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Cadmium is a widespread environmental and occupational pollutant and quercetin is a dietary flavonoid, which is reported to modulate the effects of many mutagens and carcinogens. We investigated the ability of cadmium chloride to induce DNA damage in human lymphocytes in the presence of quercetin using the alkaline comet assay. Cadmium chloride (5-150 muM) evoked dose-dependent DNA damage and quercetin at 50 muM decreased the extent of the damage. The lymphocytes exposed to cadmium chloride were able to remove their DNA damage within a time period shorter than 120 min. The cells treated with quercetin at 50 muM prior to exposure to cadmium required shorter periods of time to recover. Quercetin could chelate cadmium ions, scavenge free radicals produced by cadmium or regenerate cellular DNA-repair enzymes.
Mortality connected with tobacco smoke-associated laryngeal cancer in Poland markedly exceeds the relevant epidemiological data from other European countries. The main groups of genotoxic agents considered as potential carcinogens present in tobacco smoke are polycyclic aromatic hydrocarbons, aromatic amines, N-nitro-soamines and reactive oxygen species. Aromatic DNA adducts, N7-alkylated guanosines and oxidative DNA damage derived from tobacco smoke exposure were detected in laryngeal and oral (tumour and non-tumour) biopsies, and white blood cells of cancer subjects. Further, DNA lesions were analysed to estimate the significance of such confounders as intensity of smoking, subject's sex, age, topography of larynx, cancer staging and genetic factor. The number of cigarettes smoked per day was found to be the main determinant of an individual's DNA adduct level. The occurrence of DNA lesions was established as a reliable marker of former exposure to tobacco smoke genotoxicants. On the other hand, a comparison of DNA lesion levels in various regions of larynx indicates limited usefulness of DNA adduct analysis as an estimate of cancer risk. For a better risk estimation one has to take into account DNA lesions in proto-oncogenes and tumour suppressor genes and the efficacy of DNA repair. Altogether, DNA adducts formation and removal has to be considered as a single stage in the multistep carcinogenesis.
The results of epidemiological studies published in the 70s suggested that diets of certain Central European states might contain unique chemo- preventive components. We investigated the antioxidative properties of traditional foods such as sauerkraut or beet roots, typical for diets in Central Europe and less common in other parts of the world. Some of the foods tested displayed free radical scavenging activity comparable to green tea used as a positive control. Particularly effective in this regard were beet root concentrate, sauerkraut juice and fermented cucumbers. Additionally, comet assay showed that sauerkraut juice protected cultured HL-60 cells against oxidative DNA damage in a dose-dependent manner.
The relationship between HPV (Human Papillomavirus) and cervical carcinoma is very strong, causal, constant, specific and universal. HPV infection precedes the occurrence of preinvasive carcinoma and the evidence for biological probability of such relationship does not leave any doubt. Out of high-risk HPV types, the most often occurring in cervical infections types 16 and 18 were placed in the first group of human carcinogens. However, HPV infection is not the only factor of malignant neoplasm development. Neuroplastic transformation of cervical epithelium requires the presence of cofactors. In recent years, higher role is ascribed to the influences of steroid hormones, including estradiol and progesterone. In normal and neoplastic cells of uterine cervix, the occurrence of receptors for steroid hormones was described. In the regulatory region of oncogenic HPV hormone response elements were found. Receptors combined with steroid hormones functioning as transcription factors may influence the expression of viral oncogens E6 and E7 through stimulation of proliferation and transformation. The aim of this study was to analyse the influence of 17ß-estradiol on the amount of proteins E6 and E7 HPV 18 and to evaluate the transcript E6/E7 after incubation of HeLa line cells with 17ß-estradiol. The results revealed stimulating influence of the hormone on the expression of proteins and mRNA at concentrations of lx10-7M. At high concentrations of 1x10- 4M, estradiol was inhibiting transcription and expression of oncoproteins.
Polycyclic aromatic hydrocarbons (PAHs) are DNA adducts forming carcinogens. The DNA adducts, especially 8-oxoguanine (8-oxoG), are the established biomarker of environmental genotoxic compounds exposure. The aim of this study was to determine the 8-oxoG presence in peripheral blood lymphocytes of 27 coke oven workers and 21 rural area inhabitants comparison. The study and control groups were not significantly different according to %8-oxoG-positive lymphocytes. In the control group the difference between smokers and non-smokers was significant (p<0,01). %8-oxoG in non-smokers from the study and control groups were significantly different (p<0,009). The occupational exposure and smoking cigarettes are important oxidative DNA damage causing factors.
Tobacco smoking during pregnancy is associated with a variety of negative consequences not only for the mother, but also for the developing fetus. Many studies have shown that carcinogens contained in tobacco smoke permeate across the placenta, and are found in fetus. The aim of the study was to determine the prenatal exposure to tobacco-specific carcinogenic N-nitrosamines on the basis of measurements of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine of smoking and second-hand smoke (SHS) exposed women and in the first urine of their newborns. A questionnaire documenting demographics and socio-economical data, smoking habits and exposure to SHS was completed by 121 delivering women near or at term. Maternal concentrations of cotinine and NNAL were measured in urine of the mother and the first urine of her newborn infant by liquid chromatography tandem mass spectrometry (LC/MS/MS). The mean concentration of cotinine was 439.2 ng/mg creatinine and NNAL concentration in urine of smoking women was 74.0 pg/mg creatinine, and for her newborn 78.6 pg/mg creatinine. Among mothers exposed to SHS, cotinine and NNAL mean concentration were 23.1 ng/mg creatinine, and 26.4 pg/mg creatinine. In newborns of SHS exposed mothers during pregnancy the mean concentration of NNAL was 34.1 pg/mg creatinine, respectively. Active tobacco smoking as well as passive exposure to smoking during pregnancy is an important source of tobacco specific N-nitrosamines to the fetuses as evidenced by increased concentrations of this carcinogen. Determination of NNAL in maternal urine samples can be a useful biomarker of prenatal exposure of newborn to carcinogenic nitrosamines.
Genotoxic carcinogens, able to damage DNA by alkylation reactions, represent a very diverse class of agents which are capable of producing a wide range of DNA modifications. The mechanisms leading to genetic changes as a result of exposure to alkylating agents (AAs) have been studied in male germ cells of Drosophila using a structure-activity relationship approach (SAR). The analytical tools available concern both genetic and molecular assays. The genetic tests enable to quantify excision repair and clastogenic potency of the AA after treatment of post-meiotic male germ cells and to determine the degree of germ-cell specificity, i.e., the mutagenic effectiveness in post-versus premeiotic cell stages. For a selected group of alkylating agents the molecular spectra have been studied in post-meiotic cell stages. On the basis of these descriptors clear SAR's between genotoxic activity in germ cells and physico-chemical parameters (s-values and O6/N7-alkylguanine adducts) and carcinogenic potency in rodents became apparent, resulting in five distinct classes of alkylating agents so far. These classes are: 1) SN2-type monofunctional AAs, 2) SN1-type monofunctional AAs, 3) polyfunctional AAs, 4) agents able to form etheno-DNA adducts, and 5) aflatoxin B1 (AFB1) a bulky-adduct forming agent. The recent finding that the molecular data obtained with Drosophila and data of the specific locus tests in male mice show remarkable similarities for most genotoxic agents supports the view that Drosophila is a useful model system for the study of transgenerational damage.
A total of 50 samples of poultry feed mixtures of Slovak origin were analysed for fumonisin B1 and B2 (FB1, FB2) and moniliformin (MON) using SAX-clean up procedure being detected by high pressure liquid chromatography with mass spectrometry (HPLC-MS) and diode array detection (HPLC-DAD), respectively. The samples were also simultaneously investigated for Fusarium species occurrence, and for the capability of Fusarium isolates recovered to produce FB1 and MON in vitro. FB1 was detected in 49 samples (98%) in concentrations ranging from 43 to 798 µg.kg-1, and FB2 in 42 samples (84%) in concentrations ranging from 26 to 362 µg.kg-1. MON was detected in 26 samples (52%) in concentrations that ranged from 42 to 1,214 µg.kg-1. Only two Fusarium populations were encountered, namely F. proliferatum and F. subglutinans, of which the former was the most dominant and frequent. All 86 F. proliferatum isolates tested for FB1-production ability proved to be producers of the toxin although none of them produced MON. On the contrary, MON production was observed in a half out of 16 F. subglutinans isolates tested, yet no FB1 production was detected in this case. Despite the limited number of samples investigated during this study, it is obvious that poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in central Europe. This is the first reported study dealing with fumonisin and moniliformin contamination of poultry feeds from Slovakia.
The c-H-ras-1 gene of an B6C3F1 mouse was isolated and nucleotide sequence determined. Our study has revealed that this c-H-ras-1 gene consists of four exons, separated by three introns ranging in size from 150 to 649 bp. The coding parts of the sequence of mouse c-H-ras-1 gene show no important differences as compared with those of the rat, hamster and human gene. More numerous changes were found in introns. The identity of mouse c-H-ras-1 gene with rat, hamster and human ones at the nucleotide level is 86.40%, 80.04% and 67.87%, respectively. Comparison of amino acids in protein sequence of c-H-ras gene of mouse, rat, hamster and human points to high degree of conservation of the gene.
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