Cytogenetic analysis of peripheral blood lymphocytes in workers exposed to vinyl chloride

In the present study, the method of cytogenetic analysis of peripheral blood lymphocytes was used to examine 43 workers exposed to vinyl chloride monomer (average exposure 11.2 years) and 22 subjects selected from the same locality (control group). A total number of 8650 metaphases were analysed. All cytogenetic parameters examined were increased in the exposed group as compared to the control group and 3 parameters, chromatid breaks, percentage of aberrant cells and breaks per cell, were significantly increased (P less than 0.001).     

Cytogenetic analysis of peripheral blood lymphocytes in workers exposed to benzene

In the present study, the method of cytogenetic analysis of peripheral blood lymphocytes was used to investigate 66 workers exposed to benzene, and 20 individuals selected from general population from the same locality, not exposed to particular mutagenic or carcinogenic agents (control group). Altogether, 8,600 metaphases were analysed. Frequencies of aberrant cells, including chromatide and chromosomal breaks, and chromatide and chromosomal exchanges, were scored in both groups. A very slight increase in aberrant cell frequencies (2.152% aberrant cells) was observed in the professional exposure group as compared to the control group (1.6% aberrant cells). Increased frequencies of aberrant cells were found in smokers of both the benzene-exposed and the control group. The differences were however not significant. In addition to cytogenetic examination, the workers underwent a general examination of their health condition (preventive examination). Benzene exposure seemed to have no injurious effect on the state of health of exposed workers. Biochemical and haematological tests gave normal values.     

Occupational exposure to bidi tobacco increases chromosomal aberrations in tobacco processors

In India, workers engaged in processing of tobacco for the manufacture of bidis (the indogenous substitute for cigarettes) are chronically exposed to tobacco flakes and dust via the cutaneous and nasopharyngeal routes. Hence, workers in a tobacco processing factory were monitored for chromosomal aberrations (CA) using peripheral blood lymphocytes as the test system. Cytogenetic analysis revealed a significant increase in deletion fragments and chromatid gaps in the exposed group. The frequency of aberrant metaphases and the proportion of individuals with CA were significantly higher in workers than in controls, indicating that occupational exposure to tobacco imposes considerable genotoxicity among tobacco processors.     

The Database for Analysis of Quantitative Characteristics of Chromosome Aberration Frequencies in the Culture of Human Peripheral Blood Lymphocytes

The data on spontaneous chromosome aberration rates in cultures of human peripheral blood lymphocytes obtained in the past 30 years have been collected to form a database. The database contains the results of analysis of more than 330 000 metaphases in lymphocytes from more than 1200 subjects. The frequency of aberrant metaphases in the control group has been estimated at 0.0213 ± 0.00085. No differences between sexes have been found with respect to either the total chromosome aberration rate or the rates of individual aberration types. The total chromosome aberration rate did not depend on age; however, it has been found that the number of fragments increased and the number of exchanges decreased with age. Smoking has been found to increase the frequency of chromosome aberrations in individuals with occupational hazards, but not in those who are not occupationally exposed to radiation or chemicals. Alcohol consumption increased the frequency of paired fragments, whereas the frequencies of other aberrations did not differ from the control values.     

The database for analysis of quantitative characteristics of chromosome aberration frequencies in the culture of human peripheral blood lymphocytes]

The data on spontaneous chromosome aberration rates in cultures of human peripheral blood lymphocytes obtained in the past 30 years have been collected to form a database. The database contains the results of analysis of more than 330,000 metaphases in lymphocytes from more than 1200 subjects. The frequency of aberrant metaphases in the control group has been estimated at 0.0213 +/- 0.00085. No differences between sexes have been found with respect to either the total chromosome aberration rate or the rates of individual aberration types. The total chromosome aberration rate did not depend on age; however, it has been found that the number of fragments increased and the number of exchanges decreased with age. Smoking has been found to increase the frequency of chromosome aberrations in individuals with occupational hazards, but not in those who are not occupationally exposed to radiation or chemicals. Alcohol consumption increased the frequency of paired fragments, whereas the frequencies of other aberrations did not differ from the control values.     

Chromosomal aberrations in humans as genetic endpoints to assess the impact of pollution

The chromosomal aberration assay with peripheral blood lymphocytes has been used routinely during the last three decades to survey exposure of humans to various genotoxic agents. A large number of biomonitoring studies are based on this genetic endpoint. A great deal of data exists on occupational, life-style or medical exposure situations but less evidence of the validity of the assay is available with regards to environmental exposure. In the present paper we report our investigations on the impact of pollution in two different populations using chromosomal aberrations in human peripheral blood lymphocytes as a biomarker of chronic exposure to heavy metals and dioxins/furans for a long period and as a biomarker of acute exposure to accidentally released vinyl chloride in the air. In order to study genotoxic effects (chromosomal aberrations) of heavy metals and dioxins/furans, 52 exposed individuals from a polluted area were compared to 51 matched controls from a distant non-industrialized area. A statistically significant increase was observed in the frequency of chromosomal aberrations in peripheral blood lymphocytes from the exposed population (1.90% aberrant cells vs. 1.11% for the controls). In the case of the vinyl chloride accident, chromosomal aberrations were analyzed in peripheral blood lymphocytes from 29 potentially exposed and 29 non-exposed individuals (matched controls). The exposed group showed a statistically significant increase in the frequency of aberrant cells (1.47% vs. 1.07% for the controls).     

Analysis of sister-chromatid exchanges, cell kinetics and mitotic index in lymphocytes of smoking pesticide sprayers

Whole blood of 50 smokers who were exposed to pesticides was set up in RPMI 1640 medium, and observed for sister-chromatid exchanges (SCEs), cell kinetics (CK) and mitotic index (MI). As controls, blood samples were collected from 20 non-smokers (control I) and 27 smokers (control II) who were not exposed to pesticides. A significant increase in SCEs was observed as the duration of exposure increased. The frequency of M1 metaphases increased significantly whereas M2 and M3+ metaphases decreased in the exposed group. The mitotic index increased in control II and in the exposed population while it showed a decrease at 11-25 years' exposure.     

Follow-up study of the genetic damage in lymphocytes of pharmacists and nurses handling antineoplastic drugs evaluated by cytokinesis-block micronuclei analysis and single cell gel electrophoresis assay

A follow-up study was carried out 4 years after an initial evaluation of the micronucleus frequency in 10 healthy individuals who had been occupationally exposed to antineoplastic drugs in a Brazilian hospital. Upon the first evaluation, these 10 exposed individuals were compared with 10 non-exposed individuals matched for age, sex and smoking habits; the results revealed that the frequency of micronucleated lymphocytes in individuals exposed to antineoplastic drugs was significantly higher (P=0.038) than in controls. The frequency of dicentric bridges was also increased, although not significantly (P=0.0545). After the first analysis, the workers handling antineoplastic drugs were advised to modify their work schedule to limit exposure, and the number of workers in the group was increased from 10 to 12 individuals. In the follow-up study, 12 individuals from the same work area were assessed. In addition to micronucleus frequency, alkaline single cell gel electrophoresis was also used to monitor genetic hazard. This exposed group was compared to 12 non-exposed workers from the same hospital, matched for age, sex and smoking habits. In the follow-up study, no statistical difference was found between exposed workers and controls in terms of micronucleus and dicentric bridge frequency with the Mann--Whitney U-test (P=0.129 and 0.373, respectively). However, the mean value of SCGE analysis was significantly higher in the exposed group than in the controls (P=0.0006). Although the micronucleus analysis seems to be less sensitive to assess DNA damage, it detects chromosome aberrations and not just repairable DNA breakage and alkali-labile sites. Combination of the alkaline single cell gel electrophoresis and cytokinesis blocked micronucleus assay appears to be commendable to monitor populations chronically exposed to genotoxic agents.     

Quantitative characteristics of chromosomal aberration frequency in the residents of the regions with different levels of cancer incidences

We present the results of cytogenetic monitoring of the districts in Kemerovo region, which differ in standardized indices of cancer incidence. It has been shown that residents of the districts with high incidence of malignancies had higher average frequency of metaphases with chromosomal aberrations than the control group (4.06 ± 0.12% and 2.76 ± 0.13%, respectively). This difference is caused primarily by single or paired fragments. The increase in the frequency of aberrant metaphase incidence in the districts with elevated cancer frequency was observed both in the male and female groups as well as both in adults and children.     

Long-term cytogenetic effects in children prenatally-exposed to radiation as a result of the accident at the Chernobyl Atomic Energy Station

Forty-two children exposed to ionizing radiation in prenatal period and 15 children of control group were examined in the remote terms after the accident using the method of differential G-staining of chromosomes in lymphocytes of peripheral blood. It was found that the average group rate of aberrant cells and chromosome aberrations was reliably higher in the children exposed in utero compared to control. Long-term cytogenetic consequences of the pre-natal exposure were characterized by prevalence of aberrations of a chromosome type, mainly stable chromosome lesions. At chronic exposure to low doses of ionizing radiation the increase in the rate both stable and unstable chromosome aberrations.     

Telomeric associations in cigarette smokers exposed to low levels of X-rays

Telomeric association (TA), i.e. fusion of chromosomes by their telomeres, predisposes a cell to genetic instability. Because of this we investigated the effect of X-rays exposure and cigarette smoking on the frequency of TA in peripheral blood lymphocytes of exposed individuals, in order to determine if TA can be a chromosomal marker in populations exposed to these carcinogens and if there is an synergistic effect between both agents. We found that the exposed groups show a greater percentage of TA when compared with the control group (P<0.001). However, although the percentage of metaphases with TA in the group with combined exposure (12.6%) was greater than in the others exposed groups (P<0.05), this value was less than the sum of the two individual effects (15.1%). Our results suggest that probably there is not an additive or synergistic effect between X-rays and smoking, and that TA may be a useful cytogenetic marker for evaluating populations exposed to mutagens.     

Correlation of human lymphocyte SCE frequency with smoking history

The relationship between the level of occupational exposure to epichlorohydrin (ECHH) and the clastogenic effect was studied on a group of 33 workers. The effect of ECHH was assessed by differences in the frequency of chromosome aberrations in peripheral blood lymphocytes in ECHH-exposed and control groups. In the group exposed to the average ECHH concentration, 0.384 mg X m-3, during the last 6 months, the cytogenetic analysis revealed 2.00 +/- 0.23% AC (aberrant cells) (0.0203 B/C, breaks per cell) as compared with 1.68 +/- 0.23% AC (0.0172 B/C) in the matching controls. These results indicate that an average concentration lower than 0.40 mg X m-3 ECHH in the working atmosphere has no significant clastogenic effect on human peripheral lymphocytes.     

Cytogenetic study of a group of workers exposed to thinner

The frequency of sister-chromatid exchanges (SCE) was evaluated in a group of 24 workers exposed to thinner in a luminous advertisements factory and in three workshops for painting sheet metal in Mexico City. 50 metaphases for each exposed individual and each control were analysed; air samples of the working places were also studied; it was observed that among all the components of thinner, only benzene exceeded advisable limits. The cytogenetic data were subject to one-way analyses of variance indicating that no significant differences existed between both groups; also, there are no significant differences among workers with a long exposure time and those with fewer than 5 years of exposure. Nevertheless, use of tobacco increased significantly the SCE frequencies among the exposed group, but did not increase SCE in the control group.     

Chromosome aberrations in the lymphocytes of persons working with the use of polychloropinene in agriculture

A cytogenetic examination of persons exposed to the action of polychlorpinene under agriculture conditions was carried out. The frequency of aberrant cells in the group of persons under examination accounted for 5.8 %, which was much higher than that of aberrations in the control group (1.6%). The average frequency of aberrant cells and character of aberrations under repeated cytogenetic examination 11 months after were similar to those described above.     

Quantitative characteristics of the frequency of chromosomal aberrations in a group of residents of a large western Siberian industrial region

The results of a 15-year study of chromosome aberration frequency in cultured peripheral blood lymphocytes from subjects living in the Kuzbass industrial region are presented. The database for the analysis of the main parameters of chromosome aberrations contains data on 925 subjects, with the total number of cells examined being 92,900. It has been found that the total frequency of aberrant metaphases in the database is 3.73 +/- 0.1%, whereas this frequency for the sample of subjects from industrial areas of this region (the basic control group) is significantly lower (2.86 +/- 0.26%). It has been demonstrated that the sex and age of the subjects do not affect substantially the frequency of any type of chromosome aberrations. Tobacco smoking is associated with a slight increase in chromosome damage frequency; however, the difference between smokers and nonsmokers is insignificant even among subjects exposed to occupational hazards. The possible causes that have determined the increased basic and background chromosome aberration frequencies in the population of the Kemerovo oblast compared to these parameters for the European part of Russia and the CIS are discussed.     

Liquid-holding-mediated enhancement of the frequency of chromosomal aberrations induced by ethyleneimine in barley embryos cultivated in vitro

Barley embryos, deprived of endosperm, were treated with ethyleneimine (EI) and subjected to liquid holding in various media for 4–24 h. Liquid holding in media lacking a 1% sucrose provision led to manifold increase of the frequency of aberrant metaphases in root-tip meristems of EI-treated embryos, cultivated in vitro in a maximal medium after the cessation of the liquid-holding period. The enhancement of the chromosomal damage was dependent on the duration and temperature of the liquid holding. Autoradiographic studies showed that no semi-conservative DNA replication took place under liquid-holding conditions leading to an enhancement of the frequency of aberrant metaphases in EI-treated embryos.     

Chromosome painting for cytogenetic monitoring of occupationally exposed and non-exposed groups of human individuals

The suitability of a three-color fluorescence in situ suppression hybridization technique was examined for monitoring five different groups of individuals: 30 occupied in radiology, 26 occupied in nuclear medicine or radiation physics, 32 patients with breast cancer, 26 occupied with military waste disposal, all presumably exposed to low doses of radiation or chemical mutagens and a non-exposed control group (N=29). The average frequency of breaks constituting the various aberrations did not significantly differ between the groups of medical radiation appliers and the control group. However, breast tumor patients and military waste disposers, as groups, showed a higher aberration rate than did healthy controls. Stable rearrangements mainly characterized the groups of controls, tumor patients, and radiation appliers, while a higher proportion of unstable aberrations was found in the chemically exposed individuals. Individuals with an increased frequency of aberrations could be detected within each examined group, which clearly determined the average values of the whole group. With respect to interchromosomal distribution of the breakpoints constituting the found aberrations and the involvement of the labeled chromosomes in rearrangements, the observed values were very close to the expected ones in the controls. A rather similar trend of deviations from expectation was observed in all other groups. Chromosome 4 was slightly over-affected, while chromosome 2 was slightly underrepresented in all analyzed groups (except tumor patients). Rearrangements of the labeled chromosomes with the unlabeled ones exceeded expectation. In conclusion, chromosome painting if included in further attempts of human population monitoring will broaden the basis of argumentation with respect to health risks introduced by mutagen exposure.     

Chromosome aberrations in lymphocytes of residents living in buildings constructed with radioactively contaminated rebars

Using the G-banding technique, we examined lymphocytes from 90 individuals (43 males and 47 females, median age 31 years) living in buildings constructed with radioactively contaminated rebars. Forty-five nonexposed control subjects (22 males and 23 females, median age 30 years), matched to the radiation-exposed individuals by sex and age, were selected for comparison. At least 500 metaphases were checked for each individual. All recognizable structural aberrations of chromosomes or chromatids were recorded. After adjusting for age and smoking status, both the percentage of cells with aberrant chromosomes (PCAC) and the number of aberrant chromosomes per 100 cells (NAC) were found to be significantly higher in the radiation-exposed females than in the control females (p < 0.05 for PCAC and NAC). This difference, however, was not observed in the comparison of radiation-exposed and control males. This suggests a possible interaction between sex and radiation exposure in their effects on chromosome aberrations.     

Lymphocyte proliferation kinetics and genotoxic findings in a pilot study on individuals chronically exposed to arsenic in Mexico

In the search for early biological markers to detect genetic damage, a pilot study on a hydroarsenicism-exposed group was designed. Blood and urine samples were taken from 11 individuals chronically exposed and from 13 individuals with lower exposure to the metal. Lymphocyte cultures for cytogenetic studies and HGPRT assay were done with coded peripheral blood samples, while arsenic levels and the “rec assay” in B. subtilis were determined in urine samples. The highly exposed group excreted greater amounts of As, nevertheless the rec assay showed negative results. An interesting finding is that the cell-cycle kinetics exhibited a significant difference between the groups studied, the average generation time (AGT) was longer in the highly exposed group. The percentages of chromosomal aberrations and the frequencies of sister-chromatid exchanges were similar in both populations, although complex aberrations were more frequent in the highly exposed group, which also showed a higher average variation frequency in the HGPRT assay, but the 2 latter observations were not statistically significant. The lag in lymphocyte proliferation could mean an impairment of the immune response due to arsenic exposure.     

Chromosome aberrations in individuals occupationally exposed to ethylene oxide, and in a large control population

Chromosome aberration frequencies in 61 employees potentially exposed to ethylene oxide (ETO) were compared with those in unexposed control groups. We studied 3 worksites with differing historical ambient levels of ETO. Within worksites, groups were classified as high potential exposed, low potential exposed, or controls. Further control groups including an off-site community control group were added to give a total of 304 control individuals. Blood samples were drawn several times over a 24-month period. Aberrations were analyzed in 100 cells per sample after culture for 48-51 h. Worksites I, II and III respectively represented increasing levels of potential ETO exposure. At worksites I and II, no consistent differences in aberration frequencies were found among groups. At worksite III aberration frequencies in potentially exposed individuals were significantly increased compared with controls. The frequencies of cells with aberrations were 5.6% for the 2 individuals in the high potential exposure category and 2.6% for 23 persons in the low potential exposure group. The overall frequency of cells with aberrations in the matched control individuals was 1.4%. In the total control group of 304 individuals we found significant increases in aberrations associated with smoking and with increasing age. We have also reported previously an association between sister-chromatid exchange (SCE) frequency and ETO exposure (Stolley et al., 1984). When aberration frequencies were compared with levels of SCEs there was only a weak overall association. The correlation was found in potentially exposed but not in control groups, and for any individual, one observation could not be used to predict the other.