A study of excitation contraction coupling in rabbit cardiomyocytes

Excitation-Contraction coupling is the process that links the depolarisation of the myocardium to its contraction. We have utilised an adenoviral- mediated strategy to overexpress sorcin, calsequestrin (CSQ), FKBP 12.6 and the sodium/calcium exchanger (NCX) in isolated rabbit cardiomyocytes over a period of 24hrs in cell culture. The effects of overexpression of the above-mentioned proteins on excitation-contraction coupling were subsequently investigated using whole cell voltage clamp. The loading of cardiomyocytes with FURA-2 enabled simultaneous measurement of intracellular calcium concentration. Cardiomyocytes transfected with an adenovirus that caused overexpression of the enzyme beta-galactosidase were employed as a control. Sorcin overexpression was found to significantly reduce both the amplitude of the intracellular Ca2+ transient and the size of the SR Ca2+ load under conditions of whole cell voltage clamp when compared to control. It had no effect on the L-type Ca2+ current but significantly enhanced the activity of the sodium/calcium exchanger. Subsequently, overexpression of NCX was shown to affect excitation contraction coupling in a similar manner to sorcin overexpression. CSQ overexpression was shown to significantly increase the size of L-type Ca2+ current, the intracellular Ca2+ transient amplitude and the SR Ca2+ load under conditions of whole cell voltage clamp when compared to control. When the L-type Ca2+ current in the CSQ overexpressing cardiomyocytes was normalised using nifedipine the SR Ca2+ load remained increased while the amplitude of the intracellular Ca2+ transient was similar to that observed in control cardiomyocytes. Finally, FKBP 12.6 overexpression significantly increased both the amplitude of the intracellular Ca2+ transient and the size of the SR Ca2+ load under conditions of whole cell voltage clamp when compared to control. It had no effect on the L-type Ca2+ current. Over expression of an FKBP 12.6 mutant that lacked a calcineurin binding site had little effect on the size of the intracellular Ca2+ transient but still significantly increased the SR Ca2+ load under conditions of whole cell voltage clamp when compared to control. Experiments previously carried out in the laboratory using confocal microscopy have demonstrated that both sorcin and FKBP 12.6 overexpression alter the occurrence of spontaneous Ca2+ sparks in permeabilised cardiomyocytes. Conversely, CSQ overexpression was shown to have no effect on the occurrence of Ca2+ sparks

Linkage design effect on the reliability of surface-micromachined microengines driving a load

The reliability of microengines is a function of the design of the mechanical linkage used to connect the electrostatic actuator to the drive. The authors have completed a series of reliability stress tests on surface micromachined microengines driving an inertial load. In these experiments, the authors used microengines that had pin mechanisms with guides connecting the drive arms to the electrostatic actuators. Comparing this data to previous results using flexure linkages revealed that the pin linkage design was less reliable. The devices were stressed to failure at eight frequencies, both above and below the measured resonance frequency of the microengine. Significant amounts of wear debris were observed both around the hub and pin joint of the drive gear. Additionally, wear tracks were observed in the area where the moving shuttle rubbed against the guides of the pin linkage. At each frequency, they analyzed the statistical data yielding a lifetime (t{sub 50}) for median cycles to failure and {sigma}, the shape parameter of the distribution. A model was developed to describe the failure data based on fundamental wear mechanisms and forces exhibited in mechanical resonant systems. The comparison to the model will be discussed

Measuring Black Hole Spin using X-ray Reflection Spectroscopy

I review the current status of X-ray reflection (a.k.a. broad iron line) based black hole spin measurements. This is a powerful technique that allows us to measure robust black hole spins across the mass range, from the stellar-mass black holes in X-ray binaries to the supermassive black holes in active galactic nuclei. After describing the basic assumptions of this approach, I lay out the detailed methodology focusing on "best practices" that have been found necessary to obtain robust results. Reflecting my own biases, this review is slanted towards a discussion of supermassive black hole (SMBH) spin in active galactic nuclei (AGN). Pulling together all of the available XMM-Newton and Suzaku results from the literature that satisfy objective quality control criteria, it is clear that a large fraction of SMBHs are rapidly-spinning, although there are tentative hints of a more slowly spinning population at high (M>5*10^7Msun) and low (M<2*10^6Msun) mass. I also engage in a brief review of the spins of stellar-mass black holes in X-ray binaries. In general, reflection-based and continuum-fitting based spin measures are in agreement, although there remain two objects (GROJ1655-40 and 4U1543-475) for which that is not true. I end this review by discussing the exciting frontier of relativistic reverberation, particularly the discovery of broad iron line reverberation in XMM-Newton data for the Seyfert galaxies NGC4151, NGC7314 and MCG-5-23-16. As well as confirming the basic paradigm of relativistic disk reflection, this detection of reverberation demonstrates that future large-area X-ray observatories such as LOFT will make tremendous progress in studies of strong gravity using relativistic reverberation in AGN.Comment: 19 pages. To appear in proceedings of the ISSI-Bern workshop on "The Physics of Accretion onto Black Holes" (8-12 Oct 2012). Revised version adds a missing source to Table 1 and Fig.6 (IRAS13224-3809) and corrects the referencing of the discovery of soft lags in 1H0707-495 (which were in fact first reported in Fabian et al. 2009

Coronal mass ejections as expanding force-free structures

We mode Solar coronal mass ejections (CMEs) as expanding force-fee magnetic structures and find the self-similar dynamics of configurations with spatially constant \alpha, where {\bf J} =\alpha {\bf B}, in spherical and cylindrical geometries, expanding spheromaks and expanding Lundquist fields correspondingly. The field structures remain force-free, under the conventional non-relativistic assumption that the dynamical effects of the inductive electric fields can be neglected. While keeping the internal magnetic field structure of the stationary solutions, expansion leads to complicated internal velocities and rotation, induced by inductive electric field. The structures depends only on overall radius R(t) and rate of expansion \dot{R}(t) measured at a given moment, and thus are applicable to arbitrary expansion laws. In case of cylindrical Lundquist fields, the flux conservation requires that both axial and radial expansion proceed with equal rates. In accordance with observations, the model predicts that the maximum magnetic field is reached before the spacecraft reaches the geometric center of a CME.Comment: 19 pages, 9 Figures, accepted by Solar Physic

The effect of frequency on the lifetime of a surface micromachined microengine driving a load

Experiments have been performed on surface micromachined microengines driving load gears to determine the effect of the rotation frequency on median cycles to failure. The authors did observe a frequency dependence and have developed a model based on fundamental wear mechanisms and forces exhibited in resonant mechanical systems. Stressing loaded microengines caused observable wear in the rotating joints and in a few instances led to fracture of the pin joint in the drive gear

The celebrity entrepreneur on television: profile, politics and power

This article examines the rise of the ‘celebrity entrepreneur’ on television through the emergence of the ‘business entertainment format’ and considers the ways in which regular television exposure can be converted into political influence. Within television studies there has been a preoccupation in recent years with how lifestyle and reality formats work to transform ‘ordinary’ people into celebrities. As a result, the contribution of vocationally skilled business professionals to factual entertainment programming has gone almost unnoticed. This article draws on interviews with key media industry professionals and begins by looking at the construction of entrepreneurs as different types of television personalities and how discourses of work, skill and knowledge function in business shows. It then outlines how entrepreneurs can utilize their newly acquired televisual skills to cultivate a wider media profile and secure various forms of political access and influence. Integral to this is the centrality of public relations and media management agencies in shaping media discourses and developing the individual as a ‘brand identity’ that can be used to endorse a range of products or ideas. This has led to policy makers and politicians attempting to mobilize the media profile of celebrity entrepreneurs to reach out and connect with the public on business and enterprise-related issues

Long-term trends, current status, and transitions of water quality in Chesapeake Bay

Coincident climatic and human effects strongly influence water-quality properties in estuarine-coastal ecosystems around the world. Time-series data for a number of ecosystems reveal high spatio-temporal variability superimposed on secular trends traceable to nutrient over-enrichment. In this paper, we present new analyses of long-term data for Chesapeake Bay directed at several goals: (1) to distinguish trends from spatio-temporal variability imposed by climatic effects; (2) to assess long-term trends of water-quality properties reflecting degradation and recovery; (3) to propose numerical water-quality criteria as targets for restoration; (4) to assess progress toward attainment of these targets. The bay has experienced multiple impairments associated with nutrient over-enrichment since World War II, e.g., low dissolved oxygen (DO), decreased water clarity, and harmful algal blooms (HAB). Anthropogenic eutrophication has been expressed as increased chlorophyll-a (chl-a) driven by accelerated nutrient loading from 1945 to 1980. Management intervention led to decreased loading thereafter, but deleterious symptoms of excess nutrients persist. Climatic effects exemplified by irregular “dry” and “wet” periods in the last 30+ years largely explain high inter-annual variability of water-quality properties, requiring adjustments to resolve long-term trends. Here, we extend these analyses at a finer temporal scale to six decades of chl-a, Secchi depth, and nitrite plus nitrate (NO 2 + NO 3 ) data to support trend analyses and the development of numerical water-quality criteria. The proposed criteria build on a conceptual model emphasizing the need to distinguish climatic and human effects in gauging progress to reverse eutrophication in estuarine-coastal ecosystems

Seasonal to Inter-Annual Variability of Primary Production in Chesapeake Bay: Prospects to Reverse Eutrophication and Change Trophic Classification

Estuarine-coastal ecosystems are rich areas of the global ocean with elevated rates of organic matter production supporting major fisheries. Net and gross primary production (NPP, GPP) are essential properties of these ecosystems, characterized by high spatial, seasonal, and inter-annual variability associated with climatic effects on hydrology. Over 20 years ago, Nixon defined the trophic classification of marine ecosystems based on annual phytoplankton primary production (APPP), with categories ranging from “oligotrophic” to “hypertrophic”. Source data consisting of shipboard measurements of NPP and GPP from 1982 to 2004 for Chesapeake Bay in the mid-Atlantic region of the United States supported estimates of APPP from 300 to 500 g C m−2 yr−1, corresponding to “eutrophic” to “hypertrophic” categories. Here, we developed generalized additive models (GAM) to interpolate the limited spatio-temporal resolution of source data. Principal goals were: (1) to develop predictive models of NPP and GPP calibrated to source data (1982 to 2004); (2) to apply the models to historical (1960s, 1970s) and monitoring (1985 to 2015) data with adjustments for nutrient loadings and climatic effects; (3) to estimate APPP from model predictions of NPP; (4) to test effects of simulated reductions of phytoplankton biomass or nutrient loadings on trophic classification based on APPP. Simulated 40% decreases of euphotic-layer chl-a or TN and NO2 + NO3 loadings led to decreasing APPP sufficient to change trophic classification from “eutrophic’ to “mesotrophic” for oligohaline (OH) and polyhaline (PH) salinity zones, and from “hypertrophic” to “eutrophic” for the mesohaline (MH) salinity zone of the bay. These findings show that improved water quality is attainable with sustained reversal of nutrient over-enrichment sufficient to decrease phytoplankton biomass and APPP

Esophageal foreign-body impactions: epidemiology, time trends, and the impact of the increasing prevalence of eosinophilic esophagitis

The epidemiology of esophageal foreign-body impaction (EFBI) is poorly described, and the impact of the increasing prevalence of eosinophilic esophagitis (EoE) on this is unknown

Long-Term Trends of Nutrients and Phytoplankton in Chesapeake Bay

Climate effects on hydrology impart high variability to water-quality properties, including nutrient loadings, concentrations, and phytoplankton biomass as chlorophyll-a (chl-a), in estuarine and coastal ecosystems. Resolving long-term trends of these properties requires that we distinguish climate effects from secular changes reflecting anthropogenic eutrophication. Here, we test the hypothesis that strong climatic contrasts leading to irregular dry and wet periods contribute significantly to interannual variability of mean annual values of water-quality properties using in situ data for Chesapeake Bay. Climate effects are quantified using annual freshwater discharge from the Susquehanna River together with a synoptic climatology for the Chesapeake Bay region based on predominant sea-level pressure patterns. Time series of water-quality properties are analyzed using historical (1945–1983) and recent (1984–2012) data for the bay adjusted for climate effects on hydrology. Contemporary monitoring by the Chesapeake Bay Program (CBP) provides data for a period since mid-1984 that is significantly impacted by anthropogenic eutrophication, while historical data back to 1945 serve as historical context for a period prior to severe impairments. The generalized additive model (GAM) and the generalized additive mixed model (GAMM) are developed for nutrient loadings and concentrations (total nitrogen—TN, nitrate + nitrate—NO2 + NO3) at the Susquehanna River and water-quality properties in the bay proper, including dissolved nutrients (NO2 + NO3, orthophosphate—PO4), chl-a, diffuse light attenuation coefficient (KD (PAR)), and chl-a/TN. Each statistical model consists of a sum of nonlinear functions to generate flow-adjusted time series and compute long-term trends accounting for climate effects on hydrology. We present results identifying successive periods of (1) eutrophication ca. 1945–1980 characterized by approximately doubled TN and NO2 + NO3 loadings, leading to increased chl-a and associated ecosystem impairments, and (2) modest decreases of TN and NO2 + NO3 loadings from 1981 to 2012, signaling a partial reversal of nutrient over-enrichment. Comparison of our findings with long-term trends of water-quality properties for a variety of estuarine and coastal ecosystems around the world reveals that trends for Chesapeake Bay are weaker than for other systems subject to strenuous management efforts, suggesting that more aggressive actions than those undertaken to date will be required to counter anthropogenic eutrophication of this valuable resource