Mutation Research/Genetic Toxicology最新文献

{"title":"Interlaboratory validation of a new assay for DNA-protein crosslinks","authors":"Max Costa ,&nbsp;Anatoly Zhitkovich ,&nbsp;Michael Gargas ,&nbsp;Dennis Paustenbach ,&nbsp;Brent Finley ,&nbsp;Jim Kuykendall ,&nbsp;Ruth Billings ,&nbsp;Timothy J. Carlson ,&nbsp;Karen Wetterhahn ,&nbsp;Jian Xu ,&nbsp;Steven Patierno ,&nbsp;Matthew Bogdanffy","doi":"10.1016/S0165-1218(96)90043-9","DOIUrl":"10.1016/S0165-1218(96)90043-9","url":null,"abstract":"<div><p>In 1992, a simple and sensitive assay for detecting DNA-protein crosslinks was developed [1]. In an effort to facilitate the greater use of the assay, a number of studies were conducted to evaluate its reliability and reproducibility. During this work, the assay was used to assess whether various metals and other compounds could induce crosslinks in cultured human lymphocytes (Epstein-Barr virus-transformed Burkitt's Lymphoma cell line). Potassium permanganate, mercury chloride, lead nitrate, magnesium perchlorate, aluminum chloride, and cadmium chloride did not induce DNA-protein crosslinks at either cytotoxic or non-cytotoxic levels. Copper sulfate, arsenic trioxide, and potassium chromate induced DNA-protein crosslinks only at cytotoxic concentrations. Acute lethality of the cells was assessed immediately after exposure to metals by trypan blue exclusion while long-term lethality was assessed by cell proliferation and trypan blue exclusion following an incubation period of 5 days after exposure to the metal compound. All metals exhibited more toxicity in the long-term lethaloty assay compared to the short-term assay. The cultured human lymphocytes treated with various doses of lead acetate, cadmium chloride, arsenic trioxide and copper sulfate, as well as <em>cis</em>-platinum and chromate, were sent to four different laboratories to compare the reliability and reproducibility of the DNA-protein crosslink assay. Depending on the chemical studied, there were quantitative differences in the results observed among the various laboratories using the assay. However, all laboratories generally showed that <em>cis</em>-platinum, chromate, arsenic trioxide and copper sulfate induced DNA-protein crosslinks at levels that produced acute cytotoxicity, whereas cadmium chloride and lead acetate did not.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 13-21"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90043-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19675227","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Base analog 6-N-hydroxylaminopurine mutagenesis in the yeast Saccharomyces cerevisiae is controlled by replicative DNA polymerases","authors":"Polina V. Shcherbakova,&nbsp;Vladimir N. Noskov,&nbsp;Michael R. Pshenichnov,&nbsp;Youri I. Pavlov","doi":"10.1016/S0165-1218(96)90045-2","DOIUrl":"10.1016/S0165-1218(96)90045-2","url":null,"abstract":"<div><p>Genetic control of mutagenesis by the base analog 6-<em>N</em>-hydroxylaminopurine (HAP) was studied in a set of isogenic yeast strains carrying null or point mutations in DNA repair and replication genes. Null alleles of the <em>PMS1, RAD6, REV3</em> and <em>RAD52</em> genes did not affect HAP mutagenesis. Defects in 3′- &gt; 5′ exonucleases associated with DNA polymerases ϵ and δ led to 2- to 3-fold increases in HAP-induced forward Can^r mutant frequency. A similar increase was observed for FOA^r mutants but only in the strain with a defective exonuclease of the polymerase ϵ (mutation <em>pol2-4</em>). The polymerase ϵ mutations, <em>pol2-9</em> and <em>pol2-18</em>, which lead to temperature-sensitivity, and <em>pol2-1</em> (insertion of <em>URA3</em> at the position coding for amino acid 1134 in the <em>POL2</em> gene) substantially reduced HAP mutagenesis. The polymerase δ mutation, <em>cdc2-2</em>, slightly reduced HAP mutagenesis. Enhanced proofreading was not the cause of the antimutator effect in the <em>pol2-18</em> bearing strain, inasmuch as antimutator effect was observed in the <em>pol2-4,18</em> mutant strain lacking proofreading. From the data obtained, we conclude that both DNA polymerase ϵ and δ participate in mutation generation by HAP.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 33-44"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90045-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19675229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Retrospective dose estimates in Estonian Chernobyl clean-up workers by means of FISH","authors":"F. Granath ,&nbsp;F. Darroudi ,&nbsp;A. Auvinen ,&nbsp;L. Ehrenberg ,&nbsp;T. Hakulinen ,&nbsp;A.T. Natarajan ,&nbsp;M. Rahu ,&nbsp;T. Rytömaa ,&nbsp;M. Tekkel ,&nbsp;T. Veidebaum","doi":"10.1016/S0165-1218(96)90042-7","DOIUrl":"10.1016/S0165-1218(96)90042-7","url":null,"abstract":"<div><p>The present study deals with retrospective estimation of radiation doses, among Estonian Chernobyl clean-up workers, by means of scoring stable translocations using the FISH technique. All persons investigated in this study were sent to the area in 1986 and the majority of them were selected to be among those with the presumably highest exposure doses. In spite of the selection the estimated average dose is between 0.2–0.3 Gy, thus not clearly above the officially permitted dose level of 0.25 Gy. Due to unforseen conditions during transport of the blood samples, both the number of persons available for analysis and the number of metaphases available for scoring were substantially reduced. However, unless this selection is linked with the potential aberration frequency of the cells involved, no bias is expected.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 7-12"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90042-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19673581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of chemically induced DNA breakage in cellular and subcellular systems using the comet assay","authors":"Toshio Kasamatsu,&nbsp;Kohfuku Kohda,&nbsp;Yutaka Kawazoe","doi":"10.1016/S0165-1218(96)90041-5","DOIUrl":"10.1016/S0165-1218(96)90041-5","url":null,"abstract":"<div><p>The alkaline comet assay, employing a single cell gel electrophoresis, is a rapid, simple and sensitive technique for visualizing and measuring DNA damage leading to strand breakage in individual mammalian cells. In this report, we describe a modified version of this assay which we used to assess DNA damage as a result of treating lysed cells with genotoxic and antimetabolic agents. By means of this modified assay, DNA is no longer held under the regulation of any metabolic pathway or membrane barrier. Using 3 direct-acting agents, hydrogen peroxide, <em>N</em>-methyl-<em>N</em>-nitrosourea, and bleomycin, we were able to induce increased DNA migration by both the standard and modified comet assays. In contrast, with 4-nitroquinoline 1-oxide, 5-fluorouracil, and methotrexate, which require cellular enzymatic activity to induce DNA damage, we succeeded in inducing increased DNA migration using the standard comet assay conditions only. In some cases, the modified comet assay might be helpful in analyzing chemical and biological characteristics of genotoxic agents when performed in combination with the standard comet assay.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 1-6"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90041-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19674557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cytogenetic effects of Metalaxyl on human and animal chromosomes","authors":"P. Hrelia ,&nbsp;F. Maffei ,&nbsp;C. Fimognari ,&nbsp;F. Vigagni ,&nbsp;G. Cantelli-Forti","doi":"10.1016/S0165-1218(96)90051-8","DOIUrl":"10.1016/S0165-1218(96)90051-8","url":null,"abstract":"<div><p>The purpose of this study was to assess the cytogenetic effects in vitro and in vivo of a commonly used fungicide, Metalaxyl. Chromosome damage in vitro, quantified by cultured human peripheral blood lymphocytes, demonstrated dose-related effects not associated with mitotic inhibition or cell death. Significant induction of chromosomal aberrations was observed with between 300 and 1000 μg/ml Metalaxyl in the absence of microsomal activation. Incubation in the presence of S9 mix produced less cytogenetic damage. Single i.p. injections of 75–300 mg/kg Metalaxyl had no effect on the frequency of micronuclei, detected in murine polychromatic erythrocytes. Micronuclei results were not compromised by direct evidence of cytotoxicity in the bone marrow of treated animals. The results in the present study indicated that genotoxicity of Metalaxyl was detected only in vitro and not in vivo. Available data reported that Metalaxyl was non-carcinogenic and gave negative results in a battery of genotoxicity tests. So, clastogenicity of Metalaxyl may not be evidence for DNA reactivity, but it may indicate alterations in cell homeostasis which are well implicated in the process of carcinogenesis.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 81-86"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90051-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19673583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Organ variation in the mutagenicity of MeIQ in Big Blue® lacI transgenic mice","authors":"Takayoshi Suzuki ,&nbsp;Makoto Hayashi ,&nbsp;Masako Ochiai ,&nbsp;Keiji Wakabayashi ,&nbsp;Toshikazu Ushijima ,&nbsp;Takashi Sugimura ,&nbsp;Minako Nagao ,&nbsp;Toshio Sofuni","doi":"10.1016/S0165-1218(96)90046-4","DOIUrl":"10.1016/S0165-1218(96)90046-4","url":null,"abstract":"<div><p>2-Amino-3,4-dimethylimidazo[4,5-<em>f</em>]quinoline (MeIQ), which is a heterocyclic amine found in food and the potent mutagen in <em>S. typhimurium</em> TA98, was examined for its genotoxic potential using <em>lacI</em> transgenic mice (Big Blue®, C57BL/6N lineage). Female mice, at 7 weeks of age, were given a diet containing 0.03% MeIQ for 1, 4 and 12 weeks, and mutant frequencies (MF) were analyzed in the bone marrow, liver, forestomach, colon and heart. The MF increased in a feeding period-dependent manner. Relative to untreated mice, the MF after a 12-week-feeding of MeIQ was 38 times higher in the colon, 5.8 times higher in the bone marrow, 4.6 times higher in the liver, and 2.6 times higher in the forestomach. No increase in MF was detected in the heart, where no tumors develop.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 45-49"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90046-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19675230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection of neocarzinostatin-induced translocations in human sperm chromosomes using fluorescence in situ hybridization of chromosome 2","authors":"Hirokazu Kusakabe,&nbsp;Kohji Yamakage,&nbsp;Noriho Tanaka","doi":"10.1016/S0165-1218(96)90047-6","DOIUrl":"10.1016/S0165-1218(96)90047-6","url":null,"abstract":"<div><p>Mature sperm and late spermatid are known to be sensitive stages to clastogens in mammalian spermatogenesis. Certain types of chromosomal damage induced in these stages will pass to successive generations as heritable translocations. In the present study, we employed whole chromosome 2 painting with the fluorescence in situ hybridization (FISH) technique to detect the chemically induced translocations in human sperm. Mature human sperm were treated in vitro with an antitumor drug, neocarzinostatin (NCS), and fertilized in vitro with golden hamster oocytes. Sperm pronuclear chromosomes slides were prepared at the first cleavage metaphase. To compare the characteristics of translocations between somatic and germ cells, human lymphocytes in peripheral blood treated with NCS in vitro were analyzed at first round metaphase after PHA-stimulation. From the analysis of translocations by whole chromosome 2 painting, frequencies of the haploid genomic translocations (<em>F</em>_hG) were predicted for both sperm and lymphocytes At 1.0 μg/ml, the actual percentages of sperm and lymphocytes with chromosome 2 translocations were almost identical (11.9% and 12.0%). At the same dose, however, the <em>F</em>_hG of the sperm (1.15) was considerably higher than that of the lymphocytes (0.58), indicating that complex translocations having two or more rearranged sites were induced by NCS more frequently in sperm than in lymphocytes.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 51-58"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90047-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19675231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cytogenotoxicities of sublimed urethane gas to the mouse embryo","authors":"Taisei Nomura ,&nbsp;Satonori Tanaka ,&nbsp;Nobuo Kurokawa ,&nbsp;Keiko Shibata ,&nbsp;Hiroo Nakajima ,&nbsp;Akihiro Kurishita ,&nbsp;Tadashi Hongyo ,&nbsp;Yutaka Ishii","doi":"10.1016/S0165-1218(96)90048-8","DOIUrl":"10.1016/S0165-1218(96)90048-8","url":null,"abstract":"<div><p>Urethane (ethyl carbamate) which has long been used for commonly used drugs and has proven to be useful in the formation of products in every-day use, is volatile, and small amounts sublime spontaneously. Pregnant ICR mice were maintained in the vinyl chamber (45 liter) which was ventilated 4 times per hour. To inhale urethane gas, air was passed first through a glass bottle containing 500 g of crystalline urethane and then into the vinyl chamber. Concentration of the sublimed urethane gas in the chamber was 1.28 ± 0.08 mg/l, and sublimed urethane gas produced significantly high incidence of chromosomal aberrations in the cells of whole embryo, when mice inhaled it for 48 h from day 9 to day 11 of pregnancy. High and significant incidence of chromosomal aberrations (36.0%) was detected in the embryo 3 h after urethane gas inhalation, but decreased to 5.3% at 24 h after exposure and showed no significant differences from controls after 48 h, while the incidence in bone marrow cells from the adult (pregnan) mice was lower (21.5%) at 3 h after exposure but a significant increase remained until 72 h after exposure. A majority of chromosomal aberrations was chromatid types. As a consequence of cellular damages by urethane gas inhalation during pregnancy, significantly high incidence of fetal deaths and congenital malformations (cleft palate, polydactyl, tail anomaly etc.) was induced in the offspring. Thus, we must be aware of the risk of volatile chemicals, because it is difficult to perceive and avoid hazardous exposure via respiration.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 59-64"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90048-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19673579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative in-vivo genotoxicity of antiviral nucleoside analogues; penciclovir, acyclovir, ganciclovir and the xanthine analogue, caffeine, in the mouse bone marrow micronucleus assay","authors":"P. Haynes,&nbsp;T.R. Lambert,&nbsp;I. de G. Mitchell","doi":"10.1016/S0165-1218(96)90049-X","DOIUrl":"10.1016/S0165-1218(96)90049-X","url":null,"abstract":"<div><p>Three purine nucleoside analogues, penciclovir (PCV), acyclovir (ACV) and ganciclovir (GCV), were assessed for in-vivo genotoxicity in the mouse bone marrow micronucleus assay, together with the xanthine (purine) analogue, caffeine (CAF). All these compounds exhibit anti-viral properties and the first three are marketed anti-viral drugs. All have been shown to be genotoxic in separate in-vitro and in-vivo studies. Because of their widespread use, we considered it important to directly compare their relative in-vivo genotoxic potencies as an aid to assessing their relative genotoxic risk to humans. Accordingly, two-dose (0 and 24 h)/ single sample mouse micronucleus assays were performed on all four compounds. PCV and ACV appeared to give essentially arithmetic increases in induction of micronucleated polychromatic erythrocytes (MNPCE) with arithmetic increases in dose with apparent thresholds at approx. 1078 μmol/kg per day and 316 μmol/kg per day, respectively. The dose-response curve for GCV appeared more exponential, without a threshold, but with a no-effect dose of around 150 μmol/kg per day. With CAF, systemic toxicity allowed the assessment of only very weak effects, such that our estimate of a no-effect dose of 388 μmol/kg per day is subject to large errors. Taking into account magnitude of response, slope of dose-response curve and no-effect doses, the order of potency was GCV &gt; ACV &gt; (CAF?) &gt; PCV. The relevance of these findings in terms of risk is uncertain.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 65-74"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90049-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19673580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition by quercetin and luteolin of chromosomal alterations induced by salted, deep-fried fish and mutton in rats","authors":"Shaheen Taj,&nbsp;B. Nagarajan","doi":"10.1016/S0165-1218(96)90053-1","DOIUrl":"10.1016/S0165-1218(96)90053-1","url":null,"abstract":"<div><p>Previous studies from our laboratory have shown the clastogenic effects of long-term feeding on deep-fried fish and mutton in rat bone marrow cells. We report the chemopreventive action of two flavanoids, quercetin (Qn) and luteolin (Ln) against the induced mutagenicity by fish and mutton extracts. Groups of rats were treated with flavanoids through pre-simultaneous- and post-treatment regimens and killed at the end of treatment. The bone marrow was removed and analysed for the presence of micronuclei (MN) and chromosome aberrations (CA). Pre-treatment showed most effectively a good inhibition of mutagenicity at every dose tested. Luteolin was a better protective agent than quercetin. It protected the cells against genetic damage to 93% in the micronucleus assay and to 95% in the chromosome aberrations induced by fish extract (<em>p</em> &lt; 0.001 in both the groups). Mutton extract-induced micronuclei and chromosome aberrations were protected 85% and 90%, respectively, by luteolin and 79% and 76%, respectively, by quercetin. Our results tend to suggest that quercetin and luteolin are potential chemopreventive compounds.</p></div>","PeriodicalId":100938,"journal":{"name":"Mutation Research/Genetic Toxicology","volume":"369 1","pages":"Pages 97-106"},"PeriodicalIF":0.0,"publicationDate":"1996-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0165-1218(96)90053-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19673585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}