Frog α- and β-Ryanodine Receptors Provide Distinct Intracellular Ca2+ Signals in a Myogenic Cell Line

Background In frog skeletal muscle, two ryanodine receptor (RyR) isoforms, α-RyR and β-RyR, are expressed in nearly equal amounts. However, the roles and significance of the two isoforms in excitation-contraction (E-C) coupling remains to be elucidated. Methodology/Principal Findings In this study, we expressed either or both α-RyR and β-RyR in 1B5 RyR-deficient myotubes using the herpes simplex virus 1 helper-free amplicon system. Immunological characterizations revealed that α-RyR and β-RyR are appropriately expressed and targeted at the junctions in 1B5 myotubes. In Ca2+ imaging studies, each isoform exhibited caffeine-induced Ca2+ transients, an indicative of Ca2+-induced Ca2+ release (CICR). However, the fashion of Ca2+ release events was fundamentally different: α-RyR mediated graded and sustained Ca2+ release observed uniformly throughout the cytoplasm, whereas β-RyR supported all-or-none type regenerative Ca2+ oscillations and waves. α-RyR but not β-RyR exhibited Ca2+ transients triggered by membrane depolarization with high [K+]o that were nifedipine-sensitive, indicating that only α-RyR mediates depolarization-induced Ca2+ release. Myotubes co-expressing α-RyR and β-RyR demonstrated high [K+]o-induced Ca2+ transients which were indistinguishable from those with myotubes expressing α-RyR alone. Furthermore, procaine did not affect the peak height of high [K+]o-induced Ca2+ transients, suggesting minor amplification of Ca2+ release by β-RyR via CICR in 1B5 myotubes. Conclusions/Significance These findings suggest that α-RyR and β-RyR provide distinct intracellular Ca2+ signals in a myogenic cell line. These distinct properties may also occur in frog skeletal muscle and will be important for E-C coupling

Parallel finite element method utilizing the mode splitting and sigma coordinate for shallow water flows

Abstract Parallel ®nite element method for the analysis of quasi-three dimensional shallow water¯ow is presented. The mode splitting technique and the sigma coordinate (generalized coordinate) are employed to use parallel computers effectively. Parallel implementation of the unstructured grid-based formulation is carried out on the Hitachi parallel-super computer SR2201. The tidal¯ow of Tokyo Bay is simulated for a numerical example. The speed-up ratio and the ef®ciency of the parallelization are investigated. The present method is shown to be a useful and powerful tool for the large scale computation of shallow water¯ows. Introduction The shallow water¯ow analysis is usefully applied to thē ows in oceans, lakes and rivers. A number of numerical methods for the shallow water¯ow based on the two dimensional model have been presented in the past. The present authors have been presented a parallel ®nite element method to solve the large scale computations of shallow water¯ows, such as the storm surges and tidal ows, using the ®ne mesh which represents the geography accurately This paper presents a parallel ®nite element modeling utilizing the mode-splitting and sigma coordinate for the quasi-three dimensional shallow water analysis. The three dimensional¯ow ®eld is divided into two ®elds; the horizontal¯ow ®eld and the vertical¯ow ®eld. The quasi-three dimensional¯ow analysis can be achieved by solving both ®eld mutually. The ®nite element method is employed for the horizontal¯ow ®eld and the ®nite difference method is employed for the vertical distribution of¯ow ®eld. Parallel implementation of the unstructured-grid-based formulations are carried out on the Hitachi parallel-super computer SR2201. The effect of parallelization on the ef®ciency of the computations are examined. Governing equations The shallow water equations can be obtained from the conservation of momentum and mass, assuming a hydrostatic pressure distribution

Quantum Back Reaction to asymptotically AdS Black Holes

We analyze the effects of the back reaction due to a conformal field theory (CFT) on a black hole spacetime with negative cosmological constant. We study the geometry numerically obtained by taking into account the energy momentum tensor of CFT approximated by a radiation fluid. We find a sequence of configurations without a horizon in thermal equilibrium (CFT stars), followed by a sequence of configurations with a horizon. We discuss the thermodynamic properties of the system and how back reaction effects alter the space-time structure. We also provide an interpretation of the above sequence of solutions in terms of the AdS/CFT correspondence. The dual five-dimensional description is given by the Karch-Randall model, in which a sequence of five-dimensional floating black holes followed by a sequence of brane localized black holes correspond to the above solutions.Comment: 13 pages, 10 figure

A Search for Late-time Radio Emission and Fast Radio Bursts from Superluminous Supernovae

We present results of a search for late-time radio emission and fast radio bursts (FRBs) from a sample of type-I superluminous supernovae (SLSNe-I). We used the Karl G. Jansky Very Large Array to observe 10 SLSN-I more than 5 yr old at a frequency of 3 GHz. We searched fast-sampled visibilities for FRBs and used the same data to perform a deep imaging search for late-time radio emission expected in models of magnetar-powered supernovae. No FRBs were found. One SLSN-I, PTF10hgi, is detected in deep imaging, corresponding to a luminosity of 1.2 × 10²⁸ erg s⁻¹. This luminosity, considered with the recent 6 GHz detection of PTF10hgi in Eftekhari et al., supports the interpretation that it is powered by a young, fast-spinning (~ms spin period) magnetar with ~15 M⊙ of partially ionized ejecta. Broadly, our observations are most consistent with SLSNe-I being powered by neutron stars with fast spin periods, although most require more free–free absorption than is inferred for PTF10hgi. We predict that radio observations at higher frequencies or in the near future will detect these systems and begin constraining properties of the young pulsars and their birth environments

Late-Time Radio and Millimeter Observations of Superluminous Supernovae and Long Gamma Ray Bursts: Implications for Obscured Star Formation, Central Engines, and Fast Radio Bursts

We present the largest and deepest late-time radio and millimeter survey to date of superluminous supernovae (SLSNe) and long duration gamma-ray bursts (LGRBs) to search for associated non-thermal synchrotron emission. Using the Karl G. Jansky Very Large Array (VLA) and the Atacama Large Millimeter/submillimeter Array (ALMA), we observed 43 sources at 6 and 100 GHz on a timescale of $\sim 1 - 19$ yr post-explosion. We do not detect radio/mm emission from any of the sources, with the exception of a 6 GHz detection of PTF10hgi (Eftekhari et al. 2019), as well as the detection of 6 GHz emission near the location of the SLSN PTF12dam, which we associate with its host galaxy. We use our data to place constraints on central engine emission due to magnetar wind nebulae and off-axis relativistic jets. We also explore non-relativistic emission from the SN ejecta, and place constraints on obscured star formation in the host galaxies. In addition, we conduct a search for fast radio bursts (FRBs) from some of the sources using VLA Phased-Array observations; no FRBs are detected to a limit of $16$ mJy ($7\sigma$; 10 ms duration) in about 40 min on source per event. A comparison to theoretical models suggests that continued radio monitoring may lead to detections of persistent radio emission on timescales of $\gtrsim {\rm decade}$.Comment: 30 pages; 12 figures; accepted to Ap

Variations in Mre11/Rad50/Nbs1 status and DNA damage-induced S-phase arrest in the cell lines of the NCI60 panel

<p>Abstract</p> <p>Background</p> <p>The Mre11/Rad50/Nbs1 (MRN) complex is a regulator of cell cycle checkpoints and DNA repair. Defects in MRN can lead to defective S-phase arrest when cells are damaged. Such defects may elicit sensitivity to selected drugs providing a chemical synthetic lethal interaction that could be used to target therapy to tumors with these defects. The goal of this study was to identify these defects in the NCI60 panel of cell lines and identify compounds that might elicit selective cytotoxicity.</p> <p>Methods</p> <p>We screened the NCI60 panel in search of cell lines that express low levels of MRN proteins, or that fail to arrest in S-phase in response to the topisomerase I inhibitor SN38. The NCI COMPARE program was used to discover compounds that preferentially target cells with these phenotypes.</p> <p>Results</p> <p>HCT116 cells were initially identified as defective in MRN and S phase arrest. Transfection with Mre11 also elevated Rad50 and Nbs1, and rescued the defective S-phase arrest. Cells of the NCI60 panel exhibited a large range of protein expression but a strong correlation existed between Mre11, Rad50 and Nbs1 consistent with complex formation determining protein stability. Mre11 mRNA correlated best with protein level suggesting it was the primary determinant of the overall level of the complex. Three other cell lines failed to arrest in response to SN38, two of which also had low MRN. However, other cell lines with low MRN still arrested suggesting low MRN does not predict an inability to arrest. Many compounds, including a family of benzothiazoles, correlated with the failure to arrest in S phase. The activity of benzothiazoles has been attributed to metabolic activation and DNA alkylation, but we note several cell lines in which sensitivity does not correlate with metabolism. We propose that the checkpoint defect imposes an additional mechanism of sensitivity on cells.</p> <p>Conclusions</p> <p>We have identified cells with possible defects in the MRN complex and S phase arrest, and a series of compounds that may preferentially target S phase-defective cells. We discuss limitations of the COMPARE program when attempting to identify compounds that selectively inhibit only a few cell lines.</p

A machine learning classifier for fast radio burst detection at the VLBA

Time domain radio astronomy observing campaigns frequently generate large volumes of data. Our goal is to develop automated methods that can identify events of interest buried within the larger data stream. The V-FASTR fast transient system was designed to detect rare fast radio bursts within data collected by the Very Long Baseline Array. The resulting event candidates constitute a significant burden in terms of subsequent human reviewing time. We have trained and deployed a machine learning classifier that marks each candidate detection as a pulse from a known pulsar, an artifact due to radio frequency interference, or a potential new discovery. The classifier maintains high reliability by restricting its predictions to those with at least 90% confidence. We have also implemented several efficiency and usability improvements to the V-FASTR web-based candidate review system. Overall, we found that time spent reviewing decreased and the fraction of interesting candidates increased. The classifier now classifies (and therefore filters) 80%–90% of the candidates, with an accuracy greater than 98%, leaving only the 10%–20% most promising candidates to be reviewed by humans

Antitumour 2-(4-aminophenyl)benzothiazoles generate DNA adducts in sensitive tumour cells in vitro and in vivo

2-(4-Aminophenyl)benzothiazoles represent a potent and highly selective class of antitumour agent. In vitro, sensitive carcinoma cells deplete 2-(4-aminophenyl)benzothiazoles from nutrient media; cytochrome P450 1A1 activity, critical for execution of antitumour activity, and protein expression are powerfully induced. 2-(4-Amino-3-methylphenyl)benzothiazole-derived covalent binding to cytochrome P450 1A1 is reduced by glutathione, suggesting 1A1-dependent production of a reactive electrophilic species. In vitro, 2-(4-aminophenyl)benzothiazole-generated DNA adducts form in sensitive tumour cells only. At concentrations >100 nM, adducts were detected in DNA of MCF-7 cells treated with 2-(4-amino-3-methylphenyl)-5-fluorobenzothiazole (5F 203). 5F 203 (1 μM) led to the formation of one major and a number of minor adducts. However, treatment of cells with 10 μM 5F 203 resulted in the emergence of a new dominant adduct. Adducts accumulated steadily within DNA of MCF-7 cells exposed to 1 μM 5F 203 between 2 and 24 h. Concentrations of the lysylamide prodrug of 5F 203 (Phortress) ≥100 nM generated adducts in the DNA of sensitive MCF-7 and IGROV-1 ovarian cells. At 1 μM, one major Phortress-derived DNA adduct was detected in these two sensitive phenotypes; 10 μM Phortress led to the emergence of an additional major adduct detected in the DNA of MCF-7 cells. Inherently resistant MDA-MB-435 breast carcinoma cells incurred no DNA damage upon exposure to Phortress (⩽10 μM, 24 h). In vivo, DNA adducts accumulated within sensitive ovarian IGROV-1 and breast MCF-7 xenografts 24 h after treatment of mice with Phortress (20 mg kg−1). Moreover, Phortress-derived DNA adduct generation distinguished sensitive MCF-7 tumours from inherently resistant MDA-MB-435 xenografts implanted in opposite flanks of the same mouse