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

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

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

In vitro evaluation of amino acid prodrugs of novel antitumour 2-(4-amino-3-methylphenyl)benzothiazoles

Novel 2-(4-aminophenyl)benzothiazoles possess highly selective, potent antitumour properties in vitro and in vivo. They induce and are biotransformed by cytochrome P450 (CYP) 1A1 to putative active as well as inactive metabolites. Metabolic inactivation of the molecule has been thwarted by isosteric replacement of hydrogen with fluorine atoms at positions around the benzothiazole nucleus. The lipophilicity of these compounds presents limitations for drug formulation and bioavailability. To overcome this problem, water soluble prodrugs have been synthesised by conjugation of alanyl- and lysyl-amide hydrochloride salts to the exocyclic primary amine function of 2-(4-aminophenyl)benzothiazoles. The prodrugs retain selectivity with significant in vitro growth inhibitory potency against the same sensitive cell lines as their parent amine, but are inactive against cell lines inherently resistant to 2-(4-aminophenyl)benzothiazoles. Alanyl and lysyl prodrugs rapidly and quantitatively revert to their parent amine in sensitive and insensitive cell lines in vitro. Liberated parent compounds are sequestered and metabolised by sensitive cells only; similarly, CYP1A1 activity and protein expression are selectively induced in sensitive carcinoma cells. Amino acid prodrugs meet the criteria of aqueous solubility, chemical stability and quantitative reversion to parent molecule, and thus are suitable for in vivo preclinical evaluation

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

The SUrvey for Pulsars and Extragalactic Radio Bursts - III. Polarization properties of FRBs 160102 and 151230

We report on the polarization properties of two fast radio bursts (FRBs): 151230 and 160102 discovered in the SUrvey for Pulsars and Extragalactic Radio Bursts (SUPERB) at the Parkes Radio Telescope. FRB 151230 is observed to be 6 ± 11 per cent circularly polarized and 35 ± 13 per cent linearly polarized with a rotation measure (RM) consistent with zero. Conversely, FRB160102 is observed to have a circular polarization fraction of 30±11 per cent, linear polarization fraction of 84 ± 15 per cent for RM = -221(6) radm-2, and the highest measured dispersion measure (2596.1±0.3 pc cm-3) for an FRB to date.We examine possible progenitor models for FRB 160102 in extragalactic, non-cosmological and cosmological scenarios. After accounting for the Galactic foreground contribution, we estimate the intrinsic RM to be -256(9) rad m-2in the low-redshift case and ~-2.4×102rad m-2in the highredshift case. We assess the relative likeliness of these scenarios and how each can be tested. We also place constraints on the scattering measure and study the impact of scattering on the signal's polarization position angle

An extreme magneto-ionic environment associated with the fast radio burst source FRB 121102

Fast radio bursts are millisecond-duration, extragalactic radio flashes of unknown physical origin(1-3). The only known repeating fast radio burst source(4-6)-FRB 121102-has been localized to a star-forming region in a dwarf galaxy(7-9) at redshift 0.193 and is spatially coincident with a compact, persistent radio source(7,10). The origin of the bursts, the nature of the persistent source and the properties of the local environment are still unclear. Here we report observations of FRB 121102 that show almost 100 per cent linearly polarized emission at a very high and variable Faraday rotation measure in the source frame (varying from + 1.46 x 10(5) radians per square metre to + 1.33 x 10(5) radians per square metre at epochs separated by seven months) and narrow (below 30 microseconds) temporal structure. The large and variable rotation measure demonstrates that FRB 121102 is in an extreme and dynamic magneto-ionic environment, and the short durations of the bursts suggest a neutron star origin. Such large rotation measures have hitherto been observed(11,12) only in the vicinities of massive black holes (larger than about 10,000 solar masses). Indeed, the properties of the persistent radio source are compatible with those of a low-luminosity, accreting massive black hole(10). The bursts may therefore come from a neutron star in such an environment or could be explained by other models, such as a highly magnetized wind nebula(13) or supernova remnant(14) surrounding a young neutron star.</p

Заболевание тазобедренного сустава у детей с наследственной предрасположенностью: концептуальная модель

На основе принципов интегративной медицины, системного подхода с использованием концептуально−логического моделирования разработана единая система представлений о заболеваниях тазобедренного сустава у детей с наследственной предрасположенностью. Показано, что предлагаемый интегративный подход может служить основой для разработки диагностических и прогностических критериев развития суставов и проведения патогенетического хирургического лечения, направленного на ликвидацию или существенное снижение частоты формирования диспластического коксартроза.Based on the principles of integrative medicine, systemic approach with the use of concept of logical modelling, a uniform system of concepts about the diseases of the hip joint in children with hereditary susceptibility was worked out. It was shown that the suggested integrative approach can be used for working out diagnostic and prognostic criteria of joint development and performing pathogenetic surgery aimed at elimination or reduction in the frequency of forming dysplastic coxarthrosis