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УДК 614.876:616-053.2-008.9:612.392.64(477.41 )
https://doi.org/10.32402/dovkil2020.04.021
rinEPrOMOUHCTEIHEMIH y flITEH TA niCOBI nQWEWI y HOPHOEHnbCbKIH 3OHI BIflHyWEHHH
BaHfla^eBCbKMM 10.1., AyôoBa H.O.
HYPERHOMOCYSTEINEMIA IN CHILDREN AND FOREST FIRES IN THE CHORNOBYL EXCLUSION ZONE
1 BAN DAZ H EVS KY Yu.I., 2DUBOVA N.F.
1Ecology and Health Coordination and Analytical Centre, Ivankiv
2National P.L. Shupyk Medical Academy of Postgraduate Education, Kyiv
elevance of the problem.
Studies conducted during the implementation of projects of the European Commission and the Rhône-Alpes Region (France) have found increased blood levels of homocysteine - a metabolic product of the essential amino acid methion-ine - in a significant number of examined adolescent children living in raions located near the Chornobyl nuclear power plant [1].
Given that hyperhomocys-teinemia is associated with the development of severe pathological processes, including cardiovascular diseases and cancers, it is important to determine the cause of its occurrence [2, 3].
Hyperhomocysteinemia is known to be linked to genetic defects of folate metabolism, primarily to the MTHFR:C677T polymorphism [4].
Among environmental factors that can cause hyperhomocys-teinemia, one should also men-
tion products of burning of wood and peat in the form of finely dispersed particles with a size of 0.1-2.5 Mm (PM2.5), including black carbon, that can penetrate into the respiratory tract, reaching the bronchioles and alveoli.
Long-term exposure to PM2.5 can cause the development or exacerbation of car-diometabolic disorders [6].
An association has been found between wood burning and increased blood homocys-teine levels in the group of elderly men [7].
It should be noted that fires in the Chornobyl exclusion zone (ChEZ) are quite common. The largest forest and meadow fire in Ukraine over the past 25 years on an area of 10127 hectares occurred on April 2629, 2015.
The maximum contamination density of the territory in the areas of ground forest fires in separate sections of Lubianka Forest Division was 1040
Г1ПЕРГОМОЦИСТЕ1НЕМ1Я У Д1ТЕЙ ТА Л1СОВ1ПОЖЕЖ1 У ЧОРНОБИЛЬСЬКЙ ЗОН1В1ДЧУЖЕННЯ
1 Бандажевський Ю.1., 2Дубова Н.Ф.
1 Координатйний аналтичний центр «Екологя iздоров'я», м. 1ванюв, Укра/на 2Нащональна медична академ1я пiслядипломноí освти iM. П.Л. Шупика, м. Ки/в, Укра/на
Мета досл'щження. Визначити роль геноти-niB фолатного циклу у виникненнi ппергомо-цистеíнемií у дтей пiсля л'юово)' пожежi у Чорнобильсьюй зон в'щчуження (ЧЗВ).. Методи: iмунохiмiчний, ПЛР у режимi Realtime, математико-статистичний. Результати. Обстеження 84 дтей п^тково-го вку i3 Полiського району Ки)'всько)' облает дозволило виявити у них змни метаболiчних проце^в у вигляд пдвищеного продукування гомоцисте/ну у зв 'язку з леовою пожежею у ЧЗВ 26-29 квтня 2015 року. Проведений
пор'!вняльний анал'з питомоI ваги генотипiв фолатного циклу у п'щгрупах дтей з пперго-моцистеíнемieю до / пiсля л 'юово)' пожежi показав непричетнiсть до цього явища ендо-генного фактора у вигляд генетичного апара-ту, що контролюе синтез ферменлв фолатного циклу.
Висновки. Пожежi л'юу \ торфовищ у ЧЗВ е однею з основних причин пщвищення р'вня гомоцисте1ну у кров'1 дтей з прилеглих рай-он'в незалежно в'щ стану генетично1 системи фолатного циклу. Контроль над рiвнем гомо-цисте1ну у кров '1 дтей \ дорослих, як перебу-вають п'щ впливом продуктв горiння дереви-ни, що мiстить радюактивнi елементи, е основним компонентом програми профлак-тики онколопчних / серцево-судинних захво-рювань.
Ключов'1 слова: фолатний цикл, гомоцисте/н, пдлтки, л'сов'! пожежi, Чорнобильська зона вдчуження.
© Бандажевський Ю.1., Дубова Н. Ф. СТАТТЯ, 2020.
21 Environment & Health № 4 2020
kBq/sq.m for i37Cs; 368 kBq/sq.m for 90Sr; 11.4 kBq/sq.m for 238-24opu and 14.4 kBq/sq.m for 241Am [8].
Thus, along with a genetic predisposition, forest fires in the ChEZ can be viewed as a source of hyperhomocysteine-mia in residents of the districts adjacent to it.
In order to find out the main cause of hyperhomocysteine-mia among the population of the districts located near the ChEZ, it is necessary to conduct a comparative assessment of homocysteine levels in the same group of subjects before and after a forest fire, taking into account the proportions of folate metabolism genotypes.
The aim of this study was to determine the role of folate metabolism genotypes in the occurrence of hyperhomocys-teinemia in children after a forest fire in the ChEZ.
In this case, it is necessary to carry out a comparative assessment of proportions of folate metabolism genotypes in subgroups with hyperhomo-
cysteinemia before and after a forest fire in the ChEZ in the same group of children.
Material and methods.The
study was conducted within projects of the European Commission and the RhoneAlpes Regional Council (France) implemented in Ukraine in 2013-2017.
The examined group comprised 84 children (39 boys and 45 girls) living on the territory of Ukraine after the Chornobyl accident (Polessky district, Kiev region) with a 137Cs soil contamination level of <2 Curie/sq.km and bordering the ChEZ [9].
The children's average age at the time of examination was (15.5±0.1) years (95% CI 15.415.7 years).
In order to further determine homocysteine concentrations and genotype of folate metabolism (FM), each child had blood drawn from the ulnar vein after fasting in the morning twice on 02/04/2015 and 18/12/2015.
The blood samples were analysed at a laboratory certi-
Table 1
Dynamics of cases of hyperhomocysteinemia in groups
of children
Date of examination of children Number of cases of hyperhomocysteinemia in groups of children
Total (n = 84) Boys (n = 39) Girls (n = 45)
Abs. number % Abs. number % Abs. number %
02/04/2015 48 57.1 23 59.0 25 55.6
18/12/2015 67 79.8* 35 89.7** 32 71 1***
Note: * - statistical differences between hyperhomocysteinemia
rates in the total group of children: t = 3.26; p = 0.001481.
** - statistical differences between hyperhomocysteinemia rates
in the group of boys: t = 3.30; p = 0.001689.
*** - statistical differences between hyperhomocysteinemia
rates in the group of girls: t = 1.54; p = 0.128839.
Table 2
Frequency of polymorphic alleles of folate metabolism genes in children with blood homocysteine levels of over 10 Mmol/L during the measurement on 02/04/2015 (n = 48)
Gene, polymorphism «Neutral» allele Risk allele, heterozygous mutation form Risk allele, homozygous mutation form
Abs. number % Abs. number % Abs. number %
MTR:A2756G 35 72.9 11 22.9 2 4.2
MTHFR:A1298C 26 54.2 15 31.3 7 14.5
MTHFR:C677T 20 41.6 19 39.6 9 18.8
MTRR:A66G 7 14.6 20 41.7 21 43.7
fied under international quality standards with the agreement of the parents.
Blood homocysteine concentrations were measured using a chemiluminescent immunoassay (CLIA) method. Analyser and test kit: Architect 1000 (ABBOT Diagnostics (USA)). Blood homocysteine levels of over 10 Mmol/L were defined as hyperhomocysteinemia.
The following allelic variants were identified during genetic analysis of FM: C677T and A1298C of the MTHFR gene (methylenetetrahydrofolate reductase), A2756G of the MTR gene (B12-dependent methion-ine synthase) and A66G of the MTRR gene (methionine synthase reductase). A real-time PCR method was used. Analyser and test kit: DT-96 detecting thermocycler, DNA-Technology (Russia).
The statistical processing of the results obtained was performed using the IBM SPSS Statistics 22 software (USA). Arithmetic mean (M), ± standard error of mean (m), confidence interval for a mean value (95% CI), median (Me), interquartile range (IR), minimum and maximum parameter values and percentiles were calculated for the variables analysed.
The statistical significance of variables was assessed by determining a significance level for p with the help of the statistical software programme. The Student's t-test was used to compare relative values.
The critical level of significance for the null hypothesis (p) was set at 0.05.
Results and discussion. A statistically larger number of cases of hyperhomocysteine-mia was found in the total group of children and a group of boys during the measurement on 02/04/2015 compared with 18/12/2015 (table 1). There were no significantly statistical differences between hyperho-mocysteinemia rates during two measurements in the group of girls (table 1).
The comparison of proportions of similar genotypes of FM
polymorphisms in a total group of children, groups of boys and girls with hyperhomocysteine-mia reported on 02/04/2015 and 18/12/2015 showed no statistical differences (tables 27). A similar result was obtained when proportions of the MTHFR:677 CT/MTHFR:1298 AC heterozygous association were compared (table 8).
The results obtained indicate that a significant increase in the number of cases of hyperho-mocysteinemia was observed in the group of adolescent children living in the district bordering the ChEZ during the period between the measurements of this metabolite in the blood (02/04/2015-18/12/2015). Statistically significant differences between relative coefficients were reported in the total group of children and in the group of boys.
Statistical analysis showed that an endogenous factor in the form of a genetic apparatus regulating the synthesis of FM enzymes is not involved in this phenomenon. The proportions of genotypes including risk alleles did not differ statistically in these groups of children with hyperhomocysteinemia reported on 02/04/2015 and 18/12/2015.
Forest and peat fires officially reported in Sections 306-308 of Lubyanka Forest Division in the ChEZ during the periods 26-27/04/2015 and 2829/04/2015 on the territory with an area of 10127 hectares [8] and during the period from 29/06/2015 to 05/07/2015 on the territory with an area of 130 hectares should be probably considered as one of the causes of this phenomenon [10]. The maximum contamination density of the territory amounted to 1040 kBq/sq.m for 137Cs; 368 kBq/sq.m for 90Sr; 11.4 kBq/sq.m for 238-24opu and 14.4 kBq/sq.m for 241Am [8].
At the same time, finely dispersed burning particles with a size of 0.1-2.5 Mm (PM2.5), including black carbon, as well as radionuclides, being spread with air currents over long distances from the fire site were penetrating into the
respiratory tract of residents of localities situated near the ChEZ, reaching the bronchioles and alveoli [5].
Oxidative stress occurring with the involvement of homo-cysteine may be one of the main links in the pathogenesis of cardiovascular diseases [11], which are widespread among the population of the districts affected by the Chornobyl accident [12].
Thus, the negative effect of burning of forests containing radioactive elements on methionine metabolism resulted in an increase in blood homocysteine levels even in those children who did not have a genetic predisposition to this.
nPOB^EMM HOPHOBM^fl =
Experimental studies using laboratory animals conducted during the first years after the Chornobyl nuclear power plant accident also confirm the negative effect of incorporated radioactive agents on methion-ine metabolism.
Daily feeding of white rats for 28 days with whole oats containing 137Cs with the activity concentration of 445.7 Bq/kg and 90Sr with the activity concentration of 15.5 Bq/kg (animals of a control group received daily as food whole oats where 137Cs concentration was 44.2 Bq/kg and 90Sr concentration amounted to 1.7 Bq/kg) led to a significant decrease in methionine concentrations in the myocardium
Table 3
Frequency of polymorphic alleles of folate metabolism genes of children with blood homocysteine levels of over 10 Mmol/L during the measurement on 18/12/2015 (n = 67)
Gene, polymorphism «Neutral» allele Risk allele, heterozygous mutation form Risk allele, homozygous mutation form
Abs. number % Abs. number % Abs. number %
MTR:A2756G 47 70.1 18 26.9 2 3.0
MTHFR:A1298C 41 61.2 19 28.4 7 10.4
MTHFR:C677T 31 48.3 25 37.3 11 16.4
MTRR:A66G 13 19.4 29 43.3 25 37.3
Table 4 Frequency of polymorphic alleles of folate metabolism genes of boys with blood homocysteine levels of over 10 Mmol/L during the measurement on 02/04/2015 (n = 23)
Gene, polymorphism «Neutral» allele Risk allele, heterozygous mutation form Risk allele, homozygous mutation form
Abs. number % Abs. number % Abs. number %
MTR:A2756G 17 73.9 5 21.7 1 4.4
MTHFR:A1298C 10 43.5 10 43.5 3 13.0
MTHFR:C677T 9 39.1 9 39.1 5 21.8
MTRR:A66G 3 13.0 6 26.1 14 60.9
HYPERHOMOCYSTEINEMIA IN CHILDREN AND FOREST FIRES IN THE CHORNOBYL EXCLUSION ZONE 1 Bandazhevsky Yu.I., 1Dubova N.F.
1 Ecology and Health Coordination and Analytical Centre, Ivankiv 2National P.L. Shupyk Medical Academy of Postgraduate Education, Kyiv
Objective: We determined a role of folate metabolism genotypes in the occurrence of hyperhomocysteinemia in children after a forest fire in the Chornobyl exclusion zone (ChEZ). Material and methods: In the study, we applied immunochemical, Real-time PCR, mathematical and statistical methods. Results: A survey of 84 adolescents from the Poliske district, Kiev region revealed changes in their metabolic processes in the form of increased homocysteine production in connection with a forest fire in the ChEZ which was recorded during April 26-29, 2015. A compara-
tive analysis of the specific gravity of the folate cycle genotypes in the subgroups of the children with hyperhomocysteinemia before and after a forest fire showed that an endogenous factor in the form of a genetic apparatus, con-troling synthesis of folate cycle enzymes, was not involved in this phenomenon. Сonclusions: Forest and peat fires in the Chernobyl exclusion zone are one of the main causes for the elevation in blood homocysteine levels in children from the adjacent districts, regardless of the state of folate metabolism genetic system. Monitoring of the blood homocysteine levels in children and adults living under conditions of the exposure to wood combustion gases, containing radioactive elements, is a key component of the programmer for the prevention of cancer and cardiovascular diseases.
Keywords: folate metabolism, homocysteine, adolescents, forest fires, Chоrnobyl exclusion zone.
and skeletal muscles tissues comparing to the control group [13]. In our opinion, it is linked to the disturbance of its resynthesis from homocysteine.
The findings indicate that there is a necessity to monitor blood homocysteine levels in children affected by air currents that include products of burn-
ing of forests and peats in the ChEZ.
Conclusions
1. Forest and peat fires in the Chernobyl exclusion zone are one of the main causes for the elevation in blood homocys-teine levels in children from the adjacent districts, regardless of the state of folate metabolism geneticsystem.
Table 5
Frequency of polymorphic alleles of folate metabolism genes of boys with blood homocysteine levels of over 10 Mmol/L during the measurement on 18/12/2015 (n = 35)
Gene, polymorphism «Neutral» allele Risk allele, heterozygous mutation form Risk allele, homozygous mutation form
Abs. number % Abs. number % Abs. number %
MTR:A2756G 24 68.6 10 28.6 1 2.8
MTHFR:A1298C 17 48.6 13 37.1 5 14.3
MTHFR:C677T 17 48.6 13 37.1 5 14.3
MTRR:A66G 7 20.0 13 37.1 15 42.9
Table 6 Frequency of polymorphic alleles of folate metabolism genes of girls with blood homocysteine levels of over 10 Mmol/L during the measurement on 02/04/2015 (n = 25)
Gene, polymorphism «Neutral» allele Risk allele, heterozygous mutation form Risk allele, homozygous mutation form
Abs. number % Abs. number % Abs. number %
MTR:A2756G 18 72.0 6 24.0 1 4.0
MTHFR:A1298C 16 64.0 5 20.0 4 16.0
MTHFR:C677T 11 44.0 10 40.0 4 16.0
MTRR:A66G 4 16.0 14 56.0 7 28.0
2. Monitoring blood homocysteine levels in children and adults living under conditions of exposure to wood burning products containing radioactive elements is the key component of the programme for the prevention of cancer and cardiovascular diseases. .niTEPATyPA
1. Bandazhevsky Yu.I., Dubovaya N.F. Folate metabolism gene polymorphisms and homocysteinemia in children from families continuously living in an area affected by the Chornobyl nuclear power plant accident. Collection of Scientific and Practical Articles «Chornobyl: Ecology and Health»: Under general editorship of Yu.I. Bandazhevsky. Issue 3. Ivankiv : PI Coordination and Analytical Center «Ecology and Health». Dnipropetrovsk : Serednyak T.K., 2015. P. 16-25.
2. Lentz S.R., Haynes W.G. Homocysteine: is it a clinically important cardiovascular risk factor? Clin. J. Med. 2004. Vol. 71. P. 729-734.
3. Bandazhevsky Yu.I., Dubova N.F. Assessment of folate metabolism functioning in terms of prevention of breast cancer in the population living in areas affected by the Chornobyl nuclear power plant accident. 36ipKa HayKOBUx
ГИПЕРГОМОЦИСТЕИНЕМИЯ У ДЕТЕЙ И ЛЕСНЫЕ ПОЖАРЫ В ЧЕРНОБЫЛЬСКОЙ ЗОНЕ ОТЧУЖДЕНИЯ 1Бандажевский Ю.И., 2Дубовая Н.Ф.
1 Координационный аналитический центр «Экология и здоровье», г. Иванков, Украина Национальная медицинская академия последипломного образования им. П.Л. Шупика, г. Киев, Украина
Цель исследования. Определить роль генотипов фолатного цикла в возникновении гипергомоцистеинемии у детей после лесного пожара в Чернобыльской зоне отчуждения (ЧЗО).
Методы: иммунохимический, ПЦР в режиме Real-time, математико-статистический. Результаты. Обследование 84 детей подросткового возраста из Полесского района Киевской области позволило выявить у них изменения метаболических процессов в виде повышенной продукции гомоцистеина в связи с лесным пожаром в ЧЗО, который регистрировался в течение 26-29 апреля
2015 года. Проведенный сравнительный анализ удельного веса генотипов фолатного цикла в подгруппах детей с гипергомоци-стеинемией до и после лесного пожара показал непричастность к данному явлению эндогенного фактора в виде генетического аппарата, контролирующего синтез ферментов фолатного цикла.
Выводы. Пожары леса и торфяников в ЧЗО являются одной из основных причин повышения уровня гомоцистеина в крови детей из прилежащих районов вне зависимости от состояния генетической системы фолатного цикла. Контроль за уровнем гомоцистеина в крови детей и взрослых, проживающих в условиях воздействия продуктов горения древесины, содержащей радиоактивные элементы, является основным компонентом программы профилактики онкологических и сердечно-сосудистых заболеваний.
Ключевые слова: фолатный цикл, гомоцистеин, подростки, лесные пожары, Чернобыльская зона отчуждения.
праць с^вробтниюв НМАРО ¡м. П.Л. Шупика. 2017. Вип. 27. С. 215-225.
4. Bandazhevskyi Yu.I., Dubova N.F. MTHFR:677TT genotype and state of hyper-homocysteinemia in the children of regions suffered as a result of the accident at the Chornobyl nuclear power plant. Dovkillia ta zdorovia (Environment & Health). 2018. № 2 (86). P. 10-15.
5. Колпакова А.Ф., Шарипов Р Н., Волкова О.А. Влияние загрязнения атмосферного воздуха взвешенными веществами на сердечно-сосудистую систему. Сибирский медицинский журнал. 2015. Т. 30, № 3. С. 7-12.
6. Pope C.A., Turner M.C., Burnett R. et al. Relationships between fine particulate air pollution, cardiometabolic disorders and cardiovascular mortality. Circ. Res. 2015. Vol. 116, № 1. P.108-115.
7. Park S.K., O'Neill M.S., Vokonas P.S. et al. Traffic related particles are associated with elevated homocysteine. The VA Normative Aging Study. Am. J. Respir. Crit. Care Med. 2008. Vol. 178. P. 283-289.
8. Кашпаров В.А., Миро-нюк В.В., Журба М.А. и др. Радиологические послед-
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9. Загальнодозиметрична паспортиза^я та результати ЛВЛ-моыторингу у населе-них пунктах УкраУни, яК зазнали радюактивного
забруднення топя ЧорнобильськоУ катастрофи. Дан за 2011 рк. Збiрка 14. К.: МОЗ УкраУни, 2012. 99 с.
10. Богорад В.И., Литвинская Т.В., Шевченко И.А., Дыбач А.М., Слепченко А.Ю. Радиационные последствия пожара в зоне отчуждения Чернобыльской АЭС. Ядерна
Table 7
Frequency of polymorphic alleles of folate metabolism genes of girls with blood homocysteine levels of over 10 Mmol/L during the measurement on 18/12/2015 (n = 32)
Gene, polymorphism «Neutral» allele Risk allele, heterozygous mutation form Risk allele, homozygous mutation form
Abs. number % Abs. number % Abs. number %
MTR:A2756G 23 71.9 8 25.0 1 3.1
MTHFR:A1298С 24 75.0 6 18.7 2 6.3
MTHFR:С677Т 14 43.8 12 37.5 6 18.7
MTRR:A66G 6 18.7 16 50.0 10 31.3
Table 8
Proportions of the MTHFR:677 CT/MTHFR: 1298 AC heterozygous associationin groups of children with hyperhomocysteinemia reported on 02/04/2015 and 18/12/2015
Date of examination of children Proportions of a MTHFR:677 СТ/ MTHFR:1298 АС heterozygous association
Total group Boys Girls
Abs. % Abs. % Abs. %
02/04/2015 9 18.8 5 21.7 4 16.0
18/12/2015 8 11.9 5 14.3 3 9.4
та рад'ацйна безпека. 2016. Т. 69. № 1. С. 64-68.
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12. Bandazhevsky Yu.I., Dubovaya N.F., Bandazhevska G.S. et al. Assessment of the cardiovascular system in children from Polesie and Ivankiv districts. Collection of Scientific and Practical Articles «Chornobyl: Ecology and Health»: Under general editorship of
Yu.I. Bandazhevsky. Issue 2. Ivankiv : PI Coordination and Analytical Center «Ecology and Health»; Dnipropetrovsk : Serednyak Т.К., 2015. P. 22-35.
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Hagitfwno go pega^iï 15.06.2020
