 1.4 x 10(-8)). In the present study the association of the TRAF1/C5 region with the susceptibility to JIA was investigated. A case-control association study was performed in 338 Caucasian patients with JIA and 511 healthy individuals. We genotyped the single nucleotide polymorphism rs10818488 as a marker for the TRAF1/C5 region. The A allele was associated with the susceptibility to rheumatoid factor-negative polyarthritis with an 11% increase in allele frequency (OR 1.54, 95% CI 1.09 to 2.18; p = 0.012). This association was stronger when combining subtypes with a polyarticular phenotype (OR 1.46, 95% CI 1.12 to 1.90; p = 0.004). In addition, we observed a trend towards an increase in A allele frequency in patients with extended oligoarthritis versus persistent oligoarthritis (49%, 38% respectively); p = 0.055. Apart from being a well replicated risk factor for RA, TRAF1/C5 also appears to be a risk factor for the rheumatoid factor-negative polyarthritis subtype of JIA and, more generally, seems to be associated with subtypes of JIA characterised by a polyarticular cours

Influence of Incubation Conditions on Hydrolysis Efficiency and Iodine Enrichment in Baker’s Yeast

The influence of incubation conditions, enzyme type, hydrolysis time, and potassium iodide concentration on hydrolysis and iodine enrichment were studied in supernatant and pellets of Saccharomyces cervisiae hydrolysates. The type of enzyme used and incubation time significantly influence hydrolysis efficiency and protein concentration in supernatant and pellet. The highest protein hydrolysis efficiency was obtained by 24-h incubation with papain. Significantly lower values were observed for pepsin and autolysis. The potassium iodide concentration influences the iodine content of supernatant and pellet, but not hydrolysis. Iodide enrichment of supernatant and pellet depends on the concentration of iodide using during incubation. High concentration of iodide and long incubation times were the conditions for optimal iodide enrichment and high-protein hydrolysates. The optimal hydrolysis efficiency and iodine enrichment were obtained during 24-h incubation with papain in a 4.5-mM potassium iodide medium. The efficiency reached 98.22% with iodine concentrations of 2,664.91 and 9,200.67 μg/g iodine in pellet and supernatant, respectively

Humiliated fury is not universal: the co-occurrence of anger and shame in the United States and Japan

It has been widely believed that individuals transform high-intensity shame into anger because shame is unbearably painful. This phenomenon was first coined “humiliated fury,” and it has since received empirical support. The current research tests the novel hypothesis that shame-related anger is not universal, yet hinges on the cultural meanings of anger and shame. Two studies compared the occurrence of shamerelated anger in North American cultural contexts (where shame is devalued and anger is valued) to its occurrence in Japanese contexts (where shame is valued and anger is devalued). In a daily-diary study, participants rated anger and shame feelings during shame situations that occurred over one week. In a vignette study, participants rated anger and shame in response to standardised shame vignettes that were generated in previous research by either U.S. or Japanese respondents. Across the two studies, and in line with previous research on humiliated fury, shame predicted anger for U.S. participants. Yet, neither in the daily diary study nor for the Japanese-origin vignettes, did we find shame-related anger in Japanese participants. Only when presented with U.S.-origin vignettes, did Japanese respondents in the vignette study report shame-related anger. The findings suggest that shame-related anger is a culture-specific phenomenon