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The 'comet' assay for detection of potential genotoxicity of polluted water

100%

Kosz-Vnenchak M. , Rokosz K.

Folia Biologica

1997

tom 45

153-156

The aim of this study was to determine the potential genotoxic activity of polluted water samples taken from wastewater from selected industrial plants in Krakow: 1. the Thermal-electric Power Station 2. the Institute of Metal Cutting. The recently developed single cell gel assay (SCG or comet assay), which is a quick and simple technique for the evaluation of DNA damage and repair in individual cells, was used. The assay was carried out on human hepatoma cells (Hep G2) as target cells. A greater number of cells with comets was observed in those treated in vitro with the polluted water samples (70%-88%) than in those in the control (22%, 33%). These preliminary results indicate that comet assay can have an application in biomonitoring studies for determining the potential genotoxicity of water pollutants. s.

Physical state of human papillomavirus type 16 in cervical cell DNA

80%

Szostek S. , Klimek M. , Zawilinska B. , Kosz-Vnenchak M.

Folia Biologica

2008

tom 56

nr 3-4

269-271

Multiplex PCR with specific primers for E2/E6 genes was used to assess the viral integration status of HPV-16 in women with low and high grade squamous intraepithelial lesions (LSIL and HSIL, respectively) in comparison to cervical cancer patients. Women with confirmed HPV-16 infection were examined: 30 with LSIL, 12 with HSIL and 23 with cervical cancer. The PCR products were separated electrophoretically in agarose gels and densitometric analysis was performed using Bio-Rad Quantity One software. E2 and E6 sequences of HPV-16 were detected in 91% of the women. The free episomal viral genome was not detected in the cervical carcinoma group. Twenty six percent of the samples obtained from this group harboured the integrated form, whereas the remaining samples possessed a mixture, i.e. episomal and integrated forms of viral DNA. The free episomal form dominated in women with LSIL and HSIL. In 6 cases the episomal and integrated forms were detected simultaneously. HPV-16 integration occurred in a subset of LSILs and HSILs, not only in the cervical cancer patients and correlated with progression of cytological changes. The assessment of the status of HPV-16 may be the molecular factor preceding the morphological features leading to malignancy.