Simulated Microgravity and Hypergravity Affect the Expression Level of Soluble Guanylate Cyclase, Adenylate Cyclase, and Phosphodiesterase Genesin Rat Ventricular Cardiomyocytes.

Ion channels activity is regulated through soluble guanylate cyclase (sGC) and adenylate cyclase (AC) pathways, while phosphodiesterases (PDE) control the intracellular levels of cAMP and cGMP. Here we applied RNA transcriptome sequencing to study changes in the gene expression of the sGC, AC, and PDE isoforms in isolated rat ventricular cardiomyocytes under conditions of microgravity and hypergravity. Our results demonstrate that microgravity reduces the expression of sGC isoform genes, while hypergravity increases their expression. For a subset of AC isoforms, gene expression either increased or decreased under both microgravity and hypergravity conditions. The expression of genes encoding 10 PDE isoforms decreased under microgravity, but increased under hypergravity. However, under both microgravity and hypergravity, the gene expression increased for 7 PDE isoforms and decreased for 3 PDE isoforms. Overall, our findings indicate specific gravity-dependent changes in the expression of genes of isoforms associated with the studied enzymes.

Simulated Microgravity Changes the Number of Mechanically Gated and Mechanosensitive Ion Channels Genes Transcripts in Rat Ventricular Cardiomyocytes.

The mechanoelectrical feedback in the heart is based on the work of mechanically gated (MGCs) and mechanosensitive (MSCs) channels. Since microgravity alters the heart's morphological and physiological properties, we hypothesized that the expression of both MGCs and MSCs would be affected. We employed RNA transcriptome sequencing to investigate changes in the gene transcript levels of MGCs and MSCs in isolated rat ventricular cardiomyocytes under control conditions and in a simulated microgravity environment. For the first time, our findings demonstrated that simulated microgravity induces alterations in the gene transcript levels of specific MGCs, such as TRPM7, TRPV2, TRPP1, TRPP2, Piezo1, TMEM63A, TMEM36B, and known MSCs, including K2P2.1, K2P3.1, Kir6.1, Kir6.2, NaV1.5, CaV1.2, KV7.1. However, other voltage-gated channels and channels lacking a voltage sensor remained unaffected. These findings suggest that the altered expression of MGCs and MSCs could lead to changes in the net currents across the membrane, ultimately impacting the heart's function.

Hypergravity Increases the Number of Gene Transcripts of Mechanically Gated and Mechanosensitive Ion Channels in Rat Ventricular Cardiomyocytes.

Since hypergravity changes the morphological and physiological properties of the heart, it was assumed that the expression of ion channels that respond to cell stretching or compressing, mechanically gated channels (MGC) and mechanosensitive channels (MSC), would be affected. Using RNA transcriptome sequencing, the change in the number of transcripts for MGC and MSC genes was studied in isolated rat ventricular cardiomyocytes under 4g hypergravity for 5 days. It was shown for the first time that hypergravity induces changes in the number of transcripts of MGC genes: an increase for TRPC1, TRPC3, TRPM7, TRPP1 (PKD1), TRPP2 (PKD2), TMEM63A, TMEM63B, but a decrease for TRPV2, Piezo1, Piezo2. The number of MSC gene transcripts increases: TREK-1, Kir6.2, Nav1.5, Cav1.2, Cav1.3, Kv7.1, and Kv1.2. This potentially leads to an increase in the expression of MGC and MSC proteins leading to an increase in the net current and, as a result, pathological changes in the heart function.

Gadolinium as an Inhibitor of Ionic Currents in Isolated Rat Ventricular Cardiomyocytes.

The whole-cell patch-clamp technique was used to examine the effect of gadolinium Gd3+ (a non-specific blocker of mechanically gated current IMGCh, a component of late current IL) on ionic currents in insolated rat ventricular cardiomyocytes alone and in combination with the blockers of L-type calcium currents (ICaL) nifedipine (10 µM) or verapamil (1 µM). In K+in/K+out or Cs+in/Cs+out media, blockade of ICaL produced no effect on IL at negative potentials, but inhibited IL at positive ones. In K+in/K+out medium, Gd3+ (5 µM) decreased the net persistent current (Inp) at -45 mV from 198.6±6.4 to 96.7±9.5 pA over 15 min. Gd3+ alone or in combination with ICaL blockers shifted the reversal potential of IL to more negative values. At negative potentials, Gd3+ decreased IK1 and inward current including IMGCh. At positive potentials, Gd3+ alone or in combination with ICaL blockers decreased IL. When applied for 15 min in Cs+in/Cs+out medium at -45 mV, Gd3+ produced no effect on net current and inward and outward components of IL. Thus, Gd3+ can be viewed as a specific blocker of IMGCh only in Cs+ medium.

Participation of PKG and PKA-related pathways in the IFN-γ induced modulation of the BKCa channel activity in human cardiac fibroblasts.

This study was devoted to elucidating the interferon (IFN)-γ-induced signaling pathway and the interaction between protein kinase G (PKG) and protein kinase A (PKA) through large-conductance Ca(2+)-activated K(+) channels in human cardiac fibroblasts. The IK currents were recorded using a whole-cell patch clamp method. A large depolarization (+50 mV) and a high Ca2+ concentration (pCa 6.0) were used in the internal pipette solution to activate only the KCa channels. Iberiotoxin (Ibtx), which selectively inhibits BKCa channels at a concentration of 100 nmol/l, caused a significant reduction of basal IK. Adding IFN-γ in the presence of Ibtx had no effect on IK. Application of the IFN-γ caused a significant reduction in total K+ current amplitude, recorded with a 500 ms depolarizing pulse duration, to +50 mV from a holding potential of -80 mV. We tested the involvement of the sGC/cGMP/PKG signaling pathway by using specific PKG inhibitor KT 5823, potent sGC inhibitor NS 2028, and specific sGC agonist BAY 41-8543. The obtained data confirmed that only sGC participated in the IFN-γ-mediated BKCa channel modulation, which was mediated further by PKA. This study represents first evidence about the participation of the IFN-γ in the mechanisms responsible for BKCa modulation in HCFs. We also believe that this process occurs via negative crosstalk between the PKG- and PKA-associated pathways.

Metabolic Restructuring in the Liver under Conditions of Endogenous Intoxication.

Significant metabolic alterations in the liver were observed in dogs with modeled acute peritonitis. These changes significantly impaired detoxification function of the liver, which was seen from the increase in the titer of toxic products in the early post-surgery period not only in the lymph, but also in the blood plasma. The key pathogenic mechanism leading to acute liver failure is destabilization of cell membrane resulting from LPO, phospholipase activity, and tissue hypoxia. Activation of LPO and increase in phospholipase activity in the liver tissues were observed within 12 h after peritonitis modeling.

Effect of Successive Administration of Vancomycin and Amikacin on Auditory Function of Immature Animals.

Effect of successive administration vancomycin and amikacin in therapeutic doses on immature auditory organ was compared to single administration of the same drugs in chronic experiments on immature rabbits by recording of short-latency auditory brainstem response (ABR) and distortion product otoacoustic emission (DPOAE). Drug administration always increased significantly the ABR peak I threshold. Ototoxic antibiotics did not change DPOAE, but selectively affected activity of outer hair cells. No enhancement of the ototoxic effects was observed after successive administration of the two antibiotics.

Antiproliferative Effects of Modified Bioflavonoid in Ex Vivo Model.

We studied the effect of modified bioflavonoid and reference drug quercetin dihydrate on proliferation of mononuclears triggered by T- and B-cell mitogens. Lymphocytes were in vivo pretreated with the examined agents followed by their explantation and in vitro activation with T- and B-cell mitogens in cell culture. Intraperitoneal injection of modified bioflavonoid and quercetin dihydrate produced a dose-dependent inhibition of proliferation of the in vitroactivated splenocytes; modified bioflavonoid demonstrated higher antiproliferative activity.

[Hemoglobin oxygen transport capacity in surgical endotoxicosis ].

In surgical endointoxication hemoglobin oxygen transport capacity of red blood cells (hemoglobin affinity ligands: the ability to bind and release ligands) is reduced and is associated with the severity of endogenous intoxication. Violation of oxygen transport function of hemoglobin at endogenous intoxication is associated with conformational changes of a biomolecule, and its possible influence on reactive oxygen species, which confirmed in experiments in vitro: under the influence of oxygen-iron ascorbate ability of hemoglobin deteriorates. Largely similar structural and functional changes in hemoglobin occur in patients with surgical endotoxicosis.

Evaluation of the Effect of Modified Bioflavonoid and Quercetin Dihydrate on Cytokine Secretion by Mitogen-Activated Mononuclear Cells.

An experimental study revealed the effect on modified bioflavonoid on the inhibition of secretion of IFN-γ and IL-2 by ConA-stimulated mononuclear cells. These changes were accompanied by an increase in the secretion of IL-17 and IL-6. Our results suggest the differentiation of CD4(+) T helper cells into Th1 and Th17 subpopulations. The reference drug quercetin dihydrate induced an insignificant change in the level of IL-2 and IL-6 and small increase in IFN-γ content. The content of IL-17 was shown to decrease above the detection limit.

Auditory Function in Immature Animals after Two Consecutive Courses of Ototoxic Antibiotics.

In chronic experiments on immature rabbits receiving therapeutic courses of vancomycin, gentamicin, and consecutive administration of vancomicin and gentamicin by the scheme used in neonatology, hearing function was evaluated by the methods of auditory evoked potentials (auditory brainstem response, ABR) and distortion product otoacoustic emission (DPOAE). Comparison with the control group revealed ototoxic effects of all studied antibiotics that manifested in increased sound tolerance and more rapid shortening of latencies in 30-100 dB range. Higher thresholds were found only after gentamicin administration. Vancomycin administration significantly reduced the responses at 4 kHz. Subsequent gentamicin course did not potentiate this effect.