Micronucleus assay in genotoxicity assessment

Three different pesticide formulations were tested for micronuclei production and cell cycle arrest. Micronucleus test represents a suitable method for assessing chromosome damage because both chromosome breakage (clastogenicity) and chromosome loss (aneugenicity) can be measured simultaneously. In the experiments for 24 and/or 48 h exposure, no significant increase (p < 0.05) in MN frequency was found in comparison with negative control. On the contrary, the values of CBPI (cytochalasin blocked proliferative index) were significantly reduced in a dose dependent manner (p < 0.05, p < 0.01, p < 0.001) for both exposure time. Our results indicated an expressive cytotoxic effect of various pesticides in cultivated bovine peripheral lymphocytes

Detection of Mutations in Selected Proto-Oncogenes of Canine Lymphoma

Lymphomas belong among the most frequently diagnosed tumours of the haematopoietic system in dogs. The clinical manifestations and genetic and molecular basis of canine lymphoma resembles those of human non-Hodgkin lymphoma and therefore it can serve as a suitable model for the study of this disease. Neoplastic diseases are the consequence of a number of genetic and epigenetic changes in somatic cells. One of such changes are gene mutations that can subsequently cause changes in the activity of proto-oncogenes and tumour suppressor genes. The aim of our study was to detect potential mutations in selected exons of proto-oncogenes in DNA isolated from samples of lymphoma obtained from two donors - a Bernese Mountain Dog and a female mongrel. On the basis of literary data descriptions of human and canine haematopoietic neoplastic diseases, our investigations of potential changes in DNA focused on proto- oncogenes C-KIT - exons 8, 17; NRAS - exons 1, 2;FLT3 - exons 14, 15 and 20. The investigated samples were amplified by polymerase chain reaction (PCR) and subjected to sequencing. The DNA sequences were compared with reference sequences in the database Ensembl. The comparison of sequences of the C-KIT gene revealed an A/G transition at the 35th nucleotide of exon 8 in the mongrel. It involved a synonymous exchange of the nucleotide in the codon that did not cause a change in the amino acid. In the same sample we recorded several point mutations in the intron regions surrounding the exons 14 and 20 of the FLT3 gene. Changes in the intron regions can affect the expression of genes and thus can play an important role in the origin and development of tumours. No genetic mutations were detected in any gene regions of the Bernese Mountain Dog. In the case of the NRAS gene, no changes were observed in any sample collected from the donors

Analysis of Sister Chromatid Exchanges and Proliferation of Human Peripheral Blood Lymphocytes Exposed to Epoxiconazole

The potential genotoxic/cytotoxic effect of epoxiconazole was evaluated by means of sister chromatid exchanges (SCE) following the 24 and 48 h in vitro exposure of human peripheral blood lymphocytes to epoxiconazole at concentrations of: 5, 10, 25, 50 and 100 μg. ml–1. Dimethyl sulphoxide (DMSO), used as an epoxiconazole solvent, was used as a negative control and mitomycine (MMC) as a positive control. After the 24-hour exposure, we failed to observe a significant increase in SCE frequencies in comparison with the negative control, however, the concentrations of 10—100 μg.ml–1 caused a significant decrease in the proliferation index (PI; P < 0.001). Also, the 48-hour exposure produced no significant alterations in the SCE frequencies in comparison with the control. At epoxiconazole concentrations ranging from 10 to 50 μg.ml–1 we recorded a moderate to strong, dose-dependent inhibition of PI (P < 0.05; P < 0.01; P < 0.001), while at the highest dose (100 μg.ml–1) the reduction in PI compared to the control was less pronounced (P < 0.05). The reduction in PI at the concentration range of 10—100 μg.ml–1 depended on the number of cells in the M1, M2 and M3 phases of the cell cycle per total number of 100 evaluated metaphases. Our results indicated a significant cytotoxic or cytostatic effect on human peripheral blood lymphocytes

Effect of N-metylcarbamate pesticide bendiocarb on cattle lymphocytes after in vitro exposure

Bendiocarb is a carbamate broad-spectrum insecticide used to control disease vectors such as mosquitoes and flies, as well as household and agricultural pests. Nowadays, only few papers reporting cytogenetic or possible genotoxic effect of this insecticide on mammalian cells are available. In the present study 24-hour exposure to bendiocarbamate at concentrations ranging from 20 to 160 μg/ml was used for investigation of unstable chromosomal aberrations (CA), sister chromatid exchanges (SCE) and stable chromosomal aberration induction in cultured bovine peripheral lymphocytes. The slight but no significant increase of chromatide breaks frequency was observed after the exposure of lymphocytes to 80 μg/ml of bendiocarb. At the highest concentration added to the cell cultures (160 μg/ml) mitotic index decrease was shown in both donors (p < 0.05; p < 0.01). Both statistically significant elevation of SCEs (p < 0.05) and a reduction of proliferative indices (PI) (p < 0.01) were shown at a dose of 80 μg/ml. By means of two fluorescent-labelled whole chromosome-painting probes, stable aberrations such as bovine chromosome 1 and 5 translocation as well as numerical aberrations (polyploidies, heteroploidies) were visualised under fluorescent microscope in some examined metaphases