Sample records for azaguanine

  1. Induction of 8-azaguanine resistant mutants in human cultured cells exposed to 31 MeV protons

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    Fuhrman Conti, A.M.; Francone, G.; Volonte, M.; Gallini, R.E.


    The authors report results on the induction of 8-azaguanine (8-AG)-resistant mutants in cultured human cells (EUE) exposed to 31 MeV protons. The spontaneous frequency of mutants was 5.6 +- 0.7 x 10/sup -6/ per viable cell. Gamma rays were taken as reference radiation. Expression times giving the highest frequency of mutants after 31 MeV protons and gamma irradiation were found to be about 10 days for both radiations. The dose-response relationship for mutant induction by protons, as determined at the optimal expression time, was compared to that obtained after gamma rays. The relative biological effectiveness (RBE) is 2.4 +- 0.5, this value being higher than the RBE value determined for cell survival.

  2. Differentiation of pathogenic and saprophytic leptospira strains. (United States)

    Bazovská, S; Kmety, E; Rak, J


    Comparative studies of 249 pathogenic and 80 saprophytic leptospira strains, including 2 strains of the illini type, using the 8-azaguanine test, growth at 13 degrees C and growth on trypticase soy broth revealed their good differentiating potency if the recommended conditions were carefully observed. The same results were obtained by a simple hemolytic test using sheep and rat blood cells, having the advantage of providing results within 24 h. This test is suggested to replace the 8-azaguanine and the growth test at 13 degrees C. In these investigations, the first European strain of the illini type was recognized.

  3. 77 FR 20721 - 2-Ethyl-1-hexanol; Exemption From the Requirement of a Tolerance (United States)


    ... assay, an 8-azaguanine resistance assay, a mouse micronucleus test, a mouse lymphoma assay, a Rec-assay.../40tab_02.tpl . To access the OCSPP test guidelines referenced in this document electronically, please go to and select ``Test Methods and Guidelines.'' C. How can I file...

  4. Evaluation of the mutagenic activity of leucinostatins, a novel class of antibiotic peptides produced by Paecilomyces marquandii, in the modul Aspergillus nidulans. (United States)

    Crebelli, R; Carere, A; Conti, G; Conti, L; Rossi, C; Tuttobello, L


    Leucinostatins A, B, C, D, E, G, H, and K were thoroughly investigated for their genotoxic activity using the modul Aspergillus nidulans as the test organism. The results of assays for gene mutation (8-azaguanine resistance and methionine suppressors), gene conversion, mitotic crossing-over and mitotic aneuploidy induction suggest that these peptide antibiotics lack significant mutagenicity and that non-genotoxic mechanism(s) underlie their cytotoxic properties.

  5. Characterisation of methionine adenosyltransferase from Mycobacterium smegmatis and M. tuberculosis

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    Knodel Marvin H


    Full Text Available Abstract Background Tuberculosis remains a serious world-wide health threat which requires the characterisation of novel drug targets for the development of future antimycobacterials. One of the key obstacles in the definition of new targets is the large variety of metabolic alterations that occur between cells in the active growth and chronic/dormant phases of tuberculosis. The ideal biochemical target should be active in both growth phases. Methionine adenosyltransferase, which catalyses the formation of S-adenosylmethionine from methionine and ATP, is involved in polyamine biosynthesis during active growth and is also required for the methylation and cyclopropylation of mycolipids necessary for survival in the chronic phase. Results The gene encoding methionine adenosyltransferase has been cloned from Mycobacterium tuberculosis and the model organism M. smegmatis. Both enzymes retained all amino acids known to be involved in catalysing the reaction. While the M. smegmatis enzyme could be functionally expressed, the M. tuberculosis homologue was insoluble and inactive under a large variety of expression conditions. For the M. smegmatis enzyme, the Vmax for S-adenosylmethionine formation was 1.30 μmol/min/mg protein and the Km for methionine and ATP was 288 μM and 76 μM respectively. In addition, the enzyme was competitively inhibited by 8-azaguanine and azathioprine with a Ki of 4.7 mM and 3.7 mM respectively. Azathioprine inhibited the in vitro growth of M. smegmatis with a minimal inhibitory concentration (MIC of 500 μM, while the MIC for 8-azaguanine was >1.0 mM. Conclusion The methionine adenosyltransferase from both organisms had a primary structure very similar those previously characterised in other prokaryotic and eukaryotic organisms. The kinetic properties of the M. smegmatis enzyme were also similar to known prokaryotic methionine adenosyltransferases. Inhibition of the enzyme by 8-azaguanine and azathioprine provides a starting

  6. Cytotoxicity and mutagenicity of Kevlar: an in vitro evaluation. (United States)

    Wening, J V; Marquardt, H; Katzer, A; Jungbluth, K H; Marquardt, H


    Toxicity and mutagenicity of Kevlar 49 (PPPT; poly-para-phenylene-terephthalamide) was tested in six strains of Salmonella typhimurium (Ames test; TA97, TA98, TA100, TA102, TA1535, TA1537) with and without an external metabolic activation system (S9), as well as in a mammalian cell mutagenesis assay using V79 Chinese hamster cells. For the Ames test, liquid preincubation, which is considered particularly sensitive, was used. The cells were incubated for 24 h at a temperature of 37 degrees C either directly with Kevlar49 or with ethanol- or chloroform-extracted Kevlar49. The experiments were performed at least twice. The Ames test with six different Salmonella typhimurium strains featuring either base pair substitution or frameshift mutations revealed no cytotoxic or mutagenic activity of Kevlar49. In the mammalian cell mutagenesis assay, using 8-azaguanine (AG) as a selective agent, Kevlar49 was also devoid of cytotoxic or mutagenic activity. Both tests have to be regarded as an initial exploratory screening due to the chosen testing conditions and should be supplemented by tests at different temperatures.

  7. [A study on inclusion complexes of cyclodextrin with three anticancer xanthines by fluorescence]. (United States)

    Wei, Yan-li; Dong, Chuan


    The inclusion complexes of beta-Cyclodextrin (beta-CD) and HP-beta-Cyclodextrin (HP-beta-CD) with 6-Mercaptopurine (6-MP), Azathioprine (BAN) and 8-Azaguanine (Azan) were investigated by fluorescence. Various factors affecting the formation of inclusion complexes were discussed in detail including formation time and pH effect. The formation constants of their inclusion complexes were determined. The results indicated that their inclusion was affected significantly by laying time and pH. The formation time of beta-CD inclusion complexes is much longer than that of HP-beta-CD. The optimum pH is about pH = 7.7-12. Their maximum excitation wavelengths are all in the range of 276-285 nm and the maximum emission wavelengths are all in the range of 328-353 nm. The fluorescence signals are intensified with increasing concentration of CD. The stoichiometries of the inclusion complexes of CD with these three anticancer xanthines are all 1:1 and the formation constants are calculated.

  8. [Eremothecium ashbyii mutants resistant to 2,6-diaminopurine]. (United States)

    Stepanov, A I; Beburov, M Iu; Zhdanov, V G


    3 groups of Eremothecium ashbyii mutants resistant to 5-10(-3) M 2,6-diaminopurine (DAP) ahve been obtained. The mutants of the 1st group (Dap-r) are selected from the initial susceptible strain by the ability to grow in the presence of 5-10(-3) M DAP. The mutants of the 2nd group (Azg-Dap-r) are selected in the selective background of two analogues of 5-10(-3) M DAP and 10(-4) M 8-azaguanine (AG). The mutants of the 3rd group (Azg-r - DAP-r) are isolated from the mutant Azg-r 34 resistant to 10(-4) M AG. The results of studying cross-resistance of mutants to DAP, AG and 8-azaadenine (AA) show that Dap-r and Azg-Dap-r mutants in contrast to Azg-r - Dap-r, have common phenotypic properties and can grow only on the analogues of adenine. DAP, but not AA, eliminates the inhibitory effect of AG on the growth of these mutants. This effect is probably due to deaminating DAP to guanine. Mutants Azg-r - Dap-r retain the initial resistance to 10(-4) M AG, but are susceptible to higher concentrations of AG and in this case DAP does not eliminate the inhibitory effect of AG. In all mutants obtained the effectiveness of the incorporation of 14C-adenine (but not 14C-guanine) is sharply reduced, thus indicating the absence of adenosine-monophosphate pyrophosphorylase activity. The mutants do not excrete purine-like compounds into the medium. In the course of the continuous growth of mutants in the presence of DAP but not of guanine the red intracellular pigment is formed which seems to be a complex of riboflavin with DAP. A disturbance in the synthesis of adenosine monophosphate pyrophosphorylase does not influence practically the level of the synthesis of riboflavin in E. ashbyii.

  9. Metabolism of mutagenic polycyclic aromatic hydrocarbons by photosynthetic algal species. (United States)

    Schoeny, R; Cody, T; Warshawsky, D; Radike, M


    Polycyclic aromatic hydrocarbons (PAH) known to produce carcinogenic and mutagenic effects have been shown to contaminate waters, sediments and soils. While it is accepted that metabolites of these compounds are responsible for most of their biological effects in mammals, their metabolism, and to a large extent their bioactivity, in aquatic plants have not been explored. Cultures of photosynthetic algal species were assayed for their ability to metabolize benzo[a]pyrene (BaP), a carcinogenic PAH under conditions which either permitted (white light) or disallowed (gold light) photooxidation of the compound. Growth of Selenastrum capricornutum, a fresh-water green alga, was completely inhibited when incubated in white light with 160 micrograms BaP/l medium. By contrast concentrations at the upper limit of BaP solubility in aqueous medium had no effect on algal growth when gold light was used. BaP quinones and phenol derivatives were found to inhibit growth of Selenastrum under white light incubation. BaP phototoxicity and metabolism were observed to be species-specific. All 3 tested species of the order Chlorococcales were growth-inhibited by BaP in white light whereas neither the green alga Chlamydomonas reinhardtii nor a blue-green, a yellow-green or an euglenoid alga responded in this fashion. Assays of radiolabeled BaP metabolism in Selenastrum showed that the majority of radioactivity associated with BaP was found in media as opposed to algal cell pellets, that the extent of metabolism was BaP concentration dependent, and that the proportion of various metabolites detected was a function of the light source. After gold light incubation, BaP diols predominated while after white light treatment at equal BaP concentrations, the 3,6-quinone was found in the highest concentration. Extracted material from algal cell pellets and from media was tested for mutagenicity in a forward mutation suspension assay in Salmonella typhimurium using resistance to 8-azaguanine for