Genotoxicity assessment of human peripheral Lymphocytes induced by thallium(I) and thallium(III)

Thallium is a non-essential metal with a wide range of industrial uses. However, thallium is also a potential pollutant with high potential toxicity to humans. In the present study, we analyzed and compared the cellular and genotoxic effects of thallium in two main oxidation states by applying chromosome aberration assays to human peripheral lymphocytes. We observed that thallium(I) sulfate reduced the mitotic index at all tested concentrations (0.5, 1, 5, 50 and 100 μg/mL), whereas thallium(III) chloride was toxic at concentrations ≥1 μg/mL. Thallium(I) and thallium(III) treatment significantly increased structural chromosomal aberrations, with and without gaps, and increased the percentage of aberrant cells without gaps. Furthermore, satellite associations and numerical chromosomal aberration tests showed significant differences at a few of the tested concentrations. The satellite association test is related to aneuploidy. Thallium salts increased satellite associations when hyperploid cells were observed. Our results indicated that the two oxidation states of thallium induced toxicity in vitro – i.e. cyto/genotoxic (clastogenic and aneuploidogenic) effects.     

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.     

Glutathione S-transferase gene polymorphism,cigarette smoking and DNA damage

Several factors such as smoking habits, diet, occupational, and environmental exposure to chemical carcinogens influence the overall level of DNA damage. In 69 healthy adult volunteers’ polymorphisms of glutathione S-transferase (GST), an enzyme which participates in the metabolism of a broad range of carcinogens and endogenous compounds were determined. The level of DNA damage was assessed by comet assay and classified according to GSTT1/M1 genotype and smoking habits. GSTM1 null genotype was recognized in 48% of studied subjects and GSTT1 null genotype in only two cases (3%). In subjects carrying GSTT1/M1 alleles a significantly lower degree of DNA damage, determined as % DNA in the comet's tail, than in null individuals was noted. However, the results obtained did not indicate that in studied subjects an elevated endogenous level of DNA damage may be significantly related to smoking habits.     

Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.     

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes

The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 μg mL^?1 for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 μg mL^?1 (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 μg mL^?1 in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.     

Methylenetetrahydrofolate reductase deficiency: Prenatal diagnosis and family studies

Prenatal diagnosis of methylenetetrahydrofolate reductase (MTHFR) deficiency and family studies were performed because of a severely affected first child in this family. The fetus at risk was found to be heterozygous as confirmed by the enzymatic activity assay performed several times after birth. In the father, MTHFR activity was normal in lymphocytes and decreased in fibroblasts, whereas in the asymptomatic mother, the activity was not detectable in fibroblasts and was very low in lymphocytes. The absence of any clinical symptoms in the mother despite a clear MTHFR deficiency and hyperhomocystinemia emphasizes the heterogeneity of this disease.     

A new RFLP marker,SP282, at the btk locus for genetic analysis in X-linked agammaglobulinaemia families

X-linked agammaglobulinaemia is an inherited recessive disease in which the primary defect lies in the failure of pre-B cells to develop into mature circulating B cells, due to a defective B-cell cytoplasmic tyrosine kinase (btk). For this study we introduced a new RFLP marker, SP282, which is tightly linked to the XLA locus. In conjunction with the marker DXS178, SP282 was used to identify a carrier female and predict her male offspring to be normal. Subsequently the fetus was shown to have a normal number of circulating B cells, and at 2·5 years of age, the non-affected phenotype of the child was confirmed.     

低剂量浓缩铀对脾淋巴细胞DNA合成和UDS效应

研究机体摄入浓缩铀²³⁵UO₂F₂时,对外周免疫器官脾淋巴细胞DNA合成和UDS的影响。观察发现,当摄入低剂量浓缩铀0.1μg/kg体重时,可刺激T淋巴细胞的转化增殖,刺激指数增升。低剂量位缩铀摄入水平在0.1～20μg/kg体重时,可使紫外线诱导的脾淋巴细胞UDS呈现明显刺激效应,增强对脾淋巴细胞DNA切除修复能力。当浓缩铀内污染水平进一步加大时,可使脾T、B淋巴细胞的DNA合成以及UDS都呈现明显抑制。     

Micronuclei Frequency in Lymphocytes and Antioxidants in the Blood of Traditional Limited-Resource Farm Workers Exposed to Pesticides

Chronic low-level exposure to synthetic pesticides is implicated in many health conditions that result from the induction of oxidative stress, including cytogenetic damage. The objective of this study was to assess the risk of genotoxicity using micronuclei (MN) formation in lymphocytes and to determine changes in blood antioxidants superoxide dismutase (SOD) in erythrocytes (E) and glutathione (GSH) in E and plasma (PL) in farm workers for six months during a growing season. Blood and urine samples were collected once a month for six months (June to November 2003) from farm workers (n = 15) and urban unexposed controls (n = 10). Lymphocytes from blood were separated by density gradient centrifugation using Histopaque and cultured using the standard technique. There was no significant difference in the cytokinesis blocked proliferation index (CBPI) of lymphocytes between the farm workers and the control group, but there was a 76% increase in average MN frequency in lymphocytes of the farm worker group (P ≤ 0.05). In addition, MN frequency peaked during August as compared to the other months and the controls (P ≤ 0.05). An 18% decline was observed in the activity of E-SOD in the farm worker group (P ≤ 0.05). GSH in E and PL were similar in both groups. These data suggest that the farm workers may be at a greater risk of developing genotoxicity due to continued exposure to pesticides, especially during the intensive growing season.     

Prenatal diagnosis and outcome of congenital splenic cyst: report of two cases

Congenital splenic cyst (CSC) is a rare condition, and only a few cases with prenatal diagnosis and no associated malformation have been reported. Spontaneous regression is possible in case of mild CSC (under 40 mm), but enlargement or secondary complications may lead to surgical treatment. We report, herein, two cases of isolated mild CSC with complete spontaneous postnatal regression. Copyright © 2006 John Wiley & Sons, Ltd.