Search Results (19)

Several ion currents in the mammalian ventricular myocardium are substantially regulated by the sympathetic nervous system via β-adrenergic receptor activation, including the slow delayed rectifier K⁺ current and the L-type calcium current. This study investigated the downstream mechanisms of β-adrenergic receptor stimulation by isoproterenol (ISO) on the inward rectifier (I_K1) and the rapid delayed rectifier (I_Kr) K⁺ currents using action potential voltage clamp (APVC) and conventional voltage clamp techniques in isolated canine left ventricular cardiomyocytes. I_K1 and I_Kr were dissected by 50 µM BaCl₂ and 1 µM E-4031, respectively. Acute application of 10 nM ISO significantly increased I_K1 under the plateau phase of the action potential (0–+20 mV) using APVC, and similar results were obtained with conventional voltage clamp. However, β-adrenergic receptor stimulation did not affect the peak current density flowing during terminal repolarization or the overall I_K1 integral. The ISO-induced enhancement of I_K1 was blocked by the calcium/calmodulin kinase II (CaMKII) inhibitor KN-93 (1 µM) but not by the protein kinase A inhibitor H-89 (3 µM). Neither KN-93 nor H-89 affected the I_K1 density under baseline conditions (in the absence of ISO). In contrast, parameters of the I_Kr current were not affected by β-adrenergic receptor stimulation with ISO. These findings suggest that sympathetic activation enhances I_K1 in canine left ventricular cells through the CaMKII pathway, while I_Kr remains unaffected under the experimental conditions used. Full article

Groundwater flow systems are influenced by the changes in surface waters as well as climatic factors. These teleconnections significantly increase in cases of extreme weather conditions. To prepare and mitigate the effect of such phenomena, the background factors that create and influence natural processes must be recognized. In the present study, 94 shallow groundwater (SGW) wells’ water level time series were analyzed in the inner delta of the River Danube (Europe) the Szigetköz region to explore which factors contribute to the development of diurnal periodicity of SGW and what its drivers are. The relationship between surface meteorological processes and SGW dynamics in the Szigetköz region was investigated using hourly data from monitoring wells. Hourly water temperature data exhibited weak correlations with meteorological parameters. However, daily averaged data revealed stronger correlations, particularly between SGW levels and air temperature and potential evapotranspiration. Diurnal periodicity in SGW fluctuations correlated strongly with potential evapotranspiration. The study also demonstrated the role of capillary fringe dynamics in linking surface evapotranspiration with SGW fluctuations. Changes in groundwater levels, even small, can significantly affect soil moisture, vegetation, and ecosystem functioning, highlighting the sensitivity of the unsaturated zone to SGW fluctuations driven by surface processes. Full article

Autotransplantation of a mature tooth usually leads to pulpal necrosis. Root canal treatment is recommended to prevent related inflammatory complications a few weeks after surgery. Extraoral root-end resection may facilitate reperfusion and obviate root canal treatment, but cannot be pictured with conventional dental radiography at this point in time. In the case of a lower mature transplanted molar, contrast-enhanced magnetic resonance imaging proved to be a feasible method for visualizing pulp revascularization just 4 weeks after autotransplantation. Consequently, root canal treatment was obviated. Nevertheless, the tooth had to be extracted 18 months postoperatively due to external cervical root resorption, probably caused by the extraction trauma. This allowed the histological processing and examination of the newly generated intracanal tissue. Uninflamed fibrovascular connective tissue was found, while odontoblasts or cementoblast-like cells were absent. These findings indicated that it was most likely stem cells from the bone marrow and the periodontal ligament that drove the regeneration. Full article

The present study was designed to test the hypothesis that the selectivity of blocking the late Na⁺ current (I_NaL) over the peak Na⁺ current (I_NaP) is related to the fast offset kinetics of the Na⁺ channel inhibitor. Therefore, the effects of 1 µM GS967 (I_NaL inhibitor), 20 µM mexiletine (I/B antiarrhythmic) and 10 µM quinidine (I/A antiarrhythmic) on I_NaL and I_NaP were compared in canine ventricular myocardium. I_NaP was estimated as the maximum velocity of action potential upstroke (V⁺_max). Equal amounts of I_NaL were dissected by the applied drug concentrations under APVC conditions. The inhibition of I_NaL by mexiletine and quinidine was comparable under a conventional voltage clamp, while both were smaller than the inhibitory effect of GS967. Under steady-state conditions, the V⁺_max block at the physiological cycle length of 700 ms was 2.3% for GS967, 11.4% for mexiletine and 26.2% for quinidine. The respective offset time constants were 110 ± 6 ms, 456 ± 284 ms and 7.2 ± 0.9 s. These results reveal an inverse relationship between the offset time constant and the selectivity of I_NaL over I_NaP inhibition without any influence of the onset rate constant. It is concluded that the selective inhibition of I_NaL over I_NaP is related to the fast offset kinetics of the Na⁺ channel inhibitor. Full article

Late sodium current (I_Na,late) is an important inward current contributing to the plateau phase of the action potential (AP) in the mammalian heart. Although I_Na,late is considered as a possible target for antiarrhythmic agents, several aspects of this current remained hidden. In this work, the profile of I_Na,late, together with the respective conductance changes (G_Na,late), were studied and compared in rabbit, canine, and guinea pig ventricular myocytes using the action potential voltage clamp (APVC) technique. In canine and rabbit myocytes, the density of I_Na,late was relatively stable during the plateau and decreased only along terminal repolarization of the AP, while G_Na,late decreased monotonically. In contrast, I_Na,late increased monotonically, while G_Na,late remained largely unchanged during the AP in guinea pig. The estimated slow inactivation of Na⁺ channels was much slower in guinea pig than in canine or rabbit myocytes. The characteristics of canine I_Na,late and G_Na,late were not altered by using command APs recorded from rabbit or guinea pig myocytes, indicating that the different shapes of the current profiles are related to genuine interspecies differences in the gating of I_Na,late. Both I_Na,late and G_Na,late decreased in canine myocytes when the intracellular Ca²⁺ concentration was reduced either by the extracellular application of 1 µM nisoldipine or by the intracellular application of BAPTA. Finally, a comparison of the I_Na,late and G_Na,late profiles induced by the toxin of Anemonia sulcata (ATX-II) in canine and guinea pig myocytes revealed profound differences between the two species: in dog, the ATX-II induced I_Na,late and G_Na,late showed kinetics similar to those observed with the native current, while in guinea pig, the ATX-II induced G_Na,late increased during the AP. Our results show that there are notable interspecies differences in the gating kinetics of I_Na,late that cannot be explained by differences in AP morphology. These differences must be considered when interpreting the I_Na,late results obtained in guinea pig. Full article

Loess-paleosol bluffs can be unstable, but in the course of urbanization, houses may be built in such locations to take advantage of the view. One factor affecting the stability of such bluffs is water, the role of which in mass movements is well established. In this study, the connection of mass movements to meteorological conditions, such as rainfall and subsequent water level changes, was researched using new statistical methods. The periodicity of the water level of the Danube was analyzed using wavelet spectrum analyses, while changepoint analysis was used to determine variations in the quantity of precipitation. These results were compared to the chronology of six mass movements in Kulcs, Hungary. This study also focused on the changes in geochemical properties of loess in different weather conditions (dry periods, wet periods, and flooding). The results showed that only two mass movements were connected to hydrological conditions, and in the other case human activity and geochemical changes may have been factors. The results of geochemical models created using PHREEQC showed calcite and kaolinite precipitation, and albite and dolomite dissolution as the main mineral changes over the course of a year. Albite was found to dissolve only in wet periods, and kaolinite precipitation was significant during flood periods. Full article

ABT-333 (dasabuvir) is an antiviral agent used in hepatitis C treatment. The molecule, similarly to some inhibitors of hERG channels, responsible for the delayed rectifier potassium current (I_Kr), contains the methanesulfonamide group. Reduced I_Kr current leads to long QT syndrome and early afterdepolarizations (EADs), therefore potentially causing life-threatening arrhythmias and sudden cardiac death. Our goal was to investigate the acute effects of ABT-333 in enzymatically isolated canine left ventricular myocardial cells. Action potentials (APs) and ion currents were recorded with a sharp microelectrode technique and whole-cell patch clamp, respectively. Application of 1 μM ABT-333 prolonged the AP in a reversible manner. The maximal rates of phases 0 and 1 were irreversibly decreased. Higher ABT-333 concentrations caused larger AP prolongation, elevation of the early plateau potential, and reduction of maximal rates of phases 0, 1, and 3. EADs occurred in some cells in 3–30 μM ABT-333 concentrations. The 10 μM ABT-333-sensitive current, recorded with AP voltage clamp, contained a late outward component corresponding to I_Kr and an early outward one corresponding to transient outward potassium current (I_to). ABT-333 reduced hERG-channel-mediated ion current in a concentration-dependent, partially reversible manner with a half-inhibitory concentration of 3.2 μM. As the therapeutic plasma concentration of ABT-333 can reach the low μM range, ABT-333 application carries a risk of cardiac side effects especially in case of coadministration with strong inhibitors of CYP2C8. Full article

Cardiac diseases are the leading causes of death, with a growing number of cases worldwide, posing a challenge for both healthcare and research. Therefore, the most relevant aim of cardiac research is to unravel the molecular pathomechanisms and identify new therapeutic targets. Cardiac ryanodine receptor (RyR2), the Ca²⁺ release channel of the sarcoplasmic reticulum, is believed to be a good therapeutic target in a group of certain heart diseases, collectively called cardiac ryanopathies. Ryanopathies are associated with the impaired function of the RyR, leading to heart diseases such as congestive heart failure (CHF), catecholaminergic polymorphic ventricular tachycardia (CPVT), arrhythmogenic right ventricular dysplasia type 2 (ARVD2), and calcium release deficiency syndrome (CRDS). The aim of the current review is to provide a short insight into the pathological mechanisms of ryanopathies and discuss the pharmacological approaches targeting RyR2. Full article

Vagal nerve stimulation (VNS) has a meaningful basis as a potentially effective treatment for heart failure with reduced ejection fraction. There is an ongoing VNS randomized study, and four studies are completed. However, relatively little is known about the effect of acetylcholine (ACh) on repolarization in human ventricular cardiomyocytes, as well as the effect of ACh on the rapid component of the delayed rectifier K⁺ current (I_Kr). Here, we investigated the effect of ACh on the action potential parameters in human ventricular preparations and on I_Kr in human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs). Using standard microelectrode technique, we demonstrated that ACh (5 µM) significantly increased the action potential duration in human left ventricular myocardial slices. ACh (5 µM) also prolonged repolarization in a human Purkinje fiber and a papillary muscle. Optical mapping revealed that ACh increased the action potential duration in human left ventricular myocardial slices and that the effect was dose-dependent. Perforated patch clamp experiments demonstrated action potential prolongation and a significant decrease in I_Kr by ACh (5 µM) in hiPSC-CMs. Computer simulations of the electrical activity of a human ventricular cardiomyocyte showed an increase in action potential duration upon implementation of the experimentally observed ACh-induced changes in the fully activated conductance and steady-state activation of I_Kr. Our findings support the hypothesis that ACh can influence the repolarization in human ventricular cardiomyocytes by at least changes in I_Kr. Full article

Transient receptor potential melastatin 4 is a unique member of the TRPM protein family and, similarly to TRPM5, is Ca²⁺-sensitive and permeable to monovalent but not divalent cations. It is widely expressed in many organs and is involved in several functions by regulating the membrane potential and Ca²⁺ homeostasis in both excitable and non-excitable cells. This part of the review discusses the pharmacological modulation of TRPM4 by listing, comparing, and describing both endogenous and exogenous activators and inhibitors of the ion channel. Moreover, other strategies used to study TRPM4 functions are listed and described. These strategies include siRNA-mediated silencing of TRPM4, dominant-negative TRPM4 variants, and anti-TRPM4 antibodies. TRPM4 is receiving more and more attention and is likely to be the topic of research in the future. Full article

Transient receptor potential melastatin 4 (TRPM4) is a unique member of the TRPM protein family and, similarly to TRPM5, is Ca²⁺ sensitive and permeable for monovalent but not divalent cations. It is widely expressed in many organs and is involved in several functions; it regulates membrane potential and Ca²⁺ homeostasis in both excitable and non-excitable cells. This part of the review discusses the currently available knowledge about the physiological and pathophysiological roles of TRPM4 in various tissues. These include the physiological functions of TRPM4 in the cells of the Langerhans islets of the pancreas, in various immune functions, in the regulation of vascular tone, in respiratory and other neuronal activities, in chemosensation, and in renal and cardiac physiology. TRPM4 contributes to pathological conditions such as overactive bladder, endothelial dysfunction, various types of malignant diseases and central nervous system conditions including stroke and injuries as well as in cardiac conditions such as arrhythmias, hypertrophy, and ischemia-reperfusion injuries. TRPM4 claims more and more attention and is likely to be the topic of research in the future. Full article

Enhancement of the late sodium current (I_NaL) increases arrhythmia propensity in the heart, whereas suppression of the current is antiarrhythmic. In the present study, we investigated I_NaL in canine ventricular cardiomyocytes under action potential voltage-clamp conditions using the selective Na⁺ channel inhibitors GS967 and tetrodotoxin. Both 1 µM GS967 and 10 µM tetrodotoxin dissected largely similar inward currents. The amplitude and integral of the GS967-sensitive current was significantly smaller after the reduction of intracellular Ca²⁺ concentration ([Ca²⁺]_i) either by superfusion of the cells with 1 µM nisoldipine or by intracellular application of 10 mM BAPTA. Inhibiting calcium/calmodulin-dependent protein kinase II (CaMKII) by KN-93 or the autocamtide-2-related inhibitor peptide similarly reduced the amplitude and integral of I_NaL. Action potential duration was shortened in a reverse rate-dependent manner and the plateau potential was depressed by GS967. This GS967-induced depression of plateau was reduced by pretreatment of the cells with BAPTA-AM. We conclude that (1) I_NaL depends on the magnitude of [Ca²⁺]_i in canine ventricular cells, (2) this [Ca²⁺]_i-dependence of I_NaL is mediated by the Ca²⁺-dependent activation of CaMKII, and (3) I_NaL is augmented by the baseline CaMKII activity. Full article

Atrial fibrillation (AF) is the most prevalent cardiac arrhythmia in the clinical practice. It significantly contributes to the morbidity and mortality of the elderly population. Over the past 25–30 years intense effort in basic research has advanced the understanding of the relationship between the pathophysiology of AF and atrial remodelling. Nowadays it is clear that the various forms of atrial remodelling (electrical, contractile and structural) play crucial role in initiating and maintaining the persistent and permanent types of AF. Unlike in ventricular fibrillation, in AF rapid ectopic firing originating from pulmonary veins and re-entry mechanism may induce and maintain (due to atrial remodelling) this complex cardiac arrhythmia. The present review presents and discusses in detail the latest knowledge on the role of remodelling in AF. Special attention is paid to novel concepts and pharmacological targets presumably relevant to the drug treatment of atrial fibrillation. Full article

Transient receptor potential melastatin 4 (TRPM4) plays an important role in many tissues, including pacemaker and conductive tissues of the heart, but much less is known about its electrophysiological role in ventricular myocytes. Our earlier results showed the lack of selectivity of 9-phenanthrol, so CBA ((4-chloro-2-(2-chlorophenoxy)acetamido) benzoic acid) was chosen as a new, potentially selective inhibitor. Goal: Our aim was to elucidate the effect and selectivity of CBA in canine left ventricular cardiomyocytes and to study the expression of TRPM4 in the canine heart. Experiments were carried out in enzymatically isolated canine left ventricular cardiomyocytes. Ionic currents were recorded with an action potential (AP) voltage-clamp technique in whole-cell configuration at 37 °C. An amount of 10 mM BAPTA was used in the pipette solution to exclude the potential activation of TRPM4 channels. AP was recorded with conventional sharp microelectrodes. CBA was used in 10 µM concentrations. Expression of TRPM4 protein in the heart was studied by Western blot. TRPM4 protein was expressed in the wall of all four chambers of the canine heart as well as in samples prepared from isolated left ventricular cells. CBA induced an approximately 9% reduction in AP duration measured at 75% and 90% of repolarization and decreased the short-term variability of APD₉₀. Moreover, AP amplitude was increased and the maximal rates of phase 0 and 1 were reduced by the drug. In AP clamp measurements, CBA-sensitive current contained a short, early outward and mainly a long, inward current. Transient outward potassium current (I_to) and late sodium current (I_Na,L) were reduced by approximately 20% and 47%, respectively, in the presence of CBA, while L-type calcium and inward rectifier potassium currents were not affected. These effects of CBA were largely reversible upon washout. Based on our results, the CBA induced reduction of phase-1 slope and the slight increase of AP amplitude could have been due to the inhibition of I_to. The tendency for AP shortening can be explained by the inhibition of inward currents seen in AP-clamp recordings during the plateau phase. This inward current reduced by CBA is possibly I_Na,L, therefore, CBA is not entirely selective for TRPM4 channels. As a consequence, similarly to 9-phenanthrol, it cannot be used to test the contribution of TRPM4 channels to cardiac electrophysiology in ventricular cells, or at least caution must be applied. Full article