Frequency Of Sister Chromatid Exchanges In Lymphocytes Research Articles

Derivation of a biological limit value (BLV) for inorganic lead based on lead-induced genotoxicity in workers using the benchmark dose approach (BMD)

BackgroundSafe exposure levels for industrial chemicals are a prerequisite to the safety of workers in occupational settings. MethodsThe application areas of the benchmark dose (BMD) approach have tremendously increased in recent years. Quantitative analyzes of dose-response relationships of genotoxic chemicals are among these application areas too. This study investigates the applicability of sister chromatid exchange (SCE) frequencies in lymphocytes of lead-exposed workers to derive a biological limit value (BLV) for inorganic lead. To this end, our previously published occupational lead exposure data were pooled and re-examined by using the BMD approach. Subsequently, we determined the point of departure (POD) value for lead-induced SCE frequencies in lymphocytes. Accordingly, the blood lead concentrations and SCE frequencies in lymphocytes of 26 control and 108 lead-exposed male voluntary workers employed in a battery manufacturing plant were subjected to BMD modeling to determine the BMD confidence interval (BMD-CI). ResultsAccording to our data set, benchmark dose lower bound (BMDL) of 6.55 μg Pb/dL blood for the SCE frequencies in lymphocytes of lead-exposed workers has been obtained. ConclusionThe blood lead concentration of 6.55 μg/dL might be proposed as the BLV for lead and its inorganic lead compounds based on our dataset.

Comparison of the frequency of sister chromatid exchange in pan chewers and oral submucous fibrosis patients

Sister chromatid exchange (SCE) test is a sensitive, biomarker of genotoxic substances. The frequency of SCE in lymphocytes of ten pan chewing patients, oral submucous fibrosis (OSF) patients and age matched healthy controls were investigated. The frequency of mean SCE/cell was found to be 10.428 ± 0.755 in OSF patients, 8.752 ± 0.383 in case of pan chewers as compared to 5.912 ± 0.310 in controls. These values show a significant increase in frequency of SCE/cell in OSF patients and pan chewers when compared with that of healthy controls. There is a positive correlation co-efficient of SCE/cell with frequency, quantity, duration, intensity and period of exposure of pan-parag to oral mucosa in pan chewers and OSF patients indicating genotoxic effect of pan. Thus SCE could be used as a biomarker in chewers also to assess the level of genomic damage and to advocate efficient control measures.

Increased lymphocyte sister chromatid exchange frequency in workers with exposure to low level of ethylene dichloride

The genotoxicity of low-level exposure to ethylene dichloride (EDC) and vinyl chloride monomer (VCM) in humans is not clear. We used lymphocyte sister chromatid exchange (SCE) frequency as a parameter to investigate the genotoxicity of low level EDC and VCM in VCM-manufacturing workers. The SCE frequency was determined for 51 male workers with exposure to VCM and/or EDC and for 20 male workers devoid of such exposure. Epidemiological data were obtained by questionnaire, and included history of smoking, drinking, and any medication taken, as well as a detailed occupational history. Personal- and area-sampling and analysis were conducted in order to calculate the time-weighted average (TWA) contaminant-exposure level corresponding to different job categories. Moderate EDC exposure around 1 ppm corresponded to a significantly greater SCE frequency than was the case for the low EDC exposure group ( p<0.01). However, VCM exposure of similar level was not associated with increased SCE. We conclude that EDC may cause genotoxicity at a relatively low level of exposure.

Increased sister chromatid exchange frequency in peripheral lymphocytes of hepatocellular carcinoma patients

AbstractHepatocellular carcinoma (HCC) has been documented to be associated with a multifactorial aetiology including hepatitis B virus (HBV), hepatitis C virus, exposure to aflatoxin, tobacco and alcohol. With the specific aim to examine the relationship between HCC and frequency of sister chromatid exchange (SCE) in peripheral lymphocytes as an indicator of integral carcinogen exposure, a total of 14 newly diagnosed untreated HCC patients and 13 healthy controls were studied. Study subjects were matched on age, sex, cigarette smoking and alcohol intake. Peripheral blood samples were collected using heparinized vacuum syringes. Lymphocytes were cultured in RPMI 1640 medium containing fetal calf serum and phytohaemagglutin. After lymphocytes were cultured for 24 h, 5‐bromo‐2‐deoxy uridine was added to the medium and colcemid was added after 70 h. Cells were harvested and fixed in methanol‐acetic acid (3 : 1) solution after 72 h. Cells were first stained with 33258 Hoechst and restained with Giemsa. Photographs were taken from at least 32 lymphocytes at the second metaphase of each study subject. The frequency of SCE was scored by two readers blindly. There was a significantly increased SCE frequency in peripheral lymphocytes of HCC patients than healthy controls, with a mean ± standard deviation of 15.1 ± 4.4 and 8.9 ± 2.7 SCE per metaphase II, respectively. The increased SCE frequency in lymphocytes of HCC patients may be due to the chronic HBV infection, exposure to chemical hepatocarcinogens or both.

Cytogenetic studies of mice exposed to styrene by inhalation

The data for the in vivo genotoxicity of styrene (STY) are equivocal. To evaluate the clastogenicity and sister-chromatid exchange (SCE)-inducing potential of STY in vivo under carefully controlled conditions, B 6C 3F 1 female mice were exposed by inhalation for 6 h/day for 14 consecutive days to either 0, 125, 250 or 500 ppm STY. One day after the final exposure, peripheral blood, spleen, and lungs were removed and cells were cultured for the analysis of micronucleus (MN) induction using the cytochalasin B-block method, chromosome breakage, and SCE induction. Peripheral blood smears were also made for scoring MN in erythrocytes. There was a significant concentration-related elevation of SCE frequency in lymphocytes from the spleen and the peripheral blood as well as in cells from the lung. However, no statistically significant concentration-related increases were found in the frequency of chromosome aberrations in the cultured splenocytes or lung cells, and no significant increases in MN frequencies were observed in binucleated splenocytes or normochromatic erythrocytes in peripheral blood smears.

Passive smoking and sister-chromatid exchanges in lymphocytes

The object of this study was to determine whether exposure to environmental tobacco smoke is associated with DNA damage reflected by the frequency of sister-chromatid exchange (SCE) in lymphocytes. Within a cross-sectional design, 106 male non-smoking adults, employees of two administrative companies, were divided on the basis of self-reported exposure into high and low passive smoking groups. The high exposed subjects (passive smokers, n = 50) lived with smokers, worked with smokers and were exposed to tobacco smoke for an average of 70 h/week. The low exposed non-smokers ( n = 56) were exposed for an average of 5 h/week. Plasma cotinine levels for the passive smokers ranged between 0.4 and 9.0 ng/ml (median 1.4 ng/ml), and for the low exposed group between 0.0 and 1.9 ng/ml (median 0.4 ng/ml) ( p < 0.0001; Mann-Whitney test). No difference was observed between the two groups in the frequency of SCEs in lymphocytes: 4.66 ± 0.05 for passive smokers and 4.68 ± 0.04 for low exposed non-smokers (mean ± SEM) ( p = 0.80; t-test). Reclassification of subjects on the basis of plasma cotinine levels did not change the results substantially. These results are in accordance with observations that the increase in cancer risk due to passive smoking is small in comparison with the increase due to active smoking. The SCE test may be too insensitive to be useful for the evaluation of possible cytogenetic effects related to passive smoking.

Biological and biological-effect monitoring of workers exposed to 4,4'-methylene-bis(2-chloroaniline).

4,4'-Methylene-bis(2-chloroaniline) (MOCA), a curing agent used in polyurethane manufacture, is a genotoxic and carcinogenic amine. This study aimed to assess occupational exposure to MOCA using as indices: (1) the post-work urinary output of MOCA; (2) urinary thioethers, assuming that conjugation with glutathione might be a significant pathway for the elimination of putative electrophilic metabolites of MOCA; and (3) sister-chromatid exchange (SCE) frequency in peripheral lymphocytes as an indicator of genetic damage. Process workers at a polyurethane production unit were found to have up to 38 mumol MOCA mol-1 creatinine in their urine at the end of a work shift. Smaller quantities were found in the urine of laboratory and supervisory staff, but none was detected in the urine of a group of office and sales staff from an unrelated industry, who served as unexposed controls. There was no evidence of MOCA-related urinary thioether output. There was a graded increase in SCE frequency from controls to process workers, consistent with their apparent exposure to MOCA. Administration of MOCA to rats (5 daily i.p. injections of 125 or 250 mg kg-1 resulted in dose-related increases in MOCA excretion and in lymphocyte SCE frequency, but there was no change in thioether output. These results indicate that urinary thioether excretion is inappropriate for monitoring MOCA exposure, but that where MOCA exposure can be demonstrated, by the presence of MOCA in urine, this is associated with genetic damage in both man and in the rat.

Sister-chromatid exchanges in lymphocytes from styrene-exposed boat builders

Sister-chromatid exchanges (SCE) were measured in peripheral blood lymphocytes from 40 workers in the boat-building trade. Twenty of these workers were exposed to significant amounts of styrene. The mean air concentration of styrene in the breathing zone of the boat builders was 209 mg/m 3 in the 7 exposed current smokers and 230 mg/m 3 in the 13 exposed non-smokers. Urinary styrene metabolites were also measured and the mean mandelic acid/creatinine ratios in the exposed, smokers was 275 mg/g, and in the exposed, non-smokers 323 mg/g. The SCE frequency in lymphocytes from the styrene-exposed group did not differ from that in the controls, although smoking significantly induced SCE in these workers.

Induction of nuclear aberrations by smokeless tobacco in epithelial cells of human oral mucosa

Cytologic and cytogenetic studies were performed to assess the prevalence of somatic cell genetic damage in 48 young adults equally divided to represent users and nonusers of smokeless tobacco. Exposure was ascertained by measuring saliva cotinine using capillary gas chromatography. Squamous epithelial cells sampled from the oral mucosa demonstrated significant cytologic alterations associated with tobacco exposure. The frequency of micronucleated cells was significantly (P less than .01) higher in the labial mucosa of exposed (2.22%) compared to unexposed (0.27%) individuals. The frequency of micronuclei varied widely between exposed subjects but was higher in heavily (2.48%) compared to lightly (1.29%) exposed individuals as measured by saliva cotinine levels. Morphologic classification of epithelial cell nuclei showed that the frequency of cells with normal nuclear structure was significantly (P less than .01) reduce in exposed individuals. Analysis of oral epithelial cells of five additional nonusers of smokeless tobacco but wearers of orthodontic appliances to stimulate abrasion demonstrated no difference from the nonexposed control group. Unlike the case with cigarette smokers, peripheral lymphocyte sister-chromatid exchange frequency was not affected by exposure to smokeless tobacco. The oral cytology data, however, support an interpretation of exposure-dependent nuclear alterations, including micronuclei, in the oral epithelium associated with the use of smokeless tobacco. Altogether, results suggest that use of smokeless tobacco may cause genetic damage to cells in the oral epithelium.

Genotoxicity of coal fly ash, assessed in vitro in Salmonella typhimurium and human lymphocytes, and in vivo in an occupationally exposed population

Fly ash as a product of coal combustion is known to contain various mutagenic substances, but genotoxic properties, especially of the particular (larger-size) fly ash fraction which is electrostatically precipitated (ESP) in the energy plant, have hardly been investigated. While smaller-size fly ash particles escape through the stack during powder coal combustion, the ESP fraction is collected and used for the manufacturing, for instance according to the Lytag process, of secondary products which can serve several construction purposes. Since fly ash as well as fly ash products are generally introduced into the human environment, a study of possible genotoxic effects to human DNA is indicated. Mutagenic properties of ESP fly ash, as well as of the Lytag product, were investigated by means of the Salmonella microsome assay. The capacity to cause human chromosome damage of both ESP fly ash and Lytag dust was studied in vitro by application of the sister-chromatid exchange (SCE) test using human lymphocytes. Furthermore, effects of ESP fly ash/Lytag dust on the incidence of SCE in peripheral lymphocytes in vivo were measured in an occupationally exposed, male population, using individually matched employees from a flour-processing industry as the control population. It is demonstrated that ultrasonically treated DMSO extracts of ESP fly ash are slightly mutagenic to Salmonella tester strains TA97 and TA102. Lytag dust is effective in inducing reversions in all tester strains. Furthermore, it appeared that both compounds significantly increase the SCE frequency of human lymphocytes after incubation in vitro in comparison to non-exposed cells. Also, peripheral lymphocytes of the occupationally exposed population show a considerably higher incidence of SCE than the control population. Major disturbing factors in assessing the effects of occupational exposure to fly ash/Lytag dust on lymphocyte SCE frequency appeared to be smoking behavior and alcohol consumption. It is concluded that exposure to fly ash from powder coal combustion implies a moderate genotoxic risk to man.

Variation in the human lymphocyte sister-chromatid exchange frequency: Results of a long-term longitudinal study

The variation in lymphocyte sister-chromatid exchange (SCE) frequency as a function of time was investigated in nonsmokers and smokers. The smokers were divided into 3 groups depending on their smoking status. The group termed ‘smokers’ participated in a program to stop smoking but did not reduce or eliminate their use of tobacco; ‘smoke enders’ successfully completed the smokending program and remained free of tobacco for the duration of the study, while the ‘variable’ group stopped smoking for a limited time but then resumed smoking. 8 or more blood samples per person were obtained over a period of at least 12 months. The SCE frequencies for each of these groups were compared with each other and with those of two previous longitudinal study groups from our laboratory. The proportion of high-frequency cells (HFCs) was also determined for each sample. The results confirm our previous finding that SCE frequencies and the proportion of HFCs observed in separate samples from the same individual are more likely to be different as the time between samples increases. We also show that smokers have significantly more SCEs and HFCs than do nonsmokers, that SCE frequencies in smokers do not decline for at least 12 months when smoking is stopped, and that among smokers, significant seasonal variation in the SCE frequency occurs. These results provide useful information concerning the effects of smoking upon SCE frequencies, and will be helpful in designing and interpreting the results of long-term human population cytogenetic studies.

Metabolic activation of α-naphthoflavone by 2,3,7,8-tetrachlorodibenzodioxin-induced rat liver microsomes

Previous studies in our laboratory had demonstrated that addition of α-naphthoflavone (ANF) to lymphocytes from smokers or polychlorinated biphenyls (PCB)s-exposed individuals caused an increase in sister chromatid exchange (SCE) frequency whereas lymphocytes from controls were relatively unaffected. In order to investigate the mechanism responsible, metabolism of ANF by uninduced and 2,3,7,8-tetrachlorodibenzodioxin (TCDD)-induced microsomes was studied as a function of microsomal protein concentration and incubation time. Nonpolar metabolites were analyzed and the amount of conjugated (polar) and protein-bound metabolites determined. The initial ANF-metabolism rate was 10-fold higher in TCDD-induced microsomes (4.9 ± 0.6 nmol/min per mg TCDD-induced microsomal protein vs. 0.5 ± 0.2 nmol/min per mg uninduced microsomal protein) than in uninduced microsomes. Moreover, uninduced microsomes no longer metabolize ANF after 30–40 min while TCDD-induced microsomes metabolize ANF for longer than 2 h or until all the ANF is gone. In addition to the metabolites formed by uninduced microsomes [7,8-dihydro-7,8-dihydroxy-ANF (7,8-dihydrodiol); 5,6-dihydro-5,6-dihydroxy-ANF (5,6-dihydrodiol); 5,6-oxide-ANF and 6-hydroxy-ANF], TCDD-induced microsomes from unidentified metabolites. When TCDD-induced microsomes and 40 μM ANF were added to Chinese hamster ovary (CHO) cells, we found a correlation between the concentration of 5,6-oxide-ANF and clastogenicity to CHO cells. However, purified 5,6-oxide-ANF did not induce SCEs in CHO cells in the absence or presence of TCDD-induced microsomes. However, a minor metabolite (identified as the 9,10-dihydro-9,10-dihydroxy-ANF by acid dehydration) formed with TCDD-induced microsomes produces clastogenicity in CHO cells. These data indicate that a minor metabolite of ANF is a potent clastogen which suggests that this metabolite may be responsible for the ANF-mediated increases in SCE frequency in lymphocytes from smokers or PCB-exposed individuals.

Cadmium sulfate does not induce sister chromatid exchanges in human lymphocytes in vitro

Mutagenic properties of cadmium sulfate were analyzed by the method of sister chromatid exchanges (SCEs) in lymphocytes of human peripheral blood taken from 4 donors. No significant increase was found in the mean frequency of SCEs in lymphocytes exposed to 1.6 × 10 −3 mM, 3.1 × 10 −3 mM and 6.2 × 10 −3 mM CdSO 4 in culture.

Variation in the human lymphocyte sister chromatid exchange frequency as a function of time: results of daily and twice-weekly sampling.

The variation in lymphocyte sister chromatid exchange (SCE) frequency was investigated in healthy nonsmokers who were not taking any medication. Two separate studies were undertaken. In the first, blood was drawn from four women twice a week for 8 weeks. These donors recorded the onset and termination of menstruation and times of illness. In the second study, blood was obtained from two women and two men for 5 consecutive days on two separate occasions initiated 14 days apart. One donor participated in both studies. Analysis of the mean SCE frequencies in each study indicated that significant temporal variation occurred in each donor, and that more variation occurred in the longer study. Some of the variation was found to be associated with the menstrual cycle. Peaks in the SCE frequency were observed at times that corresponded approximately to ovulation and to the beginning and end of menstruation. In the daily study, most of the variation appeared to be random, but occasional day-to-day changes occurred that were greater than those expected by chance. To determine how well a single SCE sample estimated the pooled mean for each donor in each study, we calculated the number of samples that encompassed that donor's pooled mean within 1 or more standard errors. For both studies, about 75% of the samples encompassed the pooled mean within 2 standard errors. An analysis of high-frequency cells (HFCs) was also undertaken. The results for each study indicate that the proportion of HFCs, compared with the use of Fisher's Exact test, is significantly more constant than the means, which were compared by using the t-test. These results coupled with our previous work suggest that HFC analysis may be the method of choice when analyzing data from human population studies.

Dioxin treatment of rats results in increased in vitro induction of sister chromatid exchanges by α-naphthoflavone: An animal model for human exposure to halogenated aromatics

Recent reports have shown that α-naphthoflavone (α-NF) in vivo enhances the sister chromatid exchange (SCE) frequency in lymphocytes from human populations exposed to cigarette smoke or polychlorinated biphenyls and dibenzofurans. In this study, female Sprague-Dawley rats (9–11 weeks old) were administered a single oral dose of 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) and killed 6 days after treatment. Blood cultures were established with or without α-NF. The baseline and α-NF-induced SCE frequencies were assessed in lymphocytes after a 72-hr culture period. No effect on the SCE baseline frequency (cultures without α-NF) was detected in rats exposed to 0–30 μg TCDD/kg. However, the SCE frequencies from cultures incubated in the presence of α-NF were significantly higher in lymphocytes from rats treated with TCDD. Moreover, ΔSCE values (SCE α-NF minus SCE baseline) were significantly higher in lymphocytes from rats treated with TCDD than in controls. A dose-dependent increase in ΔSCE values was observed between 0 and 3 μg TCDD/kg, followed by a plateau at higher doses. This induction pattern closely resembled the induction of the liver microsomal aryl hydrocarbon hydroxylase activity by TCDD. In contrast to TCDD, phenobarbital treatment of rats (75 mg/kg/day) had no effect on α-NF-induced SCE frequencies in lymphocytes. Liver microsomes from TCDD-treated rats metabolized α-NF at a rate much faster than that of control microsomes. These studies indicate that TCDD-exposed rats provide a useful model to investigate the mechanism of enhanced in vitro induction of SCE frequency in lymphocytes from humans exposed to toxic halogenated aromatics or cigarette smoke.

Cytogenetic effects of methyl isocyanate exposure in Bhopal.

Among human survivors following the methyl isocyanate (MIC) gas tragedy the major complaints have been related to deep-seated suffocation, terrible pain in breathing, and severe ocular irritations. In order to assess the possible genetic effects we have used lymphocyte cultures and screened chromosomes by two techniques; one by looking for chromosomal aberrations and the other by estimating sister-chromatid exchange (SCE) frequencies. Both these parameters are good indicators of genetic damage in chromosomal DNA. SCE frequencies in lymphocytes have been increased more than three times in MIC-exposed persons. The results were compared to two groups of controls (one group comprising persons present in the same house; the second group of persons were chosen from distant places, 20-50 km away from the incident). Chromosomal breaks have been observed in 10 out of 14 MIC-affected people (71.4%) studied while only 6 out of 28 (21.4%) controls had chromosomal breaks. Some MIC-exposed persons had chromatin bodies in addition to the normal 46 chromosomes. These observations suggest that chromosomal DNA has been damaged.

Sister-chromatid exchange frequencies in lymphocytes of controls and patients with connective tissue diseases

It has been considered by some workers that sister-chromatid exchange (SCE) frequencies are elevated in patients with scleroderma and systemic lupus erythematosus (SLE). However, these observations were based on limited numbers of patients. Other have postulated the presence of a defect in DNA repair in cells from patients with various connective tissue diseases, including scleroderma and SLE. We report our findings from a large survey of SCE frequencies in patients with connective tissue diseases. Their diagnoses are scleroderma, SLE, rheumatoid arthritis, juvenile chronic arthritis, Behcet's syndrome and polyarteritis nodosa. These patients had never received cytotoxic drugs. Healthy individuals, hospital patients with diagnoses other than connective tissue diseases and relatives of patients with scleroderma have been used as controls. The results have been analysed by generalized linear modelling, and we have shown that patients with SLE and Behcet's syndrome and controls with viral infections have elevated SCE frequencies, both before and after adjustments have been made for the effect on SCEs of an individual's age, smoking habits, sex and race. The SCEs of patients with scleroderma and their relatives were normal. SCE frequencies increased with age by 4% per decade and the SCE frequencies of smokers were approximately 12% higher than those of nonsmokers of similar age. The sec of an individual did not significantly affect SCEs but individuals from the Middle East were found to have lower counts than those originating from other parts of the world.

No increase in sister-chromatid exchange frequency in lymphocytes of chromium platers

To detect mutagenic effects of hexavalent chromium (Cr) in vivo, sister-chromatid exchange (SCE) frequency was analyzed in lymphocytes of 24 Cr platers occupationally exposed to hexavalent Cr and 24 matched controls. There were no significant differences in SCE frequency between the two groups. Smokers, both Cr platers and controls, had a significantly higher SCE frequency than non-smokers.

Spontaneous and induced sister chromatid exchanges in lymphocytes of healthy persons

Spontaneous and mitomycin C-induced sister chromatid exchanges (SCEs) in lymphocytes were analyzed in 46 healthy persons. Our results suggest that the frequency of spontaneous SCE in lymphocytes is useful for evaluation of the biological effects of environmental mutagens, and that the induced-SCE test with a mutagen and lymphocytes may be used to detect persons with a high sensitivity to the mutagen.

Effects of smoking on spontaneous and induced sister chromatid exchanges in lymphocytes

Spontaneous and mitomycin C-induced sister chromatid exchanges (SCEs) in lymphocytes were analyzed in 24 non-smokers and 24 sex- and age-matched smokers. Mean spontaneous SCE frequency for non-smokers was 9.8 SCEs/cell, and that for smokers was 11.5 SCEs/cell. The difference was statistically significant ( P < 0.001 by t-test). These results suggest that spontaneous SCE frequency in lymphocytes is useful for evaluation of biological effects of environmental mutagens. However, we could not find any effects of smoking on the sensivities of lymphocytes to mitomycin C in vitro. The effects of mutagens on humans may be independent of one another.