Sister Chromatid Exchanges In Lymphocytes Research Articles

High frequency of sister chromatid exchanges in lymphocytes of the patients with Behcet's disease

Behcet's disease (BD) (OMIM 109650) is an immunogenetically based multisystem disease, characterized by iridocyclitis, arthritis, orogenital ulcerations and pustular skin lesions. Viral and autoimmune etiologies have been suggested and HLA-B5 has been found to predominate in BD. The disease is most seen in Turkey and Japan. Although familial cases have been reported, the mode of inheritance is not clear. To determine the genetic instability in BD, sister chromatid exchange (SCE) analysis has been performed on peripheral lymphocytes in 23 patients and 20 healthy controls. We found significantly higher SCE rates in the patient group ( p<0.0001). Our results may indicate that genetic impairment and genetic instability may play an important part in the etiology of BD.

Sister-Chromatid Exchanges in Lymphocytes of Medical Students Exposed to Formaldehyde

Sister-Chromatid Exchanges in Lymphocytes of Medical Students Exposed to Formaldehyde

Sister chromatid exchanges in human lymphocytes treated in vitro with cadmium in G o and S phase of their cell cycles

Sister chromatid exchanges (SCEs) were analyzed in human phytohemagglutinin-activated peripheral lymphocyte cultures exposed to varying concentrations (10 −7–10 −3 M) of cadmium chloride in vitro at two different stages of the cell cycle, G o and early S phase. When cadmium chloride was administered at the G o phase, no increase in the SCEs were observed for the doses 10 −6 and 10 −5 M. Concentrations equal to or larger than 10 −4 M cadmium chloride were lethal to human lymphocytes in our experimental conditions. A highly statistically significant increase was observed in the SCE frequency with increasing cadmium chloride concentration (10 −7–10 −4) when cadmium was administered at the early S phase, which was 24 h after culture initiation. The increase in SCE frequency was higher when the cultures were terminated at 54 h, compared to termination at 72 h. In order to examine the effects of cadmium administered at the S phase on SCE frequency in different individuals, 10 −5 M concentration was used and the cultures were terminated at 54 h after culture initiation. A 2- to 3-fold increase in the SCE frequency was observed in all six individuals examined. A progressive decrease in the proliferative index was also observed by increasing cadmium chloride concentration. These results demonstrate that the genotoxicity of cadmium chloride may be changed depending on the stage of the cell cycle in human lymphocytes. This may be one of the reasons of contradictory findings in the literature.

Induction of sister chromatid exchanges by pyrimethamine in human lymphocyte cultures

This study examined the effect of pyrimethamine (PYR) on sister chromatid exchanges (SCEs) in human lymphocytes. PYR is a folic acid antagonist used for the treatment of malaria and toxoplasmosis. PYR was added to peripheral blood lymphocyte cultures at three different doses: 0.05, 0.1, and 0.2 mg/ml. The results of the cytogenetic and statistical evaluations showed that the frequency of SCEs increased significantly in a dose-dependent manner. Thus, it is shown that PYR has a genotoxic effect on human chromosomes.

Sister Chromatid Exchanges in Sheep Peripheral Lymphocytes after in vitro Exposure to Metal-Containing Emission

D i a nov s k}' J .. K. S i v i kova: Sister Chrommid Exchanges ill Sheep Peripheral Lmzplwcytes ajier in \'itro E.ll'oslire to Metal·Containing Emissioll. Acta vet. Bmo 1998.67: 183·188. Industrial pollutants. originated from aluminum processing factory. were tested for induction of sister chromatid exchanges (SCE). Experiments were carried out using sheep peripheral lymphocytes under in vitro assays. The emission tested contained AI. Cd. As. Mn. Pb. Cu. Zn. Fe. Na. Ca. and Mg; the majority of them in the form of sulphides. sulphates. oxides or t1uorides. The emission was dissolved and neutralized according to a standard method. The mean of SeE was determined for three concentrations the emitted material (30.60 and 90 J-lg lOll) with presence and absence of the metabolic activation (S9 mix). The lowest concentration used corresponded to daily oral intake by sheep grazing on contaminated area. Results from both assays were similar; the significant increase in SeE was observed at the concentration of 60 J-lg/ml (p < 0.0 I). More conspicuous results were observed without of S 9. No significant decrease in the induction of proliferation index (PI) was found. A dose related effect was observed for SeE induction but not for inhibition of proliferation kinetics. Pasture confaminants. gellot()xicity. metabolic (lctit'afioll, in1'itro

Sister chromatid exchanges of human peripheral blood lymphocytes induced by N,N-diethylaniline in vitro

N,N-Diethylaniline is a reagent used in organic synthesis and is an important intermediate in the manufacturing of dyes. To evaluate its genotoxicity, we examined whether it can induce sister chromatid exchanges (SCEs) in human lymphocytes. We found that N,N-diethylaniline significantly increased the frequency of SCEs both in the absence and presence of S-9 mix. The SCEs from cultures treated by N,N-diethylaniline in the presence of S-9 mix displayed a marked increase which was about 5-fold greater than the control. ANOVA analyses indicated that there is a dose–response relationship between doses of N,N-diethylaniline and the frequency of SCEs, especially in the presence of S-9 mix. The results suggested that N,N-diethylaniline has genotoxicity.

Increased sister chromatid exchanges in peripheral lymphocytes of patients with Crohn's disease

A cytogenetic study was performed using Crohn's disease patients to determine whether the presence of chromosome instability is related to Crohn's disease, a chronic inflammatory bowel disease. Sister chromatid exchange (SCE) frequencies in peripheral blood lymphocyte cultures from 22 Crohn's disease patients and an equal number of healthy controls matched for sex and age were analyzed. The mean of SCE frequency in Crohn's disease patients was 11.64±0.42 (SEM) per cell, which was significantly higher than the value of 8.38±0.22 per cell in the matched controls ( p<0.0001). The Crohn's disease patients showed significantly increased high frequency cells (HFC) as compared to those among the matched controls. There was a significant correlation between HFC frequencies of the Crohn's disease patients and the severity of their disease as determined by the number of relapses per year and the degree of chronic activity after adjusting for the smoking status ( r=0.54, p=0.011). In both smokers and non-smokers, the mean SCE and HFC frequencies of the patients were significantly higher than those of the controls. These results suggest that Crohn's disease is a condition with increased chromosome instability characterized by a high level of SCE frequencies which are associated with the inflammatory condition itself.

Erythrocytes modulate cell cycle progression but not the baseline frequency of sister chromatid exchanges in pig lymphocytes

The effect of co-culturing varying concentrations of pig and human red blood cells (RBCs) on the baseline frequency of sister chromatid exchanges (SCEs) and cell-cycle progression in pig plasma (PLCs) and whole blood leukocyte cultures (WBCs) was studied. No variation in SCE frequency was observed between pig control WBC and PLC. Addition of pig and human RBCs to pig PLCs did not modify the baseline frequency of SCEs. On the other hand, cell proliferation was slower in PLCs than in WBCs. The addition of pig or human RBCs to PLCs accelerated the cell-cycle progression of pig lymphocytes. When RBCs were added to PLCs the concentration and time sequence of RBC incorporation affected the cell-cycle progression of swine lymphocytes. When doses of pig or human RBCs equivalent to those present in WBCs were added immediately after PLC stimulation, the cell-cycle kinetics were similar to those of WBCs. Shorter co-incubation periods or a reduction in the dose of RBCs made cell-cycle progression intermediate between PLC and WBC values. Thus, pig and human RBCs modulated the in vitro cell-cycle progression of pig lymphocytes in a time- and dose-dependent manner, and the low baseline frequency of SCEs of pig lymphocytes is independent of the presence or absence of erythrocytes in culture

`Spontaneous' genetic damage in man: evaluation of interindividual variability, relationship among markers of damage, and influence of nutritional status

The `spontaneous' frequency of genetic damage (normal background) and the possible relationship of this damage to nutritional variables in humans were investigated in 22 subjects using several indices of genetic damage. The subjects were chosen, out of 122 initially analyzed, for being at the extremes of the highest and lowest values of one index of genetic damage, the frequency of micronucleated erythrocytes in peripheral blood. This index reflects chromosomal damage and loss in bone marrow erythropoietic cells. The assay for micronuclei is convenient but is restricted to splenectomized individuals because the human spleen removes micronucleated cells. The initial 122 subjects were splenectomized, but all were normal and healthy at the time of this study and none had a previous history of neoplastic disease. Factors investigated were stability of micronucleus frequency as a function of time, correlations among multiple markers of genetic damage, and influence on damage indices of nutritional variables, including blood levels of folate, B 12 and antioxidant vitamins. Among different individuals, the range of values was 10-fold or more in the erythrocyte micronucleus, glycophorin A, plasma ascorbate and urinary 8-hydroxydeoxyguanosine (oxo 8dG) assays, was approximately 6-fold in the lymphocyte micronucleus assay, and was 2-fold in the lymphocyte sister chromatid exchange (SCE) assay. Red blood cell folate and plasma folate, B 12 and α-tocopherol values varied by up to 10-fold among individuals. Micronucleus frequencies in erythrocytes and peripheral blood lymphocytes ranged from <0.3 to 16.9/1000 in mature red blood cells, <1 to 33/1000 in reticulocytes, and 2.5 to 15/1000 in binucleate lymphocytes. Frequencies of glycophorin A variant erythrocytes ranged from 5.6 to 77.3×10 6 N/0 cells and 3.2 to 16.2×10 6 N/N cells, and oxo 8dG excretion varied from 32 to 397 pmol/kg/day. Although a wide range of values was observed in each genetic endpoint, the extreme values for various endpoints of genetic damage were not observed in the same individuals. The frequency of micronucleated erythrocytes varied over time within individuals and indicated that individuals with the highest levels of damage exhibit greater variability than those with lower levels. In some subjects, frequencies of micronucleated erythrocytes changed dramatically over an interval of 2–3 years: four subjects with initial micronucleated reticulocyte frequencies of 20.4, 5.9, 6.4 and 33/1000 changed to 2.5, 20.5, 18.5 and 12/1000, respectively. Among more than 150 individuals we have studied, including the 64 individuals studied by Everson et al. [(1988) J. Natl. Cancer Inst., 80, 525–529] and Smith et al. [(1990) Cancer Res., 50, 5049–5054], the seven individuals with the highest observed frequencies of micronucleated erythrocytes all had exceptionally low values of plasma folate, red cell folate, or plasma B 12, suggesting that folate and B 12 status are the major determinants of the types of damage that lead to spontaneous micronucleus formation in erythrocytic cells.

Symptoms, lung and liver function, blood counts, and genotoxic effects in coastal tanker crews.

The deck crew on tankers can be exposed to high concentrations of benzene and other chemicals during loading, unloading and tank-cleaning operations. The objective of this study was to investigate whether genotoxic or other early health effects of cargo vapour exposure could be detected in coastal tanker crews. The association between exposure to cargo vapours and clinical symptoms and signs, spirometry, blood cell count, blood test for liver function, and the frequency of micronuclei and sister chromatid exchanges in peripheral lymphocytes was studied in a cross-sectional investigation of 107 male crew members (66 deck crew and 41 others) on ten coastal tankers. Seven of the tankers had automatic cargo level gauging systems but some of the ships still had open hatches during loading and unloading operations. Acute symptoms such as headache, nausea, vertigo, fatigue and dizziness after loading or tank-cleaning operations were reported by 56 of the 66 deck crew members (85%). Irritation of the mucous membrane in eyes and upper respiratory tract by cargo vapours were also common in this group. Obstructive symptoms were more common in the group with the highest exposure to cargo vapours but persistent effects on lung function (vital capacity and forced expiratory volume in 1 s), nervous system, liver enzymes or blood counts were not found. The frequency of micronuclei after mitotic stimulation with phytohaemagglutinin was higher among the deck crew (mean 4.2 SEM 0.40) than in other crew members (mean 3.6, SEM 0.35). although the difference was not statistically significant. We found no association between exposure and the frequency of sister chromatid exchanges or micronuclei after stimulation with pokeweed mitogen. This study indicates that exposure to cargo vapours in coastal tanker crews may cause symptoms in the respiratory and nervous systems.

Variation in arsenic-induced sister chromatid exchange in human lymphocytes and lymphoblastoid cell lines

This study was undertaken to compare the genotoxic effects of arsenite in cultured human lymphocytes and lymphoblastoid cell lines from a group of normal human volunteers. The goal was to determine whether, as found with other genotoxins, subgroups might exist which showed relative high or low sensitivity to induction of sister chromatid exchanges (SCEs) by this metal. Primary lymphoblast cultures were established by treatment with phytohemagglutinin (PHA-L). Lymphoblastoid cell lines were established by transformation with Epstein-Barr virus. Cultures were exposed for 40 h to sodium arsenite (AsIII) and SCEs assayed by 5-bromo-2'-deoxyuridine incorporation and staining by fluorescence plus Giemsa. SCEs were increased by arsenite in a dose-dependent manner over the concentration range of 10(-7)-10(-5) M. SCEs could not be scored above 10(-5) M because of cytotoxicity. Comparison of SCE frequency in primary lymphocyte cultures among individuals showed substantial variation in sensitivity to arsenite, with some showing no significant effect while others showed a 2-3-fold increase in SCE frequency. In one lymphoblastoid cell line especially sensitive to arsenite, arsenic acid (AsV) or dimethylarsinic acid (DMA) at concentrations up to 10(-5) M did not increase the SCE frequency suggesting that AsIII is the active form of arsenic. When pooled data from the primary lymphocytes was compared to that obtained with the lymphoblastoid cells, the slopes of the dose-response curves for ASIII-induced SCEs were similar. The sensitivity of the majority of the individual primary lymphocyte cultures to SCE induction by arsenite was correlated with the sensitivity of the lymphoblastoid cultures established from the same individual. However, in three individuals no correlation was found. Individual lymphoblastoid cell lines retained their As sensitivity after cryopreservation and subsequent revival. Whether the genotoxic response to As is genetically controlled or the result of phenotypic selection is being explored in these stable lymphoblastoid cell lines.

DNA single strand breakage, DNA adducts, and sister chromatid exchange in lymphocytes and phenanthrene and pyrene metabolites in urine of coke oven workers.

OBJECTIVES: To investigate the specificity of biological monitoring variables (excretion of phenanthrene and pyrene metabolites in urine) and the usefulness of some biomarkers of effect (alkaline filter elution, 32P postlabelling...

Sister chromatid exchanges at different times after repeated injections of thiophosphamidein vivo

The frequency of sister chromatid exchanges in rabbit peripheral blood lymphocytes was estimated after three intravenous injections of thiophosphamide in doses of 0.5 and 2 mg/kg body weight at 2-week intervals. It is demonstrated that 24 h after the first injection the frequency is significantly higher than after subsequent injections.

Lack of effect of long-term fluoride ingestion on blood chemistry and frequency of sister chromatid exchange in human lymphocytes

Two studies were conducted to assess the potential for adverse physiologic and genotoxic effects of long-term fluoride ingestion in adults residing in three communities with varying water fluoride levels (0.2 ppm, 1.0 ppm, and 4.0 ppm). All were long-time (> or = 30 years) residents of their respective communities. Plasma and urine samples were collected for fluoride analyses. Additional plasma was collected to determine blood chemistry, and plasma lymphocytes were examined to determine the frequency of sister chromatid exchange. Significant differences in urine (P = 0.001) and plasma (P = 0.0001) fluoride levels were found in the three communities. Seven of the blood parameters were statistically different among the communities, although all were within the defined normal range of the pathology laboratory. Sister chromatid exchange frequency was statistically higher in the 4.0 ppm fluoride community as compared to the other communities. Because of the higher SCE frequency in the 4.0 ppm fluoride community, a second study was performed to determine if the increased frequency was potentially related to the fluoride level of the communal water supply. Of the 58 adults recruited; 30 had used city water and 28 had used well water (< or = 0.3 ppm fluoride) as their principal water source for 30 years. Data analyses showed that the sister chromatid exchange frequency did not differ between the groups, indicating that the increased sister chromatid exchange frequency was not related to the fluoride level of the communal water. The investigation provided evidence that the long-term ingestion of water containing 4.0 ppm fluoride did not have any clinically important physiologic or genotoxic effects in healthy adults.

Problems in the biological monitoring of chromium(VI) exposed individuals

The usefulness of currently available techniques for the biological monitoring of chromium(VI) exposed individuals is reviewed. Chromium levels in body fluids, such as urine and blood plasma, are reliable markers of exposure to chromium in oxidation states (VI) and (III) and provide a measure of the internalized dose of chromium. These markers are sufficiently sensitive to be useful in most occupational settings encountered today. In contrast, the majority of cytogenetic surveillance studies among chromium platers, ferrochromium workers and stainless steel welders using the manual metal arc (MMA) method have yielded negative or inconclusive results. As a marker for genotoxicity, the number of sister chromatid exchanges in blood lymphocytes proved to be relatively insensitive towards exposure to chromium(VI). There were however significant increases in rare chromosome aberrations among MMA stainless steel welders, although the reported levels of all aberrations combined were similar to those observed among control groups of many other studies. The relative lack of success of cytogenetic surveillance studies using blood lymphocytes is surprising in view of the strong genotoxicity of chromium(VI). A possible explanation comes from recent studies which showed that the differences in chromium lymphocyte levels between exposed and controls were disproportionately small. Another factor which complicates attempts to correlate genotoxic effects in lymphocytes with the processes giving rise to cancers of the respiratory system is the toxicokinetics of inhaled chromium(VI). Only small fractions of the total inhaled dose are distributed in the body while the bulk of chromium(VI) deposited in the lungs remains there for very long periods of time. The vast majority of lymphocytes will therefore come into contact with chromium(VI) not while travelling through the supporting tissues of the lungs but during their migration through the blood. There they take up chromium(VI) that has leached from the lungs. Blood lymphocytes therefore seem to be inappropriate for the monitoring of the biologically effective dose, and of early biological effects arising from exposure to chromium(VI). Thus there is an urgent need to develop techniques which would allow the non-invasive monitoring of internalized doses of chromium in the lung.

Chromosomal changes in rheumatoid arthritis patients treated with CPH82.

Chromosomal changes were assessed in 19 patients with rheumatoid arthritis (RA) treated with CPH82, a benzylidated podophyllotoxin glycoside, for up to one year. The frequency of chromosomal aberrations (CA) and sister chromatid exchanges (SCE) in peripheral lymphocytes increased significantly after 12 weeks of treatment and remained elevated after 48 weeks treatment in peripheral lymphocytes. The number of CA and SCE were significantly increased in CPH82 treated patients compared with the RA patients treated with other disease modifying anti-rheumatic drug (sulphasalazine, gold, D-penicillamine, azathioprine, methotrexate, cyclophosphamide). Only two patients treated with cyclophosphamide and azathioprine had changes of comparable levels. The results of this study suggest a mutagenic potential of CPH82 similar to that described for other immunosuppressive drugs and the newer podophyllotoxin derivatives, etoposide and teniposide.

Dose dependent of sister chromatid exchanges in humans lymphocytes induced by in vitro alpha-particle irradiation.

Exposure of human G0 lymphocytes to high-LET particles under different conditions has been seen, unlike low-LET radiations, to be substantially effective in the induction of sister chromatid exchanges (SCE). However, whereas for fast neutrons a linear dose response of SCE has been determined, there is no sign of a dose-response relationship for alpha-particles. A likely reason for this lack of dose dependence may be the irradiation procedure. Therefore, a technique developed in our laboratory to ensure uniformity of irradiation with alpha-particles was used in the present study. Monolayers of 3 h-stimulated lymphocytes were exposed with alpha-particles from 241Am. Underdispersion was found for the cell-to-cell variance of the number of SCE. The dose response of SCE was linear, with a yield of 3.4 SCE per cell and per Gray.

Effect of chemical mutagenic agent on chromosomal pattern of Egyptian Kwashiorkor infants

Genetic polymorphisms that affect xenobiotic metabolism or cellular response to DNA damage can modulate individual sensitivity to genotoxins. Information on the effects of such polymorphisms on the level of chromosome damage may facilitate the identification of risk groups and increase the sensitivity of cytogenetic endpoints as biomarkers of genotoxic exposure and effect. Glutathione S-transferase M1 (GSTM1) is an important detoxification enzyme which, due to a homozygous gene deletion (null genotype), is lacking from about 50% of Caucasians. A higher level of DNA adducts and chromosome damage has been detected in lymphocytes of tobacco smokers and bus drivers who lack the GSTM1 gene. Other polymorphic glutathione S-transferases include GSTM3, GSTP1, and GSTT1. The GSTT1 null genotype (10 – 20% of Caucasians) has been associated with an increased “baseline” level of sister chromatid exchanges (SCEs) in lymphocytes. N-acetyltransferase 2 (NAT2), metabolizing xenobiotics with primary aromatic amine and hydrazine structures, is another important polymorphic phase II enzyme. Subjects having the NAT2 slow acetylator genotype appear to show an increased baseline frequency of lymphocyte CAs in the absence of identified environmental exposure. Besides human biomonitoring studies, genetic polymorphisms may be important in explaining individual variation in genotoxic response observed in genetic toxicology tests with human cells. Several studies have suggested that blood cultures from GSTT1 null and GSTM1 null individuals have increased in vitro sensitivity to various genotoxins. The best-known example is probably the diepoxybutane sensitivity of GSTT1 null donors. Recently discovered polymorphisms affecting DNA repair may be expected to be of special importance in modulating genotoxic effects; the first available studies have suggested that the exon 10 Arg399Gln polymorphism of XRCC1 gene (X-ray repair cross-complementing group 1) could affect individual genotoxic response. In conclusion, the genetic polymorphism of GSTM1 influences the frequency of chromosome damage in exposed humans, while that of GSTT1 and NAT2 affect the “baseline” level of such damage. Both GSTM1 and GSTT1 genotypes may shape the in vitro genotoxic response of human lymphocytes. The significance of DNA repair polymorphisms is presently unclear.

Sister chromatid exchanges in human peripheral blood lymphocytes after ingestion of high doses of arsenicals

Sister chromatid exchanges in human peripheral blood lymphocytes after ingestion of high doses of arsenicals

Effect of melatonin on mitotic and proliferation indices, and sister chromatid exchange in human blood lymphocytes

Cells from human peripheral blood were cultured in vitro in the presence of 0.05 to 1.00 mM melatonin, 10 −7 M mitomycin C (positive control) and 0.5% ethanol (solvent control) for 72 h at 37 ± 1°C. Lymphocytes were examined for mitotic and proliferation indices, and for the incidence of sister chromatid exchange. The results indicate that the lymphocytes which were cultured in the presence of ≥ 0.20 mM concentrations of melatonin exhibited a significant and concentration-dependent decrease in mitotic index and alteration in proliferation kinetics. This was demonstrated by an increase in the frequency of lymphocytes in their first division, with a concomitant decrease in the second and third or later division cells. The incidence of sister chromatid exchange was similar in the lymphocytes exposed to 0.05 to 1.00 mM melatonin and untreated controls. Exposure of the cells to ethanol, the solvent used, did not alter either the mitotic or proliferation indices, or the frequency of sister chromatid exchange. The lymphocytes treated with mitomycin C showed the expected decrease in mitotic and proliferation indices, and an increased incidence of sister chromatid exchange. These observations indicate that melatonin, when continuously present in the cultures for 72 h at the concentrations tested, while not genotoxic as indicated by the sister chromatid exchange assay, inhibits the proliferation of mitogen stimulated (and proliferating) human blood lymphocytes at supraphysiological concentrations.