Junctional sarcoplasmic reticulum motility in adult mouse ventricular myocytes.

Excitation-contraction (EC) coupling is the coordinated process by which an action potential triggers cardiac myocyte contraction. EC coupling is initiated in dyads where the junctional sarcoplasmic reticulum (jSR) is in tight proximity to the sarcolemma of cardiac myocytes. Existing models of EC coupling critically depend on dyad stability to ensure the fidelity and strength of EC coupling, where even small variations in ryanodine receptor channel and voltage-gated calcium channel-α 1.2 subunit separation dramatically alter EC coupling. However, dyadic motility has never been studied. Here, we developed a novel strategy to track specific jSR units in dissociated adult ventricular myocytes using photoactivatable fluorescent proteins. We found that the jSR is not static. Instead, we observed dynamic formation and dissolution of multiple dyadic junctions regulated by the microtubule-associated molecular motors kinesin-1 and dynein. Our data support a model where reproducibility of EC coupling results from the activation of a temporally averaged number of SR Ca2+ release units forming and dissolving SR-sarcolemmal junctions. These findings challenge the long-held view that the jSR is an immobile structure and provide insights into the mechanisms underlying its motility

Coverage with evidence development: applications and issues

Copyright © Cambridge University Press, 2010OBJECTIVES: The aim of this study was to describe the current issues surrounding Coverage with Evidence Development (CED). CED is characterized by restricted coverage for a new technology in parallel with targeted research when the stated goal of the research or data collection is to provide definitive evidence for the clinical or cost-effectiveness impact of the new technology. METHODS: Presented here is information summarized and interpreted from presentations and discussions at the 2008 Health Technology Assessment International (HTAi) meeting and additional information from the medical literature. This study describes the differences between CED and other conditional coverage agreements, provides a brief history of CED, describes real-world examples of CED, describes the areas of consensus between the stakeholders, discusses the areas for future negotiation between stakeholders, and proposes criteria to assist stakeholders in determining when CED could be appropriate. RESULTS: Payers could interpret the evidence obtained from a CED program either positively or negatively, and a range of possible changes to the reimbursement status of the new technology may result. Striking an appropriate balance between the demands for prompt access to new technology and acknowledging that some degree of uncertainty will always exist is a critical challenge to the uptake of this innovative form of conditional coverage. CONCLUSIONS: When used selectively for innovative procedures, pharmaceuticals, or devices in the appropriate disease areas, CED may provide patients access to promising medicines or technologies while data to minimize uncertainty are collected.The development of the manuscript was funded by Medicines Australi

Extensive differential protein phosphorylation as intraerythrocytic Plasmodium falciparumschizonts develop into extracellular invasive merozoites

Pathology of the most lethal form of malaria is caused by Plasmodium falciparum asexual blood stages and initiated by merozoite invasion of erythrocytes. We present a phosphoproteome analysis of extracellular merozoites revealing 1765 unique phosphorylation sites including 785 sites not previously detected in schizonts. All MS data have been deposited in the ProteomeXchange with identifier PXD001684 (http://proteomecentral.proteomexchange.org/dataset/PXD001684). The observed differential phosphorylation between extra and intraerythrocytic life-cycle stages was confirmed using both phospho-site and phospho-motif specific antibodies and is consistent with the core motif [K/R]xx[pS/pT] being highly represented in merozoite phosphoproteins. Comparative bioinformatic analyses highlighted protein sets and pathways with established roles in invasion. Within the merozoite phosphoprotein interaction network a subnetwork of 119 proteins with potential roles in cellular movement and invasion was identified and suggested that it is coregulated by a further small subnetwork of protein kinase A (PKA), two calcium-dependent protein kinases (CDPKs), a phosphatidyl inositol kinase (PI3K), and a GCN2-like elF2-kinase with a predicted role in translational arrest and associated changes in the ubquitinome. To test this notion experimentally, we examined the overall ubiquitination level in intracellular schizonts versus extracellular merozoites and found it highly upregulated in merozoites. We propose that alterations in the phosphoproteome and ubiquitinome reflect a starvation-induced translational arrest as intracellular schizonts transform into extracellular merozoites

LGBTQ Emerging Adults: Their Experiences with Homelessness

Using a qualitative method, this study explored the experiences of emerging adults (ages 18-24) who are LGBTQ and homeless in the Charlotte/Mecklenburg area of North Carolina. Emerging adults who are homeless and identify as LGBTQ are shown to have different health and safety concerns compared to peers. Because of these unique factors, researchers suggest these individuals require specific social services that cater to their needs. Nine face-to-face interviews were conducted which resulted in a total of 9 hours of collected data. In order to ensure that research results are as accurate as possible, grounded theorists suggest that one have at least fifty hours of collected data (Charmaz, 2006). Therefore, this study uses a grounded theory method as a guide to explore the experiences of emerging adults who identify as LGBTQ and are homeless. Questions in this study were designed to explore participants’ experiences with community-based outreach services and their overall experience with homelessness. This paper addresses three major themes found within the data: unsafe shelter conditions for LGBTQ individuals, couch surfing, and resiliency. Participants who had access to organizations such as RAIN (REGIONAL AIDS INTERFAITH NETWORK) which provided social support systems, were in college, and living a healthy lifestyle. Data suggests that those with a strong social capital are more likely to be successful (in their definition of the word)

“I Don’t Fit in a Box; No One Does:” Intersectionality and Gay Male Identity

Using an intersectionality framework, this qualitative study explores how stigma affects identity development and how intersecting identities can compound to either foster resiliency or create health concerns for 11 men who are emerging adults (18-29), same sex identified, African American, HIV +, and homeless. Semi-structured one-on-one interviews were conducted through RAIN (Regional Aids Interfaith Network) in Charlotte, NC. Questions were formulated to understand how participants view themselves and perceived stigmas, current/past health conditions, and their five to ten year prospects. This study uses grounded theory as a guide to analyze and interpret data. Themes explored include: risks (acquiring HIV through homeless status), biographical disruptions, and self-acceptance as a foundation to resiliency through self-empowerment. All participants in this study displayed resilient behaviors post-diagnosis to present, even those who experienced the worse “biographical disruptions.” Participants all spoke about being a survivor of sorts. I found that, for my participants, owning one’s identity created a buffer against the adverse effects of stigma

The malaria parasite cyclic GMP-dependent protein kinase plays a central role in blood-stage schizogony

A role for the Plasmodium falciparum cyclic GMP (cGMP)-dependent protein kinase (PfPKG) in gametogenesis in the malaria parasite was elucidated previously. In the present study we examined the role of PfPKG in the asexual blood-stage of the parasite life cycle, the stage that causes malaria pathology. A specific PKG inhibitor (compound 1, a trisubstituted pyrrole) prevented the progression of P. falciparum schizonts through to ring stages in erythrocyte invasion assays. Addition of compound 1 to ring-stage parasites allowed normal development up to 30 h postinvasion, and segmented schizonts were able to form. However, synchronized schizonts treated with compound 1 for ≥6 h became large and dysmorphic and were unable to rupture or liberate merozoites. To conclusively demonstrate that the effect of compound 1 on schizogony was due to its selective action on PfPKG, we utilized genetically manipulated P. falciparum parasites expressing a compound 1-insensitive PfPKG. The mutant parasites were able to complete schizogony in the presence of compound 1 but not in the presence of the broad-spectrum protein kinase inhibitor staurosporine. This shows that PfPKG is the primary target of compound 1 during schizogony and provides direct evidence of a role for PfPKG in this process. Discovery of essential roles for the P. falciparum PKG in both asexual and sexual development demonstrates that cGMP signaling is a key regulator of both of these crucial life cycle phases and defines this molecule as an exciting potential drug target for both therapeutic and transmission blocking action against malaria

The effect of a scanning flat fold mirror on a CMB B-mode experiment

We investigate the possibility of using a flat-fold beam steering mirror for a CMB B-mode experiment. An aluminium flat-fold mirror is found to add $\sim$0.075% polarization, which varies in a scan synchronous way. Time-domain simulations of a realistic scanning pattern are performed, and the effect on the power-spectrum illustrated and a possible method of correction applied.Comment: 8 pages, 5 figures. Accepted for publication in Rev Sci Ins

Does Money Talk Equate to Class Talk? Audience Responses to Poverty Porn in Relation to Money and Debt

This chapter focuses on transcripts collected for the Benefits Street project at Sheffield Hallam University, which elicited audience responses to clips of poverty porn programming. We conducted four focus groups with members of the public from different social backgrounds across the north and Midlands of England and asked our participants what they thought of the representations of the working class that were shown on screen. Using techniques from corpus linguistics (specifically the use of semantic tagging software) and discourse analysis, we focus here on how our participants used terms associated with money and debt. Our analysis aims to ascertain whether talk of money in relation to benefits claimants actually equates to talk about their social class

Galliformes science and species extinctions: what we know and what we need to know

Extinciones de especies de Galliformes y conocimientos científicos: lo que sabemos y lo que necesitamos saber A principios de 2010, las 193 partes que habían firmado el Convenio sobre la Diversidad Biológica reconocieron que no habían cumplido el objetivo que ellas mismas habían fijado en 1992 de reducir de forma significativa las extinciones de especies en 2010. Al final del año establecieron un objetivo nuevo y más ambicioso que consistía en evitar las extinciones de especies en 2020. Lograr dicho objetivo requerirá una utilización mucho más eficiente de los recursos y la investigación tiene un papel fundamental en hacer que esto ocurra. Existen 290 especies de Galliformes, de las cuales el 26% se considera en peligro de extinción, en comparación con el 12% del total de las 10.000 especies de aves. Al mismo tiempo, hay numerosos estudios publicados sobre el grupo que abarcan décadas para algunas especies. Por consiguiente, es oportuno analizar si es posible aumentar la eficiencia y las repercusiones a escala mundial de la investigación sobre aves de caza, de forma que, con la planificación meticulosa que conlleva más orientación estratégica e intercambio de experiencias, los biólogos especializados en este tipo de aves puedan desempeñar una función destacada en la consecución del objetivo de 2020 para las especies aprobado por el Convenio sobre la Diversidad Biológica. Los ámbitos específicos que necesitan este planteamiento de intercambio de experiencias son la estimación de la población y la evaluación de las amenazas, el análisis de la explotación y la determinación de la base ecológica de las intervenciones que hayan obtenido buenos resultados

Rock Response in a 12-M Tunnel through a Zone of Low Strength

At the Rocky Mountain Pumped Storage Project a 12 meter diameter power tunnel was excavated through sedimentary rock for 760 meters. Approximately 10% of this tunnel was through Pennington shale that is described as a dark gray massive organic shale. This paper will describe the methods of testing .and rock characterization, the results of instrumentation and monitoring, and the post-construction testing program for the excavation, and conclude with a discussion of the observed rock response in relation to the measured strength and deformation properties. This particular zone of the tunnel required the addition of longer rockbolts, and a discussion of that supplemental rock reinforcement will be included