[CHANGING OF PHYSICO-CHEMICAL PARAMETERS OF NON-CONTACT (ELECTROCHEMICAL) ACTIVATED DRINKING WATER IS ASSOCIATED WITH INDUCTION OF GENOMIC INSTABILITY OF CULTIVATED HUMAN BLOOD LYMPHOCYTES].

In the article there are presented data which are the fragment of large multidisciplinary study of genetic safety of non-contact electrochemically activated water (NAW). The aim of this study was the analysis of the relation of impacts of genomic instability (micronucleus test with cytochalasin B) detected in human blood cells, cultured in medias prepared on the base of these NAWs, with physical and chemical properties of these NaWs. In experiments there were used catholytes and anolytes obtained by activation of osmotic, tap and dining bottled water As a result of such activation, all waters were shown to acquire the ability to induce genomic instability in cellular cultures. Notably in cell cultures on catholytes and anolytes these effects differed between themselves and have been associated with different physical and chemical properties of the NAWs.

[Buccal micronucleus cytome assay in complex genetic-hygienic study of preschool children].

There are presented results of the cytome analysis of epithelium of buccal mucosa in 167 children aged of 5-7 years, permanently residing in the city of Magnitogorsk and visiting municipal kindergartens. Frequencies of the main indices of genotoxic and toxic effects were: cells with micronuclei - 0.29±0.04 %, with protrusions ofnuclei - 1.47±0.17 %, with double nuclei -1.96±0.13 %, with two nuclei - 3,28±0,16 %, with perinuclear vacuole - 9.66±0.86 %, with pycnosis of nucleus - 4.60±0.31 per 1000 cells, with karyorrhexis - 0.86±0.15per 1000 cells, with lysis of nucleus - 54.34±3.90 per 1000 cells, the condensed chromatin cells - 10.59±1.35 per 1000 cells. Most of these indices didn't differ from the same detected earlier in buccal epithelial cells from children of the same age in Moscow, except cells with perinuclear vacuole, which frequency in Moscow cohort was twice higher. Frequencies of cells with these anomalies were associated with the content of 19 out of 326 revealed components of contamination of the total-winter snow samples, taken on the territories of the kindergartens, which were visited by examined children. There was shown lack of differences from basic values of main indices previously establishedfor the children of the same age in Moscow, with the exception of cells with perinuclear vacuole, the rate of which in Moscow was twice higher. There were revealed confounding factors associated with test indices: gender of the child, the annual number of acute respiratory diseases, health group, income per one person in family, alcohol intake by parents.

[The effect of treatment of chrysotile with chloride iron (III) on the effect of genomic instability in cultured human lymphocytes].

All forms of asbestos are carcinogenic to humans, that caused the prohibition of its use in Europe, America, etc. However, the unique physical and chemical properties of asbestos and many opportunities for its applications in various industries and human activities require the creation of new technologies to. Analysis of genomic instability in human lymphocytes in a large range of doses (micronucleus test with cytochalasin B showed a decline of core indicators of genome instability (frequency of dividing cells with cytogenetic damage, asymmetry in the distribution of the genetic material in mitosis, proliferative activity) incubation of cells with asbestos modified chloride iron, as compared to the initial sample. These data are consistent with the results of analysis of the intensity of neutrophil chemiluminescence of luminol by exposure of human blood samples studied. Based on the findings made some practical recommendations to the protocol cytogenetic analysis of genomic instability of people exposed to asbestos.

[Chrysotile-asbestos induces cytogenetic effects in the rat's mesothelium in vitro and in vivo].

Under cultivation of rat peritoneal mesothelial cells in vitro in them there are appeared signs of the genomic instability, evidencing their transformation: increasing of the number of both binucleated cells with micronuclei and polynuclear cells and the increase of sizes and polymorphism. Asbestos greatly accelerates this process. Asbestos-induced carcinogenesis in vivo is accompanied in pleural mesothelium in the rats there also revealed with similar signs of genomic instability and cellular transformation.

[Instability and sensitivity of the genome of healthy children in Magnitogorsk].

Problem of the influence of factors of the industrial city on the hereditary apparatus of its residents has not been fully resolved, because of traditionally in such studies only the pollution of environment components is taken into account. However the existence of a set of contributing socialfactors that modify the genotoxic effects ofpollution, requires the creation of a new methodology for genetic and toxicological studies. For this purpose, in Magnitogorsk, where one of Russia's largest steel plants is located, we conducted a comprehensive survey, whose tasks included the analysis of the influence of the complex of exogenous and endogenous factors on the genome of children. In this publication there are presented the results of the fifth fragment of this work - the analysis of instability and individual sensitivity of the genome of 166 children of 5-7 years, residing in two districts of Magnitogorsk: around the steel plant and on the opposite bank of Ural river, where there are no large-scale industrial enterprises. The study was conducted in the micronucleus test on peripheral blood lymphocytes cultured with cytochalasin B. For assessment of individual sensitivity of genome blood cultures were exposed to standard N-methyl-N-nitro-N-nitrosoguanidine (MNNG) mutagen. Cytogenetic analysis was performed in binucleated cells accordingly to international protocol, as well as with the use of an extended protocol including 32 indices. Average group frequency of binuclear cells with micronuclei (0.5-0.7%) were found not differ from the levels defined in children residing in Europe, and not differ between areas of the town. However the extended protocol of cytogenetic analysis discovered that the real frequency of dividing cells with lesions in blood cultures of children was 1,49-1,66%. Higher spontaneous proliferative activity of the cells and the frequency of dividing cells with injuries were found in blood cultures of children residing in settlements around the Magnitogorsk steel plant, while the individual sensitivity of the genome in these children was lower than that of their peers from the opposite bank of the Ural river A similar conclusion, but already in relation to the state of the systems for adaptation of entire body has been made from results of other fragments of the study, published earlier.

[Factors of anxiety and autonomic tonus in senior preschool children from Magnitogorsk].

In the paper there are presented the results of a study of anxiety and balance ofparts of autonomous nervous system in healthy children 5-7 years old, residing in different parts of Magnitogorsk. It is shown that state of heightened and high alert was shown to be more common among children living on the left bank of the Urals river around the Magnitogorsk Metallurgical Integrated Plant. In these children an imbalance in the work of the parts of the autonomic nervous system was detected more frequently, at that shifts were observed mainly in the direction to ergotropic tone. At the same time balanced work of the parts of the autonomic nervous system was observed more frequently in children living on the right bank of the Urals river. Discovered psychosomatic features of examined children turned out to be associated with both the social characteristics of family lifestyle and the emotional stress of parents, and the contents of some organic compounds in total snow samples collected in the territories of kindergartens which they attended. One ofthe most significant results ofthe work we consider the detection of a correlation relationship between emotional stress of parents and activity of key enzymes in their children, reflecting the protective and adaptive reactions of the organism. On the basis of these and previously obtained data, we suggest that social and psychological factors of the family are not only a potential source of maladaptation of the child, but, probably, can have an impact on the stability and sensitivity of the genome of children.

[Carcinogenic and mutagenic activity of chrysotile, processed with iron chloride (III)].

This work is devoted to the study of the role of iron ions in the carcinogenic and mutagenic activity of chrysotile. For this aim natural chrysotile was treated with ferric chloride (III), washed, crushed and intratracheally introduced into Wistar rats. When administered to rats intact chrysotile induced mesotheliomas in 27,9 + 4,6% of cases, and chrysotile modified with ferric chloride - in 1,3 +/- 1,3%. Mutagenicity of the same samples was studied in the micronucleus test when administered intraperitoneally to mice Fl (CBA x S57Bl6). Polychromatic erythrocytes in the bone marrow were investigated 24 hours after intraperitoneal administration. The frequency of polychromatic erythrocytes with micronuclei was decreased from 7,4 +/- 0,18 by 1000 due to the action of chrysotile, from 2,8 +/- 0,42 for 1000 after the introduction of a modified sample. It is hypothesized that the ferric chloride modifies the surface of asbestos fibers that reduces the induction of free radicals which are the primary cause of and carcinogenic effects of chrysotile.

[To a question on the impact of nanoparticles of metals present in the aquatic environment, on bacteria and continuous cell lines HEp-2 and BGM].

Promising application of nanoparticles and nanomaterials is the creation of sanitary hygienic means of new generation used for disinfection of water and indoor surfaces of mass use, furniture, sanitary technical equipment by virtue of modifying traditional materials to bring them effective biocidal properties, and for the development of methods in vitro for assessment their toxicity In this paper the possibility of the use various forms of silver, copper and aluminum as disinfectant for bacterial test organisms in the aquatic environment and assess their toxicity on biological models of continuous culture of BGM cells (a stable line of African green monkey kidney cells) and HEp-2 (Human epithelial type 2 (HEp-2) cells, derived from a human laryngeal carcinoma) is considered.

[Harmonization with international approaches to guidance documents on methods of assessment of the mutagenic properties of chemical environmental factors].

Evaluation of mutagenic activity--an indispensable element of the toxicological characteristics of chemicals in their hygienic regulation. In the article there is performed an analysis of the adopted in the Organization for Economic Cooperation and Development (OECD) and in Russia guidance documents on methods of assessment of the mutagenic properties of chemicals. In the OECD there are 17 manuals, each of which describes a single method, in Russia there were approved five guidance documents for specific groups of chemicals (drugs, pesticides, nanomaterials, substances normalized in the water and the air), which represent the basic and auxiliary methods of testing mutagens. Basic methods aimed to the evaluation of gene, chromosomal and genomic mutations include: assessment of gene mutations in bacteria, methods for estimating mutations in cell cultures of human and mammals in vitro, methods for inspecting mutations in somatic and germ cells of mammals in vivo. Analysis of Russian documents shows that the protocols of basic tests of assessment of the mutagenic activity are close to the protocols of the OECD. It is necessary to publish guidance documents on tests of assessment of the mutagenic activity of chemicals harmonized with the OECD documents.

[Detection of potentially carcinogenic compounds persisting in atmospheric air and having high priority for hygienic regulation].

The algorithm for the choice of potentially carcinogenic compounds (PCS) among emitted into air and results of the study, undertaken for realization of this algorithm are presented. The investigation was carried out in Magnitogorsk - Russian town of black metallurgy - in frames of the other study, aimed to evaluation of the influence of atmospheric pollution on children's health and genomic instability. The 11 PCS for further profound study of biological activity were selected out of more than 300 PCS, persisting in the air. The carcinogenic activity for 6 compounds out of these 11 ones was already have been found out before, that testifies correctness of the created approach and its adequacy for detection PCS in atmospheric air.

[Role of repair gene polymorphism in estimating the sensitivity of human genome to radon in concentrations exceeding maximum permissible level].

The paper gives the results of investigating chromosome aberrations in human peripheral blood lymphocytes due to DNA repair genes, such as hOGG1, ADPRT, APE1, XRCC1, XpG, XpC, XpD, and NBS1, upon long-term exposure to excess indoor radon concentrations. The frequency of chromosome aberrations was found to be significantly lower in the carriers of the genotype hOGG1 326 Ser/Ser (versus the variant Ser/Cys), APE1 148 Asp/Asp (versus Val/Ala and Ala/Ala). The study polymorphic systems were shown to be of value in giving rise to individual types of chromosome aberrations (single fragments and chromosome exchanges).

[Taking into account the emotional tension of adults and children in the evaluation of the influence of environmental factors on genomic instability and sensitivity: results and prospects].

The paper summarizes the results of 8 genetic and psychological examinations in adults and children, which are aimed at evaluating the genotoxic effects of environmental factors on cultured lymphocytes. Human emotional tension was proved to be directly related to the level of genome damage, to genomic sensitivity to in vitro exposures, and to lymphocyte proliferative activity and to be inversely related to the level of apoptosis. There was an association of the degree of anxiety in children with their morbidity. High emotional tension in adult family members was found to negatively affect the degree of anxiety and the level of genomic instability in a child.

[Effect of titanium dioxide nano- and microparticles on the values of the micronucleus test using human blood lymphocytes in culture].

The activity of the nano- and microparticles of titanium dioxide (TD) given in doses of 1.6, 40.0, and 500 microg/ml was studied in the cytochalasin B block micronucleus test with blood cells from two donors. It was shown that nano-DT in doses of 1.6 and 40 microg/ml was more active than micro-DT in inducing micronuclei and nucleoplasmic bridges, asymmetric cell division, and accelerated cell proliferation. Some differences in individual sensitivity to TD between donor blood cultures were due to an inverse trend in the association between the indicators of genomic damage and apoptosis. The new nuclear morphometry test that had developed to screen genotoxic effects and used in this study for the first time revealed a nano-DT dose-depended increase in the asymmetry of nuclei in the binucleated cells in the cultures of both blood donors, the morphometric effects being directly correlated with the values of cytogenetic analysis.

[Morbidity among senior preschool children in Magnitogorsk].

The publication is the third fragment of the multiparameter study. The level and causes of genome instability and sensitivity are assessed in children living in Magnitogorsk, a city with one of the largest mills in Russia. The city is in the list of the world's 35 most polluted ones. A transverse retrospective analysis found no differences in primary morbidity among 5-7-year-old children who had been living in different districts of Magnitogorsk since birth and were going to municipal kindergartens located adjacent to their house. Contrary to the expectations, the prevalence of morbidity and diseases that were markers for an industrial town was significantly lower among the children residing in the settlements situated around the mills than that in the city's other districts. There were 9 organic compounds that had no hygienic standards, the content of which in the snow samples collected in the areas of the examined kindergartens correlated with the prevalence of the children's morbidity. Family social and living conditions were shown to affect the children's morbidity.

[Hygienic evaluation of the total mutagenic activity of snow samples from Magnitogorsk].

The paper gives the results of 4-year monitoring of the total mutagenic activity of snow samples from different Magnitogork areas in a test for induction of dominant lethal mutations (DLM) in the gametes of Drosophila melanogaster. An association was first found between the rate of DLM and the content of some chemical compounds in the ambient air and snow samples; moreover all the substances present in the samples, which had found genotoxic effects, showed a positive correlation with the rate of DLM. Furthermore, direct correlations were first established between the rate of DLM and the air pollution index and morbidity rates in 5-7-year-old children residing in the areas under study. The findings allow the test for induction of dominant lethal mutations (DLM) in the gametes of Drosophila melanogaster to be recommended due to its unique informative and prognostic value for monitoring ambient air pollution and for extensive use in the risk assessment system.

[Toxic and genetic risk assessments in children of Gornaya Shoria].

This comprehensive study assessed a number of environmental factors that were potentially able to induce genotoxic effects in man. A set of radiological, physicochemical, and bioindication techniques was used to estimate the quality of water, air, and soil in the places of residence and education of children and adolescents from the boarding school of the town of Tashtagol and schoolchildren from the village of Krasnoye, Kemerovo Region. Excess radon levels in the air of living spaces and classes, a small excess of the maximum allowable concentration of gross forms of heavy metals in individual soil samples, and high toxic effects in the Drosophila gametes exposed to air samples were revealed in the Tashtagol children having higher genotoxic effects in the lymphocyte than those in the Krasnoye village ones. The findings suggest that the marked genotoxic effects recorded over a long time in the lymphocytes of children and adolescents living in Tashtagol may be associated with the complex influence of the above factors. It is north supplementing further investigations by an extended chemical analysis of water, air, snow, and soil samples taken in the places of residence and education of children.

[Estimation of adaptive capacities in Magnitogorsk children from the activity of some detoxification enzymes].

The paper provides the first fragment of a multiparameter study analyzing the influence of environmental pollution, the social and psychological features of a family, and some endogenous factors on genome stability and sensitivity in a developed ferrous metallurgy town. It also gives data on the urine and serum activity of the lysosomal enzyme N-acetyl-b-D-glucosaminidase (NAG) and the serum activity of catalase in an organized contingent of apparently healthy children (n = 178; 6 kindergartens) aged 5-7 years, who live permanently in Magnitogorsk at different distances from the metallurgical works. More than 70% of children selected for examination were found to have average normal levels of activity of the enzymes studied. According to the average levels of enzyme activity, there were only 2 kindergartens (both from the left-bank region). In the children from the left-bank area, enzyme activities varied more greatly, which suggests the higher prevalence of tense adaptation. Correlation analysis revealed association between the children's serum activity of enzymes and some components of snow pollution. It is anticipated that the found changes in serum activities of N-acetyl-beta-D-glucosaminidase and catalase may be determined by individual differences in a child's response to ambient air pollutants.

[Evaluation of epidemic risk from pathogenic and opportunistic bacteria isolated from water of its various use types].

The investigation was concerned with wild gram-positive and gram-negative microorganisms (E. coli spp., Klebsiella spp., Salmonella spp., and the nonfermentative bacteria Pseudomonas spp.) isolated from the waters of different types, as well as museum cultures (E. coli strain 1257, E. coli strain 675, Salmonella enteritidis ATCC 5765, Staphylococcus aureus 906, and Pseudomonas aeruginosa ATCC 10145). The wild strains were isolated from water when conducting experimental and field studies; these are able to survive in the waters disinfected by various procedures (a guanidine-containing disinfectant in non-toxic concentrations; photo-activated decontamination with sensitizers; exposure to magnetic and ultrasound waves). The cytotoxic, adhesive, and invasive activities of the bacteria isolated from environmental water objects increased on their cultivation on nutrient growth media, by simulating their possible effects in man. The developed experimental approach makes it possible to estimate the hazard of potentially pathogenic bacteria in one test trial, by applying the BGM cells and may be used to assess the microbial risk.

[Use of micronuclear test to assess drinking waters].

The effect of drinking waters with their altered structure and charge state (a total of 9 samples) on the epitheliocytes of the proventriculus, liver, and kidney was studied in rats after 6- and 12-month water consumption. It was found that in the absence of changes in the animals' morphofunctional status, the degree of water structuredness was associated with the frequency of epitheliocytes with nuclear protrusion into the proventricular mucosa following 6-month water consumption and with the manifestations of adipose degeneration, hepatocytic alterations, and stromal overgrowth into the liver after 12-month water use. The biological implication of little studied (additional) types of anomalies of the proventricular epitheliocytic nucleus: the necrotic nature of a perinuclear vacuole, the predominant implication of nuclear pyknosis and rrhexis to apoptosis and the association of binucleated patterns with the restorative proliferation of hepatic and renal tissues, was specified.

[Proliferative activity parameters and their correlation with genetic damage of blood lymphocytes during cultivation under the conditions of cytokinetic block].

The subjects of the study were 15 volunteers aged 22 to 25 years, who underwent 25 air ionization sessions. The effects of genome instability were evaluated, and correlations between indicators of genome damage (lesions of micronuclei and nucleoplasmatic bridges) and parameters of proliferative and replicative activity (mitotic index, proliferative pool, the fraction of rapidly dividing cells, and replication index) of blood lymphocytes in the culture were studied. In order to establish the associations between the parameters, the parallel cultures were exposed to 0.07 mM of the standard mutagen MNNG during 5 hours. The study showed that the course of air ionization did not induce the micronuclei and nucleoplasmatic bridges in binuclear cells, but increased proliferative cell activity. This effect was accompanied by an increase in the fraction of rapidly dividing cells among all the dividing cells, and an increase in the dispersion of all proliferation parameters. MNNG induced a constant level of micronuclei in binuclear cells during the whole course, but not before the beginning of air ionization. The changes in the parameter "the fraction of dividing cells" (proliferative pool) were the most prominent manifestation of the suppression of proliferation by MNNG. MNNG loading inhibited the formation of binuclear cells most of all. The results demonstrate a non-random character of the correlation between the level of micronuclei in binuclear cells and proliferative activity parameters during cell cultivation under the conditions of cytokinetic block.