Genotoxic evaluation of the organophosphorous pesticide temephos

The chemical compound temephos (0,0,0',0'-tetrametyl-0,0'-thiodi-p-phenylene phosphorothioate) is an organophosphorous pesticide that has been used in Brazil since 1967 in control campaigns against the mosquito Aedes aegypti, the vector of dengue and yellow fever. We used single cell gel electrophoresis (SCGE), SOS/umu and Ames/Salmonella assays to test the toxicity and mutagenicity of temephos. Temephos was genotoxic in the SCGE assay, inducing severe DNA lesions (type IV lesions) at doses above 1.34 micro M. It was mutagenic, but not toxic, in the SOS/umu assay to Escherichia coli strain PQ37, but not to PQ35, at concentrations above 1.33 micro M, particularly when the S9 mixture was not used in the assay. Temephos was not mutagenic in the Ames assay with S. typhimurium strains TA97, TA98, TA100 and TA102, both with and without metabolic activation. However, temephos at concentrations above 3.33 micro M was mutagenic to TA98NR, YG7104 and YG7108, both with and without metabolic activation. In conclusion, temephos was genotoxic and mutagenic in all the three tests used, and in two of them at concentrations similar to those routinely used to combat Aedes aegypti.     

Increased frequency of 6-thioguanine-resistant lymphocytes in peripheral blood of workers employed in cyclophosphamide production

The frequency of 6-thioguanine-resistant peripheral blood lymphocytes has been determined by autoradiography in a control population and a population of cyclophosphamide-exposed individuals. The mean variant frequency in a non-exposed population was found to be 2.76 ± 1.48 × 10⁻⁵. Subpopulations of smokers and non-smokers revealed statistically significantt differences in the variant frequencies, i.e. 3.52 ± 1.55 × 10⁻⁵ and 2.07 ± 1.05 × 10⁻⁵ respectively. In 20 out of a total of 23 individuals employed in cyclophosphamide synthesis and manufacturing, the variant frequency of 6-thioguanine-resistant lymphocytes was found to be higher than the maximum individual frequency found in the control population. The mean variant frequency in the cyclophosphamide-exposed population was 13.64 ± 13.56 × 10⁻⁵, a statistically significant increase as compared to the mean control frequency. There was no correlation between variant frequency and duration of employment suggesting that this test reflects the actual exposure and not a cumulative effect.     

Genotoxic effects of pesticide fipronil in somatic and generative cells of mice

The pronounced genotoxic effect of fipronil in all used doses (4.75, 9.50, 19.00, and 31.70 mg/kg) at a single exposure in the liver, lungs and spleen was ascertained by the Comet assay. Organ specificity of genotoxic effects of the pesticide was revealed. The liver was the most sensitive to fipronil. Fipronil at a dose of 9.50 mg/kg in a single and repeated exposure (within 10 days) induced aberrations in mouse bone marrow cells with the frequency exceeding the spontaneous mutation rate (p < 0.01 and p < 0.001, respectively). Fipronil also showed genotoxic activity in the germ cells of the experimental animals, causing abnormalities of the structure of synaptonemal complexes in the spermatocytes.     

Risk of cerebrovascular disease incidence in the cohort of Mayak production association workers first employed during 1948-1958

Incidence of cerebrovascular diseases (CVD) has been studied in a cohort of 12210 workers first employed at one of the main plants (reactors, radiochemical or plutonium) of the Mayak nuclear facility during 1948-1958 and followed up to the end of 2000. Information on external gamma doses is available for virtually all (99.9%) of these workers; the mean (+/- one standard deviation) total gamma dose was 0.91 +/- 0.95 Gy (99% percentile 3.9 Gy) for men and 0.65 +/- 0.75 Gy (99% percentile 2.99 Gy) for women. Plutonium body burden was measured only for 30.0% of workers. Amongst those monitored, the mean (+/- standard deviation) cumulative liver dose from plutonium alpha exposure was 0.40 +/- 1.15 Gy (99% percentile 5.88 Gy) for men and 0.81 +/- 4.60 Gy (99% percentile 15.95 Gy) for women 4418 cases (first diagnosis) of CVD were identified in the studied cohort. A statistically significant increasing trend in CVD incidence with total external gamma dose was revealed after adjustment for non-radiation factors and internal exposure from incorporated plutonium-239. Excess relative risk per Gy was 0.464 (95% confidence interval 0.360-0.567). Incidence of CVD was statistically significantly higher for the workers chronically exposed to external gamma rays at a dose above 1.0 Gy A statistically significant increasing trend in CVD incidence with internal liver dose from plutonium alpha exposure was observed after adjustment for non-radiation factors and external exposure. ERR per Gy was 0.155 (95% confidence interval 0.075-0.235). CVD incidence was statistically significantly higher among workers with a plutonium liver dose above 0.1 Gy, although the trend estimates differed between workers at different plants. The incidence risk estimates for external radiation are generally compatible with estimates from the study of Chernobyl clean-up workers, although the incidence data point to higher risk estimates compared to those from the Japanese A-bomb survivors.     

Genotoxic evaluation of Ara-C by multiple parameters

The cytogenetic effects of the antimetabolite, cytosine arabinoside (Ara-C) are evaluated using in vivo and in vitro test systems and applying multiple parameters. The in vivo assay was carried out on 8-10-week-old inbred Swiss albino male mice using bone marrow as the somatic test system and the cells of testis as the meiotic test system. In vitro human leukocyte cultures were also employed. In vivo experimental doses were computed on surface area basis within the therapeutic dose range and injected intraperitoneally and for in vitro they were calculated on blood volume basis. Evaluation of somatic chromosome mutations included conventional screening for chromosome aberrations, variations in mitotic index and sister-chromatid exchanges (SCEs) by in vivo and in vitro methods besides studies on meiotic test systems using conventional screening for chromosome and sperm-head abnormalities. The quantitative data were subjected to statistical analysis by applying appropriate tests to evaluate their significance. The results of in vivo and in vitro experiments reveal the chromosome mutational activity of the compound. This is further supported by data on SCEs from both systems. However, a comparison of both demonstrated a differential mutagenic response of the drug, more in vivo than in vitro. This is also true for SCEs. Even though the mechanisms involved in causing chromosome aberrations and SCEs are different, the data on both corroborate each other on induction of chromosome mutations.     

Genotoxic evaluation for the estrogenic mycotoxin zearalenone

Genotoxic effects of the mycotoxin Zearalenone (ZEN) were evaluated on albino mice. The investigation was assessed using 4 criteria: chromosome aberrations in bone marrow and spermatocytes of adult male mice; chromosome analysis and teratological effects of mice embryos. Zearalenone was administrated to both adult males and pregnant females with 2 doses level (5 microg x kg(-1) and 10 microg x kg(-1) ZEN). Zearalenone was found to reduce the mitotic activity in treated males and the embryos proving that it is a cytotoxic substance. In treated males and females, it induced some chromosome abnormalities with no significant increase over the control at the doses investigated, except for some few figures. Similar results were observed for the teratological study. The results in general could consider zearalenone as a toxic mycotoxin for both adult animals and embryos. It is highly recommended that a great attention should be paid towards the toxicity of zearalenone to mono-gastric animals and human, especially it contaminate corn that is widely used in human and animal feeding.     

Radiologic diagnosis of morphologic and functional changes in the bronchopulmonary system in workers employed in production and processing of ethylene oxide]

A total of 3475 subjects, engaged in such production for 2-20 years, were annually and before starting work examined by clinical and x-ray methods; 2427 of these were regularly exposed to ethylene oxide (the main group). X-Ray examinations included roentgenomorphologic (total fluorogram of the thoracic cavity organs and long-distance roentgenograms, tomograms, bronchograms, if necessary) and roentgenofunctional (roentgenopneumopolygraphy) methods of examination of the bronchopulmonary system. Chronic bronchitis was detected in 407 workers of the main group and in 79 controls; in 76.2% of the main group patients and in 90% of controls it was chronic nonobstructive functionally unstable bronchitis, in the rest it was chronic obstructive bronchitis.     

Genotoxic evaluation in two amphibian species from Brazilian subtropical wetlands

Biomarkers analysis serves as an early warning system for the presence of pollutants because their responses appear before irreversible damage to the ecosystem takes place. The genotoxic effects of pollutants may occur at cellular pollutant concentrations that are well below levels that would cause gross cytotoxicity, making this a useful tool to detect early effects of toxic environmental agents. Combining the importance of Brazilian wetlands to the conservation of amphibian biodiversity with the potential negative impacts of irrigated rice fields in the surrounding areas, the aim of the present study was to evaluate genotoxic damage in two amphibian species, Pseudis minuta, and Leptodactylus gr latrans, from the southern Brazilian wetlands. Adult specimens from both Anuran species were captured from preserved (Taim Ecological Station = TAIM) and non-preserved (Senandes) wetlands. Nuclear abnormalities were quantified in erythrocytes, and the results were compared using the Mann–Whitney U test. There was a higher incidence of micronucleated erythrocytes in P. minuta, and of notched nuclei in L. gr latrans that were collected in TAIM when compared to those that were collected in Senandes, despite the fact that TAIM is a conservation unit. These findings indicate that Anurans are coping with genotoxic substances in their habitats, and underscore the need to implement monitoring programs in TAIM to determine which compounds or mixtures might be causing cell damage and to investigate the effects of such compounds on other anuran species and animal groups.     

Effect of dithiocarbamate pesticide zineb and its commercial formulation, azzurro. III. Genotoxic evaluation on Chinese hamster ovary (CHO) cells

The in vitro genotoxicity exerted by the dithiocarbamate fungicide zineb, and its commercial formulation azzurro, were studied in Chinese hamster ovary (CHO) cells by the analysis of the sister chromatid exchange (SCE), cell-cycle progression and single cell gel electrophoresis (SCGE) assays. Both zineb and azzurro activities were tested within the range of 0.1-100.0 microg/ml. Concentrations of 0.1-25.0 microg/ml of zineb or azzurro induced a significant dose-dependent increase in SCE frequency over control values. For both test compounds, while doses ranging from 0.1 to 1.0 microg/ml did not alter the rate of cell proliferation, a significant delay in cell-cycle progression was observed within the 5.0-25.0 microg/ml dose-range. A regression test showed that either the proliferative replication index or the mitotic activity of cultures decreased as a function of the pesticide concentration within the 1.0-25.0 microg/ml dose-range. Doses higher than 50.0 microg/ml were cytotoxic. SCGE assay revealed an increase in zineb-induced DNA damage by enhancing the proportion of slightly damaged cells in the 25.0-100.0 microg/ml dose-range and by increasing in a dose-dependent manner the proportion of damaged cells within the 1.0-100.0 microg/ml dose-range. Overall, image analysis showed statistically significant positive relationships between zineb concentration and DNA damage (expressed by image length and width) and between length and width of the damaged cells. In azzurro-treated cells, only when 100.00 microg/ml was employed a significant increase in the frequency of damaged cells over control values affecting the totality of the cells was observed only when 100.0 microg/ml was employed. When lower doses were employed, no DNA damage was revealed. Based on these results, the evaluation of zineb as a genotoxic/non-genotoxic compound for human health should be reconsidered. Even though we demonstrate that the pesticide induces large DNA alterations in vitro, does no necessarily mean that the chemical should be considered clastogenic.notoxic     

Psychological symptoms among workers employed in companies undergoing privatization in postwar Bosnia and Herzegovina

In Central and Eastern European countries, after abandoning communism, significant political, economic and social changes occurred, followed by the increase in income inequality and social disparity. The goal of this study was to examine the relationship between psychological symptoms and monthly income of employees in companies undergoing privatization. The study included 258 workers from seven companies undergoing privatization in the Tuzla Canton region. For the study purposes, the Brief Symptom Inventory (BSI) and a general questionnaire with questions about socio-demographic characteristics, income, and workplace, were used. Monthly income of the majority of workers (207 or 80.2%) was below the monthly income in Bosnia and Herzegovina. Workers with salaries below the average salary for Bosnia and Herzegovina have pronounced somatization, anxiety, paranoia, interpersonal sensitivity and hostility. The BSI scale yielded significant negative correlation between the level of monthly salary and the expression of psychological symptoms (r = -0.184, p = 0.002) and between the level of family income and the expression of psychological symptoms (r = -0.123, p = 0.024). Based on the study results, it was determined that socio-economic factors such as the level of salary and total family income and job insecurity, educational level, marital status and gender may be predictors of psychological symptoms.     

Genotoxic evaluation of chronic fluoride exposure: sister-chromatid exchange study

As part of a continuing investigation, this study was conducted to examine the genotoxic effects of chronic exposure to sodium fluoride (NaF) in drinking water on the frequency of sister-chromatid exchange (SCE) in the bone-marrow cells of male Chinese hamsters. Animals at about 3 weeks of age were randomly assigned to 6 groups, each with at least 3 hamsters, and were maintained on a low fluoride diet (less than 0.2 ppm F) throughout the experiment. At 4 weeks of age, the animals in groups I-V began to receive drinking water containing fluoride at concentrations of 0, 1, 10, 50 and 75 ppm, respectively. Group VI was treated with cyclophosphamide and served as the positive control. The animals were sacrificed at 24 weeks of age by cervical dislocation. The humeri and plasma were analyzed for fluoride content, which was found to increase with the increase in fluoride concentration in drinking water. Slides of chromosomes from bone-marrow cells were prepared and blindly examined for the frequency of SCE. The mean scores of SCE for the hamsters receiving drinking water containing F concentrations up to 75 ppm for 21 weeks ranged from 4.28 to 6.28 per cell, and were not significantly different from those of the negative controls (4.60-5.44/cell). The results indicated that chronic fluoride exposure had no effect on the frequency of SCE in Chinese hamster bone-marrow cells under the conditions of the present investigation.     

Genotoxic evaluation of bupivacaine and levobupivacaine in the Drosophila wing spot test

Bupivacaine and levobupivacaine are amino amide local anesthetics commonly used in medical practice. Although bupivacaine consists of a racemic mixture of S (–)-bupivacaine and R (+)-bupivacaine enantiomers, levobupivacaine is comprised of pure S (–)-bupivacaine. It has been known that levobupivacaine is preferable to bupivacaine since it may cause cardiovascular and nervous system toxicity. For determining genotoxicity of these anesthetics, we used the wing somatic mutation and recombination test in Drosophilamelanogaster. Three-day-old trans-heterozygous larvae were treated with bupivacaine and levobupivacaine. Analysis of the standard crosses indicated that bupivacaine and levobupivacaine did not exhibit mutagenic or recombinogenic activity until toxic doses have been reached at the larval stage. When we examined bupivacaine and levobupivacaine in the HB cross, bupivacaine did not exhibit any genotoxicity at high concentrations (500 µg/mL), but levobupivacaine did exert genotoxicity at high concentrations (1000 µg/mL)—depending on the substantial recombinogenic effect.     

Identification of markers of depression and neurotoxicity in pesticide exposed agriculture workers

Earlier, we reported that chronic exposure to pesticides causes a reduction in the acetylcholinesterase activity and hematological and biochemical alterations in agriculture workers. In continuation with that, the present study aimed to investigate the pesticide‐induced neurochemical imbalance and its association with behavior alterations in agricultural workers. A significant increase in depressive symptoms, assessed by the Beck Depression Inventory‐II was observed in pesticide exposed workers as compared to the unexposed. A decrease in the level of dopamine in plasma and levels of dopamine, 3,4‐dihydroxyphenylacetic acid, homovanillic acids, norepinephrine, serotonin, and hydroxyindoleacetic acid in urine was also observed. An increase in the levels of MAO‐A and MAO‐B has also been observed in these individuals. The decreased levels of neurotransmitters in the blood and urine have been linked with increased levels of MAO and pesticide residues in plasma and urine. Furthermore, these changes were associated with a higher incidence of depression in agricultural workers.     

Cerebrovascular diseases in the cohort of workers first employed at Mayak PA in 1948-1958

The incidence of and mortality from cerebrovascular diseases (CVD) have been studied in a cohort of 12,210 workers first employed at one of the main plants of the Mayak nuclear facility during 1948-1958 and followed up to 31 December 2000. Information on external γ-ray doses is available for virtually all of these workers (99.9%); the mean total γ-ray dose (± SD) was 0.91 ± 0.95 Gy (99th percentile 3.9 Gy) for men and 0.65 ± 0.75 Gy (99th percentile 2.99 Gy) for women. In contrast, plutonium body burden was measured only for 30.0% of workers; among those monitored, the mean cumulative liver dose from plutonium α-particle exposure (± SD) was 0.40 ± 1.15 Gy (99th percentile 5.88 Gy) for men and 0.81 ± 4.60 Gy (99th percentile 15.95 Gy) for women. A total of 4418 cases of CVD, including 665 cases of stroke, and 753 deaths from CVD, including 404 deaths from stroke, were identified in the study cohort. Having adjusted for non-radiation factors, there were statistically significant increasing trends in CVD incidence but not mortality with both total external γ-ray dose and internal liver dose. Much of the evidence for increased incidence in relation to external dose arose for workers with cumulative doses above 1 Gy. Although the dose response is consistent with linearity, the statistical power to detect non-linearity at external doses below 1 Gy was low. CVD incidence was statistically significantly higher among workers with a plutonium liver dose above 0.1 Gy. There was a statistically significant increasing trend in incidence with increasing internal dose, even after adjusting for external dose, although the trend estimates differed between workers at different plants. The risk estimates for external radiation are generally compatible with those from other large occupational studies, although the incidence data point to higher risk estimates compared to those from the Japanese A-bomb survivors.     

Genotoxic evaluation of the antimicrobial drug, trimethoprim, in cultured human lymphocytes

The antimicrobial drug, trimethoprim, was evaluated for genotoxicity in human peripheral blood lymphocyte cultures set-up from two healthy donors. Sister-chromatid exchanges (SCE) and micronuclei (MN) were scored as genetic endpoints. The treatment was done using different trimethoprim concentrations ranging from 1 to 100 μg/ml. From our results, we can conclude that this drug is able to induce both cytotoxic and moderate genotoxic effects, as revealed by the increases seen in SCE and MN frequencies in cultures from the two donors and, at least, at one of the concentrations tested.