Long-term follow-up cytogenetic survey and biological dosimetry in persons evacuated from 30-km Chernobyl NPP zone

The paper presents the results of the follow-up cytogenetic survey and biological dosimetry carried out in inhabitants of Pripiat' town and nearby villages, who were departured from the Chernobyl NPP 30-km exclusive zone during first days after the Chernobyl catastrophe. The unstable chromosome aberration level in inhabitants were significantly increased above control in terms up to 1 year after evacuation and declined gardually during next 14 years. In early period the cytogenetic damage frequency in evacuees showed no dependence on gender. The chromosome type aberration level appeared to be lower in young persons comparing with adults. The dicentrics plus centric rings yield had a positive correlation with duration of staying at Chernobyl zone. The average doses of protracted exposure were calculated from the dicentrics and centric rings yields; the dose estimations appeared to be 1.4 times higher in persons evacuated 3-11 days after the accident than that of in persons with shorter departure time. Uing the Bayesian analysis the probabilistic distribution of biological doses was constructed for the studied evacuees group. This distribution was characterized by a mean dose of 360 mGy, the modal doses of 200-450 mGy and 80% of probability density within the dose range 0-1000 mGy, that seems to be sufficient for considering the increased risk of late somatic radiation effects for this cohort.     

Level of chromosome aberrations in peripheral blood lymphocytes in evacuees from the 30-kilometer zone of the Chernobyl Atomic Energy Station and residents of radioactively-polluted territories in the long term after the Chernobyl accident

Eighteen Ukrainian evacuees from the Chernobyl exclusive zone, twenty one inhabitants of radioactively contaminated areas of Belarus and twelve control donors age-matched to the exposed persons were investigated 14-15 years after the Chernobyl accident for chromosomal aberration yields detected in blood lymphocytes by fluorescence in situ hybridisation technique. Unstable aberration yields measured in both Chernobyl cohorts were close to the background frequencies. Positive age-dependence trends in control donors were determined for the all type stable aberration levels. In evacuees the tendency for diminishing the difference between them and controls for stable aberration levels with persons' age increasing was found. The total stable chromosome exchange yields in evacuees 46-55 years old and inhabitants of areas with low contamination level didn't exceed the control values, but for younger evacuees and inhabitants of sufficiently contaminated regions the statistical increase above the age relevant background meanings was detected for this end-point. The advantages of using the FISH-detectable stable aberrations and particularly the total level of stable chromosome exchanges as the end-points for retrospective biological indication of past radiation exposure in Chernobyl cohorts were discussed.     

Chromosome alterations in cleanup workers sampled years after the Chernobyl accident

Cytogenetic analysis performed 4-13 years after the Chernobyl accident showed an elevated frequency of acentrics, chromatid exchanges, dicentrics and rings in Chernobyl cleanup workers compared to the control group. Cytogenetic data were analyzed according to the information on exposure to radiation (the year when the cleanup workers worked at the Chernobyl station, doses rates, time elapsed since exposure to radiation, and cytogenetic examination) and some lifestyle factors. The data obtained suggested that some types of chromosome aberrations could be influenced by the action of different environmental factors or lifestyle factors. The frequency of acentrics was correlated with the age of the cleanup workers, and the increased chromatid exchange frequency was attributed to smoking. The numbers of dicentrics and rings suggested a genotoxic effect of ionizing radiation that is still present over 13 years after the exposure.     

Possibilities and limitations of fluorescence in situ hybridization technique in retrospective detection of low dose radiation exposure in post-chernobyl human cohorts

Cytogenetic analysis using the fluorescence in situ hybridisation (FISH) technique was performed late time after the Chernobyl accident in groups of liquidators, evacuees from 30 km exclusive zone, residents of radioactively contaminated areas and control donors age-matched to exposed persons. Stable and unstable chromosome type exchanges were recorded using a hybrid conventional-PAINT nomenclature. The mean yield of stable chromosome exchanges in liquidators did not correlate with registered radiation doses but had a clear negative dependence on the duration of liquidators' staying in Chernobyl zone, that was in a good agreement with early data based on conventional dicentrics plus rings analysis. The overspontaneous excess for stable chromosome exchange level appeared to be higher in evacuees 16-40 years old than that of senior persons, whereas no age-dependent difference occurred for initially induced dicentrics plus rings yields in this cohort. The stable chromosome exchange yield, as well as combined yield of dicentrics plus rings and potentially unstable incomplete translocations in residents of radioactively contaminated areas showed a reasonable positive correlation with levels of 137Cs contamination. The observed yields of stable chromosome exchanges in all three exposed groups appeared to be somewhat lower than those of expected from unstable exchange-based doses which were referred to an in vitro dose response of stable exchanges outcome in human lymphocytes. Thus, FISH analysis can be successfully applied for qualitative cytogenetic indication of past and chronic radiation exposure to low doses but further refinement of FISH-based system for quantitative dose assessment is still required. Some practical approaches of solving this task are discussed.     

Time-effect relationship for unstable chromosome exchange levels in Chernobyl clean-up workers

The post-irradiation changes of dicentric and centric ring levels were studied in Chernobyl liquidators using the data of 507 individual chromosomal surveys of persons sampled at different time after their activities at Chernobyl NPP accident zone. The time-effect relationship within 0-10.5 years after exposure was displayed as exponential decline of the mean chromosome exchange frequency with average decay half-time 2.2 y. During 10.5-13 years after exposure the increasing and stabilization of chromosome exchange yield on the level 2-3-times higher than control was observed. In the first few months after irradiation the dicentric and centric ring frequency in liquidators had the clear reverse correlation with the duration of person's duties at the Chernobyl zone. The parameters of unstable chromosome exchange elimination were independent on the initially induced aberration yield, that resulted in earlier reaching the subcontrol level in persons who had more protracted duration of duties at the Chernobyl accident zone.     

The role of cytogenetic examination for prognosis of remote consequences of irradiation

Correlation between the level of somatic pathology and cytogenetic characteristics of blood was analyzed in a group of liquidators of the accident at the Chernobyl Nuclear Power Plant (ChNPP). A statistically significant correlation was found between the occurrence of cardiovascular diseases and the level of chromosome aberrations (total frequency of chromosome aberrations, frequency of dicentrics and centric rings, frequency of chromatid aberrations). The results obtained are of great importance for the substantiated prediction of the development of postradiation pathologies.     

Results of cytogenetic studies of the consequences of the Chernobyl accident

The selected results of the cytogenetic studies of the Chernobyl accident consequences were summarised. The chromosomal aberrations were used as a method of biodosimetry for a dose assessment for victims during the initial period after the Chernobyl accident. A good correlation between doses calculated based on chromosomal aberrations (dicentrics) and severity of acute radiation syndrome observed in clinic was found. The biodosimetry based on conventional cytogenetic technique (dicentrics) has been unsuccessful for various groups (rehabilitation workers, evacuees, inhabitants of contaminated areas) sampled long time after the Chernobyl accident. The possible reasons of the failure are analysed. The original results of multiaberration cell yield observed in different cohorts of the Chernobyl victims are presented. The problems related to the phenomena are discussed.     

Increased risk of cancer in radon-exposed miners with elevated frequency of chromosomal aberrations

In spite of the extensive use of cytogenetic analysis of human peripheral blood lymphocytes in the biomonitoring of exposure to various mutagens and carcinogens, the long-term effects of an increased frequency of chromosomal aberrations in individuals are still uncertain. Few epidemiologic studies have addressed this issue, and a moderate risk of cancer in individuals with an elevated frequency of chromosomal aberrations has been observed.In the present study, we analyzed data on 1323 cytogenetic assays and 225 subjects examined because of occupational exposures to radon (range of exposure from 1.7 to 662.3 working level month (WLM)). Seventy-five subjects were non-smokers. We found 36 cases of cancer in this cohort.Chromatid breaks were the most frequently observed type of aberrations (mean frequency 1.2 per 100 cells), which statistically significantly correlated with radon exposure (Spearman's correlation coefficient R=0.22, P<0.001). Also, the frequency of aberrant cells (median of 2.5%) correlated with radon exposure (Spearman's correlation coefficient R=0.16, P<0.02). Smoking and silicosis were not associated with results of cytogenetic analyses.The Cox regression models, which accounted for the age at time of first cytogenetic assay, radon exposure, and smoking showed strong and statistically significant associations between cancer incidence and frequency of chromatid breaks and frequency of aberrant cells, respectively. A 1% increase in the frequency of aberrant cells was paralleled by a 62% increase in risk of cancer (P<0.000). An increase in frequency of chromatid breaks by 1 per 100 cells was followed by a 99% increase in risk of cancer (P<0.000). We obtained similar results when we analyzed the incidence of lung cancer and the incidence other than lung cancer separately.Contrary to frequency of chromatid breaks and frequency of aberrant cells, the frequency of chromatid exchanges, and chromosome-type aberrations were not predictive of cancer.     

The problem of the transgeneration phenomenon of genome instability in sick children of different age groups after the accident at the Chernobyl Nuclear Power Plant

A complex genetic examination of children which belong to two cohorts and their parents were carried out. The first cohort included children and constantly living on territories contaminated with radionuclides (Novozybkov district, Bryansk region). They were subdivided in groups according to the ontogenetic age periods of development of their parents at the time of the Chernobyl accident. In the children born in 1986-1995 the level of aberrant genomes is significantly higher as compared to the control (p < 0.001). In children born in 1998-2002 the differences are insignificant (p > 0.05). The frequency of aberrant genomes had a tendency to decrease with the period of time between the birth date of a child and the moment of the accident. Analysis of the results of cytogenetic investigation for the same living on territories with different densities of radioactive contamination (zone I-- 627-688 kBq/m2, 137Cs and zone II-- 135-402 kBq/m2, 137Cs) revealed insignificant differences in the spectrum and average frequencies of chromosome aberrations. The second cohort included children born in 1987-1991 and 1993-2002 from irradiated fathers (Chernobyl clean-up workers) and unirradiated mothers living on territories without radionuclide contamination. These children also displayed increased frequencies of aberrant genomes as compared to the control (p < 0.001). The analysis of the dynamics years of birth of cytogenetic disturbances in the same cohorts of children showed the average frequencies of aberrant genomes remain higher than the control level. In most of the children of both cohorts the repair synthesis of genome DNA by gamma- and UV-radiation is reduced as compared to one in the children from the control group.     

The retrospective cytogenetic dosimetry using the results of conventional chromosomal analysis in Chernobyl clean-up workers

The paper presents the results of the cohortal biodosimetry carried out in 435 Chernobyl clean-up workers, who were surveyed with the conventional cytogenetic technique in terms from several days to 10 years after the end of their duties in the Chernobyl accident exclusive zone. An empirical model of the aberrant cell dynamics was utilized for the calculation of mean initial yields of dicentrics and centric rings in groups with different terms and duration of staying in the Chernobyl zone. Corresponding protracted irradiation doses estimated from aberration levels ranged from 79 to 670 mGy. The probabilistic distribution of the radiation doses was constructed by the applying the Bayesian analysis to initial individual chromosome exchange yields extrapolated to the exposure termination moment. This distribution was characterized by the mean dose about 460 mGy and maximum of probability density in the interval of 50-300 mGy. For the late somatic risk assessment in clean-up workers the probabilistic distribution of equivalentally-acute radiation doses was proposed; that had the mean value about 270 mGy, modal classes of 250-350 mGy and 99.8% of the probability density concentrated within the dose range from 0 to 1000 mGy.     

The cytogenetic effect in a group of settlers from a 30-kilometer area of right of way

One of the critical human group of potential risk is the group of population from 30-km zone around the Chernobyl nuclear power plant. The cytogenetical examination of 12 persons (8 man, 4 woman, 30-60 years old) from 5 villages situated in north-west and south-east directions from the ChNPP in the frame of this zone was conducted. The data of the conventional analysis showed relatively low cytogenetic effects comparable with the effects in exposed people out of the zone (the mean level of unstable cytogenetic markers in different villages was 0.21-0.35 per 100 cells). The level of stable translocations in one case (woman, 59 years old) determined using FISH (16.62 per 100 cells for the whole genome) exceeded their spontaneous aged frequency and confirmed the high sensitivity of this method for the evaluation of real radiation exposure.     

The significance of cytogenetic investigation for the estimation of Chernobyl accident consequences

The results of the cytogenetic investigation of people, which were exposed to radiation in the result of the Chernobyl NPP accident, were presented. Also the possibilities of the application of cytogenetic findings for dose estimations and for the prediction of the radiation influence consequences were examined. During the period of time since 1986 till 2004 the cytogenetic investigations of 1724 liquidators participating in the liquidation works after the Chernobyl accident were carried out. The radiation dose estimated by the frequency of dicentrics in 1986 was about 0.16 Gy. The doses for liquidators were determined by the frequency of translocations (FISH method) during the period from 1992 till 1995. For liquidators who worked in Chernobyl only in 1986 the average dose of radiation was about 0.19 Gy and for liquidators who worked repeatedly during the period from 1986 till 1995 - 0.39 Gy. There was shown that during the whole period of investigation (1986-2004) the frequency of dicentrics in peripheral blood lymphocytes was significantly higher than the control level. The cytogenetic investigation of Bryansk region inhabitants which was carried out in 1992-1994 discovered heightened value in 5 times than the control one. Findings are of great importance for the prediction of ill effects of radiation and for the development of sensitive criterions for early exposure disturbances in state of health.     

Doses of Ukrainian female clean-up workers with diagnosed breast cancer

The Chernobyl reactor accident in 1986 has caused significant exposure to ionizing radiation of the Ukrainian population, in particular clean-up workers and evacuees from the exclusion zones. A study aiming at the discovery of radiation markers of the breast cancer was conducted from 2008 to 2015 within a collaborative project by HZM, LMU, and NRCRM. In this study, post-Chernobyl breast cancer cases both in radiation-exposed female patients diagnosed at age less than 60 from 1992 to 2014 and in non-exposed controls matched for residency, tumor type, age at diagnosis, TNM classification as well as tumor grading were investigated for molecular changes with special emphasis to copy number alterations and miRNA profiles. Cancer registry and clinical archive data were used to identify 435 breast cancer patients among female clean-up workers and 14 among evacuees from highly contaminated territories as candidates for the study. Of these, 129 breast cancer patients fit study inclusion criteria and were traced for individual reconstruction of the target organ (breast) doses. The doses were estimated for 71 exposed cases (clean-up workers and evacuees from which biomaterial was available for molecular studies and who agreed to participate in a dosimetric interview) by the use of the well-established RADRUE method, which was adjusted specifically for the assessment of breast doses. The results of 58 female clean-up workers showed a large inter-individual variability of doses in a range of about five orders of magnitude: from 0.03 to 929 mGy, with median of 5.8 mGy. The study provides the first quantitative estimate of exposures received by female clean-up workers, which represent a limited but very important group of population affected by the Chernobyl accident. The doses of 13 women evacuated after the accident who did not take part in the clean-up activities (from 4 to 45 mGy with median of 19 mGy) are in line with the previous estimates for the evacuees from Pripyat and the 30-km zone.     

Chromosome instability revealed in children of fathers irradiated during Chernobyl accident

Four children groups with and without thyroid pathology born to fathers exposed to ionizing irradiation in 1986 during Chernobyl accident as liquidators as well as residents of territory with radioactive contamination have been cytogenetically observed. The frequency and spectrum of chromosome aberrations in peripheral blood lymphocytes have been studied using two-termed cultivation (during 48 and 144 hours). Under the short-term incubation the observed groups did not significantly differ on the mean-group integral cytogenetic parameters which corresponded to age norm, but in progeny of parents from radionuclide contaminated territory the increased level of chromosome type exchanges has been revealed. In long-termed lymphocyte cultures of children with chronic thyroiditis the significantly increased cytogenetic effects of both chromatid (single fragments) and chromosome types (abnormal monocentrics, centric rings) have been established. The data received testified the reality of the transmissible chromosome instability phenomenon in progeny of irradiated parents and confirmed the possibility of its expression in consequent mitoses.     

Chromosome aberrations and rogue cells in lymphocytes of Chernobyl clean-up workers

A cytogenetic analysis was performed on peripheral blood lymphocytes from 183 Chernobyl clean-up workers and 27 control individuals. Increased frequencies of chromosome aberrations were associated with exposure to radiation at Chernobyl, alcohol abuse and a history of recent influenza infection. However, only approximately 20% of Chernobyl clean-up workers had an increased frequency of dicentric and ring chromosomes. At the same time, an increased frequency of acentric fragments in lymphocytes of clean-up workers was characteristic. The use of multivitamins as dietary supplement significantly decreased the frequency of chromosome aberrations, especially of chromatid breaks. Rogue cells were found in lymphocytes of 28 clean-up workers and 3 control individuals. The appearance of rogue cells was associated with a recent history of acute respiratory disease (presumably caused by adenoviral infection) and, probably, alcohol abuse. Dicentric chromosomes in rogue cells were distributed according to a negative binomial distribution. Occurrence of rogue cells due to a perturbation of cell cycle control and abnormal apoptosis is suggested.     

Results of dynamic cytogenetic study of children and teenagers living in areas, radioactive by contaminated after the Chernobyl accident

Results of multiyear cytogenetic study of children and teenagers living in areas, radioactive by contaminated after Chernobyl accident, were adduced. Mean density of 137Cs contamination in two compared living areas were 111 and 200 kBq/m2 and mean external doses accumulated for 1986-2001 were 6.7 and 11.4 mGy correspondingly. Averaged thyroid doses receives by subjects of all age groups in the second area were approximately 1.5 times higher than in the first area; in the youngest group (0-1 year) the doses were 114.3 and 174.3 mGy. During 17 years cytogenetic investigation approximately from 30% to 60% of examined persons were observed the increased level of chromosome aberrations in lymphocytes of peripheral blood. Average frequency of unstable aberrations (acentrics, dicentrics and centric rings) constituted about 0.4 per 100 cells (0.22 per 100 cells in controls) during all period of observation. Level of marker aberrations (dicentrics and centric rings) was increased almost all times of study and varied within 0.04-0.19 per 100 cell (0.03 in control group). The parallel investigation of frequency of stable aberrations by FISH method showed up their level about 3 times exceeding observed dicentrics level. Comparably higher indexes of cytogenetic disturbances were revealed in group exposed in utero during period of accident.     

Cytogenetic aberrations in peripheral blood mononuclear cells in acute Lyme borreliosis patients

Cytogenetic aberrations are known to be caused by biological mutagenic factors, including both intracellular parasites, such as viruses, and some extracellular infectious agents, including bacteria, protozoa, and helminthes. The present study is aimed at investigating the cytogenetic status of acute Lyme borreliosis (LB) patients. As a result of the present study, a peripheral blood mononuclear cell culture of acute LB patients demonstrated a significantly higher frequency of lymphocytes with cytogenetic aberrations of the aneugenic type, such as polyploidy, hypoploidy, or endoreduplication, as well as of the clastogenic type, such as chromatid and chromosomal breaks accompanied by the inhibition of proliferative responses to mitogens and an increase in the elimination of genetically abnormal cells by apoptosis compared to the control.     

Species specificity of karyotype instability under conditions of radionuclide contamination (the Chernobyl Nuclear Power Plant) in the voles, Micrototus oeconomus, Micrototus arvalis, and Clethrionomys glareolus

The comparative analysis of frequencies of different type cytogenetic anomalies in voles Microtus oeconomus, Microtus arvalis, Clethrionomys glareolus trapped in territories with different levels of radionuclide contamination inside the 30-km zone of estrangement around the Chernobyl NPP was carried out. Animals with constitutive chromosome abnormalities were not revealed. The frequency of lympocytes with micronuclei was the most universal and sensitive character to chronic low-dose radiation in investigated species. The species-specific relation was observed between level of radionuclide contamination in trapping zone and increase in frequencies of cytogenetic abnormalities: aneuploidy in Microtus arvalis and metaphase plates with asynchronous centromere fission in Microtus oeconomus. Common vole appears to be the most sensitive species of investigated ones to chronic low-dose ionizing irradiation.     

20 years after Chernobyl accident--is it a lot or not for the estimation its characteristics ?

The results of long standing researches in the zones, which suffered from Chernobyl accident, on cultured cells, mice, blood lymphocytes of inhabitants of polluted regions (adults and children) are brought in the paper. The conclusion is that residence in polluted territories results in genomic instability, that is manifested in many effects on cytogenetic, cellular and organisms levels. It was shown, that in late descendants of cells after exposition in Chernobyl zone the increase of cell death, the micronuclei level, the frequency of giant cells, the enhancement of radiosensitivity and the absence of the adaptive response was observed. In the culture of embryonic fibroblasts, that was obtained from the exposed mice, the increase of cells with aberrations of chromosome and the frequency of multiaberrant cells was noticed. In mice exposed in Chernobyl zone the decrease of amounts of the endothelial cells in the different parts of the brain, the enhancement of mice radiosensitivity was observed. All effects were discovered in the late descendants of cells and can be the result of genomic instability induced by low level irradiation in polluted by Chernobyl accident regions.     

Restoration of pine plantations after effect of ionizing radiation in the region of the accident at the Chernobyl Atomic Energy Station

The main results of the 12-year radiation-genetic monitoring of radiobiological, cytogenetic, and genetic parameters in the Pinus sylvestris L. forest plantations from the zone of the accident at the Chernobyl nuclear power plant presented. Acute ionizing irradiation at doses > 1 Gy was shown to induce the formation of morphoses and depressed growth; at doses > 2 Gy, the reproductive ability of the trees declined. The radiobiological parameters showed a linear (or close to linear) dose-effect relationship. Acute irradiation at a dose of 0.5 Gy induced cytogenetic and genetic effects that were significantly higher than the corresponding control values. The relationship between the cytogenetic effects and the absorbed dose was exponential. The dependence of the mutation frequency at specific loci on the absorbed dose was described by a nonlinear curve. The results of cytogenetic analysis of sprouts obtained from seeds annually (1986-1998) collected in zones of slight, moderate, and strong damage of Pinus sylvestris L. are presented.