THE ROLE OF HEREDITARY PREDISPOSITION (POLYMORPHIC MARKERS OF GLUTATHIONE-S-TRANSFERASE, CATALASE, ENDOTHELIAL NITROGEN OXIDE SYNTHASE GENES) AND SOME ADVERSE ENVIRONMENTAL FACTORS IN THE DEVELOPMENT OF BRONCHO-OBSTRUCTIVE PATHOLOGY IN CHILDREN LIVING IN RADIOACTIVELY CONTAMINATED AREAS. / РОЛЬ СПАДКОВОЇ СХИЛЬНОСТІ (ПОЛІМОРФНИХ МАРКЕРІВ ГЕНІВ ГЛУТАТІОН-S-ТРАНСФЕРАЗ, КАТАЛАЗИ, ЕНДОТЕЛІАЛЬНОЇ СИНТАЗИ ОКСИДУ АЗОТУ) ТА ДЕЯКИХ НЕСПРИЯТЛИВИХ ЧИННИКІВ ДОВКІЛЛЯ У ВИНИКНЕННІ БРОНХООБСТРУКТИВНОЇ ПАТОЛОГІЇ У ДІТЕЙ, МЕШКАНЦІВ РАДІОАКТИВНО ЗАБРУДНЕНИХ ТЕРИТОРІЙ.

OBJECTIVE: summarizing the results of many years of research by the authors on the influence of gene polymorphisms encoding xenobiotic biotransformation enzymes (GSTТ1, GSTM1, GSTР1), antioxidant protection (С^262Т of the catalase gene), endothelial nitric oxide synthase (4a/4b VNTR polymorphism of the eNOS gene), and some environmental factors on the occurrence of broncho-obstructive disorders and the development of bronchial asthma in children, residents of radioactively contaminated areas. MATERIALS AND METHODS: The examined school-aged children were residents of radioactively RCA who had no clinical signs of respiratory pathology. Deletion polymorphism of catalase gene (CAT C^262T), polymorphism of glutathione-S-transferase gene (GSTТ1, GSTM1, GSTР1) and the polymorphism in the 4th intron (4a/4b) of the eNOS gene were studied in the molecular genetics laboratory of the State Institution «Reference Center for Molecular Diagnostics of Public Health Ministry of Ukraine¼. Molecular genetic studies were performed by polymerase chain reaction. The study of the ventilation lung capacity was carried out by the method of computer spirometry based on the data of the «flow-volume¼ loop analysis. A pharmacological inhalation test with a bronchodilator drug which affects the ß2-adrenergic receptors of the lungs was used to detect early changes in the ventilatory lung capacity - bronchial hyperreactivity. RESULTS AND CONCLUSIONS: One of the leading mechanisms, due to which the implementation of hereditary predisposition to bronchial asthma in children living in radioactively contaminated areas is the polymorphism of certain genes of glutathione-S-transferase, catalase, endothelial nitric oxide synthase. With such polymorphic variants of the GST genes, isoforms of enzymes with reduced activity are produced, which limits their ability to effectively neutralize free radicals, which are formed in excess when free radical oxidation processes are activated due to the constant intake of radionuclides with a long half-life into the body of children. Unfavorable factors that increase the risk of developing broncho-obstructive disorders and the likelihood of their implementation in the form of bronchial asthma in children, residents of radioactively contaminated areas, have been identified. It has been established that among them the leading role is played by hereditary predisposition to this disease. On the part of the child, such negative factors were unfavorable conditions of intrauterine development, the presence of signs of exudative-catarrhal diathesis, manifestations of allergies and frequent respiratory diseases from the first months of life.

CATALASE CRS-262T GENE POLYMORPHISM AND CHANGES IN VENTILATION LUNG CAPACITY IN CHILDREN-RESIDENTS OF RADIOACTIVELY CONTAMINATED TERRITORIES. / POLIMORFIZM SRS-262T GENA KATALAZY I ZMINY VENTYLIaTsIINOÏ SPROMOZhNOSTI LEGENIV U DITEI - MEShKANTsIV RADIOAKTYVNO ZABRUDNENYKh TERYTORII.

OBJECTIVE: to determine the association of catalase С-262Т gene polymorphism with the presence of bronchial hyper-reactivity in children living in radioactively contaminated territories. MATERIALS AND METHODS: There were examined school-age children-residents of radioactively contaminated territories (RCT), who did not have clinical signs of respiratory pathology. Catalase (CAT) С-262Т gene deletion polymorphism was studied in the molecular genetic laboratory of the State Institution «Reference Center for Molecular Diagnostic of Public Health Ministry of Ukraine¼. Determination of the polymorphic variant by the catalase С-262Т gene was performed by Polymerase Chain Reaction (PCR) using specific oligonucleotide primers, followed by Restriction Fragment Length Polymorphism (RFLP) analysis. The CAT С-262Т gene polymorphism in children living in RCT was compared with that in the reference group of practically healthy individuals. Ventilation lung capacity was performed by computer spirometry according to the analysis of the loop «the flow-volume¼. A pharmacological inhalation test with a bronchodilator that acts on ß2-adrenergic receptors of the lungs was used to detect early changes in the ventilatory capacity of the lungs - bronchial hyperreactivity. RESULTS: Comparative analysis showed that in the presence of bronchial hyperreactivity in children living in RCT, the CT genotype was more common than in children without bronchial hyperreactivity, and the frequency of the CC genotype was correspondingly reduced. There was a trend towards a decrease in the frequency of the TT genotype. An analysis of the frequency distribution of allelic variants of the CAT С-262Т gene polymorphism in children living in the RCT revealed a tendency to increase in the frequency of the T-allele and according to the decrease in the frequency of C-allele in the presence of bronchial hyperreactivity.Сonclusions. Thus, among children living in RCT, CT-homozygotes of CAT С-262Т gene polymorphism had bronchial hyperreactivity probably more often than CC-heterozygotes. In the presence of bronchial hyperreactivity, there was a trend towards an increase in the frequency of the T-allele and, accordingly, a decrease in the frequency of the C-allele.

FUNCTION OF EXTERNAL RESPIRATION AND MARKERS OF OXIDATIVE STRESS IN CHILDREN-RESIDENTS OF RADIOACTIVELY CONTAMINATED TERRITORIES. / FUNKTsIIa ZOVNIShN'OGO DYKhANNIa I MARKERY OKSYDATYVNOGO STRESU U DITEY̆ ­ MEShKANTsIV RADIOAKTYVNO ZABRUDNENYKh TERYTORIY̆.

OBJECTIVE: to evaluate the influence of processes of lipid peroxidation, and antioxidant protection on the function of external respiration in children-residents of radioactive contaminated territories. MATERIALS AND METHODS: There were examined children of school age, inhabitants of radioactive contaminated ter- ritories (RCT) without respiratory and pulmonary pathology, and patients with bronchial asthma (BA). Examination of the ventilation lung function was performed by the method of pneumotachography according to the analysis of the «flow-volume¼ loop. End products of lipid peroxidation (LPO), reacting with thiobarbituric acid (malondialde- hyde), enzymes-antioxidants - catalase, superoxide dismutase, glutathione peroxidase, glutathione transferase were studied to determine the signs of oxidative stress. RESULTS: Examinations of ventilation lung function in children-residents of RCT without respiratory and pulmonary pathology showed no significant deviations of its parameters beyond the limits of physiological fluctuations; a decrease in bronchial patency at different levels of the bronchial tree was established in children with BA. An increased incidence of bronchial hyperreactivity was noted in both the patients with BA and those who did not have bronchopulmonary pathology. The absence of an increase in the content of LPO products (malondialdehyde) in blood serum of children-residents of RCT with increased activity of catalase and group of glutathione enzymes indi- cates the sufficiency of compensatory possibilities of antioxidant protection. CONCLUSIONS: Studies of correlation between the function of external respiration and markers of oxidative stress determined that the indices of bronchial patency directly correlate with the activity of enzymes-antioxidants of the glutathione group and inversely - with the content of the LPO products in children-residents of RCT. The frequency of bronchospasm inversely correlated with the activity of glutathione group antioxidants. There are inverse correla- tion of 137Cs content in the body with the activity of glutathione transferase and glutathione peroxidase.

FEATURES OF NITRIC OXIDE METABOLISM AND RISK OF DEVELOPING ENDOTHELIAL DYSFUNCTION IN CHILDREN WITH e-NOS GENE 4a/4b POLYMORPHISM UNDER LONG-TERM ENTERING 137Cs TO BODY. / OSOBLYVOSTI METABOLIZMU OKSYDU AZOTU TA RYZYK ROZVYTKU ENDOTELIAL'NOÏ DYSFUNKTsIÏ U DITEI Z 4a/4b POLIMORFIZMOM GENA eNOS PRY TRYVALOMU NADKhODZhENNI 137Cs DO ORGANIZMU.

OBJECTIVE: To determine the features of the nitrogen oxide metabolism and risk of developing endothelial dysfunc-tion in children with e-NOS 4a/4b gene polymorphism, who live under prolonged enter 137Cs to the body. MATERIALS AND METHODS: There were examined 117 children-residents of radioactively contaminated territories and50 children of control group. The level of stable metabolites was defined in blood serum (NO2- and NO3-). The ther-mographic method was used to register the endothelium dependent reaction of the vascular bed to changes in theblood supply. The ventilation capacity of the lungs was evaluated using this method of pneumotachography.Polymorphism in intron 4 of the gene e-NOS was studied by the method of polymerase chain reaction. The contentof 137Cs in the body of children was determined using a human radiation counter Skrynner M-3. RESULTS AND CONCLUSIONS: In children-residents of radioactively contaminated territories with genotype 4a/4b com-paring to children who had genotype 4b/4b, the decrease in the nitric content of in the blood serum, the increase inthe thermographic index of the recovery period of blood circulation to the baseline level after occlusion test werenoted, that is indicative of the decreased NO-synthase active of vascular endothelium in the carriers of the minorallele a in the 4th intron of gene eNOS (genotype 4a/4b), and is a risk factor for development of endothelial dysfunc-tion. It was proved a decrease in the index of lung tissue elasticity and stretchability - FVC / NFVC of the lungs com-paring to children with genotype 4b/4b, there was a reduction of integral index of respiratory tract permeability -FEV1/NFEV1. The inverse correlation dependence between the presence of allele a in the genotype and the values ofFVC/NFVC of the lungs (r = -0.259; p <0.05) and FEV1/NFEV1 (r = -0.2267; p <0.05) was found. Signs of bron-chospasm were found in the carriers of the allele a in 1.5 times more often than in children-carriers of homozy-gotes from allele b.

Efficiency of endothelial dysfunctioncorrection in children residents of radioactively contaminated areas using the method of intermittent normobaric hypoxi therapy. / EFEKTYVNIST' KOREKTsIÏ ENDOTELIAL'NOÏ DYSFUNKTsIÏ U DITEY̆ MEShKANTsIV RADIOAKTYVNO ZABRUDNENYKh TERYTORIY̆ ZA DOPOMOGOIu METODU PERERYVChASTOÏ NORMOBARYChNOÏ GIPOKSYTERAPIÏ.

Objective is to evaluate the effectiveness of intermittent normobaric hypoxi therapy as the means that can posi tively influence on the different links in the pathogenesis of endothelial dysfunction in children residents of radioactively contaminated areas. MATERIALS AND METHODS: Disorders of personal and emotional sphere, symptoms of asthenic vegetative and neurot ic character, the signs of the cardiovascular and respiratory systems, abdominal and dyspeptic syndromes are attrib uted to the clinical signs of endothelial dysfunction. There were determined the biochemical parameters of the con tent of stable metabolites of NO, L arginine, lipid peroxidation, antioxidant enzymes in the blood serum; the indices of cellular and humoral immunity; the instrumental indices of vascular endothelium dependent reaction to occlu sion test, ventilating ability of the lungs, the bioelectric activity of the myocardium. Intermittent normobaric hypoxi therapy (INHT) was performed using the individual device of mountain air of the «Borei¼ type produced by State Research Medical Engineering Center «NORT¼ NAS of Ukraine (Kyiv). RESULTS AND CONCLUSIONS: The clinical examination showed that the administration of additional intermittent nor mobaric hypoxi therapy to the basic treatment of 44 children with endothelial dysfunction who live in the radioac tively contaminated areas, promoted to reduce the frequency of complaints and intensity of clinical manifestations in the mentioned symptom complex. An increase of serum L arginine in the absence of significant changes in the content of NO stable metabolites was revealed in the most examined children residents of radioactively contami nated areas by studying NO system after using the course of intermittent normobaric hypoxi therapy. Positive changes in vascular endothelium dependent reaction to occlusion test, indicating the reduction length of the recovery period for thermographic circulation index after occlusive test, were determined. The decrease in the inten sity of lipid peroxidation processes - a reduction of serum LPO end products content that react with thiobarbituric acid - was observed. The bronchospasm elimination was revealed in a large number of children with the study of the lung ventilation capacity. There were trends to reduce the imbalance of different chain of the heart rhythm man agement. The immunomodulatory effect was detected as: the increase in the functional activity of neutrophils, the normalization of humoral immunity link indices. Intermittent normobaric hypoxi therapy is an effective tool that can positively affect the condition of nitric oxide system and endothelium dependent functions of the organs and systems in children residents of radioactively contaminated areas.

Correction of endothelial dysfunction in children residents of radioactively contaminated areas by nitric oxide donator. / KOREKTsIIa ENDOTELIAL'NOÏ DYSFUNKTsIÏ U DITEY̆ MEShKANTsIV RADIOAKTYVNO ZABRUDNENYKh TERYTORIY̆ ZA DOPOMOGOIu DONATORA OKSYDU AZOTU.

OBJECTIVE: to determine the effectiveness of NO L arginine donator for correcting endothelial dysfunction in chil dren residents of radioactively contaminated areas. MATERIALS AND METHODS: A group of children residents of radioactively contaminated areas with the presence of clinical and paraclinical signs of endothelial dysfunction was selected to assess the effectiveness of correcting endothelial dysfunction by NO L arginine donator according to selective screening. Biochemical parameters for the content of stable NO metabolites, serum L arginine; instrumental parameters of endothelium dependent vascular reaction on occlusion test, the ventilation capacity of the lungs were determined. A course of NO BetarhinТ dona tor, which active substance is L arginine, was received by examined children. RESULTS AND CONCLUSIONS: A group of children residents of radioactively contaminated areas with the presence of clinical and paraclinical signs of endothelial dysfunction was selected according to selective screening. The pro nounced positive therapeutic effect, good tolerability, absence of side effects were observed in children with endothelial dysfunction, who were received a course of NO L arginine donator. It has been established that L argi nine use contributed to increasing the content of serum arginine, nitrite, nitrate, and amounts of NO metabolites in children with endothelial dysfunction. Bronchospasm elimination was noted after L arginine use in the significant part of examined patients. An increase in the diameter of the brachial artery and its space in the test with post occlusion reactive hyperemia after the use of L arginine course was revealed by an evaluation of indicators for endothelium dependent vasodilatation of the large vessels using ultrasound method visualizing the brachial artery lumen. This indicates an increase in endothelial vasomotor capacity.

Influence of eNOS gene 4a/b VNTR polymorphism on development of endothelial dysfunction and respiratory system disorders in children - residents of radioactively contaminated areas. / VPLYV VNTR (4a/b) POLIMORFIZMU GENA eNOS NA ROZVYTOK ENDOTELIAL'NOI DYSFUNKTsII TA PORUShEN' SYSTEMY DYKhANNIa U DITEY - MEShKANTsIV RADIOAKTYVNO ZABRUDNENYKh TERYTORIY.

UNLABELLED: Objective is to study the role of еNOS gene polymorphism in the development of endothelial dysfunction and func tional disorders of the respiratory system in children - residents of radioactively contaminated areas. MATERIALS AND METHODS: Total of 184 children were examined to determine the possible associative links between еNOS gene intron 4m polymorphism and indices characterizing the functional state of the endothelium and the bronchopulmonary system. Of them the main group of children consisted of 135 children residents of radioactively contaminated areas, the control group - of 49 healthy children who lived in "clean" regions by the radioactive con tamination and were not belonging to victims of the Chernobyl accident contingents. The molecular genetic study determining the еNOS gene intron 4 polymorphism was performed. An examination of lung ventilation capacity was evaluated using the pneumotachography method. The thermographic method was used for registering the endothe lium dependent reaction of the vascular bed to changes in blood supply conditions. The levels of nitrogen com pounds (NO2 and NO3 ) were determined by the standard method using Hriss reagent. The level of body incorporat ed 137Cs was determined using human radiation counter Skrynner 3M (Institute of Human Ecology). RESULTS: Children of the main group did not differ from that of control and literature data by the frequency of geno types 4a/b and еNOS gene allele polymorphism. An increase in the duration of circulation restoration up to the ini tial level after occlusion test was noted in children of the main group with genotype 4a/4b comparing to children with genotype 4b/4b; a decrease in the nitrite content was observed in blood serum. The integral indices of elastic ity and tensility of the lung tissue, respiratory permeability were decreased; the signs of bronchospasm were record ed 1.5 times more often. These adverse effects were associated with the presence of genotype allele 4a, the signs of endothelial dysfunction and the level of the incorporated 137Cs. CONCLUSIONS: More pronounced changes in vascular endothelial reaction to occlusion and the significant decrease of serum nitrite level in allele 4a carriers, especially in children who live in adverse ecological conditions, give the right to include them into the risk group for the development of endothelial dysfunction, and to conside pulmonary dysfunction as one of its manifestations.

Changes in stable metabolites of nitric oxide in children-residents of radioactively contaminated territories. / Zminy stabil''nyh metabolitiv oksydu azotu u ditejmeshkanciv radioaktyvno zabrudnenyh terytorij.

Objective. An evaluation of the changes in nitric oxide system by the level of its stable metabolites in the blood serum and their comparison with content of 137Сs in the body of children-residents of radioactively contaminated territories. Materials and methods. The children of school age, the residents of radioactively contaminated territories were examined. The levels of nitrogen compounds (NO2- and NO3-) were determined by the standard method using Grissreagent. 137Сs content was determined in the children body by whole-radioactive body counter. Results. Studies of nitric oxide metabolite content in the blood serum revealed the decrease in nitrite level under an absence of the significant changes in nitrate content and sum of NO metabolites in the children-residents of radioactively contaminated territories. An increase in 137Cs content in the body was accompanied by a decrease of nitrite level in the blood serum of a significant part of children.