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Laser interferometry is a measurement technique used in physical sciences, with a potential for new applications in microbiology. Our previously studies, focused on the quantitative analysis of antibiotics diffusion through membranes or their releasing from gel structure, indicate that this method might be useful in analysis of substances diffusion across the bacterial biofilms. As antibiotic - biofilm interaction model, we tested above method for determination of ciprofloxacin or gentamicin diffusion through Proteus mirabilis O18 biofilm. Laser interferometry analysis of antibiotics diffusion showed that the amount of ciprofloxacin transported through mature biofilm is 1.9 times higher than gentamicin. It was correlated with lower level of gentamicin in compare to the level of ciprofloxacin in biofilm, which amounts were predicted in biofilm during diffusion process by laser interferometry method. We suggest that the analysis of antibiotic diffusion in biofilm might by helpful in evaluation of effectiveness of antibacterial agents.
Using the alkaline comet assay, we showed that bleomycin at 0.1-5 μg/ml induced DNA strand breaks and/or alkali-labile sites, measurable as the comet tail moment, in human colonic mucosa cells. This DNA damage was completely repaired during a 120-minute post-treatment incubation of the cells. Post-treatment of the bleomycin-damaged DNA with 3-methyladenine-DNA glycosylase II (AlkA), an enzyme recognizing alkylated bases, gave rise to a significant increase in the extent of DNA damage, indicating that the drug could induce alkylative bases in DNA. We did not observe any change in the comet tail moment in the presence of catalase. Vitamin E ((+)-α-tocopherol) decreased DNA damage induced by bleomycin. The results obtained suggest that hydrogen peroxide might not be involved in the formation of DNA lesions induced by bleomycin in the colonic mucosa cells, and that vitamin E may exert protective effects on these cells.
The pathogenesis of stomach cells can be associated with their susceptibility to exogenous dietary irritants, like nitrosamines such as dimethylnitrosamines (DMNA), and to the effects of non-dietary factors, including Helicobacter pylori infection. We used N-methyl-N'-nitro N-nitrosoguanidyne (MNNG) as a surrogate agent that induces a spectrum of DNA damage similar to DMNA. Using the alkaline comet assay, we showed that antioxidants--vitamins C and E, quercetin, and melatonin--reduced the genotoxic effect of MNNG in H. pylori-infected and non-infected human gastric mucosa cells (GMCs). To compare the sensitivity of the stomach and the blood, the experiment was also carried out in peripheral blood. We observed a higher level of DNA damage induced by MNNG in H. pylori-infected than in noninfected GMCs. We did not note any difference in the efficacy of the repair of the damage in either type of GMC. H. pylori infection may play an important role in the pathogenesis of GMCs, as it can modulate their susceptibility to dietary mutagens/carcinogens, thus contributing to gastric cancer.
Introduction and objective. According to the cancer epidemiology databases, cancer is the second leading cause of death in developing countries. Moreover, the WHO predicts a continuing increase in the incidence of cancer, extending this trend well into the next several decades. Hence, it seems obvious that the prediction of cancer susceptibility and early diagnosis is an important goal for modern biomedical sciences. The aim of this study is to clarify the value of chromosomal damage, capacity for the repair of double-strand breaks (DSBs), polymorphisms in DNA repair genes, and apoptosis as prognostic markers for prostate and cervical cancer. Materials and methods. 30 prostate cancer patients and 30 cervical cancer patients were enrolled into the study. In addition, 30 healthy female donors and 30 healthy male donors served as controls. The following endpoints were investigated: frequency of micronuclei, gamma-H2AX fluorescence, XRCC1 194C>T, XRCC1 399G>A, XRCC3 IVS5–14 A>G, OGG1 326 Ser>Cys polymorphisms and apoptosis. Results. Among all tested factors, only the homozygous variant (Arg/Arg) in XRCC1 (399 Arg/Gln) was strongly associated with prostate cancer risk, and only a low apoptotic response was connected with cervical cancer risk. The presented study confirmed a positive association between the frequency of MN and increased prostate and cervical cancer risk. However, such a biomarker is not cancer specific. In addition, the information gained by analyzing the gamma-H2AX fluorescence, as well apoptosis, had no value for predicting the risk of prostate and cervical cancers. Conclusions. The final conclusion of the study is that cancer susceptibility is a complex phenotype not readily detectable in relatively small studies by functional assays or analysis of SNP in few, selected genes.
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