Genetikos praktikumas : mokymo priemonė

Vytauto Didžiojo universiteta

A preliminary assessment of singlet oxygen scavenging, cytotoxic and genotoxic properties of Geranium macrorrhizum extracts

Strong radical-scavenging activity of Geranium macrorrhizum extracts isolated by using various solvent systems has been reported previously. This study aimed at expanding the knowledge on the bioactivities of antioxidatively active G. macrorrhizum butanol fraction, which was isolated from ethanolic extract (EB), and water fraction, which was isolated from water extract (WW) by measuring their singlet oxygen scavenging properties, as well as preliminary assessment of cytotoxicity and genotoxicity toward mammalian cells. The cytotoxicity (necrosis induction) of the extracts in bovine leukemia virus-transformed lamb kidney fibroblasts (line FLK) was partly prevented by antioxidants and stimulated by the prooxidant BCNU (N,N'-bis(2-chloroethyl)-N-nitrosourea). This indicates that the cytotoxicity of G. macrorrhizum extracts is at least partly attributed to their prooxidant action, presumably due to the formation of quinoidal products of their (auto)oxidation. The latter was evidenced by the nature of the peroxidase-catalyzed oxidation products, which supported DT-diaphorase-catalyzed oxidation of NADPH and participated in conjugation reactions with reduced glutathione. The genotoxic properties were studied using chromosome aberration (CA) and sister chromatid exchange (SCE) tests in human lymphocytes in vitro and Drosophila melanogaster somatic mutation and recombination test (SMART) in vivo. In the CA test, only the highest doses of both fractions significantly increased chromosome aberration frequency. In the SCE test, both fractions induced SCEs in a clear dose-dependent manner. G. macrorrhizum extracts were not genotoxic in the SMART test in vivo. Our data indicate that in spite of the possible beneficial (antioxidant) effects of Geranium extracts, the possibilities of their use as ingredients of functional foods and/or food supplements should be further examined due to their cyto- and genotoxic effects resulting mainly from the action of quercetin-derived components abundant in the extracts

Evaluation of the Biological Activity of Naturally Occurring 5,8-Dihydroxycoumarin

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Genotoxic properties of Betonica officinalis, Gratiola officinalis, Vincetoxicum luteum and Vincetoxicum hirundinaria extracts

article no. 110815Genotoxicity of B. officinalis, G. officinalis, V. luteum and V. hirundinaria extracts, which demonstrated strong antioxidant capacity, was tested using chromosome aberration, sister chromatid exchange (SCE), cytokinesisblock micronucleus and alkaline single-cell gel electrophoresis (comet) assays in human lymphocytes in vitro and Ames Salmonella/microsome test. All tested extracts were not mutagenic in S. typhimurium strains TA98 and TA100 with and without metabolic activation and did not induce chromosome aberrations in human lymphocytes in vitro. Extract from G. officinalis was the only one, which induced significant increase in micronuclei, indicating possible aneugenic effect. All investigated plant extracts induced DNA damage evaluated by the comet assay, while B. officinalis and V. luteum extracts induced slight increase in SCE values. The determined variation in response might be due to the plant extract tested and donor susceptibilityKauno technologijos universitetasVilniaus universitetasVytauto Didžiojo universiteta

Evaluation of biochemical, genotoxic and microbiological features of different viburnum genotypes

Biologijos katedraBotanikos sodasGamtos tyrimų centrasVilniaus universitetasVytauto Didžiojo universiteta

Biochemical, genotoxic and microbiological properties of different Viburnum genotypes

Less common plant species are of special interest extending list of horticultural plants. Use of Viburnum fruits containing variety of biologically active compounds requires more detailed evaluations from the microbiological and toxicological point of view [1]. Till now taxonomy of Dipsacales remains among the most problematic hot spots in phylogeny of Angiosperm groups [2,3]. The present work was conducted with the purpose to differentiate accessions of Viburnum grown in Kaunas Botanical garden according to system of molecular markers, also biochemical, microbiological and genotoxic characters (Study was sponsored by Research Council of Lithuania, project No SVE04/2011). Eight out of eleven tested primer pairs were used for molecular analyses at simple sequence repeat (SSR / microsatellites) loci. Totally 97 fragments were generated at 8 SSR loci. For each loci number of DNA fragments was varying from 2 to 10, and length of the fragments ranged between 80 and 150 bp (base pairs). The genotoxic potential of lyophilized juices of Viburnum opulus and V. sargentii fruits was evaluated using the Ames Salmonella / microsome mutagenicity test, alkaline single-cell gel electrophoresis (comet) assay and cytokinesis-block micronucleus test in human lymphocytes in vitro. All investigated genotypes were negative results according to the Ames test, indicating that Viburnum juices do not produce reverse mutations in bacterial cells. In human lymphocytes, no increase of micronuclei frequency and primary DNA damage was determined. Hereby, lyophilized juices of V. opulus and V. sargentii fruits are not genotoxic under the conditions of the present study. Agar diffusion method was employed to evaluate antimicrobial properties of the juice. Bioassays showed that the fruit juices prepared from all studied genotypes of Viburnum demonstrated bacteriocidic and bacteriostatic activity against 8 bacteria species. [...]Biologijos katedraBotanikos institutasBotanikos sodasVilniaus universitetasVytauto Didžiojo universiteta

Differentiation of Viburnum accessions according to their molecular, biochemical, genotoxic and microbiological features of importance to selection

In recent decades, Viburnum has been given special attention as raw material in medicine and as a food-supplementing product in the horticulture of different countries with temperate climate. The present study aimed at evaluating Viburnum varietetus using molecular markers, relates the SSR data to biochemical ones, as well as assessing the genotoxicity and antimicrobial effect of fruit juices of the species that have been selected. On the basis of the diagram of the microsatellite data, different relations between detected antocyanins and the total amount of phenols in the fruit of different accessions were discovered. In the evaluation of genotoxicity with the help of the Ames test, Viburnum accessions showed negative results. As such lyophilised juices of Viburnum fruit were not genotoxic. The results of bioassays showed that juices of different Viburnum accessions displayed bactericidal and bacteriostatic activity against all bacteria and yeast that were studied. When the juices of Viburnum berries were investigated, no inhibitory effect on microscopic fungi was foundBiologijos katedraBotanikos sodasGamtos mokslų fakultetasGamtos tyrimų centrasLietuvos agrarinių ir miškų mokslų centras, [email protected] universitetasVytauto Didžiojo universiteta