Issue 4 | December 2024
Full-length research paper
Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 166-198; doi:10.24412/2500-2295-2024-4-166-198
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The most complications of diabetes mellitus are associated with endothelial dysfunction, the nature of which is not fully understood. Oscillations of calcium (Ca2+) and nitric oxide (NO) concentrations may play a role in regulating endothelial functioning under normal and pathological conditions. We suHGested that changes of Ca2+ and NO oscillations characteristics in endothelial cells may play a role in the pathogenesis of diabetes mellitus complications. To test this hypothesis, we assessed the dependence on the glucose concentration (normo- and hyperglycemia) of the amplitude-frequency characteristics of Ca2+ and NO oscillations in endothelial cells from mouse skin microvessels at rest and during functional tests (heating), the effect of the vasodilator acetylcholine (ACh), insulin, as well as when blocking NOS and PI3K. Hyperglycemia changes the amplitude-frequency characteristics of Ca2+ and NO oscillations in endotheliocytes and the proportion of cells with oscillations. In addition, hyperglycemia changes the characteristics of Ca2+ and NO oscillations in endothelial cells in presence of ACh, L-NNA, wortmannin and insulin at rest (37 °C) and heating to 40 °C. It indicates the participation of TRPV-, NOS-, ACh- and PI3K-associated signaling pathways in the regulation of Ca2+ and NO oscillations in endoteliocytes in health and disease.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 149-165; doi:10.24412/2500-2295-2024-4-149-165
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Despite the large volume of empirical data on the effects of magnetic fields on living organisms, there are almost no studies showing the possibility of the existence of magnetic effects in processes outside the cell or in vitro. The protein folding process in a cell can be sensitive to magnetic fields. We the first time experimentally demonstrated the possibility of biochemical effects of extremely low frequency magnetic fields and hypomagnetic fields on chemical renaturation using hen egg-white lysozyme as an example. The degree of lysozyme renaturation was estimated at different magnetic conditions by fluorescence intensity and enzymatic activity. The extremely low frequency magnetic field (50 Hz, 40 μT) accelerates the renaturation compared to the control and a hypomagnetic field (less than 40 nT). The effects of hypomagnetic and extremely low frequency magnetic field on the protein fluorescence spectrum were opposite. It confirms the participation of the recently described level mixing mechanism in the implementation of magnetobiological effects. The magnetic nanoparticles abolished the effects of extremely low frequency magnetic and hypomagnetic fields fluorescence and activity of lysozyme, which indicates their ability to modulate magnetobiological effects. The results obtained expand fundamental ideas about the mechanisms of action of magnetic fields on isolated protein molecules and can be useful in the practice of using magnetic nanoparticles in biomedicine.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 138-148; doi:10.24412/2500-2295-2024-4-138-148
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Infertility is a major public health problem worldwide and its prevalence is as high as 17, 5% depending on the population studied The research is aimed to study the possibility of using miR-20a, miR-135a, miR-34b, miR-449v, miR-449c as markers of male infertility and to evaluate the dependence of the effectiveness of ART programs on the level of expression of exosomal microRNAs. The experimental group included patients entering the program of ART (assisted reproductive technologies) with the diagnosis of idiopathic male infertility (n = 30), the control group included couples with female infertility of tubal origin (n = 19). The isolation of exosomal microRNA from ejaculate was performed using the exoRNeasy Midi regent kit. The miRCURY LNA miRNA SYBR® Green PCR System was used to evaluate the expression of the following exosomal microRNAs: miR-20a, miR-135a, miR-34b, miR-449c, miR-449c and control miR-16. The expression of exosomal miR-449c and miR-135a was significantly different in the experimental group (p = 0.03 and p = 0.04, respectively). There was also a tendency to decrease the expression for such microRNAs as: miR-20a, miR-34b and miR-449c. Moreover, the expression level of miR-34b, miR-449c, and miR-449c and miR-135a has a direct correlation with the effectiveness of ART programs (p < 0.05 for miR-135a). The strength of correlation relationship of the above relationships using Cheddock scale was moderate. MicroRNA molecules selected for the study not only demonstrated their potential ability to be used as a diagnostic marker of male infertility, but also showed the ability to reflect the efficiency of fertilization and embryo formation processes.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 130-137; doi:10.24412/2500-2295-2024-4-130-137
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This study examined female rats' physiological and histological responses to cadmium chloride and lead acetate. Histological lung tissue examinations included oxidative stress and antioxidant status. Six female rat groups were studied. A group that served as a control was provided with water that had been distilled. The dosage of cadmium chloride that was administered to the second group was 5 mg/kg, whereas the dosage that was administered to the third group was 10 mg/kg. The fourth group received a dosage of lead acetate that was 50 mg/kg, whereas the fifth group received 100 mg/kg. All of the standard concentrations of lead acetate and cadmium chloride were administered to the sixth group in accordance with their protocol. Following thirty days of treatment of cadmium chloride and lead acetate to rats, the levels of oxidative indicators like MDA and 8-OHDG showed a substantial increase (P < 0.05). On the other hand, the levels of antioxidants like GSH, SOD, and CAT showed a significant drop (P ≤ 0.05 following the administration of these substances. Histological research has shown that exposure to cadmium chloride and lead acetate is associated with an increased risk of blood clots in the lungs as well as a thickening of the pulmonary alveolar wall. This is in comparison to a control group. These results demonstrate that cadmium chloride and lead acetate treatment adversely affected lung tissue's physiological and histological properties. The researchers discovered that the detrimental effect was more pronounced when the two drugs were administered concurrently to rats.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 122-129 ; doi:10.24412/2500-2295-2024-4-122-129
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The present study was carried out to evaluate different estrus synchronization protocols in mice and follow up the proportion of delivery and gender of litters. Five protocols were tested with total number of 48 adult females that divided equally into six groups including control. Total number of 24 adult males were divided equally and introduced for mating for four days. Hormonal synchronization including intraperitoneal administration of single or double dose of 0.5 µg Cloprostenol (Prostaglandin F(PGF)) and 3 µg of Progesterone (P4), with or without 5 IU of equine Chorionic Gonadotropin (eCG) in five protocols. Results showed that estrus and mating rate increase significantly after administration of PGF (P<0.05). However, low delivery rate was evident in all groups. There were no differences in the average number and the proportion of gender of litters within groups. In conclusion, synchronization of estrus in mice was not fully achieved using the current protocols. However, administration of prostaglandin increases mating rate, but the pregnancy success might fundamentally depend on other factors such as managemental.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 111-121; doi:10.24412/2500-2295-2024-4-111-121
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This study aimed to evaluate the efficacy of High-Frequency Transcutaneous Electrical Nerve Stimulation (TENS) in combination with Complex Decongestive Physical Therapy (CDPT) on lower limb lymphedema. Every participant provided informed consent for the randomized controlled trial. The trial was conducted at Ahmed Maher Teaching Hospital and High-Top Clinic in Egypt. 60 patients with secondary lower limb lymphedema, ages 35 to 65, were involved in the trial and were split into two groups at random. Group A received CDPT and High-Frequency TENS, while Group B received CDPT only. Both interventions were administered thrice weekly for eight weeks. A non-flexible tape was used to assess the limb girth as the major result; the Lymphedema Quality of Life Questionnaire (LYMQOL) was used as a secondary outcome. ANOVA tests were used in the statistical analysis to compare the pre and post-treatment outcomes within and across groups. The findings revealed that both groups had significantly lower limb girth and improved LYMQOL scores, with Group A showing more improvements than Group B (p < 0.05). The results indicate that for patients with lower limb lymphedema, the combination of High-Frequency TENS and CDPT is superior to CDPT alone in terms of limb girth reduction and improved quality of life. This combination of therapies may be a useful strategy for treating lymphedema and enhancing patient outcomes.
Invited review
Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 104-110; doi:10.24412/2500-2295-2024-4-104-110
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Cognitive impairments are among the most common types of neurological disorders that require the creation of reliable diagnostic and correction tools. Along with the development of effective drugs, in recent years a line of research has been actively developing, in which various methods of non-invasive brain stimulation with feedback from the current physiological parameters of a person, and primarily from the rhythmic components of the electroencephalogram (EEG), are successfully used to diagnose and correct cognitive impairments. The purpose of the presented work is to analyze recent publications, consider the achievements of this line of research and highlight the most promising directions for its further development. Studies using transcranial magnetic and electrical stimulation, as well as sensory types of stimulation - acoustic, photic and audiovisual stimulations, in which non-invasive stimulation is carried out on the basis of feedback signals from the patient's own bioelectrical processes, are considered. The advantages of EEG-guided light-music stimulation, developed by the authors for the correction of various cognitive disorders, are demonstrated.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 93-103; doi:10.24412/2500-2295-2024-4-93-103
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Asthma is a common chronic multifactorial respiratory disease. Genes involved in the metabolism of drugs used for asthma management play an important role in asthma development. The aim of the study was to analyze the methylation of the promoter regions of AOC1, GLCCI1 and ARG2 genes involved in the metabolism of drugs used for asthma treatment in asthma patients and controls from the Republic of Bashkortostan. DNA was extracted from peripheral blood samples of 157 asthma patients and 155 control subjects. Methylation-Sensitive High Resolution Melting analysis and sequencing of bisulfite-treated genomic DNA were applied to estimate the degree of methylation. Analysis of the methylation status of promoter region of the AOC1 gene revealed a higher frequency of full methylation (100%) of the studied region in patients with severe and moderate asthma than in controls (38.61%; p=0.002; OR=2.58; 95%CI 1.4-4.75). A significantly higher level of promoter methylation of the GLCCI1 gene was found in patients with severe and moderate asthma compared to control group (p=0.01; OR=3.1; 95%CI 1.22-7.88). A low level of promoter methylation of the ARG2 gene was determined in both analyzed groups of patients and controls. The results of MS-HRM analysis were confirmed by bisulfite sequencing of analyzed samples. Thus, this study revealed differences in the level of methylation of promoter regions of AOC1 and GLCCI1 genes between samples of asthma patients and controls. The results of the study expand general understanding of the possible contribution of DNA methylation to asthma development.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 80-92; doi:10.24412/2500-2295-2024-4-80-92
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Previous studies have demonstrated that administration of clobetasol at a concentration of 0.25 mg/kg stimulates the synthesis of neurotrophic growth factors, likely triggering the phosphatidylinositol-3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling pathways, resulting in increased levels of structural and axonal proteins necessary for the functional recovery of damaged somatic nerves. However, it has been shown that using higher concentrations of the drug (0.5 and 1.0 mg/kg) leads to a reduction in both structural and axonal proteins in both segments of the nerve conduit and exerts an inhibitory effect on the expression of neurotrophic factors responsible for cytoskeletal remodeling and axonal growth regulation.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 71-79; doi:10.24412/2500-2295-2024-4-71-79
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The liver is a vital organ involved in a wide range of processes such as detoxification, protein synthesis, metabolism, and hormone production. Liver diseases, both inherited and viral hepatitis, liver cancer and fatty degeneration are among the leading causes of death in the world. Recent advances in 3D cell culture technology include the use of pluripotent stem cells and adult stem cells that are cultured in vitro to form self-organizing systems. Organoids are self-organizing multicellular structures that reproduce the structure and function of organs and can be used to model the development, maintenance and repair of organs ex vivo. That is why the search for new methods for the formation of hepatic organoids seen as an urgent task. For the fabrication of 3D organoids we isolated hepatic duct cells from rat liver. The obtained cellular structures were analyzed using atomic force microscopy to estimate their mechanical properties and demonstrated increasing of Young Modulus in comparison with normal liver sections. Morphometric evaluation showed that extracellular matrix fibers occupy up to 60% of the organoid, while cell agglomerations up to 40% of it. We observed the spontaneously formed fibrotic liver tissue-like constructs within 21 days. So, obtained hepatic organoids characterized by a tissue-like structure with a predominance of extracellular matrix fibers in its composition similar to liver tissue affected by fibrosis.
Invited review
Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 60-70; doi:10.24412/2500-2295-2024-4-60-70
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Molecular genetic studies make it possible to determine associations of multifactorial diseases (MFDs) with many specific SNPs, which influence on MFDs etiopathogenesis is often difficult to explain. This is due to the one-sided focus of strategies in the search for mechanisms of these SNPs influence, which are mainly limited to determining the role of protein-coding genes, near or within which these polymorphisms are located. This article provides data on the mechanisms of SNP influence on MFDs etiopathogenesis due to changes in the transposable elements, which leads to their activation, dysfunction or susceptibility to exogenous viral infections. As a result, the relationship of transposable elements with specific proteins, non-coding RNA and epigenetic factors changes, which is a predisposing factor for MFDs development. Indeed, most disease-associated SNPs are located in intronic and regulatory regions of genes, and in intergenic regions. Transposable elements of the human genome are also localized in these places. Therefore, the association of specific SNPs with certain MFDs is due to the different activities of specific transposable elements. Determining the influence of SNPs on transposable elements is promising in bioinformatics studies with the construction of maps of the distribution of these elements in the genome within genes and in intergenic regions with the identification of changes in their structure under the influence of polymorphisms. Using neurodegenerative diseases as an example, it has been shown that pathological functioning and activation of retroelements due to SNPs in the regions of their location in the human genome leads to these MFDs development.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 51-59; doi:10.24412/2500-2295-2024-4-51-59
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During cardiac surgery, as well as on-pump CABG, it is crucial to monitor arterial blood parameters in order to assess the condition of patients and risks that might occur in the operating room. In some cases, the cardiac activity of patients undergoing on-pump CABG cannot be self-restored and resuscitation measures are required. Thus, the aim of the study was to analyze the differences between the two groups of patients (self-restoration of cardiac activity vs additional resuscitation measures were required) and identify arterial blood parameters potentially indicating that the patient is at risk. The data of 21 patients were analyzed with Python packages. Statistically significant differences were found between the control group (self-restoration of cardiac activity) and the test group (additional resuscitation measures were required) in the following arterial blood parameters: the levels of sodium, chloride, glucose and blood osmolality. We believe that hyponatremia and blood hypoosmolality might be a reason for cell edema which creates a greater load on the heart and leads to inability of self-restoration of cardiac activity.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 40-50; doi:10.24412/2500-2295-2024-4-40-50
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Over the past three decades, the majority of norovirus outbreaks and sporadic cases have been caused by strains of the GII.4 genotype. Notably, this period has seen a periodic change in the norovirus variants within the GII.4 genotype, with some variants acquiring epidemic distribution. Currently, six polymerase gene variants classified as P-type GII.P4 (GII.P4US95, GII.P4FarmingtonHills, GII.P4Hunter, GII.P4Yerseke, GII.P4DenHaag, and GII.P4NewOrleans) are represented in public nucleotide sequence databases, corresponding to GII.4 genotype variants of the gene encoding the main capsid protein VP1 of norovirus. In this study, the nucleotide sequence of the complete RdRp gene was obtained for a strain from Nizhny Novgorod, identified as GII.4Sydney[P4NewOrleans]. According to the results of Bayesian phylogenetic analysis based on RdRp GII.P4 gene sequences available in the GenBank database divided the analyzed strains into clusters according to polymerase variant. Within these clusters, lineages of intervariant recombinants are formed in accordance with the affiliation of the capsid protein variants associated with this polymerase variant. Analysis of the deduced amino acid sequences revealed substitutions specific to different polymerase variants that have emerged and become established within the norovirus population. It appears that the emergence of new norovirus variants within the GII.4 genotype involves selection for strains in which the polymerase and major capsid protein genes have coevolved, conferring certain advantages that facilitate further dissemination. These findings are highly relevant for advancing therapeutic strategies targeting RdRp inhibition for norovirus treatment and for development of specific prophylactic agents.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 35-39; doi:10.24412/2500-2295-2024-4-35-39
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The effect of molecular hydrogen (H2) on the functional state of spermatozoa through signaling pathways was studied. In the work, drugs that affect elements of intracellular signaling cascades were used to study the mechanisms of action of H2. We used the adenylatecyclase stimulator – forskolin, the Ca2+-channel blocker – verapamil, and the proteinkinase C inhibitor – staurosporine. We studied concentrations of MDA, ATP, the percentage of total motility and the average speed of spermatozoa under the action of H2 against the background of the action of drugs and without the action of drugs. It was shown that the concentration of MDA in all series did not change. Forskolin causes an increase in sperm motility and velocity. The use of H2 increased the effects stimulated by forskolin: sperm motility and average velocity increased. The concentration of ATP, the percentage of motility and the velocity of sperm decreased under the action of staurosporine and verapamil. The combined action of H2 and verapamil as well as the combined action of H2 and staurosporine determined an increase in sperm motility and velocity. Thus, H2 had a modulating effect through signaling pathways and caused an increase in the functional indices of sperm.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 21-34; doi:10.24412/2500-2295-2024-4-21-34
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Chronic kidney disease biomarkers are not studied well yet. Klotho and fibroblast growth factor 23 can be used as sensitive chronic kidney disease biomarkers, as well as single nucleotide polymorphisms of renin-angiotensin-aldosterone system genes may signal the risk of a genetic predisposition to this pathology. In this regard, the purpose of our work is to study the association of polymorphisms of renin-angiotensin-aldosterone system genes with physiological indicators of end-stage renal disease. Levels of fibroblast growth factor 23, Klotho, urea, creatinine, albumin, C-reactive protein, complete blood count parameters were considered in end-stage renal disease patients and control group. Also single nucleotide polymorphisms of AGT (rs4762), AGTR1 (rs5186) and AGTR2 (rs1403543) genes were determined. As a result, a statistically significant increase of fibroblast growth factor 23, urea and creatinine in the plasma of patients with end-stage renal disease was revealed. Statistically significant relationships were found between the complete blood count parameters, between albumin level and parathyroid hormone, between mean corpuscular hemoglobin and C-reactive protein. No difference of the frequency of polymorphisms of the studied genes between the groups was found. An increase of albumin and decrease of Klotho levels were recorded in carriers of the heterozygous genotype of the rs5186 polymorphism and in the case of a complete replacement for the wild allele in the FGFR2 gene (rs1403543) in the group of patients with end-stage renal disease. In the control group a relationship was found between the levels of urea, albumin and creatinine and the carriage of rs4762 and rs1403543 polymorphisms.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 13-20; doi:10.24412/2500-2295-2024-4-13-20
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This study investigates the effects of omaveloxolone, a Keap1 inhibitor, on mitochondrial network (MN) dynamics and cell survival under oxidative stress in control fibroblasts and fibroblasts from a Parkinson's disease patient with a PINK1 mutation. The PINK1-mutant fibroblasts showed increased sensitivity to hydrogen peroxide-induced stress. Omaveloxolone pre-treatment improved cell viability under stress conditions in both cell types. Under normal conditions, PINK1-mutant fibroblasts exhibited lower MN connectivity compared to control cells. Oxidative stress reduced MN density in both cell types, while omaveloxolone treatment normalized MN connectivity in PINK1-mutant cells and maintained higher MN connectivity under stress. Similar effects were observed for mitochondrial branch length. Omaveloxolone (50 nM) also increased mitophagy in both cell types under normal conditions. Our findings demonstrate that omaveloxolone exerts protective effects by maintaining mitochondrial dynamics and activating mitophagy. It enhances mitophagy under normal conditions and supports MN structure under oxidative stress, improving cell viability in both control and PINK1-mutant fibroblasts. These results highlight omaveloxolone's potential as a therapeutic agent for protecting cells in diseases associated with impaired mitochondrial dynamics, particularly Parkinson's disease linked to PINK1 mutations.
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Published ahead of print December 01, 2024; Printed December 02, 2024; OM&P 2024 Volume 11 Issue 4, pages 5-12; doi:10.24412/2500-2295-2024-4-5-12
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This study investigates the effects of the α1-adrenergic receptor agonist methoxamine at various concentrations (10−9 to 10−6 M) on the frequency and characteristics of spontaneous action potentials in right atrial preparations of newborn and adult rats. In newborn rats, methoxamine at all studied concentrations significantly increased the duration of the action potential repolarization phase, while in adult rats, decreasing it. Interestingly, in adult rats, methoxamine at a concentration of (10−6 M), exhibited a dual effect, decreasing the duration of the action potential repolarization phase in some samples while increasing it in others. Stimulation of α1-adrenergic receptors by methoxamine in working cardiomyocytes of the right atrium in both age groups led to an increase in the frequency of action potential generation; however, this effect was more pronounced in newborn rats. These findings highlight the age-dependent effects of α1-adrenergic receptor stimulation on cardiac electrical activity, suggesting that the mechanisms underlying these responses may differ significantly between newborn and adult rats.