The cytogenetic parameters of Pinus sylvestris L. under conditions of the Far North of Russia (Karelia)

The cytogenetic analysis of the Pinus sylvestris L. seed progeny in the forests of the northern taiga in the Far North of Russia conditions was carried out. Two Pinus sylvestris populations growing in Ambarnskoy and Pyaozerskoy forest divisions of Loukhsky district of Karelia Republic were studied. The number of chromosomes, the frequency and types of mitosis disturbances at the metaphase, anaphase and telophase (as a percentage of the total number of dividing cells at the same stages), and the frequency of micronuclei occurrence, as well as the laboratory seed germination, were studied. As a result of the study, it was found that 50–56% of the studied sprouts were mixoploid. Analysis of mitosis showed that the root meristem of the studied samples contained cells with various pathologies; simultaneously, the frequency of mitosis pathologies in the Ambarnskoy population was significantly higher than in the Pyaozerskoy one. It amounted to 6.8 ± 0.4 and 4.9 ± 0.4%, respectively. Nine types of abnormalities were identified; chromosome overrun and bridges prevailed. Micronuclei were registered in 35–38% of the studied Pinus sylvestris sprouts. At the same time, the proportion of cells with micronuclei in the Ambarnskoy population was 2-fold higher than in the Pyaozerskoy one and averaged 0.12 ± 0.03% and 0.07 ± 0.02%, respectively.

Genotoxicity of chromium (III) and cobalt (II) and interactions between them

Introduction. Chromium and cobalt are essential trace elements that are required only in a small amount, otherwise their excess can cause toxic effects. Aim. The aim of this study was to determine the effects of chromium (III) and cobalt (II) and their combinations on genotoxicity in human fibroblasts cells (BJ). Material and methods. In this work, comet and micronucleus assays were used. The BJ cells were exposed to chromium chloride and cobalt chloride at concentration ranges from 100 to 1400 µM. Mixtures of these elements were prepared so as to examine interactions between them. Results. The present study shows the genotoxic effects of chromium (III) and cobalt (II) and their mixtures on BJ cells. In the comet assay, no comets were observed at the lowest concentrations; in the higher, a significant increase in their percentage was observed. In the other assay (formation of micronuclei), a statistically significant increase in the number of cells with micronuclei was observed in the BJ cells spiked with cobalt chloride and chromium chloride. In the case of simultaneous incubation of chromium chloride at 200 µM and cobalt chloride at 1000 µM in the BJ line, antagonism was observed. However, the interaction of chromium chloride at the 1000 µM and cobalt chloride at 200 µM leads to synergism between the studied elements. Conclusions. Cobalt (II) and chromium (III) show genotoxic properties, they induce breaks in double and single-stranded DNA and they cause formation of AP-sites that do not have purine or pyrimidine bases.

Buccal Micronucleus Cytome Assay for the assessment health status of population living in the area of nuclear heritage

Introduction. An essential task of hygiene is assessing the health status of the population living in areas with adverse environmental and hygienic conditions. Materials and methods. A study of the health status of 50 children of three age groups was carried out in Dunay city, Primorsky Krai, located near the enterprise “The Far Eastern Center for Radioactive Waste Management, Far East Center “DalRAO.” Health status was assessed using the buccal micronucleus cytome assay (BMCA) to determine the cytogenetic and cytotoxic effects of environmental factors. Results. The frequency of buccal cells with micronuclei in groups of children from the Dunay city varied within 0.33-0.47‰, which does not exceed the control level of 1.1%, determined within the framework of the international project HMNxl. The level of cytogenetic stress is defined as acceptable (but not low). Given the index of accumulation of cytogenetic damages, it is possible to rank the studied groups according to their cytogenetic status in the direction of deterioration: kindergarten children, older and younger schoolchildren. Conclusion. The approach proposed in this study made it possible to quantitatively determine the mutagenic and cytotoxic effects of factors on the children’s population of the Dunay city. It can be recommended as an informative method for the population health status assessment during socio-hygienic monitoring, including in the territories of nuclear (legacy) sites.

Genome instability as an indicator of environmental state in the Murmansk region

The data of various researchers indicate that the combined effects of natural and anthropogenic factors directly affect the nature of the territorial morbidity of the inhabitants of the Far North. In biomonitoring of the environment, a micronucleus test on human cells is widely used to assess the degree of genotoxicity of the comparison areas. The aim of the research is to study the local situation with the accumulation of lesions in the lymphocytes of children living in the Murmansk region using a micronucleus test. As a result of assessing the cytogenetic status of the child population living in the comparison territories (Krasnoshchelye, Lovozero, Umba, Apatity), specific territorial features of the cytogenetic status of children were revealed, which are probably associated with the prevailing morbidity in the territories comparisons. This is probably due to territorial genotoxic and toxic agents. Significant differences in the frequency of occurrence of binucleated lymphocytes with micronuclei in schoolchildren were noted when all cells (mononuclear, binuclear, trinuclear, quadrenuclear cells, as well as cells containing more than 4 nuclei) were taken into account between Krasnoshchelye, Lovozero (U = 45.0, p = 0.0009), Umba (U = 91.0, p = 0.0125) and Apatity (U = 113.0, p = 0.0125). Also, significant differences were noted between the frequency of occurrence of cells with micronuclei among cells that did not respond to the mitogenic signal (mononuclear) and contain more than 2 nuclei in lymphocytes between adolescents from Krasnoshchelye and Apatity (U = 109.0, p = 0.0093) and Umba (U = 73.5, p = 0.0025). When comparing the frequency of occurrence of all types of cells containing micronuclei in the lymphocytes of adolescents, significant differences were revealed between the frequency of occurrence of such cells in adolescents from Apatity and Umba (U = 97.0, p = 0.0036).

Cytological features of buccal epithelium in patients of various ages

The article describes the potential of buccal cells investigations. The authors presented buccal epithelium application in noninvasive diagnosis of early human aging; identified common cytological features of buccal epithelium for different ages; revealed the accumulation of cytogenetic abnormalities (epithelial cells with micronuclei, protrusions of the nucleus) and degenerative-dystrophic changes (perinuclear vacuole, condensed chromatin, karyorexis, karyolysis) with age. These findings reflect the predominance of apoptosis over reparation in the process of aging.

Cytogenetic status of the residents of the Gydansky Peninsula (Gydan)

The relevance of the study on the Gydansky Peninsula lies in poor knowledge and inaccessibility of the territory, planned intensive industrial development of the Gydansky Peninsula, in 2011 there were received licenses for the exploration of license areas at the peninsula up to 2031. Industrial development will inevitably lead to certain environmental shifts, emission of the harmful substances into biosphere, the accumulation of anthropogenic pollutants in soil and water sources. The proposed development of the territory of the Peninsula Gydan sets the task of assessment of the impact of gas and oil production in conditions of the far North on health, as well indigenous persons as employees recruited to this of the region. One of the informative approaches to the assessment of population health is the assessment of the cytogenetic status with the use of noninvasive analysis of buccal epithelium. The aim of the study is to determine the cytogenetic status of the inhabitants of the village of Antipayuta of the Yamal-Nenets Autonomous Okrug for the assessment of the impact of environmental factors on the health of the population at the present stage of the industrial development of the territory. Samples of buccal epithelium of 81 alien and indigenous people of the Yamal-Nenets Autonomous district were the object of the investigation. There was performed the analysis of indices of cells of buccal epithelium of the residents living in the village in comparison with the control group. The analysis of samples was performed on a Nikon Eclipse E100 microscope. For the assessment of the cytogenetic status of the individual there was used the proposed by Sycheva L. P. (2012-Index of accumulation of cytogenetic damages (Iac). It is shown that the frequency of occurrence of micronucleus and nuclear protrusions does not exceed the performance of the control group. The index of accumulation of cytogenetic damage for the population of the village is 0.78±0.07% and corresponds to a low level. The analysis of indices of buccal epithelial cells of residents living in rural areas was performed in the comparison with the control group The prevalence rate of cells with micronuclei and protrusions were shown to account of 14.6% andfail to exceed indices in the control group. Accumulation index of cytogenetic damages for the village population is 0.78 ± 0.07 % and corresponds to the low level. The amount of cells with micronuclei and protrusions in the current study is by 14.6% lower than the average of the control group. The intensity of proliferative processes and apoptosis in the comparison groups occurs evenly.

Cytogenetic indices of buccal epithelium in schoolchildren residing in territories with different levels of the air pollution

In the article there are contained the results of the study of ambient air of the regional center, urban-type settlement, and buccal epithelium in 87 schoolchildren aged of 11-13 years. Schoolchildren residing in the regional center were shown to be more susceptible to the exposure to such toxicants as carbon monoxide, nitrogen oxide, fluorhydric acid, chlorine and its compounds, sulfur dioxide, formaldehyde contained in the ambient air. In this group of students if compared with students who live in the village, there were revealed more pronounced cytogenetic deteriorations in oral mucosal epitheliocytes: cells with micronuclei, different forms of protrusion of the core, dual-core cell mitosis, kariokynesis, apoptotic bodies significantly more common occur, that can speak about both not only of cytogenetic instability, but and the degree of environmental genotoxicity, in particular, ambient air. There was noted the tendency to increase the level of cytogenetic damage and destruction of the nucleus in the group of girls in comparison with boys.

ON THE ISSUE OF ACCELERATED HYGIENIC ASSESSMENT OF ENVIRONMENTAL GENOTOXIC CARCINOGENS

The aim of the work was assessment of the criterial significance of the complex of early immunological reactions of the organism, pathomorphological and genotoxic changes in the organs for the acceleration of testing and hygienic assessment of carcinogenic danger of chemical substances. Materials and methods: Investigations were carried out in the chronic experiment on white random-bred male mice and included 2 series of the investigations: 1 – application of benz(a)pyrene on skin in different doses (10.5μg; 2.1μg; 0.21μg); 2 - peroral administration of benz(a)pyrene and phenol (single dose - 0.1 mg). Genotoxic changes in skin and forestomach were assessed with the help of micronuclear test. Results: Under carcinogen exposure, regardless of routes of administration we determined an increase frequency of the cells with micronuclei and suppression of T-link of immune system during the first month which were characterized by a parallelism of development, unidirectional relative to carcinogenesis and presence of reliable reverse correlative connection between them. In the period between the 1t and the 3d months we observed a stabilization of the number of cells with micronuclei and deepening of immunosuppression at the expense of the suppression of humoral chain of the immunity. Conclusions: The obtained data became a basis for the development of methodic scheme of accelerated testing of the chemical substances under investigations for carcinogenicity and hygienic setting of genotoxic carcinogens.

CYTOTOXICITY AND GENOTOXICITY OF SOIL IN SHUMEN CITY PARK

Sedentary lifestyle has become a great concern to human health. City parks play a great role in the solution of this problem, but exposure to urban pollutants leads to the necessity to monitor environmental quality.Purpose of Study The aim of the study was to evaluate the possible cytotoxicity and genotoxicity of soil samples collected from Shumen city park using Allium cepa L.-test.Methods The following microscopic parameters were used: mitotic index, index of each phase of mitotic division, mitotic abnormalities and interphase cells with micronuclei or two nuclei.Findings and Results The decline of the mitotic activity and changes in the proportion of mitotic phases indicate soil cytotoxicity. Various mitotic abnormalities and binucleated cells revealed genotoxicity.Conclusions and Recommendations Established cytotoxicity and genotoxicity of soil revealed a potential health risk to park users. Further analyses should be provided, since positive results from Allium-test serve as an alarm.

EFFECT OF ANTITUMOR DRUGS IN LOW CONCENTRATIONS ON THE BIOLOGICAL, IMMUNOPHENOTYPIC AND CYTOGENETIC CHARACTERISTICS OF HUMAN COLON CANCER CELLS in vitro

Objective: To estimate the impact of the low-dose anticancer drugs (ACD) with the different mechanisms of action and human interferon (IFN) alpha 2b on the biological properties, immunophenotypic and cytogenetic characteristics of colon cancer cells in vitro. Materials and Methods: The study was performed on human colon cancer cell lines COLO 205, HT-29 and 3C-P treated with ACD and IFN in subtoxic concentrations. Expression of CD44, N-cadherin, vimentin, β-catenin, ERCC1 and Slug was assessed by immunocytochemical method. Using cytogenetic analysis, the numbers of mitoses, cells with micronuclei, apoptotic cells and cells with nuclear protrusions were studied. Results: The prolonged exposure (up to 30 days) of colon cancer cells to low-dose ACD (0.2–0.5 µg/ml cisplatin and 0.1–0.2 µg/ml irinotecan) in combination with IFN (500–1000 IU/ml) led to 37-fold decreased colony-forming activity of these cell and 10-fold reduction of the number of cells expressing mesenchymal protein markers (N-cadherin, vimentin). Also, in COLO 205 cells treated with ACD and IFN the number of SLUG- and CD44-positive cells decreased by 92 and by 85%, respectively. Long-term cultivation of HT-29 cells in the presence of cisplatin and IFN resulted in 5-fold suppression of ERCC1 expression. The cytogenetic analysis has shown that the ACD, IFN and their combinations in subtoxic concentrations caused significant genotoxic effect, suppression of cell proliferation and accumulation of cells with micronuclei. The sensitivity of colon cancer cells to ACD in standard cytotoxic concentrations did not change after prolonged low-dose exposure. Conclusion: The data showed that the prolonged action of the low doses of ACD on human colon cancer cells resulted in the suppression of cell proliferation, colony-forming activity in soft agar, expression of epithelialmesenchymal transition-associated markers and significant cytogenetic changes.