Valeriy Pokhylko

Analysis of gene variants, oxidative and nitrosative stress markers, and their interrelationships may facilitate the development of personalized approaches to the prevention and treatment of coronavirus disease 2019 (COVID-19) and other infectious diseases, particularly in regions affected by environmental pollution. The present study was conducted to evaluate the influence of MTHFR (rs1801133, rs1801131) and VDR (rs731236) gene variants, as well as oxidative and nitrosative stress markers, on the course of COVID-19 in children residing in the Poltava region, with consideration of nitrate and fluoride pollution. Methods. The study included 40 children diagnosed with COVID-19 and 17 age-matched children in the comparison group. The following parameters were assessed: variants of the MTHFR and VDR genes; urinary concentrations of nitrates, nitrites, malondialdehyde (MDA), and sialic acids; and residence in territories with elevated fluoride and nitrate pollution. Results. In children with COVID-19, a significantly higher frequency of the AA genotype of the rs1801131 variant was observed. Elevated urinary levels of nitrites, MDA, and sialic acids were identified as significant pathogenic factors associated with the disease. Among children living in fluoride- and nitrate-polluted areas, correlations were found between MTHFR gene variants and serum concentrations of bilirubin, creatinine, and MDA. Betaine-arginine supplementation was identified as a potential strategy for personalized prevention and treatment in pediatric COVID-19. Conclusion. Associations were established between MTHFR gene variants and increased susceptibility to COVID-19. The influence of fluoride- and nitrate-polluted environments was confirmed, key pathogenic factors (nitrite, MDA, and sialic acid levels) were delineated, and betaine-arginine supplementation was proposed as a potential preventive and therapeutic intervention.

Among neonates, 2 per 1,000 experience hypoxia/ischemia at birth, resulting in hypoxic-ischemic encephalopathy (HIE) and subsequent neurodevelopmental disorders, which impose a lifelong burden on families and society. Infants with stage 3 HIE exhibit a high incidence of convulsive syndrome and severe neurodevelopmental delay. Even in the absence of profound impairments, affected children may present with motor, cognitive, or behavioral abnormalities. The aim of our study was to analyze and identify maternal and neonatal risk factors associated with psychomotor developmental delay. Materials and methods of the study. The study enrolled 45 infants born to mothers with metabolic syndrome who were diagnosed with moderate or severe HIE in the early neonatal period; among them, 30 were preterm and 15 were full-term. Follow-up of infants continued until 9 months of age; сontinued observation was possible in 27 (17 preterm, 10 full-term) of the 45 infants diagnosed with moderate or severe HIE. Statistical analysis was performed using standard descriptive methods and simple and multiple logistic regression, including Poisson regression. The predictive value of maternal history, metabolic parameters, and other variables was evaluated using receiver operating characteristic (ROC) curve analysis with calculation of the area under the curve (AUC) in STATA 14.0. According to the decision of the Bioethics Commission of Poltava State Medical University No. 217 dated 12.06.2023, the study adhered to ethical standards for the humane treatment of patients, as defined by the Tokyo Declaration of the World Medical Association, the Helsinki Declaration, the Universal Declaration of Human Rights, the Council of Europe Convention on Human Rights and Biomedicine, current Ukrainian legislation, orders of the Ministry of Health, and the Code of Ethics for Physicians of Ukraine. Informed consent was obtained from all parents. The study was conducted as part of the scientific research project of the Department of Pediatrics No. 1 with Neonatology, Poltava State Medical University, No. 0120U102856: “Development of clinical and laboratory criteria and methods for predicting and preventing metabolic disorders in young children.” Research results. Most infants included in the study were diagnosed with moderate asphyxia at birth. The Apgar scores at 1 and 5 minutes were 5.83 ± 0.23 and 6.7 ± 0.14 points, respectively, in preterm infants, and 5.8 ± 0.52 and 7.2 ± 0.34 points in full-term infants. Mechanical ventilation was required immediately after birth in 16.7% of preterm infants with HIE and 46.7% of full-term infants; the duration of mechanical ventilation was 4.26 ± 0.7 days in preterm infants and 5.47 ± 1.26 days in full-term infants. The most common neurological conditions at 9 months of age were hypoxic-ischemic CNS injury and psychomotor delay. The frequency of hypoxic-ischemic CNS injury was 88.2% in preterm infants and 60% among full-term infants. The incidence of psychomotor delay was 20.0% in preterm infants and 17.6% in full-term infants. Multivariate logistic regression analysis, adjusted for gestational age at birth, was used to develop predictive models for psychomotor delay. The Apgar score and duration of mechanical ventilation were identified as the strongest prognostic factors. . The predictive model including the duration of mechanical ventilation demonstrated a reliable predictive coefficient (β = 0.31; 95% CI, 0.11–0.50) with an area under the ROC curve of 0.9180. Conclusion. Approximately 20% of infants with HIE, both preterm and full-term, exhibited delayed neurodevelopment at 9 months of age. Multivariate logistic analysis identified low Apgar scores and prolonged mechanical ventilation as the most significant prognostic factors, with the predictive model incorporating ventilation duration demonstrating the highest predictive accuracy. These findings highlight the need to optimize medical monitoring and follow-up of infants with HIE born to mothers with metabolic syndrome.

Recent studies have demonstrated a direct association between hypoxic-ischemic encephalopathy (HIE) and maternal overweight or obesity. HIE remains one of the leading causes of neonatal mortality and long-term neurological impairment in newborns. It represents a systemic condition affecting not only the central nervous system but also multiple peripheral organs, including the liver. One of the indirect biochemical indicators of hypoxic injury is elevated hepatic transaminase activity — specifically, alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Particular emphasis has been placed on the involvement of nitric oxide (NO) in the pathophysiology of HIE Objective: to analyze and identify risk factors for hypoxic-ischemic encephalopathy in full-term and premature infants born to mothers with metabolic syndrome, to build clinical prognostic models. Materials and methods. The study cohort consisted of 125 neonates. The study cohort consisted of 125 neonates. Group 1 included infants born to mothers with metabolic syndrome who were diagnosed with HIE (n = 45), while Group 2 included infants born to mothers without metabolic syndrome and without clinical manifestations of HIE (n = 79). ). Nitrite concentrations were measured by quantifying the diazo compounds formed via reaction with sulfanilic acid, followed by a subsequent reaction with the Griess-Ilosvai reagent. The bioethical principles of the study were confirmed by the Commission on Ethical Issues and Bioethics of Poltava State Medical University (protocol No. 233 dated 21 December 2024). Statistical analysis was conducted using the STATA 14.0 software package. Statistical analysis was performed using the STATA 14.0 software package. The work was carried out within the research project of the Department of Paediatrics No. 1 with Neonatology of Poltava State Medical University ’’To develop clinical and laboratory criteria, methods for predicting and preventing metabolic disorders in young children’’ (state registration number 0120U102856, term of completion 2020-2024). Results of the study and their discussion. Initial clinical assessments were performed immediately postnatally. Among term infants, the most frequently diagnosed conditions included neonatal depression syndrome, respiratory distress syndrome, heart failure, seizures, and muscular dystonia. In preterm infants, depression syndrome, muscular dystonia, and seizures were the predominant clinical manifestations. HIE in term neonates is typically characterized by multisystem involvement, whereas in preterm infants, clinical signs are predominantly neurological. A metabolic profile analysis confirmed the multisystemic nature of HIE. On day one of life, AST and ALT levels in preterm infants with HIE were significantly elevated compared to preterm infants without HIE (p = 0.055 and p = 0.049, respectively). The urinary nitrite level in preterm infants with HIE was significantly higher compared to that in preterm infants without HIE (p = 0.025). Similar findings were observed in term infants, where those with HIE exhibited elevated urinary nitrite levels (p = 0.042). Based on these data, we developed predictive models for the early neonatal onset of HIE. After adjusting for gestational age and ALT or AST activity, urinary nitrite concentration proved to be a reliable prognostic marker for HIE in both preterm and term neonates. The areas under the ROC curves for models incorporating urinary nitrite levels, gestational age, and either ALT or AST were 0.9952 and 0.9279, respectively. Conclusions. HIE in neonates is associated with multisystem involvement, affecting various organs in addition to the central nervous system. In light of the systemic nature of the condition, potential metabolic biomarkers were evaluated. Among these, serum ALT and AST levels, as well as urinary nitrite concentration, emerged as significant risk factors for early neonatal HIE in infants born to mothers with metabolic syndrome. Due to the heterogeneity of the cohort in terms of gestational age, multiple logistic regression analysis with obligatory adjustment for gestational age was employed to ensure accurate interpretation of the findings. A predictive model for HIE development was constructed, incorporating urinary nitrite and ALT levels. Newborns diagnosed with HIE and born to mothers with metabolic syndrome require close monitoring of ALT and AST levels during the early neonatal period. Furthermore, urinary nitrite concentration serves as an optimal biomarker for predicting HIE onset.

Aim - retrospective analysis of the catamnesis observation of children with eventration/relaxation of the diaphragm due to birth trauma with surgical reinnervation and plication. Clinical cases. Two children were followed up and analyzed, both born with full relaxation of the diaphragm dome on the right side, Erb-Duchenne plexitis, hydrocephalic syndrome due to hypoxic-ischemic brain injury, and pneumonia. The diagnosis was established in the early neonatal period, and the severity of respiratory disorders required respiratory support (mechanical ventilation of the lungs) and intensive therapy in the newborn intensive care unit. The clinical cases presented demonstrate a rare pathology of birth paralysis of the diaphragm dome due to damage to the phrenic nerve. The patients were treated surgically by means of reventration of the diaphragm dome using the plication method with simultaneous reinnervation. These surgical interventions had a 100% success rate among the patients. Conclusions. Diaphragmatic eventration is a rare pathology accompanied by respiratory disorders, severe respiratory failure requiring mechanical ventilation with subsequent surgical intervention to restore diaphragm function. Correction of the diaphragm eventration using the plication method with simultaneous reinnervation is the most effective surgical method of treatment in newborns. The research was carried out in accordance with the principles of the Declaration of Helsinki. Informed consent of the child and child's parents was obtained for the studies. No conflict of interest was declared by the author.

Aim: to determine the influence of maternal and neonatal variants of the eNOS (G894T, rs1799983) and IL1B (C3953T, rs1143634) genes and their intergenic interactions on the development of HIE in newborns.Material and methods: The study included a cohort of 105 newborns and their 99 mothers. Determination of variants of the genes eNOS (G894T, rs1799983) and IL1B (C3953T, rs1143634) was carried out for the patients of study groups.Results: The frequency of detection of the 894TT genotype by the eNOS gene was increased in newborns with severe asphyxia (p=0.018) and in their mothers (p=0,0057). Further analysis of intergenic interactions, performed in mother-child pairs, revealed an increased frequency of the neonatal 894GG (eNOS)/maternal 3953С (IL-1B) genotype combination in the comparison group versus the group of newborns with HIE (p=0.007).Conclusions: The significance of the intergenic maternal combination of 894GG/3953CT genotypes for the eNOS and IL1B genes and the intergenic combination of neonatal 894GG (eNOS)/maternal 3953CT (IL-1B) genotypes in the development of HIE in newborns has been proven. Associations of maternal and neonatal 894TT genotypes for the eNOS gene with the development of severe asphyxia, bradycardia, and respiratory failure were found in newborns with HIE.

Background The pro-inflammatory cytokine IL-1 plays an important role in severe COVID-19. A change in IL-1 production may be associated with a mutation in the IL1Β gene. Our study analyzed the impact of the IL1Β gene variants (rs1143634) on disease progression in patients with severe COVID-19 pneumonia, taking into account treatment strategies. Methods and results The study enrolled 117 patients with severe COVID-19 pneumonia. The IL1Β gene variants were identified using the polymerase chain reaction-restriction fragment length polymorphism method. In the group of patients, the following genotype frequencies were found based on the investigated rs1143634 variant of the IL1Β gene: CC–65.8%, CT–28.2%, and TT–6.0%. Our results showed that the group of patients with the T allele of the IL1Β gene had higher leukocyte counts (p = 0.040) and more pronounced lymphopenia (p = 0.007). It was determined that patients carrying the T allele stayed on ventilators significantly longer (p = 0.049) and required longer treatment with corticosteroids (p = 0.045). Conclusion Identifying variants of the IL1Β gene can be used as a predictive tool for assessing the severity of COVID-19 pneumonia and tailoring personalized treatment strategies. Further research with a larger patient cohort is required to validate these findings.

Background: Previous studies indicate a close relationship between the severity of COVID-19 and oxidative stress. N-acetyltransferase 2 (NAT2) is an enzyme that metabolizes a wide range of xenobiotics and plays an important role in the regulation of reactive oxygen species, consequently contributing to the development of oxidative stress. Aim: To determine the impact of NAT2 gene variants on the risk of developing and the progression of severe COVID-19-associated pneumonia in patients from the Poltava region of Ukraine. Methods: The study included 117 patients who were diagnosed with severe COVID-19 pneumonia and received treatment in the intensive care unit. The NAT2 gene variants were identified through the PCR-RFLP method. Results: The presence of the AA genotype of the A803G variant in patients with severe COVID-19 pneumonia is associated with an almost 3-fold reduction in the risk of lethal outcomes. The presence of the TT genotype of the C481T variant was associated with the need for artificial lung ventilation. Patients in the study group with a heterozygous GA genotype of the G590A variant demonstrated a notable rise in the risk of developing systemic inflammatory response syndrome and acute respiratory distress syndrome, with a nearly 2.5-fold and 3-fold increase, respectively. The G857A variant was not associated with the risk of developing the above-- mentioned complications in the examined patients. Conclusion: The obtained results suggested that the NAT2 gene variants might influence the incidence, course, and adverse consequences of COVID-19.

COVID-19 was first reported in December 2019 in Wuhan (Hubei Province, China). Later, the pandemic of this disease took the world by storm, challenging the medical community. Its clinical manifestations vary from asymptomatic to a severe course that requires hospitalization and intensive therapy with oxygen support. The mortality rate in patients with a severe course of COVID-19 can exceed 50% The majority of fatal cases of COVID-19 were associated with thrombotic events, despite the prophylactic use of anticoagulant therapy. Numerous theoretical overviews and research articles indicate the need for genetic testing in patients with COVID-19 to determine the genetic profile of proteins involved in thrombophilia. According to the researchers, the Leiden mutation (G1691A, rs6025) of the FV gene is one of the promising candidates for testing. The aim of the work was to demonstrate the clinical features of the severe course of COVID-19 in the presence of the Leiden mutation. 58 patients with COVID-19 of the intensive care unit were genotyped. The Leiden mutation in the heterozygous state was found only in one patient, who had Down syndrome. The Leiden mutation was detected with a frequency of 1.72% in the investigated group. The described clinical case clearly showed that individuals with Down syndrome, associated with hereditary thrombophilia are at risk of undesirable clinical consequences in the treatment of COVID-19. The condition of the patient with the Leiden mutation was severe when admitted to the hospital. The score according to the sequential organ failure assessment scale was 2 points. Bilateral multisegmental pneumonia was detected on the X-ray. On the second day after admission, due to the development of acute respiratory distress syndrome and multiple organ failure, the patient was transferred to the intensive care unit, where he received oxygen therapy through a facial mask. Medical treatment was carried out according to the protocol: non-steroidal anti-inflammatory drugs, antibacterial therapy, anticoagulants, sympatho¬mimetics, and glucocorticosteroids. Despite the medical measures taken, progression of respiratory failure, renal failure, and portal hypertension was noted. On the 11th day, the patient developed asystole. Resuscitation measures were unsuccessful. Thus, the described case of a severe course of COVID-19 in a carrier of a heterozygous variant of the Leiden mutation with Down syndrome confirms the recommendations regarding the need for genetic testing for thrombophilia in high-risk groups and the appointment of personalized measures to prevent complications.

Background: Pneumonia is one of the most common complications of the course of COVID-19. Interferon-γ (IFN-γ) is an essential cytokine for lung defense against intracellular and extracellular pathogens. Since the rs2430561 variant of the IFNG gene, which encodes IFN-γ, affects the production of IFN-γ, it can potentially be associated with a severe course of COVID-19. Objective: The aim of our research was to determine the impact of the rs2430561 variant of the IFNG gene on the propensity of patients to develop and experience a severe course of COVID-19-related pneumonia. Methods: The study included 117 patients with a complicated course of COVID-19- associated pneumonia, who were hospitalized in the intensive care unit. All patients received a standard clinical and laboratory evaluation and course of treatment for COVID- 19. IFNG gene variants were determined by PCR method. For comparisons, we used clinical and laboratory indicators from the inpatients’ medical records, which reflected the course of the patient's disease, taking into account the rs2430561 variants of the IFNG gene. Results: In the group of patients, the following genotype frequencies were found: TT – 18.8%, TA – 52.1%, and AA – 29.1%. The results showed that AA genotype carriers had essentially higher SpO₂/FiO₂ ratio (p=0.043). High base excess rates were present in patients with the TT genotype (p=0.047). It was found that patients with the T allele (whether homozygous or heterozygous state) had significantly higher concentrations of such electrolytes as potassium and sodium (p=0.016 and p=0.004, respectively). Сonclusion: In patients with COVID-19, the rs2430561 variant of the IFNG gene is linked to a complicated course of pneumonia. Certainly, this finding requires further research.

Objectives: COVID-19 continues to range around the world and set morbidity and mortality antirecords. Determining the role of genetic factors in the development of COVID-19 may contribute to the understanding of the pathogenetic mechanisms that lead to the development of complications and fatalities in this disease. The aim of our study was to analyze the effect of TNF-α (rs1800629), IL-6 (rs1800795) and VDR (rs731236 and rs1544410) genes variants on the development risk and the course of COVID-19 in intensive care patients. Methods: The study group included 31 patients with diagnosis “viral COVID-19 pneumonia”. All patients underwent standard daily repeated clinical, instrumental and laboratory examinations. Determination of IL-6 , TNF-α , and VDR genes variants was performed using the PCR-RFLP method. Results: It was found a significant increase in the rate of the CC genotype and C allele (38.7 vs. 12.0% and 0.6 vs. 0.4%, respectively) of the IL-6 gene in all patients of the study in comparison with population frequencies. There was a significantly higher rate of heterozygous genotypes TC and GA of the VDR gene in group of died patients. The rs1800629 variant of the TNF-α gene is associated with the need for respiratory support and its longer duration in patients with COVID-19. Conclusions: The obtained results support a hypothesis about the influence of variants of IL-6 , TNF-α and VDR genes on severity of COVID-19. However, in order to draw definite conclusions, further multifaceted research in this area are need.