Frequency Of Unstable Chromosomal Aberrations Research Articles

Chromosome damage in regions with different levels of air pollution.

Air pollution is an important environmental factor influencing human health. In this study, we compared chromosome damage in city policemen from three cities in the Czech Republic: industrial Ostrava characterized by high levels of benzo[a]pyrene, Prague with heavy traffic emitting nitrogen oxides, and relatively clean Ceske Budejovice located in an area with predominantly agricultural activity. Chromosomal aberrations in lymphocytes were evaluated by fluorescence in situ hybridization with painting probes for chromosomes 1, 2, 3, and 4 in spring and autumn. An increase in the frequency of unstable chromosome aberrations, that is, dicentric chromosomes and acentric fragments, was observed in spring samples from Ostrava (p = .014 and p = .044, respectively) and Prague (p = .002 and p = .006, respectively) in comparison with Ceske Budejovice. The difference was significant only for samples taken after the winter period, when the concentration of pollutants in the air increases due to poor dispersion conditions. An increased frequency of dicentric chromosomes was observed in spring compared to autumn in both Ostrava and Prague (p = .017 and p = .023, respectively), but not in Ceske Budejovice. More breakpoints were observed on chromosome 1 than on the other chromosomes examined (p < .001). The number of breakpoints in the heterochromatin region 1p11-q12 was lower than in other parts of chromosome 1 (p < .001), suggesting a protective function of heterochromatin against damage. Our study showed, that air pollution increased the frequency of unstable chromosome aberrations, especially dicentric chromosomes. However, we did not show an effect on stable chromosome rearrangements.

Оценка влияния хронического облучения на преждевременное старение Т-лимфоцитов человека на основе нестабильных хромосомных аберраций

For more than 60 years, residents of the villages on the Techa River have been chronically exposed to combined radiation, receiving a wide range of doses. Red bone marrow (RBM) is the critical system in the exposure conditions. This study aimed to assess the effect of chronic exposure on premature aging of T-lymphocytes based on the frequency of unstable chromosome aberrations; the subjects were the residents of the Southern Urals that have been chronically exposed to radiation. The increased frequency of occurrence of dicentrics and rings in T-cells of the exposed persons was the marker of cellular aging, with the associated doses to the red bone marrow (RBM dose) at 0.5–2.5 Gy. The participants (RBM donors), both exposed and non-exposed, were divided into three age subgroups: 40–59 years old, 60–69 years old, 70–79 years old. The differences in the RBM dose among the exposed individuals were insignificant. In the exposed group, unstable chromosome aberrations (UCA) were recorded significantly more often than in the control group (p = 0.04). The age group of 40–59 years was the one where the exposed donors had significantly more frequently occurring chromosome aberrations compared to the non-exposed participants. There were no such differences registered in other age groups. The age-associated increase of the amount of chromosome aberrations was registered in the non-exposed group only. Chronic exposure to radiation indirectly promotes premature aging of T-lymphocytes: 1) in the long term, the exposed individuals had UCA significantly more often; 2) compared to the control group, the 40–59 years age subgroup of the exposed group had increased cytogenetic index. In the context of this study, the number of dicentrics and rings was not registered as increasing in the older age subgroups of exposed individuals, which may be due to the specifics of the donor inclusion criteria, which, for the elderly, may favor radioresistant individuals.

Evaluation of the types and frequency of unstable chromosomal aberrations induced in lymphocytes of breast cancer patients before and after radiotherapy

Evaluation of the types and frequency of unstable chromosomal aberrations induced in lymphocytes of breast cancer patients before and after radiotherapy

Delayed Changes in the Frequency of Unstable Chromosomal Aberrations and the CNA-Genetic Landscape of Blood Leukocytes in People Exposed to Long-Term Occupational Irradiation

Abstract—The results of a study of delayed changes in the frequency of unstable chromosomal aberrations and the CNA- and LOH-genetic landscape of blood leukocytes in healthy employees at the main facilities of the Siberian Group of Chemical Enterprises exposed to external irradiation are presented. In 2014, 11 persons with de novo induced mosaic deletions and amplifications associated with dicentric and/or ring chromosomes (markers for individual radiosensitivity) were detected. Three years later, the frequency levels of chromosomal aberrations were compared. In 2017 the frequency of aberrant cells had increased by 1.7 times compared to 2014, the frequency of the chromosomal type aberrations had doubled, the frequency of pair fragments had a more than fourfold increase, and the frequency of dicentrics had almost doubled. The mosaic deletion in employee no. 1490 (3q12.3–13.11) identified in 2014 persisted until 2017. Mosaic amplifications identified in 2014 were preserved in five out of six employees (nos. 178, 203, 450, 1620, and 1792). Moreover, we identified that the CNstate in employee no. 450 had increased 1.5 times by 2017. The mosaic amplification of the short arm of the third chromosome in employee no. 278 had been eliminated by 2017. Other changes in the CNA and LOH-genetic landscape were not detected. Thus, these data indicate the formation of self-sustained bone marrow clones, whose descendants produce a population of aberrant cells in the blood of irradiated individuals. Further research into the preservation of chromosomal aberrations and CNA is needed, as well as studies aimed at detecting the clones of aberrant cells in the bone marrow.

Study of the Individual Radiosensitivity in Humans Based on the Assessment of the Frequency of Chromosome Aberrations and Micronuclei in Peripheral Blood T Lymphocytes

The influence of low-dose ionizing radiation on a human being is increasing in the modern world (medical diagnostics etc.). The issue of the effect of low-doses on human health is topical and controversial. It is known that in the population up to 15% of people react to radiation exposure differently from what has been expected, which, according to the published data, happens owing to their genome peculiarities. The objective of the current study was to investigate the influence of chronic lowdose rate exposure (doses to red bone marrow varied within the range 3—4600 mGy) of the Techa River residents (South Urals) on the radiosensitivity of T cells in vivo and in vitro under additional gamma exposure of peripheral blood T lymphocytes (0.5, 1, and 2 Gy). Such cytogenetic parameters as frequency of unstable chromosome aberrations and micronuclei were used as irradiation markers. Persons were considered radiosensitive if the obtained values of the analyzed parameters for them exceeded the 90% percentile. On the basis of the findings of the performed study, the following conclusions were drawn: about 10% of donors demonstrated increased radiosensitivity; moreover, the effect did not depend on the dose of chronic exposure; no age dependence of radiosensitivity in the studied age range (49–89 years) of the donors was revealed. In most cases, we did not detect a reproducible radiosensitivity effect in the same donor under different irradiation regimes.

Biodosimetry Based on γ-H2AX Quantification and Cytogenetics after Partial- and Total-Body Irradiation during Fractionated Radiotherapy.

The aim of this current study was to quantitatively describe radiation-induced DNA damage and its distribution in leukocytes of cancer patients after fractionated partial- or total-body radiotherapy. Specifically, the impact of exposed anatomic region and administered dose was investigated in breast and prostate cancer patients receiving partial-body radiotherapy. DNA double-strand breaks (DSBs) were quantified by γ-H2AX immunostaining. The frequency of unstable chromosomal aberrations in stimulated lymphocytes was also determined and compared with the frequency of DNA DSBs in the same samples. The frequency of radiation-induced DNA damage was converted into dose, using ex vivo generated calibration curves, and was then compared with the administered physical dose. This study showed that 0.5 h after partial-body radiotherapy the quantity of radiation-induced γ-H2AX foci increased linearly with the administered equivalent whole-body dose for both tumor entities. Foci frequencies dropped 1 day thereafter but proportionality to the equivalent whole-body dose was maintained. Conversely, the frequency of radiation-induced cytogenetic damage increased from 0.5 h to 1 day after the first partial-body exposure with a linear dependence on the administered equivalent whole-body dose, for prostate cancer patients only. Only γ-H2AX foci assessment immediately after partial-body radiotherapy was a reliable measure of the expected equivalent whole-body dose. Local tumor doses could be approximated with both assays after one day. After total-body radiotherapy satisfactory dose estimates were achieved with both assays up to 8 h after exposure. In conclusion, the quantification of radiation-induced γ-H2AX foci, but not cytogenetic damage in peripheral leukocytes was a sensitive and rapid biodosimeter after acute heterogeneous irradiation of partial body volumes that was able to primarily assess the absorbed equivalent whole-body dose.

Chronic radioisotope effects on residents of the Techa River (Russia) region: Cytogenetic analysis more than 50 years after onset of exposure

This paper presents the results of a cytogenetic study conducted among residents of the Techa Riverside communities (Southern Urals, Russia) exposed in the early 1950s as a result of releases of liquid radioactive wastes from the Mayak plutonium-production facility. The study was performed 50–60 years after the beginning of the exposure for those individuals who were predominantly exposed to strontium radioisotopes (89,90Sr) through drinking contaminated river water and consumption of local foodstuff.Standard cytogenetic methods were used for evaluation of the frequency of unstable chromosome aberrations in exposed persons as well as in persons from the control group who were of similar age and sex, living in similar socio-economic conditions in non-contaminated territories of the Southern Urals.The exposure doses were reconstructed for the studied donors using the Techa River Dosimetry System developed in 2009. The doses of internal exposure from ingested radionuclides were evaluated using individual or family in vivo measurements of 90Sr-body burden. Individual cumulative absorbed doses in red bone marrow (RBM) in the studied persons varied in the range of 0.01–4.4Gy.A significantly higher level of unstable chromosome aberrations (UCA) in T-cells was observed in the group of exposed individuals as compared to control group. The highest UCA level was detected in the individuals who were suspected of having chronic radiation syndrome.

Сравнительный анализ результатов цитогенетических обследований контрольных групп лиц в различных отечественных лабораториях

The incidence of unstable chromosome aberrations in peripheral blood lymphocytes from unirradiated control subjects was analyzed using cytogenetic data obtained from 9 cytogenetic laboratories located in Moscow, St.-Petersburg, Obninsk, and Dubna (Russia). The objective of this study was to estimate the level and spectrum of spontaneous chromosome aberrations in human lymphocytes. 1140 blood samples were taken from 1112 subjects (594 men and 546 women) aged 1 to 72. The total metaphase number was 466795. The uniform Giemsa method for peripheral blood lymphocyte cultures was used. After counting 466795 metaphases, 4288 chromosomal aberrations of various types were classified. The most frequent types of aberrations were acentrics and chromatid deletions. They made up 90% of the total number of aberrations. The remaining 10% were exchange aberrations. The number of chromosome exchanges (dicentrics and centric rings) was twice the number of chromatid exchanges. Overall, the portion ofcells with chromosomal or (and) chromatid aberrations was 0.89 +/- 0.01%; the frequency of acentrics was 0.29 +/- 0.01; the frequency of dicentrics was 0.046 +/- 0.003; the frequency of unstable chromosome aberrations was 0.35 +/- 0.01; and the frequency of chromatid aberrations was 0.57 +/- 0.01 per 100 cells.

Streptonigrin induces delayed chromosomal instability involving interstitial telomeric sequences in Chinese hamster ovary cells

We analyzed the induction of chromosomal aberrations in Chinese hamster ovary (CHO) cells exposed to the radiomimetic compound streptonigrin (SN), in order to determine whether interstitial telomeric sequences (ITSs) are involved in the long-term clastogenic effect of this antibiotic. CHO cells were treated with a single concentration of SN (100ng/ml), and the frequency of unstable chromosomal aberrations was determined at three times after treatment (18h, and 6 and 15 days) by using PNA-FISH with a pan-telomeric probe. Cytogenetic analysis revealed a higher frequency of aberrations at 18h and 6 days after treatment in SN-exposed cultures vs. untreated cultures. The percentage of damaged cells and the yield of SN-induced aberrations at 6 days after treatment increased on average twofold compared with the ones at 18h after treatment. Moreover, a significant decrease in the frequency of aberrations was observed in SN-exposed cells at 15 days after treatment, resulting in a frequency of aberrations significantly lower than the frequency of aberrations observed in the corresponding control cultures. These data indicate that SN induces delayed chromosomal instability in CHO cells, and that the in vitro clastogenic effect of this compound persists for at least 6 days but less than 15 days after treatment. In addition, we found that SN induces delayed ITSs instability, cytogenetically detectable as additional FISH signals and centromeric breaks involving dissociation of the telomeric signal 6 days after treatment. We propose that the delayed effect of SN on ITSs results from breakage of heterochromatic centromeric ITSs blocks and further insertion of these sequences at the sites of mono- or isochromatid breaks occurring at G2 or G1-S phases of the cell cycle, respectively, since most of the additional FISH signals were present as single or double dots, and located at interstitial sites of the involved chromosomes.

Bleomycin induces delayed instability of interstitial telomeric sequences in Chinese hamster ovary cells

We analyzed the behavior of interstitial telomeric sequences (ITSs) in the progeny of Chinese Hamster Ovary (CHO) cells exposed to the radiomimetic compound bleomycin (BLM) in order to determine if ITSs play some role in the long-term clastogenic effect of this antibiotic. To this end, CHO cells were treated with a single concentration of BLM (2.5μg/ml), and the frequency of unstable chromosomal aberrations was determined at several times after treatment (18h, and 6, 15 and 34/36 days) by using PNA-FISH with a pan-telomeric probe [(TTAGGG)n repeats]. Cytogenetic analysis revealed a higher frequency of aberrations at 18h and 6 days after treatment in BLM-exposed cultures vs. untreated cultures, although the yield of BLM-induced aberrations decreased on average five times 6 days after treatment compared with the one induced 18h after treatment. Moreover, no significant differences in the frequency of aberrations were observed between untreated and BLM-exposed cells at 15 or 34/36 days after treatment. These data indicate that, in terms of unstable aberrations, the in vitro clastogenic effect of BLM on CHO cells persists for at least 6 days but less than 15 days after exposure. In addition, we found that BLM induces ITSs instability, cytogenetically detectable as acentric fragments (18h after treatment) or additional (new) FISH signals (6 days after treatment). We propose that the delayed effect of BLM on ITSs mainly results from breakage of heterochromatic ITSs blocks and further insertion of these sequences at the sites of monochromatid breaks occurring at G2 phase of the cell cycle, since most of the additional FISH signals were present as single dots and located at interstitial sites of the involved chromosomes.

FOLLOW-UP STUDY BY CHROMOSOME ABERRATION ANALYSIS AND MICRONUCLEUS ASSAYS IN VICTIMS ACCIDENTALLY EXPOSED TO 60Co RADIATION

The goal of this study was to assess the persistence of chromosomal aberrations and micronuclei of three victims 2 y after accidental radiation exposure to Co gamma rays. Traditional chromosome aberration analysis was performed by scoring the dicentric chromosomes (dic) and rings (r) in peripheral blood lymphocytes. Micronuclei were detected using the cytokinesis block micronucleus assay. G-banding and semi-automatic karyotype analysis was used to record translocations (t), inversions (inv) and deletions (del). The frequency of unstable chromosomal aberrations (dicentrics and rings) remained at high levels 6 mo after the accident. Two years after exposure, the frequency was reduced to 4-11% in the three victims. However, stable chromosome aberrations, which were detected by G-banding and included t, inv, and del, remained at a high level and have an obvious dose-dependent relationship even 2 y post-exposure. The frequency of micronuclei decreased faster than that of chromosome aberrations, reaching almost a normal level two years after the accident, especially for the child victim. Unstable chromosome aberrations reduced gradually, but the stable aberration remained at a high level along with the time-lapse. The micronucleus assay was less valuable for assessing long-term effects after high dose irradiation.

Seventeen-year follow-up study on chromosomal aberrations in five victims accidentally exposed to several Gy of 60Co γ-rays

On 25 June 1990, a radiation accident occurred in a (60)Co source radiation unit in Shanghai, due to violations in operation regulations. This accident resulted in the exposure of seven individuals to acute high-dose and dose-rate whole-body external irradiation. Conventional chromosomal aberration analysis, G-banding automatic karyotype analysis and/or fluorescent in situ hybridization (FISH) painting methods were used to analyze chromosomal aberrations in peripheral blood lymphocytes from five of the victims 24 h to 17 years after accidental exposure to 1.9-5.1 Gy of (60)Co gamma-rays. The frequency of unstable chromosomal aberrations (dicentrics and rings) remained at constant levels 1 month after exposure. Three months after exposure, the frequency was reduced by 20-40% in three victims, while no reduction was seen in the other two victims. Twelve years after exposure, the number of dicentrics and rings decreased by more than 90%, and did not reveal a dose-dependent relationship. However, even at 12-17 years after exposure, stable chromosome aberrations, dominated by translocations, remained at a high level in a dose-dependent manner. The frequency of stable chromosomal aberrations detected by FISH showed a similar dose-dependent relationship as that detected by karyotype analysis of G-banding chromosomes. The G-banding analysis also suggested that the pattern of chromosome breakpoints is random. The FISH data showed a decreasing tendency with time for chromosome translocation frequency in the peripheral lymphocytes, and the rate of reduction varied among different individuals. It is likely that the higher dose the victim received, the lesser the translocation frequency decreased with time. The G-banding data also showed that the rate of reduction of translocations is different among individuals. From 5 to 17 years after accidental irradiation, a very small reduction (approximately 10%) of translocation frequency was observed in victims C and D, while there was about a 35% reduction (the highest among the victims) for victim G who received the smallest dose (1.9 Gy). These observations can be used to validate the existence of chromosomal aberrations in peripheral blood lymphocytes as a biological dosimeter for radiation exposures.

Mechanism of adaptive response: Estimation of human lymphocyte adaptive response to radiation using various criteria

Blood lymphocytes of 15 healthy donors have been investigated for their ability to reduce radiosensitivity after low-dose irradiation-radio-induced adaptive response (AR). The frequency of unstable chromosome aberrations was used to evaluate cell radiosensitivity after the irradiation of cells in low adaptive (5 cGy) and high challenge (1 Gy) doses in comparison with the effect of challenge irradiation only. Three indexes have been used, i.e., (A) the frequency of cells with aberrations per total analyzed cell, (B) the number of chromosome aberrations per one cell, (C) and the number of chromosome aberrations per one aberrant cell. It was found that the donors can be divided in the four following groups: 1. AR was not estimated any of the indexes used; 2. AR was estimated with indexes A and B, but not C; 3. AR was shown by indexes B and C; 4. AR was evident with all three indexes. The generally accepted AR repair model only explains the appearance of group-3 and-4 donors, but not group-2. For the purpose of understanding the AR mechanisms and the difference in AR estimations with various criteria, the metaphase distribution by the number of chromosome aberrations has been analyzed for each donor. It was shown that, in group-2 donors, the number of cells without aberrations after adaptive and challenge irradiations was significantly higher than after irradiation with a challenge dose only. Thus, in this group, AR is formed as a result of the changed frequency of cells in the 0 class (population shift). A similar shift is observed in the metaphase distribution in the donors of group 4, but not in group 3. The data obtained show that AR is probably a result of several processes, including the activation of the reparation of premutational genome damages, population shifts evident in the frequency of undamaged cells, and, possibly, the activation of apoptotic cell death. The complex character of AR is reflected to different degrees in each criterion of radiosensitivity.

BIOLOGICAL ESTIMATION OF DOSE IN HEMI-BODY IRRADIATION OF CANCER PATIENTS BY CYTOGENETIC ANALYSIS

The objective of this study was to estimate equivalent whole body dose and dose to the irradiated region in hemi-body irradiation of cancer patients using chromosome aberrations and micronuclei techniques. The frequency of unstable chromosome aberrations and micronuclei was studied in peripheral blood lymphocytes of cancer patients before and after hemi-body irradiation. The mean estimated equivalent whole body dose was 3.74 +/- 0.16 Gy and 4.07 +/- 0.15 Gy from dicentrics and micronuclei frequency in lymphocytes after hemi-body irradiation. The mean dose to the irradiated region from unstable chromosome aberrations was estimated to be 6.85 +/- 0.2 Gy and 6.79 +/- 0.26 Gy using Dolphin's contaminated Poisson dispersion analysis and the Qdr method, respectively. The mean dose to the irradiated region from micronuclei frequency was 7.04 +/- 0.25 Gy by Dolphin's method. There was a slight underestimation of mean dose to the irradiated part in vivo. The percentage of irradiated blood and fraction of the cells exposed could be estimated using the contaminated Poisson method.

Characteristics of genomic instability in clones of TK6 human lymphoblasts surviving exposure to 56Fe ions.

Genomic instability in the human lymphoblast cell line TK6 was studied in clones surviving 36 generations after exposure to accelerated 56Fe ions. Clones were assayed for 20 characteristics, including chromosome aberrations, plating efficiency, apoptosis, cell cycle distribution, response to a second irradiation, and mutant frequency at two loci. The primary effect of the 56Fe-ion exposure on the surviving clones was a significant increase in the frequency of unstable chromosome aberrations compared to the very low spontaneous frequency, along with an increase in the phenotypic complexity of the unstable clones. The radiation-induced increase in the frequency of unstable chromosome aberrations was much greater than that observed previously in clones of the related cell line, WTK1, which in comparison to the TK6 cell line expresses an increased radiation resistance, a mutant TP53 protein, and an increased frequency of spontaneous unstable chromosome aberrations. The characteristics of the unstable clones of the two cell lines also differed. Most of the TK6 clones surviving exposure to 56Fe ions showed unstable cytogenetic abnormalities, while the phenotype of the WTK1 clones was more diverse. The results underscore the importance of genotype in the characteristics of instability after radiation exposure.

Results of a Cytogenetic Follow-up Study 7.5 Years After 137Cs Exposure at the Goiania (Brazil) Radiological Accident

Ten persons exposed to 137 Cs during the radiological accident in Goiania (Brazil) were re-examined for the frequency of unstable chromosomal aberrations (dicentric chromosomes, centric rings and acentric fragments) 7.5 years after the first examination. It was found that the frequencies fell to about 5% of the initial values, for those individuals who had been exposed to moderate to high doses. For the subjects exposed to low doses, of the order of 0.2 Gy or less, the observed frequencies of chromosomal aberrations fell much more slowly.