Cytogenetic damage in peripheral blood cultures of Chaetophractus villosus exposed in vivo to a glyphosate formulation (Roundup).

Different concentrations of a glyphosate formulation, Roundup® Full II (66.2% glyphosate) were tested in culture peripheral blood of armadillo Chaetophractus villosus with cytogenetic biomarkers like mitotic index (MI), chromosomal aberrations (CA), sister chromatid exchange (SCE) and cell proliferation kinetics (CPK) by means of replication index. Adults animals of both sexes were exposed to RU at four concentrations ranging from 0.026 mL RU solution to 0.379 mL RU daily in oral treatment with the same volume (0.2 mL) during 7 days. We analyzed the induced damage at different times considering T0 as control value, one (T1), seven (T7) and 30 days (T30). One day after, only the higher concentration shows MI significant differences (p < 0.05), at T7 the frequency increases and at T30 it decreases reaching T0 values. The analysis of CA frequencies shows that only 0.106 mL RU/day exhibit significant differences vs T0 values. A great variability is expressed in the values of standard deviation (SD) and in the wide confidence intervals of the media. One day after treatments (T1) all four concentrations shows significant differences in SCE vs T0 values. Replication Index (RI) does not show significant differences. The dose-response behavior was not observed in either CA or SCE. The consistency of the findings obtained with the same biomarkers in vitro support the idea of expanding studies in order to characterize the risk doses for these mammals.

Absence of toxicity and genotoxicity in an extract of Rubus coriifolius.

Rubus coriifolius Focke is a wild plant from the Rosaceae family. It grows in both Guatemala and Mexico. The polar extract of the aerial parts of this plant has antibacterial, anti-inflammatory, and anti-protozoal activities. These properties may explain the traditional use of this plant. In vivo and in vitro assays were used to assess the genotoxic and toxic effects of an ethanol extract of the aerial parts of R. coriifolius. Three groups of rats were orally administered the R. coriifolius extract diluted in ethanol (5%) at doses of 1.89 mg/kg body weight (low dose), 4.72 mg/kg body weight (medium dose), and 9.44 mg/kg body weight (high dose) for 3 weeks. Genotoxic/cytotoxic effects induced by the R. coriifolius ethanol extract were evaluated in vivo by a micronuclei (MN) test in rat's bone marrow cells and in vitro by MN and sister chromatid exchange (SCE) in human lymphocyte cultures. In vivo genotoxicity analyses revealed that the average number of micronucleated polychromatic erythrocytes and the polychromatic erythrocyte/red blood cell ratio at all doses were not significantly different from those of the negative control. In vitro genotoxicity analyses showed that MN, SCE, and proliferative index frequencies in a human lymphocyte cell culture were not significantly different from those of the negative control. These results demonstrate that the ethanol extract of R. coriifolius aerial parts is not toxic or mutagenic (in vitro and in vivo) and does not affect cell proliferation at the concentrations analyzed.

In vitro evaluation of the mutagenic and cytostatic effect of Tamaron, Lannate and Manzate alone and in mixture.

Pesticides are often used in agriculture, especially in floriculture. They are frequently applied in binary or ternary mixtures. Nevertheless, their impact on the genetic material has been scarcely explored. In this study, the mutagenic and cytostatic effect of three widely used pesticides, alone and combined, were analyzed. Briefly, lymphocytes cultures were obtained from peripheral blood samples of five healthy donors to determine the sister chromatid exchange and the replicative index (RI). Then, lymphocytes were exposed to Tamaron (100 ppm), Lannate (200 ppm) and Manzate (300 ppm) alone and combined. For the binary mixtures, the concentrations used were 50 ppm of Tamaron, 100 ppm of Lannate and 150 ppm of Manzate. For the ternary mixtures the following concentrations were used: Tamaron (33 ppm), Lannate (70 ppm) and Manzate (100 ppm). Finally, differential staining was performed. It was found that the frequency of SCE/cell showed a significant difference (P ≤ 0.05) between the control (2.66) and the individual treatments of Tamaron (4.87), Lannate: (5.12) and Manzate (4.23). Also, the values of the SCE in the binary mixture of Tamaron+Lannate (5.57), Tamaron+Manzate (6.06) and Lannate+Manzate (6.22) and the ternary mixture (6.63) were statistically different compared to the control. In the RI there was a significant difference between the control (1.98) and the Manzate (1.87). RI differences were also statistically significant (P ≤ 0.05) in mixtures of Tamaron+Lannate (1.64), Tamaron+Manzate (1.63), Lannate+Manzate (1.69) and total mixture (1.53). Therefore, it is suggested that these pesticides alone and in mixtures have both mutagenic and cytostatic synergistic effect in human lymphocytes in vitro.

Cytogenetic findings in patients diagnosed with breast cancer having undergone adjuvant chemotherapy regimens.

The aim of the present study was to investigate DNA damage in peripheral blood lymphocytes of breast cancer (BC) patients before and after administration of chemotherapy. We analyzed the frequency of sister chromatid exchange (SCE), occurrence of micronuclei (MN), and lymphocyte proliferation rate index (PRI) as cytogenetic markers in 28 female BC patients before and after chemotherapy, and in 20 age-matched healthy female volunteers. Prior to treatment, BC patients showed significantly increased background levels of SCE and MN, and lowered PRIs compared to healthy women. In comparison with pre-treatment levels, SCE and MN frequencies were significantly elevated and PRI reduced in blood samples collected after chemotherapy. Our findings indicate that SCE, MN, and PRI may serve as sensitive biomarkers for routine detection of the genetic abnormalities that may occur following administration of antineoplastic drugs in the clinical setting, as well as for the monitoring of high-risk patients receiving chemotherapy for BC.

Chaetophractus villosus as a sentinel organism: Baseline values of mitotic index, chromosome aberrations and sister chromatid exchanges.

Sentinel species are useful tools for studying the deleterious effects of xenobiotics on wildlife. The large hairy armadillo (Chaetophractus villosus) is the most abundant and widely distributed mammal in Argentina. It is a long-lived, omnivorous, burrowing species, with fairly restricted home ranges. To evaluate the level of spontaneous genetic damage in this mammal, we determined the baseline values of several genotoxicity biomarkers. The study included 20 C. villosus adults of both sexes from eight pristine localities within its geographic distribution range. Genotoxicity analysis was performed on 72-h lymphocyte cultures, using mitomycin C as positive control. We obtained the baseline values of mitotic index (MI=10.52±0.30 metaphases/total cells, n=20), chromosome aberrations (CA=0.13±0.22, n=20), sister chromatid exchanges (SCE)=6.55±0.26, n=6) and replication index (RI=1.66, n=6). MI and CA did not show significant differences (P>0.05) among localities or between sexes. No significant differences in MI, CA, SCE, and RI (P>0.05) were found between values from the pristine localities and historical data. There were significant differences in CA, SCE, and RI (P<0.05) between lymphocyte cultures from pristine localities and those exposed to mitomycin C. We propose the large hairy armadillo as a sentinel organism for environmental biomonitoring of genotoxic chemicals due to its abundance, easy manipulation, well-known biology, the fact that it is usually exposed to different mixtures and concentrations of environmental contaminants, and the baseline values of genetic damage characterized by MI, CA, SCE and RI as biomarkers.

Aqueous extracts of Lippia turbinata and Aloysia citriodora (Verbenaceae): assessment of antioxidant capacity and DNA damage.

The aim of the present work was to make a contribution to the knowledge of aqueous extracts of Lippia turbinata and Aloysia citriodora (Verbenaceae; infusion and decoction) in relation with the establishment of its antioxidant activity and lack of DNA damage, for its potential use in therapeutics. The cytogenotoxic profile was evaluated through genotoxic biomarkers such as mitotic index, cellular proliferation kinetics, sister chromatid exchanges, single-cell gel electrophoresis assay, and micronucleus test in human peripheral blood lymphocyte cultures. No statistical differences were found (P > .05) between control and exposed cultures, even between both aqueous extracts. The total antioxidant capacity was shown to be higher in the decoction than in the infusion and both aqueous extracts protected against protein carbonylation and lipid peroxidation, the decoction being more efficient than the infusion (P < .005). These results suggest the safe use of these medicinal plants as chemoecologic agents in therapeutics.

Susceptibility to the cytogenetic effects of dichloromethane is related to the glutathione S-transferase theta phenotype.

The carcinogenicity of dichloromethane (DCM) has been demonstrated by mutagenicity studies using bacteria and yeasts and using animal bioassays. Epidemiological studies indicate that exposure to DCM increases the incidences of liver and pancreas cancers. In the present study, we determine whether DCM generates DNA damage in human peripheral blood mononuclear cells (PBMCs) and whether that process depends on glutathione S-transferase theta (GSTT)-1 activity. GSTT1 is one of the enzymes that biotransforms DCM. To this end, PBMC cultures from healthy men were treated with DCM (15-500 ppm) for 72 h. Cell cultures were harvested and processed according to classical cytogenetic techniques. The frequency of sister chromatid exchanges (SCEs), the mitotic index (MI), the cell proliferation kinetic (CPK) value, and the level of GSTT1 activity were determined. DCM exposure decreased the MI in a dose-dependent manner in all individuals tested (20). The CPK value decreased from 125 ppm DCM, and the SCEs frequency increased from 60 ppm DCM. A significantly different response was observed when the group of individuals with low GSTT1 enzymatic activity (4 individuals), the group with medium GSTT1 activity (10 individuals), and the group of individuals with high GSTT1 enzymatic activity (6 individuals) were compared (0.077 ± 0.0124, 0.325 ± 0.0269, and 7.365 ± 1.3474 nmol HCOH/min/mg protein, respectively). These differences were reflected in the amount of change for all of the evaluated cytogenetic parameters (p<0.05, ANOVA) and indicated a clear susceptibility to DCM genotoxic effects related to GSTT1 activity because the cytogenetic effects were directly related to the GSTT1-specific activity. DCM was highly cytotoxic in PBMCs, even at doses within the safety range. Due to this toxicity, a review of the maximal limits for occupational exposure to DCM is advised.

Genotoxic biomonitoring of agricultural workers exposed to pesticides in the north of Sinaloa State, Mexico.

Genotoxic damage was evaluated in 70 agricultural workers, 25 women and 45 men, exposed to pesticides in Las Grullas, Ahome, Sinaloa, Mexico, with an average of 7 years of exposure. The effect was detected through the sister chromatid exchanges (SCE) in lymphocytes of peripheral blood and micronuclei (MN) and other nuclear anomalies (NA) in buccal exfoliated cells. Also, the influence on cellular proliferation kinetics (CPK) was studied by means of the replication index (RI) and the cytotoxic effect was examined with the mitotic index (MI). The non-exposed group consisted of 70 other persons, 21 women and 47 men from the city of Los Mochis, Sinaloa, Mexico. Significant differences between the exposed and the non-exposed groups were observed in SCE, CPK, MI, MN and NA. Analysis of variance revealed that age, gender, smoking and alcohol consumption did not have a significant effect on genetic damage. However, there was a correlation between exposure time to pesticides and SCE frequency. These results could have been due to the exposure of workers to pesticides containing different chemical compounds. This study afforded valuable data to estimate the possible risk to health associated with pesticide exposure.

Antigenotoxic, antioxidant and lymphocyte induction effects produced by pteropodine.

Pteropodine is a heterohimbine-type oxindole alkaloid specifically isolated from 'Cat's claw' (Uncaria tomentosa), a plant that has shown cytostatic, anti-inflammatory and antimutagenic properties and is used in traditional medicine to cure a number of diseases. In this report, we studied the ability of pteropodine to decrease the rate of sister-chromatid exchanges and micronucleated polychromatic erythrocytes in mice administered doxorubicin. We also determined its capacity to induce lymphocyte production in mice as well as its free radical scavenging potential by applying the DPPH assay. We found pteropodine (100-600 mg/kg) to significantly decrease the frequency of sister-chromatid exchanges and micronucleated polychromatic erythrocytes in mice administered with 10 mg/kg of doxorubicin. Furthermore, we determined that pteropodine partially corrected bone marrow cytotoxicity induced by doxorubicin, as it showed an improvement in the rate of polychromatic erythrocytes. Besides, 600 mg/kg of pteropodine increased 25.8% of the production of lymphocytes over the control value along a 96-hr assay, and it exhibited a strong capacity to trap the DPPH-free radical (98.26% with 250 microg/ml). Our results establish that pteropodine is an effective antimutagen in the model used, and suggest that pteropodine deserves further research in the area of cell protective potential and its mechanism of action.

Nitroimidazole derivatives: non-randomness sister chromatid exchanges in human peripheral blood lymphocytes.

Nitroimidazole derivatives exhibited genotoxic effect in different experimental conditions. This study focuses on an evaluation of possible genomic targets, at a chromosomal level, of two 5-nitroimidazoles (ornidazole and metronidazole) using the in vitro human peripheral blood culture as experimental system. We observed that both derivatives showed a decrease in mitotic index (MI) (P < 0.001), an increase in sister chromatid exchanges (SCE) frequency (P < 0.001) and no modifications in cellular proliferation kinetics (CPK). As a null hypothesis we considered the assumption that larger chromosomes should harbor more SCE, which was viewed using a novel sequential G-band (400 band resolution)/SCE technique. The analysis showed highly significant chi square values (P < 0.001), indicating that SCE frequency per chromosome is not proportional to chromosome length. SCE could be considered an instability indicator due to the high correlation between SCEs in certain chromosomal bands and the exposure to nitroimidazole derivatives.

Analysis of spontaneous and streptonigrin-induced sister chromatid exchanges in peripheral lymphocytes of aircrew members of international flights.

In this work, we extend our previous studies concerning mutagen sensitivity in flight personnel from commercial airlines by analyzing the frequency of spontaneous and streptonigrin (SN)-induced sister chromatid exchanges (SCEs) in peripheral blood lymphocytes of 18 long-haul aircrew members from Argentina and of 18 control individuals. Statistical analysis revealed no significant differences between aircrew and controls in the background level of SCEs (p > 0.05), which suggests that chronic exposure to cosmic radiation and other occupational hazards does not affect SCEs frequency in peripheral lymphocytes of aircrews. The fact that almost no correlation was found between cumulative flight hours and the yield of spontaneous SCEs in aircrews adds further support to this assumption. Therefore, the background SCEs frequency cannot be use as a valid biomarker to determine the genotoxic effects of cosmic radiation or other occupational hazards exposure in aircrews. Following SN treatment, a significant increase in the mean frequency of SCEs was observed in the control group (p < 0.05) but not in the aircrew group (p > 0.05), suggesting that at the population level, aircrew are more resistant to the mutagenic effects of SN than controls. The reasons of this resistance remain to be determined. Since cosmic radiation had no effect on the background SCEs frequency and no relationship was found between cumulative flight hours and SCEs inducer effect by SN in aircrews, a direct effect of cosmic radiation on SN resistance should be discarded.

Genotoxic and cytotoxic effects of carbofuran and furadan on Chinese hamster ovary (CHOK1) cells.

The in vitro geno- and cytotoxicity exerted by the N-methylcarbamate pesticide carbofuran (CF) and its commercial formulation furadan (F) were studied in Chinese hamster ovary (CHO(K1)) cells by several bioassays for both genotoxicity (e.g., the sister chromatid exchange (SCE) and micronuclei (MNi) frequencies), and cytotoxicity (e.g., cell-cycle progression, mitotic index (MI), 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and neutral red (NR)). Both CF and F activities were tested within the range of 5-100 microg/ml. CF within a 10-100 microg/ml concentration-range induced a significant dependent increase of SCE frequency and MNi over control values. At the same concentration-range, F increased significantly the SCE frequencies over control values although in a non-dependent manner while only an enhanced frequency of MNi was found in those 50 microg/ml-treated cultures. No binucleated cytokinesis-block cells were found in 100 microg/ml F-treated cultures. The NDI index revealed a delay in the onset of cell-division with 50 and 100 microg/ml of CF and F, respectively. The delayed rate of nuclear division induced by 100 microg/ml of F was higher than that induced by an equal concentration of CF. CF and F induced both a significant concentration-dependent delay in cell-cycle progression and a decrease in the proliferative replication index within 5-100 microg/ml and 50-100 microg/ml concentration-range, respectively. Decreased cell viability was found in up to 26% and 47% in 100 microg/ml CF- and F-treated cultures, respectively. The NR and MTT assays revealed a clear cell growth inhibition when concentrations of 50 and 100 microg/ml of either CF or F were employed. Accordingly, the results highlight that CF by itself and F, even in a greater extend exerts both genotoxicity and cytotoxicity in mammalian cells in culture, at least in CHOK1 cells.

Genotoxicity and cell death induced by tinidazole (TNZ).

Tinidazole (TNZ), a second-generation 5-nitroimidazole compound chemically related to metronidazole (MTZ), has been widely used throughout Europe and developing countries for the treatment of amoebic and parasitic infections. Despite TNZ's increasing use in therapeutics, scarce experimental reports are available in literature on its potential genotoxicity in human cells. Therefore, the aim of the present study was to achieve a precise characterization of the cytotoxic and genotoxic activities of this nitroimidazole in cultured human lymphocytes at therapeutic concentrations (0.1, 1, 10 and 50 microg/ml of culture) and evaluate the possible cell death mechanism associated with it. The endpoints analyzed included: mitotic index (MI), replication index (RI), sister chromatid exchange (SCE) and chromosomal aberrations (CA). A significant decrease (p<0.0001) in MI as well as an increase in SCE (p<0.0001) and CA (p<0.0001) frequencies were observed. No modifications in RI were found. The results suggest a genotoxic and cytotoxic effect of TNZ related with cell death process. Therefore, we evaluated this mechanism by DNA fragmentation (laddering), fluorescence microscopy using acridine orange/ethidium bromide (AO/EB) staining and flow cytometry propidium iodide (PI). DNA extracts of TNZ-treated cells resulted in nucleosomal DNA ladder pattern after 48 h of cell treatment; meanwhile no differences were detected in untreated cells. This pattern correlated with the observed decrease in cellular viability (p<0.05), morphological evidence of apoptosis and increase in the percentage of nuclei with hypodiploid DNA content of TNZ exposed cultures compared with control (p<0.05). We concluded that TNZ is genotoxic, cytotoxic and is able to modulate cell death through apoptotic mechanisms in the experimental design employed.

Cell protection induced by beta-sitosterol: inhibition of genotoxic damage, stimulation of lymphocyte production, and determination of its antioxidant capacity.

Beta-sitosterol (BS) is a compound that has shown various activities potentially useful for human health. In the present study, we determined its antigenotoxic capacity and lymphocyte induction potential in mouse as well as its capacity to trap free radicals in vitro. BS, in doses from 200 to 1,000 mg/kg, was able to significantly reduce the frequency of sister chromatid exchanges induced by 10 mg/kg of doxorubicin (DX) in bone marrow cells. The same range of BS doses also gave rise to a strong reduction in the rate of micronucleated, polychromatic erythrocytes induced by DX. In addition, we determined an increase in the production of lymphocytes in mice administered with BS. By means of the DPPH assay, the compound was shown to trap free radicals in a concentration dependent manner as high as 78.12% using 250 mug/ml. Our research established three relevant biological activities of BS which show its potential as a chemopreventive agent.

Assessment of the genotoxic risk of Punica granatum L. (Punicaceae) whole fruit extracts.

Punica granatum L. (Punicaceae) whole fruit extracts, have been used in Cuban traditional medicine as an effective drug for the treatment of respiratory diseases. This species showed interesting anti-viral activity, e.g. aqueous or hydroalcoholic extracts of whole fruits have proved highly active against the influenza virus. However, some toxic properties of this extract have also been reported and, to date, very little is known about its genotoxic properties. In the present study, the genotoxicity of a Punica granatum (pomegranate) whole fruit extract was assessed using different in vitro and in vivo assays that detect DNA damage at different expression levels. Results from reversion and gene-conversion test in microorganisms, sister chromatid exchanges, micronuclei and sperm-shape abnormality assays in mice, clearly showed that the hydroalcoholic extract of P. granatum whole fruits is genotoxic when tested both in vitro and in vivo.

Cell proliferation kinetics and genotoxicity in lymphocytes of smokers living in Mexico City.

Genotoxicity caused by tobacco smoke was assessed in peripheral blood lymphocytes of smokers living in Mexico City by determining sister chromatid exchange (SCE), cell proliferation kinetics (CPK), replication index (RI) and mitotic index (MI). Nicotine levels, and its major metabolite cotinine, were also estimated in urine samples using gas-chromatography-mass spectrometry to quantify smoking intensity. The outcome of the analysis and the comparison of the 77-smoker group with a non-smoking control group showed that moderate and heavy smokers exhibited significant differences (P < 0.001 and P < 0.05, respectively) in CPK, with an underlying delay in the cellular cycle; similarly, RI was significantly different in these groups (P < 0.001 and P < 0.0001, respectively). There were significant correlations (P < 0.05) between age and number of years the subject had been smoking, as well as between RI and nicotine and cotinine levels and between CPK (M1, M2 and M3) and nicotine and cotinine levels. Smokers were classified for the analysis according to the nicotine levels (it is in relation to number of cigarettes smoked per day) found in urine (ng/mL) as: light (10-250), moderate (251-850) and heavy (851-4110). Significant differences in CPK were found (P < 0.05) between moderate and heavy smokers and non-smokers. Significant differences in RI were found between moderate (P < 0.001) and heavy smokers (P < 0.0001) and non-smokers, but not for the light smoking group. MI was determined in 57 of the smokers, whereas SCE frequency was only recorded in 34 smokers. Both parameters yielded no significant differences, nor correlations with any of the assessed variables. In conclusion, cytokinetic and cytostatic effects were mainly detected in heavy and moderate smokers. Cell cycle delay and RI decrease were found in all ;healthy' smokers. The nicotine and cotinine exposure (causing oxidative damage to DNA) may have implications in the decrease in cell replication due to direct damage to DNA and/or a decrease in the DNA repair mechanisms. Alternatively, nicotine and cotinine may possibly induce apoptosis.

Herbicide 2,4-dichlorophenoxyacetic acid (2,4-D)-induced cytogenetic damage in human lymphocytes in vitro in presence of erythrocytes.

The genotoxic effects of 2,4-D and its commercial derivative 2,4-D DMA were studied by measuring sister chromatid exchange (SCE), cell-cycle progression and mitotic index in human whole blood (WBC) and plasma leukocyte cultures (PLC). Concentrations of 10, 25, 50 and 100 microg herbicide/ml were used during 72 h. In WBC, a significant increase in SCE frequency was observed within the 10-50 microg 2,4-D/ml and 25-100 microg 2,4-D DMA/ml dose range. Contrarily, in PLC, none of the concentrations employed affected the SCEs frequency. A significant delay in cell proliferation was observed in WBC after treatments with 25 and 50 microg 2,4-D/ml and 50 and 100 microg 2,4-D DMA/ml. In PLC, only 100.0 microg 2,4-D/ml altered cell-cycle progression. For both chemicals, a progressive dose-related inhibition of mitotic activity was observed. The results demonstrated that the presence of erythrocytes in the culture system modulated the DNA and cellular damage inflicted by 2,4-D and 2,4-D DMA into human lymphocytes in vitro as well as both 2,4-D and 2,4-D DMA were more potent genotoxic agents in the presence of human red cells.

Inhibitory effect of mannan on the toxicity produced in mice fed aflatoxin B1 contaminated corn.

Aflatoxin B(1) (AFB(1)) is a potent mutagenic and carcinogenic agent found in numerous agricultural and dairy products consumed by humans. Therefore, we evaluated the capacity of mannan to cope with its genotoxic potential. We prepared a diet constituted of corn (90%) plus the recommended amount of other nutrients, as well as with the tested compounds (mannan and/or AFB(1)). Mice were fed this diet during 4 weeks as follows: one group with AFB(1)-contaminated corn (0.25 mg/kg of corn), three groups with mannan (50, 250, and 500 mg/kg of corn) plus AFB(1) (0.25 mg/kg), another group with only mannan (500 mg/kg), and the last group with an uncontaminated diet and no mannan added. We determined the weight, the micronucleated normochromatic erythrocyte rate (MNNE), the polychromatic/normochromatic index, and the sister chromatid exchange rate (SCE). We also examined the recovery response of mice during 4 additional weeks, when they were fed only the normal diet without AFB(1) or mannan. The results in the first period revealed the following: a) mice fed with mannan alone presented values in the range determined for the control group; b) mice fed AFB(1) had a significant weight decrease, as well as a significant increase in the rate of MNNE and SCE; and c) animals fed the combined regimen (AFB(1) plus mannan) presented a 25% weight increase with respect to the animals treated with AFB(1) alone, as well as a significant reduction in the level of MNNE and SCE with the two high doses tested. In the second (recovery) period, the control and the mannan fed groups maintained values similar to those exhibited in the previous phase, and the AFB(1) group as well as the groups fed the regimen combined with mannan showed an improvement in all evaluated parameters; the best response was that found in mice fed AFB(1) plus 500 mg/kg of mannan. Our study established an antigenotoxic effect of mannan that could be due to its adsorbent capacity.

Genotoxicity analysis of the phenoxy herbicide dicamba in mammalian cells in vitro.

The cytogenetic effects exerted by the phenoxy herbicide dicamba and one of its commercial formulations banvel (57.71% dicamba) were studied in in vitro whole blood human lymphocyte cultures. The genotoxicity of herbicides was measured by analysis of the frequency of sister chromatid exchanges (SCEs) and cell-cycle progression assays. Both dicamba and banvel activities were tested within 10.0-500.0 microg/ml doses range. Only concentrations of 200.0 microg/ml of dicamba and 500.0 microg/ml of banvel induced a significant increase in SCE frequency over control values. The highest dose of dicamba tested (500.0 microg/ml) resulted in cell culture cytotoxicity. The cell-cycle kinetics was affected by both test compounds since a significant delay in cell-cycle progression and a significant reduction of the proliferative rate index were observed after the treatment with 100.0 and 200.0 microg/ml of dicamba and 200.0 and 500.0 microg/ml of banvel. For both chemicals, a progressive dose-related inhibition of the mitotic activity of cultures was observed. Moreover, only the mitotic activity statistically differed from control values when doses of both chemicals higher than 100.0 microg/ml were employed. On the basis of our results, the herbicide dicamba is a DNA damage agent and should be considered as a potentially hazardous compound to humans.

Genotoxic effects of benzo[a]pyrene and dibenzo[a,l]pyrene in a human lung cell line.

Several studies have shown that polycyclic aromatic hydrocarbons (PAHs) produce genotoxic effects in assays performed in vivo and in vitro. This study was undertaken to investigate the ability of benzo[a]pyrene (BP) and dibenzo[a,l]pyrene (DBP) to induce DNA damage in a human lung fibroblast cell line (MRC-5), using sister-chromatid exchanges test (SCEs), the comet assay, and evaluating point mutations in codon 12 of the K-ras protooncogene by polymerase chain reaction-single-strand conformation polymorphisms (PCR-SSCPs) and restriction fragment length polymorphisms (RFLP)-enriched PCR methods. Sister-chromatid exchanges frequencies were significantly increased in cells exposed to benzo[a]pyrene and dibenzo[a,l]pyrene in relation to controls (p < .001). Using the standard alkaline comet assay, significant differences between groups were found for the variable comet moment (CM) when cells were exposed to BP (p < .001) and DBP (p < .001). Nevertheless, PCR-SSCP and RFLP-enriched PCR methods did not show any association between treatments with BP and DBP and K-ras point mutations. The data presented in this study indicated that BP and DBP induced both DNA strand breaks and sister-chromatid exchanges but not significant point mutations at codon 12 of K-ras gene in the MRC-5 cell line.