Influence of naringin on cadmium-induced genomic damage in human lymphocytes in vitro.

Cadmium is an important toxic environmental heavy metal. Generally, occupational and environmental exposures to cadmium result from heavy metal mining, metallurgy and industrial use and the manufacturing of nickel-cadmium batteries, pigments and plastic stabilizers. Cadmium induces oxidative stress and alters the antioxidant system, resulting in oxidative DNA damage and lipid peroxidation. The effect of naringin, a grapefruit flavonone, on cadmium-induced genomic damage was studied by using an in vitro system to test for chromosomal aberrations and sister chromatid exchanges. Cadmium significantly increased the total chromosomal aberrations in human lymphocytes at concentrations of 20 and 40 µM, and although naringin alone did not induce any chromosomal aberrations, it decreased those induced by cadmium. The mitotic index was not affected by either cadmium or naringin. Cadmium also induced a significant number of sister chromatid exchanges, but naringin alone did not induce sister chromatid exchanges and was unable to decrease the frequency of sister chromatid exchanges induced by cadmium. Replicative index analysis revealed that naringin and cadmium did not significantly alter replicative index frequencies. In this study, we show that plant-based flavonoids, such as naringin, may reduce the genomic damage induced by cadmium and may protect the cellular environments from free radical damage by its possible antioxidative potential.

A biomonitoring study of genotoxic risk to workers of transformers and distribution line stations.

A cytogenetic monitoring study was carried out on a group of workers from transformer and distribution line stations in the Bursa province of Turkey, to investigate the genotoxic risk of occupational exposure to extremely low frequency electric (ELF) and magnetic fields (EMF). Cytogenetic analysis, namely chromosomal aberrations (CAs) and micronucleus (MN) tests were performed on a strictly selected group of 55 workers and compared to 17 controls. CA and MN frequencies in electrical workers appeared significantly higher than in controls (p < 0.001, 0.05, respectively). The frequency of CA in exposed groups were significantly enhanced with the years of exposure (p < 0.01). The effect of smoking on the level of CA and MN was not significant in the control and exposure groups. The results of this study demonstrated that a significant induction of cytogenetic damage in peripheral lymphocytes of workers engaged to occupational exposure to ELMF in electric transformer and distribution stations.

In vivo evaluation of the toxic effects of pyrimethamine on spermatogenesis in male mice.

Pyrimethamine is known to have antimalarial activities and used clinically in the therapy of toxoplasmosis and human immunodeficiency virus-associated pneumonia. In this study we aimed to test the effects of pyrimethamine on spermatogenesis in mice. For this aim, animals were given pyrimethamine as a single application and the doses were 5, 10, 20, and 40 mg/kg. For the spermatogenic effects, the sperm shape abnormality, epididymal sperm counts, and testes weights were evaluated at the end of days 7, 14, 21, 28, and 35 after single pyrimethamine injection at the first day. Pyrimethamine increased the frequency of abnormal sperm shape for all studied weeks except the first week and its germ cell stage-specific effects correspond to spermatozoa, spermatids, and spermatocytes. It also decreased the epididymal sperm counts at the end of days 28 and 35, which corresponds to the spermatocyte stage of mouse spermatogenesis.

A comparative study on the genotoxic effect of pyrimethamine in bone marrow and spermatogonial mice cells.

Pyrimethamine is an antimalarial agent widely used in clinical therapy. We aimed to compare its mutagenic potential in mammalian spermatogonial and bone marrow cells. For studying chromosomal aberrations mice were treated acutely (single treatment) with 4 dose levels of pyrimethamine (5, 10, 20 and 40 mg/kg). Pyrimethamine was found to produce a significant increase in structural chromosomal aberrations after acute treatment in bone marrow cells of mice (p < 0.001). It also induced chromosome abnormalities in spermatogonial cells (p < 0.05) at the highest dose.

Genotoxic effects induced by fotemustine and vinorelbine in human lymphocytes.

The aim of this study was to investigate the in vitro genotoxic effects of the anticancer drugs fotemustine and vinorelbine on human lymphocytes and to determine individual and sex-related responses to these drugs. Fotemustine is a DNA-alkylating drug while vinorelbine is a semi-synthetic Vinca alkaloid. The study was carried out with twenty independent healthy donors for each drug. We have tested the ability of these drugs to induce chromosome aberrations (CAs) and sister chromatid exchanges (SCEs) as well as effect on the mitotic index (MI) in cultured human lymphocytes. Fotemustine was shown to induce CAs and SCEs at all concentrations tested (2, 4 and 8 microg/ml) in a dose-dependent manner. Additionally it also decreased the mitotic index in a similar dose-dependent manner. Vinorelbine had no effect on structural CAs, but it significantly increased the numerical CAs at all doses tested (0.5, 1 and 2 microg/ml). Vinorelbine also induced SCE events and increased the MI values. Two-way analyses of variance were used to compare the individual and gender-related susceptibilities to fotemustine and vinorelbine with respect to the CA, SCE and MI values. The results indicated that individuals in fotemustine treatment groups showed different genotoxic responses with respect to CA and SCE induction and additional findings indicated a gender-specific response in this group. Individuals in the vinorelbine test group also exhibited statistically significant numerical CA, SCE and MI responses to vinorelbine. A statistically significant gender-related SCE response to this drug was also evident. This study indicates that these drugs have potentially harmful effects on human health.

In vitro genotoxic effects of the anticancer drug gemcitabine in human lymphocytes.

This research was carried out to investigate in vitro genotoxic effects of the anticancer agent gemcitabine on the induction of chromosomal aberrations and sister-chromatid exchange in human lymphocytes. Three doses of gemcitabine (0.001, 0.002 and 0.004 microg/ml) were applied to lymphocyte cultures from 15 donors. There was a significant increase in the induction of chromosome aberrations and in the occurrence of sister-chromatid exchange in these cells. In addition, gemcitabine significantly decreased the mitotic index and replicative index for all doses. Dose-response regression lines were used to compare the individual susceptibilities to gemcitabine with respect to the chromosome aberration and sister-chromatid exchange frequencies. Our results indicate that gemcitabine is able to induce both cytotoxic and genotoxic effects in human lymphocyte cultures in vitro in a dose-dependent manner.

The investigation of the genotoxic effects of fenarimol and propamocarb in mouse bone marrow in vivo.

In this study, we aimed to evaluate the genotoxic effects of fungicides fenarimol and propamocarb which are used to protect crops from fungi. For this reason, bone-marrow micronucleus and chromosome aberration tests were carried out in Swiss albino mice. Mice were injected with four different doses of fenarimol and propamocarb intraperitoneally; 50, 100, 200 and 400 mg/kg b.w. Fenarimol did not induce any significant increase in micronucleated erythrocytes after 24, 36, and 48 h treatment but it decreased the ratio of polychromatic/normochromatic erythrocytes at all dose groups and sampling intervals. Fenarimol did not increase the number of chromosome aberrations significantly, but it reduced the mitotic index at the higher doses (P < 0.05). Propamocarb did not increase the frequency of micronucleated erythrocytes, but decreased the polychromatic/normochromatic erythrocytes ratio at all sampling intervals. Propamocarb increased only gaps in total chromosome aberrations, but when gaps were excluded, there were no significant differences in total aberrations between the control and dose groups (P > 0.05). Propamocarb also reduced the mitotic index compared with the negative control group (P < 0.001). Contributing these results, we can suggest that fenarimol and propamocarb are non-genotoxic in mouse bone marrow in vivo but have cytotoxic effects.

Genotoxicity of two anticancer drugs, gemcitabine and topotecan, in mouse bone marrow in vivo.

In this study, the genotoxic effects of gemcitabine and topotecan were investigated in mouse bone marrow cells using the micronucleus and chromosomal aberration test systems. Gemcitabine increased the frequency of micronuclei, particularly at the median dose for the 24-, 36-, and 48-h sampling intervals. It had cytotoxic effects on the bone marrow and decreased the polychromatic/normochromatic erythrocyte ratio dose-dependently for all sampling intervals. Gemcitabine significantly decreased the mitotic index at the 24-h time point. It increased the number of abnormal cells and induced a significant increase in total chromosomal aberrations. For the 6-h sampling time, gemcitabine neither induced chromosomal aberrations nor reduced the mitotic index. Topotecan also induced high levels of micronuclei, particularly for the 24- and 36-h sampling times and it decreased the polychromatic/normochromatic erythrocyte ratio for all sampling intervals, which is indicative of bone marrow cytotoxicity. The bone marrow metaphase analysis showed that topotecan significantly elevated the number of abnormal metaphases and total chromosomal aberrations at 6 and 24h, in a dose-dependent manner. It also decreased the mitotic index for both sampling intervals. In conclusion, the results of this study indicate that the two chemotherapeutics gemcitabine and topotecan have cytotoxic and genotoxic effects in mouse bone marrow.

Evaluation of genotoxicity from Nilufer Stream (Bursa/Turkey) water using piscine micronucleus test.

In the present study, the in vivo micronucleus (MN) test in fish erythrocytes was used to evaluate the genotoxic potentials of water samples collected from four different sites along the Nilufer Stream which empties into the Marmara Sea on the north-west of Turkey. Nilufer Stream receives discharges of industrial and domestic wastes resulting from industrialization and urbanization activities in Bursa city. Nile tilapias (Oreochromis niloticus) were exposed to collected water samples under laboratory conditions for 3 and 6 days. Micronuclei analyses were carried out in peripheral blood erythrocytes. In addition to micronuclei, other nuclear abnormalities (NAs) such as bi-nucleated cells and binuclei with nucleoplasmic bridge and cells with blebbed, notched and lobbed nuclei, were assessed in the erythrocytes. Chemical analyses were also carried out in the water samples to assess the presence of major pollutants. MN and NA frequencies were significantly elevated in fishes exposed to water from polluted areas compared to those exposed to clean water sample. The results of this study indicate that Nilufer Stream is contaminated with potential genotoxic chemicals and the genotoxicity is possibly related with the industrial, agricultural and domestic activities.

Inhibitory effects of ascorbic acid and folinic acid on chromosome aberrations induced by pyrimethamine in vitro.

In this study, the anticlastogenic effects of ascorbic acid and the protective effect of folinic acid against the formation of chromosomal aberrations in humans by pyrimethamine were investigated. Pyrimethamine is a folic acid antagonist used for the treatment of malaria and toxoplasmosis. In this study, 18 different healthy people, who do not drink alcohol and are non-smokers, were chosen as an experimental group; 0.025 mg/ml pyrimethamine was given to the lymphocyte culture, which had been prepared with the peripheral blood taken from this group. After that each of the following doses were given to the same culture: 20, 40, and 80 mM of ascorbic acid and 25, 50, and 100 mM of folinic acid. The results of the cytogenetic evaluation showed that the aberrations due to pyrimethamine in the chromosomes were reduced by ascorbic acid and folinic acid significantly, depending on the given dose.

Investigation of genotoxic effect of taxol plus radiation on mice bone marrow cells.

In this study, we investigated the genotoxic effect of taxol, radiation, or taxol plus radiation on highly proliferative normal tissue-bone marrow cells of Swiss albino mice. Swiss-albino mice, 3-4 months old, were used in this study. Taxol was administered bolus intravenously through the tail vein. Radiation was given by using a linear accelerator. There were four treatment categories, which had a total of 34 groups. Each group consisted of five animals. The first was the control category that had one group (n = 5). The second treatment category was taxol alone, which had three groups as per taxol dose alone (n = 15). The third treatment category was radiation alone, which had three groups as per the radiation dose (n = 15). The fourth treatment category was taxol plus radiation, which had 27 groups as per combined radiation dose plus taxol dose concentration and as per pre-treatment timing sequence of taxol before radiation (n = 135). Mice were sacrificed 24 h after taxol or radiation or combined administration using ether anesthesia. The cells were then dropped on two labeled slides, flamed, air dried, and stained in 7% Giemsa; 20-30 well-spread mitotic metaphases were analyzed for each animal; the cells with chromosome breaks, acentric fragments, and rearrangements were evaluated on x1,000 magnification with light microscope (Zeiss axioplan). The mitotic index was determined by counting the number of mitotic cells among 1,000 cells per animal. Differences between groups were evaluated with Student's t-test statistically. Taxol caused a dose-dependent increase in chromosomal aberrations (P = 0.027). Similarly, radiation caused a dose-dependent increase in chromosomal aberrations (P = 0.003) and decreased mitotic index (P = 0.002). In combination, there were a small enhancements at the 40 mg/kg taxol dose level and at 0.25 and 0.5 Gy radiation doses in the 48 h group. However, an increase in chromosomal aberrations was observed after 48 hours of taxol exposure when compared 12 or 24 h of taxol exposure (P = 0.001 and P = 0.019). These findings suggest that taxol at the high doses with low dose radiation caused radiosensitizing effect in bone marrow cells. Forty-eight-hour pretreatment of taxol exposure followed by radiation caused significant induction of chromosomal aberrations and a reduction of mitotic index when compared to other taxol timing sequence.