Modulation of Cardiac Ryanodine Receptor Channels by Alkaline Earth Cations

Cardiac ryanodine receptor (RyR2) function is modulated by Ca2+ and Mg2+. To better characterize Ca2+ and Mg2+ binding sites involved in RyR2 regulation, the effects of cytosolic and luminal earth alkaline divalent cations (M2+: Mg2+, Ca2+, Sr2+, Ba2+) were studied on RyR2 from pig ventricle reconstituted in bilayers. RyR2 were activated by M2+ binding to high affinity activating sites at the cytosolic channel surface, specific for Ca2+ or Sr2+. This activation was interfered by Mg2+ and Ba2+ acting at low affinity M2+-unspecific binding sites. When testing the effects of luminal M2+ as current carriers, all M2+ increased maximal RyR2 open probability (compared to Cs+), suggesting the existence of low affinity activating M2+-unspecific sites at the luminal surface. Responses to M2+ vary from channel to channel (heterogeneity). However, with luminal Ba2+or Mg2+, RyR2 were less sensitive to cytosolic Ca2+ and caffeine-mediated activation, openings were shorter and voltage-dependence was more marked (compared to RyR2 with luminal Ca2+or Sr2+). Kinetics of RyR2 with mixtures of luminal Ba2+/Ca2+ and additive action of luminal plus cytosolic Ba2+ or Mg2+ suggest luminal M2+ differentially act on luminal sites rather than accessing cytosolic sites through the pore. This suggests the presence of additional luminal activating Ca2+/Sr2+-specific sites, which stabilize high Po mode (less voltage-dependent) and increase RyR2 sensitivity to cytosolic Ca2+ activation. In summary, RyR2 luminal and cytosolic surfaces have at least two sets of M2+ binding sites (specific for Ca2+ and unspecific for Ca2+/Mg2+) that dynamically modulate channel activity and gating status, depending on SR voltage

Fundamentos del enfoque de sistemas para la metodología de evaluación de instalaciones de obras acuáticas en medio natural

De acuerdo con los requisitos reglamentarios y ambientales sobre el funcionamiento de las instalaciones sanitarias denominadas objetos de actividad, se debe evaluar la influencia ambiental de estas instalaciones. en términos de utilización de los recursos hídricos, que es exactamente el objetivo de este estudio. Los fundamentos de la metodología de esta evaluación se desarrollan mediante el enfoque de sistema centrado en la noción de sistema. Los resultados del estudio de la interacción del objeto de actividad (OA) con los ambientes naturales (NENV) del geosistema de cuenca se utilizan en el desarrollo de los fundamentos de la metodología de evaluación de la influencia ambiental (EIA). Los resultados del estudio se han utilizado para diseñar y construir instalaciones de ingeniería hidroeléctrica en el norte del Cáucaso en el sur de Rusi

Additive singular spectral model of a dendrochronological signal

Relevance. Allocation of structural components in a dendrochronological signal of annual rings of coniferous trees expands the possibilities of the bioindication method and allows obtaining information about changes in environmental conditions in the past for extended territories. Aim. Creation of an additive singular spectral model based on the frequency trigonometric components of the dendrochronological signal; reconstruction of changes in the total ozone content in the atmosphere in the past, affecting the level of ultraviolet radiation in the B range radiation. Objects. Time series of total ozone content in the atmosphere (data from 1932), width and density of annual rings (data from 1686–2004) on the example of the territory near the observatory in Arosa, Switzerland, time series of stable oxygen isotopes δ18Oc, France. Methods. Time series analysis (decomposition of a time series, identification of model parameters, prediction of a time series), statistical analysis (F-criterion, χ2 – Pearson criterion), experimental measurements of the percentage components of the wood of annual rings, correlation analysis, spectral singular analysis. Results. Decomposition of the dendrochronological signal of individual chronologies into trigonometric components in the Caterpillar program. Correlation analysis of the sensitivity of trees to atmospheric effects. Reconstruction of the ozone level in the stratosphere using the first trigonometric component of the dendrochronological signal. In cellulose, the first low-frequency component of the signal and the structural component of wood, a reliable response of trees to long-period fluctuations in the total ozone content is recorded. This allows assessment of the impact of the stress factor on conditions of exploitation of forest resources. The use of data on the width of the annual rings allows you to expand the territorial boundaries of the method

ПОЛУЧЕНИЕ КОМПЛЕКСНЫХ ПРЕПАРАТОВ НА ОСНОВЕ ЛИПОСОМАЛЬНОЙ ФОРМЫ СТРЕПТОКИНАЗЫ И ИХ ФАРМАКОКИНЕТИЧЕСКИЕ ХАРАКТЕРИСТИКИ

It was obtained that liposomal streptokinase (SK) with a hydrodynamic diameter of ~70 nm and a zeta-potential of –6.2 mV contains 14.1 % wt of drug. The complex formulations based on liposomal SK include “associated” and “free” SK in the ratios of 20/80, 40/60 and 50/50. The in vivo experiments on rats showed an increase in the elimination half-time from 1.8 to 31.9 min and in the time to reach the maximum concentration of streptokinase from 15 to 45 min. The decrease in the elimination rate constant by a factor of 18 compared with SK was also found. The optimal ratio of “associated” and “free” SK in the complex formulation was 40 and 60 % respectively. It was used to obtain liposomal fibrin-specific form of thrombolytic with similar physico-chemical and pharmacokinetic parameters. Получена липосомальная форма стрептокиназы (СК) с гидродинамическим диаметром ~70 нм, дзета-потенциалом –6,2 мВ и степенью включения вещества 14,1 %, на основе которой приготовлены комплексные препараты, содержащие «связанную» и «свободную» СК в соотношениях 20/80, 40/60 и 50/50. Для них в эксперименте in vivo на крысах показано увеличение периода полувыведения от 1,8 до 31,9 мин и времени достижения максимальной концентрации стрептокиназы от 15 до 45 мин, а также уменьшение константы элиминации в ~18 раз по сравнению с нативной СК. Оптимальное соотношение «связанной» и «свободной» СК в комплексном препарате составило 40 и 60 % соответственно, что было использовано для получения липосомальной фибрин-специфичной формы тромболитика, который обладает практически такими же физико-химическими и фармакокинетическими параметрами.

A Regional Reduction in Ito and IKACh in the Murine Posterior Left Atrial Myocardium Is Associated with Action Potential Prolongation and Increased Ectopic Activity.

BACKGROUND: The left atrial posterior wall (LAPW) is potentially an important area for the development and maintenance of atrial fibrillation. We assessed whether there are regional electrical differences throughout the murine left atrial myocardium that could underlie regional differences in arrhythmia susceptibility. METHODS: We used high-resolution optical mapping and sharp microelectrode recordings to quantify regional differences in electrical activation and repolarisation within the intact, superfused murine left atrium and quantified regional ion channel mRNA expression by Taqman Low Density Array. We also performed selected cellular electrophysiology experiments to validate regional differences in ion channel function. RESULTS: Spontaneous ectopic activity was observed during sustained 1Hz pacing in 10/19 intact LA and this was abolished following resection of LAPW (0/19 resected LA, P<0.001). The source of the ectopic activity was the LAPW myocardium, distinct from the pulmonary vein sleeve and LAA, determined by optical mapping. Overall, LAPW action potentials (APs) were ca. 40% longer than the LAA and this region displayed more APD heterogeneity. mRNA expression of Kcna4, Kcnj3 and Kcnj5 was lower in the LAPW myocardium than in the LAA. Cardiomyocytes isolated from the LAPW had decreased Ito and a reduced IKACh current density at both positive and negative test potentials. CONCLUSIONS: The murine LAPW myocardium has a different electrical phenotype and ion channel mRNA expression profile compared with other regions of the LA, and this is associated with increased ectopic activity. If similar regional electrical differences are present in the human LA, then the LAPW may be a potential future target for treatment of atrial fibrillation

Spatiotemporally Controlled Cardiac Conduction Block Using High-Frequency Electrical Stimulation

Background: Methods for the electrical inhibition of cardiac excitation have long been sought to control excitability and conduction, but to date remain largely impractical. High-amplitude alternating current (AC) stimulation has been known to extend cardiac action potentials (APs), and has been recently exploited to terminate reentrant arrhythmias by producing reversible conduction blocks. Yet, low-amplitude currents at similar frequencies have been shown to entrain cardiac tissues by generation of repetitive APs, leading in some cases to ventricular fibrillation and hemodynamic collapse in vivo. Therefore, an inhibition method that does not lead to entrainment – irrespective of the stimulation amplitude (bound to fluctuate in an in vivo setting) – is highly desirable. Methodology/Principal Findings: We investigated the effects of broader amplitude and frequency ranges on the inhibitory effects of extracellular AC stimulation on HL-1 cardiomyocytes cultured on microelectrode arrays, using both sinusoidal and square waveforms. Our results indicate that, at sufficiently high frequencies, cardiac tissue exhibits a binary response to stimulus amplitude with either prolonged APs or no effect, thereby effectively avoiding the risks of entrainment by repetitive firing observed at lower frequencies. We further demonstrate the ability to precisely define reversible local conduction blocks in beating cultures without influencing the propagation activity in non-blocked areas. The conduction blocks were spatiotemporally controlled by electrode geometry and stimuli duration, respectively, and sustainable for long durations (300 s). Conclusion/Significance: Inhibition of cardiac excitation induced by high-frequency AC stimulation exhibits a binary response to amplitude above a threshold frequency, enabling the generation of reversible conduction blocks without the risks of entrainment. This inhibition method could yield novel approaches for arrhythmia modeling in vitro, as well as safer and more efficacious tools for in vivo cardiac mapping and radio-frequency ablation guidance applications

Membranes with the Same Ion Channel Populations but Different Excitabilities

Electrical signaling allows communication within and between different tissues and is necessary for the survival of multicellular organisms. The ionic transport that underlies transmembrane currents in cells is mediated by transporters and channels. Fast ionic transport through channels is typically modeled with a conductance-based formulation that describes current in terms of electrical drift without diffusion. In contrast, currents written in terms of drift and diffusion are not as widely used in the literature in spite of being more realistic and capable of displaying experimentally observable phenomena that conductance-based models cannot reproduce (e.g. rectification). The two formulations are mathematically related: conductance-based currents are linear approximations of drift-diffusion currents. However, conductance-based models of membrane potential are not first-order approximations of drift-diffusion models. Bifurcation analysis and numerical simulations show that the two approaches predict qualitatively and quantitatively different behaviors in the dynamics of membrane potential. For instance, two neuronal membrane models with identical populations of ion channels, one written with conductance-based currents, the other with drift-diffusion currents, undergo transitions into and out of repetitive oscillations through different mechanisms and for different levels of stimulation. These differences in excitability are observed in response to excitatory synaptic input, and across different levels of ion channel expression. In general, the electrophysiological profiles of membranes modeled with drift-diffusion and conductance-based models having identical ion channel populations are different, potentially causing the input-output and computational properties of networks constructed with these models to be different as well. The drift-diffusion formulation is thus proposed as a theoretical improvement over conductance-based models that may lead to more accurate predictions and interpretations of experimental data at the single cell and network levels

Nonstandard Errors

In statistics, samples are drawn from a population in a data-generating process (DGP). Standard errors measure the uncertainty in estimates of population parameters. In science, evidence is generated to test hypotheses in an evidence-generating process (EGP). We claim that EGP variation across researchers adds uncertainty-nonstandard errors (NSEs). We study NSEs by letting 164 teams test the same hypotheses on the same data. NSEs turn out to be sizable, but smaller for more reproducible or higher rated research. Adding peer-review stages reduces NSEs. We further find that this type of uncertainty is underestimated by participants

Non-Standard Errors

In statistics, samples are drawn from a population in a data-generating process (DGP). Standard errors measure the uncertainty in estimates of population parameters. In science, evidence is generated to test hypotheses in an evidence-generating process (EGP). We claim that EGP variation across researchers adds uncertainty: Non-standard errors (NSEs). We study NSEs by letting 164 teams test the same hypotheses on the same data. NSEs turn out to be sizable, but smaller for better reproducible or higher rated research. Adding peer-review stages reduces NSEs. We further find that this type of uncertainty is underestimated by participants

In silico assessment of the effects of quinidine, disopyramide and E-4031 on short QT syndrome variant 1 in the human ventricles.

Aims Short QT syndrome (SQTS) is an inherited disorder associated with abnormally abbreviated QT intervals and an increased incidence of atrial and ventricular arrhythmias. SQT1 variant (linked to the rapid delayed rectifier potassium channel current, IKr) of SQTS, results from an inactivation-attenuated, gain-of-function mutation (N588K) in the KCNH2-encoded potassium channels. Pro-arrhythmogenic effects of SQT1 have been well characterized, but less is known about the possible pharmacological antiarrhythmic treatment of SQT1. Therefore, this study aimed to assess the potential effects of E-4031, disopyramide and quinidine on SQT1 using a mathematical model of human ventricular electrophysiology. Methods The ten Tusscher et al. biophysically detailed model of the human ventricular action potential (AP) was modified to incorporate IKr Markov chain (MC) formulations based on experimental data of the kinetics of the N588K mutation of the KCNH2-encoded subunit of the IKr channels. The modified ventricular cell model was then integrated into one-dimensional (1D) strand, 2D regular and realistic tissues with transmural heterogeneities. The channel-blocking effect of the drugs on ion currents in healthy and SQT1 cells was modeled using half-maximal inhibitory concentration (IC50) and Hill coefficient (nH) values from literatures. Effects of drugs on cell AP duration (APD), effective refractory period (ERP) and pseudo-ECG traces were calculated. Effects of drugs on the ventricular temporal and spatial vulnerability to re-entrant excitation waves were measured. Re-entry was simulated in both 2D regular and realistic ventricular tissue. Results At the single cell level, the drugs E-4031 and disopyramide had hardly noticeable effects on the ventricular cell APD at 90% repolarization (APD90), whereas quinidine caused a significant prolongation of APD90. Quinidine prolonged and decreased the maximal transmural AP heterogeneity (δV); this led to the decreased transmural heterogeneity of APD across the 1D strand. Quinidine caused QT prolongation and a decrease in the T-wave amplitude, and increased ERP and decreased temporal susceptibility of the tissue to the initiation of re-entry and increased the minimum substrate size necessary to prevent re-entry in the 2D regular model, and further terminated re-entrant waves in the 2D realistic model. Quinidine exhibited significantly better therapeutic effects on SQT1 than E-4031 and disopyramide. Conclusions The simulated pharmacological actions of quinidine exhibited antiarrhythmic effects on SQT1. This study substantiates a causal link between quinidine and QT interval prolongation in SQT1 and suggests that quinidine may be a potential pharmacological agent for treating SQT1 patients