Vera Garaj-Vrhovac

Cytogenetic monitoring of croatian population occupationally exposed to a complex mixture of pesticides.

This paper describes a longitudinal study of possible genetic damage in Croatian workers occupationally exposed to a complex mixture of pesticides. The methods of choice were chromosomal aberration analysis, sister chromatid exchange analysis (SCE), micronucleus assay and comet assay. In order to determine primary genotoxic effects in workers, blood samples were taken after the workers spent 8 months in the production of pesticides. During the production all subjects were simultaneously exposed to a complex mixture of pesticides containing atrazine, alachlor, cyanazine, 2,4-dichlorophenoxyacetic acid, and malathion. To detect DNA repair in lymphocytes of the same subjects the second series of blood samples was taken 8 months after the workers were removed from production. Regardless of the time sampling time the exposed workers showed an increased number of chromosomal aberrations, SCE frequency, micronucleus (MN) frequency, and values of comet assay parameters. After 8 months of non-exposure the workers showed a significantly decreased number of chromosomal aberrations, MN frequency, and DNA migration compared to the results of the first sampling, but it was still significantly higher than in controls. Furthermore, the SCE frequency in the exposed subjects did not drop after the 8 months of non-exposure, which indicates long-term exposure to a mixture of pesticides.

Sister chromatid exchange and proliferative rate index in the longitudinal risk assessment of occupational exposure to pesticides.

At present, there are more than 1,000 chemicals classified as pesticides and many reports have shown that some of them have genotoxic properties. In the present longitudinal study, possible genetic damage on a population of workers occupationally exposed to a mixture of pesticides by using sister chromatid exchange (SCE) analysis has been evaluated. As an additional cytogenetic parameter, the proportion of lymphocytes that undergo one, two or three cell divisions as well as proliferative rate index have been determined. This study was performed on the exposed group of workers employed in pesticide production, simultaneously exposed to a complex mixture of pesticides (atrazine, alachlor, cyanazine, 2,4-dichlorophenoxyacetic acid, and malathion). The blood samples of the exposed subjects were collected in three different periods: before the beginning of the new pesticide production period, after 8 months of everyday work in the pesticide production, and 8 months after the removal of subjects out of the production. In all three samplings, the mean value of SCE and number of cells with high sister chromatid exchange frequency (HFC) in the exposed group was significantly higher in the comparison with the control group. There were no differences in the proliferative rate index (PRI) between the control and exposed group, regardless of the sampling period. In both groups examined, the majority of lymphocytes were found in the second cell division, following cultivation. These results suggest that the increase in the number of SCE found in the exposed subjects is not the result of either cytotoxic or epigenetic action of pesticide mixture, but chronic occupational exposure to mixture of pesticides.

Acute cytogenetic effects of antineoplastic drugs on peripheral blood lymphocytes in cancer patients chromosome aberrations and micronuclei.

Aims and Background The aim of the present study was to evaluate the individual sensitivity of cancer patients to different antineoplastic drugs administered in standard protocols by assessing their acute cytogenetic effects on peripheral blood lymphocytes. Methods and Study Design In 12 patients undergoing cancer chemotherapy, acute cytogenetic effects on peripheral blood lymphocytes were evaluated by analysis of structural chromosome aberrations and micronuclei. All patients were given antineoplastic drugs, mainly as polychemotherapy. The frequencies of both cytogenetic biomarkers determined after the first chemotherapy cycle were compared with their pre-treatment (baseline) values. Results All chemotherapy protocols employed induced clear cytogenetic effects in both tests studied. The results obtained indicate interindividual variations between cytogenetic damage in peripheral blood lymphocytes among cancer patients. Statistically significant increases in the total number of structural chromosome aberrations and micronuclei in lymphocytes analyzed after chemotherapy compared to pre-therapy samples were observed in almost all patients studied. The highest level of chromosome damage as well as the highest incidence of micronuclei was observed following administration of the ACOP protocol (adriamycin, cyclophosphamide and vincristine). The proportions of signal-positive and signal-negative micronuclei were evaluated using DAPI staining, while silver staining revealed Ag-NOR+ and Ag-NOR− micronuclei. In some patients the incidence of signal-positive and Ag-NOR+ micronuclei after treatment was increased, indicating a more pronounced susceptibility of particular chromosomes to damage caused by antineoplastic drugs. Conclusions With regard to the results obtained we may conclude that both parameters used in the present study on peripheral lymphocytes are sensitive biomarkers and can be successfully employed for biomonitoring of acute cytogenetic effects induced by antineoplastic drugs in standard clinical protocols for cancer treatment.

Cytogenetic monitoring of cardiology unit hospital workers exposed to doppler ultrasound

We studied the effects of ultrasound on the peripheral blood lymphocytes of medical personnel from a cardiology unit working with colour Doppler ultrasonic equipment. Cytogenetic risks from ultrasound exposure were assessed by analysis of chromosome aberrations, sister chromatid exchanges (SCE), study of cell‐cycle kinetics and micronucleus assay. We found significant increases (P < 0.001) in the total number of chromosome aberrations, mainly due to chromatid breaks and acentric fragments, increases in the total number of micronuclei and SCE and disturbances in cell‐cycle kinetics in the exposed group compared to the control. In spite of their limitations, the results of the present investigation indicate that individuals occupationally exposed to ultrasound may experience an increased genotoxic risk and strongly emphasize the need for more research into the nature and extent of the biological consequences to medical personnel working with Doppler ultrasound. Copyright

Induction of micronuclei in human lymphocytes after occupational exposure to ultrasound.

Micronucleus assay combined with Giemsa and DAPI staining was performed on blood samples of subjects occupationally exposed to ultrasound. Lymphocytes were cultivated in vitro for 72 h. At 44h cytochalasin-B was added in cultures. Frequencies of micronuclei in exposed subjects statistically significant increased compared to control. The frequency of micronucleated cells and micronuclei in exposed subjects shows interindividual variability. Using DAPI staining we observed signal-positive and signal-negative micronuclei. Percentage of signal-positive micronuclei varies between 0 and 66.7% and signal-negative micronuclei between 33.3% and 100%. This study indicate harmful effects of ultrasound on human genome, but further investigations are necessary.