Genetic effects in the liquidators of consequences of Chernobyl Nuclear Power Plant accident

The purpose of the present research was the estimation of probable genetic consequences at the liquidators of the consequences of Chernobyl accident in 1986-1987. The research is made on two groups of the liquidators. The first group included the liquidators taking place on the account in the branch register and working now at the enterprises of a nuclear industry. The second group included 902 liquidators of consequences of Chernobyl accident in 1986 constantly living in the Ryazan area and which are taking place on permanent observation the account in regional hospital. For an estimation of probable genetic effects analyzed the data on frequency and outcomes pregnancy of the wives of the liquidators, on condition and on diseases of newborn, on switching intrauterine development defects (IDD). The analysis carried out depending on dozes of an irradiation: up to 5 cGy; 5-10 cGy and 10-25 cGy. Received materials testify, that at the liquidators, at a doze of an external irradiation 10-25 cGy, the determined effects--period long sterility, kept at a part them till 3 years come to light. The set of the received data, such as depending from the dose increase of frequency of spontaneous abortions and of inherent defects of development of newborn, the increase of frequency diseases of newborn and share newborn with low weight, allows to make a conclusion about an induction of genetic effects in sexual cells of the liquidators of consequences of Chernobyl accident at dozes of an external irradiation more than 10 cGy. Taking into account high biological efficiency of alpha-radiation (K = 20), and of beta-radiation (K = 2-4), the equivalent effective doze male gonads (testes) in 3-5 times is higher, than estimated only from external gamma-radiation.     

Mechanism of induction of cytogenetic damage in plant meristematic cells caused by combined effect of heavy natural radionuclides and heavy and alkaline metals

The analysis of experimental data on the combined effect of 232Th and water from melted snow containing metal ions from industrally loaded territories on Tradescantia (clone 02) and on Allium shoenoprasum meristematic root tip cells treated with heavy natural radionuclides from natural water sources in the area of radioactive wastes storehouse is presented. It was demonstrated that plant meristematic cells response to heavy metals and heavy natural radionuclides, both from two-component model system and from natural water sources, have commom features.     

The action of 232Th separately and in combination with heavy and alkaline metal salts on Tradescantia (clone 02)

The action of 232Th-nitrate in concentration 0.09, 0.18 and 0.36 mg/l (counting on 232Th ion) on water cultures of Tradescantia (clone 02) was investigated. It was found that all investigated concentrations of 232Th showed statistically significant genotoxic effect and increased level of morphological abnormal cells in the stamen hairs of Tradescantia in the absence of modifying action of other metal ions. Synergistic toxic interaction was found between the 232Th in concentration 0.18 mg/l and metal ions for all samples of thawed water. Synergistic genotoxic effect of the combined action of these factors was revealed only at the low total contents of ions of heavy and alkaline metals in thawed water samples. The observed synergistic effects of the combined action of 232Th and metal ions should be taken into account when controlling the radionuclide level in the environment.     

Cytogenetic effects of separate and combined effect of thorium-232 and potassium nitrates on Tradescantia (clone 02)

Under both separate and combined influence of 232nd-nitrate and potassium nitrate a study was made of the yield of somatic mutation frequencies, the level of morphologically abnormal cells and the loss of reproduction integrity in the stamen hairs of Tradescantia (clone 02). All investigated concentrations of 232nd-nitrate and potassium nitrate exerted statistically significant genotoxic effects and increased the level of morphological cell abnormalities in the stamen hairs of Tradescantia. Also, 0.36 mg/l 232nd and 0.51 mg/l KNO3 concentrations cause a statistically significant loss of reproduction integrity in the stamen hair. The combined action of 232nd-nitrate and potassium nitrate in all investigated concentrations and parameters causes a statistically significant effect.     

Mutagenic potential of copper compounds and modification of effects of silver iodide]

Mutagenic potential of copper compounds and its alteration in case of the interaction with silver compounds were analyzed by use of plant test systems. As test systems, Crepis capillaris L., Tradescantia clone 02, and soybean (Glycine max (L.) Merrill) were used. Mutagenic properties of copper iodide and copper sulfate were not detected. CuI, being not a mutagen by itself, remarkably enhanced mutagenic potential of AgI.     

The plant activation of m-phenylenediamine by Tradescantia clone 03 and clone 4430 cells in liquid suspension culture

In this study, we expanded the use of the genus Tradescantia to investigate the plant activation of promutagens and further refine the methodology of the plant cell/microbe coincubation assay. Liquid suspension cell cultures of Tradescantia clone 03 and Tradescantia clone 4430 were used to activate the promutagen m-phenylenediamine into a mutagenic compound which was detected by Salmonella typhimurium strain TA98 in the plant cell/microbe coincubation assay. Optimum treatment parameters were established for both plant cell lines. Optimum was defined as the lowest concentration or shortest time period that provided consistently positive results and high rates of revertants. Preliminary experiments with both cell lines defined 2.5 mumoles m-phenylenediamine per plate as the optimum concentration to be used in the determination of the optimal coincubation period and the optimal concentration of plant cells. These experiments also determined the optimal physiological stage at which both clones should be used in the coincubation assay. Differences were found in the optimal of coincubation (1h for clone 03, 2 h for clone 4430) and growth stage (mid-log for clone 03, mid- to late-log for clone 4430). Similar activation responses were seen for both clones when the concentration of plant cells (mg/ml) was varied. Under optimized conditions, clone 03 cells demonstrated an approximately 10% higher activation response than clone 4430.     

Influence of radiation exposure rate on somatic mutation frequency and loss of reproductive integrity in tradescantia stamen hairs

Inflorescences of Tradescantia clone 02 (2n = 12), hetero- or hemi-zygous for flower color, were exposed to a series of γ-ray exposures at two different exposure rates, 29.3 R/min and 0.026–0.52 R/min. Pink mutation-response curves, and survival curves based on reproductive integrity, were constructed for each of the exposure rates. Loss of reproductive intergrity was also assessed at high (256 R/min) and low (0.52–4.17 R/min) γ-ray exposure rates in T. blossfeldiana (2n = 72). All observations were made on stamen hairs.The higher exposure rate was 1.3–1.7 times more effective in inducing pink mutations in clone 02. A greater efficiency of the higher exposure rate was also found for both taxa at the loss of reproductive integrity endpoint. The D₀ values obtained at the higher exposure rates, 154 R for clone 02 and 720 R for T. blossfeldiana, were significantly lower than the corresponding values of 270 R and 1880 R obtained at the lower exposure rates. These D₀'s differ by factors of 1.75 and 2.61 for clone 02 and T. blossfeldiana, respectively. D₀'s for the two taxa were found to be inversely correlated with their interphase chromosome volumes.     

Analysis of the Mutagenic Potential of Copper Compounds and Modification of Their Effect by Silver Iodide

Mutagenic potential of copper compounds and its alteration in case of the interaction with silver compounds were analyzed by use of plant test systems. As test systems,Crepis capillarisL., Tradescantia clone 02, and soybean (Glycine max(L.) Merrill) were used. Mutagenic properties of copper iodide and copper sulfate were not detected. CuI, being not a mutagen by itself, remarkably enhanced mutagenic potential of AgI.     

Transformation of C3H 10T1/2 cells with 4.3 MeV alpha particles at low doses: effects of single and fractionated doses.

Oncogenic transformation of C3H 10T1/2 cells was determined after exposure to graded doses of 4.3-MeV alpha particles LET = 101 keV/microns. The source of alpha particles was 244Cm and the irradiation was done in an irradiation chamber built for the purpose. Graded doses in the range of 0.2 to 300 cGy were studied with special emphasis on the low-dose region, with as many as seven points in the interval up to 10 cGy. The dose-effect relationship was a complex function. Transformation frequency increased with dose up to 2 cGy; it seemed to flatten at doses between 2 and 20 cGy but increased again at higher doses. A total of 21 cGy was delivered in a single dose or in 3 or 10 equal fractions at an interval of 1.5 h. An inverse dose-protraction effect of 1.4 was found with both fractionation schemes. Measurements of the mitotic index of the population immediately before the various fractions revealed a strong effect on the rate of cell division even after very low doses of radiation. Mitotic yield decreased markedly with the total dose delivered, and it was as low as 50% of the control value after 4.2 cGy and 20% after 14 cGy with both fractionation schemes.     

Effect of the substances "Ammivit" and "cerloplasmin" on lipid peroxidation products level and of antioxidant system indicators under irradiation

It has been shown, that the total X-ray irradiation in the dozes of 0.5 and 1 Gy influences on the content of lipid peroxidation products and enzymatic activity of antioxidant system in rat spleen and thymus cells. The influence of preparations "AMMIVIT" and "Ceruloplasmin" on these processes is investigated also. So, the animals feeding by the vitamin concentrate "AMMIVIT" have lead to increase of MDA level (a final product of lipid peroxidation) and the overactivity of some antioxidant enzymes in rat spleen and thymus cells. Injection of the preparation "Ceruloplasmin" to experimental animals up 1 hour before the irradiation has normalized LPO intensity and activity of AO enzymes.     

Anticlastogenic effect of Spirulina maxima extract on the micronuclei induced by maleic hydrazide in Tradescantia

The aim of this investigation was to determine if extracts of Spirulina maxima reduce the genotoxic damage induced by maleic hydrazide (MH) using the Tradescantia biosssay. Two types of extracts from the alga were prepared: an aqueous extract with two different concentrations, 100 and 500 mg/ml, and a second one, the extract of a 1% solution of dimethyl sulfoxide (DMSO) which corresponded to 100 mg/ml of the alga. The capacity of MH to induce micronuclei (MN) was initially established by administering 0.005, 0.01, and 0.015 mg/ml of the chemical to the Tradescantia inflorescences, and observing its effect after 24 h.The results of this experiment showed a significant MN increase with the two high concentrations tested, although no dose-response effect was observed. For the anticlastogenic assay, the extracts of Spirulina were applied to the inflorescences alone or immediately before the application of MH (0.01 mg/ml) and the induced MN were observed 24 h later. We found that none of the extracts increased the MN level with respect to the untreated plants; also, that MH more or less doubled the basal micronuclei frequency, and finally, that all tested extracts reduced the genotoxic damage caused by MH. The inhibitory indices obtained for the aqueous extracts (100 and 500 mg/ml) and for the DMSO extract were respectively 59, 85, and 56.3%. These data indicate that Spirulina is an anticlastogenic agent and suggest that it is advisable to extend studies on this matter using other biological models.     

Mutagenicity of ground water (in the bore-holes) in Ararat Valley (Armenia) detected by Trad-SHM bioassay

The genotoxicity of ground water from four bore-holes of different depths (40-120m) in the Ararat valley (Armenia) used both for drinking and irrigation was investigated. The frequency of recessive somatic mutations was determined using the Tradescantia-stamen-hair-mutation (Trad-SHM) test. The Tradescantia clone 02 was used. The pink mutation events (PMEs) were increased by 3.18-6.81-fold in comparison with the control depending both on the depth of subterranean water location and the increase of Na(+) ion concentration in these water samples. The peak frequency was found in water from the 40-45m depth. The deeper the bore-holes, the lower the mutagenicity of water and the concentration of Na(+) ions. Different types of mutant sector arrangements and their frequencies changed depending on the subterranean water depth.     

Genotoxic effects of the chlorinated hydroxyfuranones 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone and 3,4-dichloro-5-hydroxy-2[5H]-furanone in Tradescantia micronucleus assays

This is the first report of clastogenic effects of chlorinated hydroxyfuranones (CHFs) in plants. Two byproducts of water chlorination, 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX) and 3,4-dichloro 5-hydroxy-2[5H]-furanone (MA) induced a dose dependent increase of micronuclei (MN) in pollen mother cells of Tradescantia when doses up to 100 μg MX and 500 μg MA were applied directly to the inflorescences. In contrast, exposure of the stems in aqueous solutions containing up to 1 mg/I MX and 10 mg/I MA did not cause a positive response.     

Enhancement of micronuclei frequency in the Tradescantia/micronuclei test using a long recovery time

The Tradescantia/micronuclei test (TRAD/MCN) is a well-validated test for monitoring environmental genotoxicants. These pollutants induce at the early meiotic stage of pollen mother cells chromosome fragments which become micronuclei at the tetrad stage. The standard test protocol requires some hours of exposure of the inflorescences and a recovery time of about 24 hours to reach the early tetrad stage. Since the recovery period represents a critical step of the TRAD/MCN, experiments were performed to establish its length in plants of clone #4430 of the hybrid T. hirsutiflora x T. subacaulis which is widely used in environmental monitoring. The aim of the present research was to ascertain the exact duration of recovery time in order to improve the sensitivity of the TRAD/MCN test. First, studies were performed to select the flowers at the beginning of the meiosis, and then anthers were sampled and studied for a period of 48-86 hours. The complete meiosis in the plants examined required about 80 hours. Second, exposure to genotoxic substances followed by different recovery times was carried out to demonstrate that effectiveness of the TRAD/MCN test is closely related to the duration of the recovery time. The test was carried out by exposing inflorescences to known mutagens (sodium azide and maleic hydrazide) for six hours followed by different recovery times (24-72 hours). The results showed that the frequency of micronuclei in the pollen mother cells increased with the length of the recovery time.     

Some approaches to evaluation of the mutagenic effect of industrial waste on the environment

The mutagenic effect of an industrial enterprise (tungsten and molybdenum factory) was studied in three stages. At the first stage, the putative impact of the industrial sewage of the factory was studied using three plant test systems: Crepis capillaris L., Tradescantia sp. clone 02, and Glycine max (L.) Merill. It was found that the sewage increased the mutation level by a factor of 11-45. At the second stage, the rate of mutation was studied in the native vegetation growing on solid waste piles of the enterprise. It exceeded the corresponding index of uncontaminated areas by a factor of 2-4.5. At the third stage, the rates of children with birth defects and miscarriages were studied in the vicinity of the enterprise. The rate of miscarriages proved to be higher than the value averaged over the autonomous republic by a factor of 2.4. No change in the rate of birth defects was detected.     

Features of action of low doses of gamma-radiation on yeast cells

Lethal effect of low doses and adaptive response to low doses of prolonged irradiation were investigated in experiments on yeast cells. The phenomenon of hypersensitivity at low dozes was not found in yeast cells at gamma-irradiation. The adaptive response was observed after exposure to low doses of prolonged irradiation, the degree of the reaction depends on a dose rate. The adaptive reaction was kept for some time after the termination of adaptive irradiation then the sensitivity of cells increased even in comparison with unirradiated ones.     

Postnatal effects in children irradiated during the intra-uterine development, as a result of failure at the Chernobyl NPP

Somatic and genetic effects at children who have undergone the influence of ionizing radiation during in utero development and at the subsequent stages ontogenesis are investigated, in view of dozes of the general irradiation and equivalent dozes of an irradiation of red bone brain. 1144 children were examined: 1st group consisted of children who have been born from women pregnant at the moment of failure, evacuated from Pripyat; 2nd group included children who have been born from women pregnant at the moment of failure, stayed to live in a zone of the strict radiation control; 3rd group was the control group--children who have been born in 1986, living in safe region of the Ukraine according to radiation conditions. The presence of direct correlation link between quantity of small anomalies of the development and the total radiation dose of the fetus (R = 0.61, p < 0.002) and inverse link with fetus age at the moment of radiation factor effect (R = -0.53, p < 0.003) has been established. Significant correlation links between the total radiation dose and the level of children health (R = 0.45, p < 0.03); the equivalent radiation dose of the red bone marrow and the frequency of damages in the chromosomal apparatus of the somatic cells (1st group--R = 0.51, p < 0.02; 2nd group--R = 0.62, p < 0.002) have been revealed.     

Comparison of cytogenic response of human lymphocytes to in vivo and in vitro exposure to low doses of gamma-rays. Unstable chromosomal exchanges detected by FPG techniques

On peripheral lymphocytes of eight cancer patients undergone whole-body therapeutic irradiation (at daily dose of 10 cGy up to total dose of 50 cGy of 60Co gamma-rays) the dose-response of unstable chromosome exchanges (dicentrics and centric rings) was studied. This dose response fitted well linear function. The lower slope of dose-response curve was found for in vivo irradiated lymphocytes as compared to the dose response curve obtained for in vitro irradiated lymphocytes of the same patients. This finding seems to provide evidence that in case of protracted irradiation of individuals an absorbed dose could be underestimated if for biological dosimetry an in vitro dose response curve for unstable chromosome aberrations is used as referent one.     

Experimental study of the radioprotective action of melanin on the somatic development during irradiation in the antenatal period of ontogenesis

Daily introduction per os of the exogenous melanin in a doze of weight of 10 mg/kg pregnant female rats Wistar eliminated the functional deficiency of somatic development revealed at posterity at chronic gamma-irradiation in a doze 1.00-1.25 Gy for all period of pregnancy. The irradiation or introduction melanin antenatal to a phase ontogenesis resulted in stimulation of the immune answer, which was determined at offspring on 3rd week after birth. On the basis of the received data it is concluded presence radioembryoprotective actions melanin in the relation embryotoxic effects of small dozes of ionizing radiation and its participation in regulation immunogenes.     

The action of low doses of gamma radiation on mammalian cells]

Effect of gamma irradiation in low doses (10, 20, 40 and 50 cGy) on HeLa cells was studied. The survival of cells exposed to the irradiation in the dose of 50 cGy was decreased while it remained unchanged in cells irradiated in the dose of 10-40 cGy and their descendants. Nonetheless, their survival following an additional treatment with a mixture of cytosine arabinoside and hydroxyurea was reduced. It was suggested that the genome stability was diminished in irradiated cells and their descendants.