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Genotoxicity in water and sediment extracts from the St. Lawrence river system, using the SOS chromotestLangevin, Robert January 1991 (has links)
Surface water and sediments from the St. Lawrence River system (Quebec region) were analysed for genotoxicity using nonlinear SOS Chromotest parameters, as well as for their chemical concentrations of polycyclic aromatic hydrocarbons and heavy metals. Additionally, sediments chlorobenzenes, polychlorinated biphenyls, organochlorinated pesticides, ammonia and nitrites concentrations were determined. Organic contaminants were extracted dichloromethane. For surface water, fifteen extracts of filtered water and seven of particulates, and for sediments, one extract of pore water and three of particulates proved to be weakly genotoxic. All but one of the genotoxic responses observed in the surface water were obtained from samples taken from the highly industrial portion of the St. Lawrence River system, with the strongest responses observed in Lake St-Louis. Surface water genotoxicants partitioning favors the particulate fraction. Bottom particulates genotoxicity was one thousand fold weaker than suspended particulates. Additionally, whole sediments were extracted with a 10% dimethylsulfoxide-saline solution. Genotoxicity of hydrophilic contaminants was detected in all extracts. The observed distributions of genotoxicity values did not correlate with observed concentrations of demonstrated SOS inducers, mutagens and/or carcinogens, nor with the presence of other toxic chemicals.
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Detection of latent heritable genetic damage in populations of aquatic snails, Lymnaea stagnalis, exposed in situ to genotoxic pollutionHum, Stanley. January 1999 (has links)
We hypothesized that populations exposed to chronic levels of genotoxins for many generations in situ will accumulate latent heritable genetic damage leading to increases in mutational load and decreased population fitness. Common pulmonate snails (Lymnaea stagnalis) were collected from three sites that differed in pollution level [Manitoulin Island (reference), Beauharnois (moderate pollution), and Varennes (highly polluted)]. These organisms have a rapid generation time and are capable of self-fertilization. Fitness indicators were clutch size, survival (hatching to day 30) and growth (length from hatching to day 90). Recessive deleterious mutations that have accumulated through time are masked as snails preferentially outcrossed, but are expressed when snails self-fertilize. Results obtained by comparing fitness components of snails derived from selfing and outcrossing experiments showed that in sites with higher levels of pollution inbreeding depression increased for clutch size and survival. Results for growth rate were similar to those for clutch size and survival in the two least polluted sites, but not for the most polluted site, due possibly to biased mortality of smaller snails at this site. After correction for size biased mortality, the growth estimates showed patterns similar for the two polluted sites when compared to the reference site. This study suggests that long-term in situ exposure to genotoxic pollution may effect population fitness due to the accumulation of latent heritable genetic damage.
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Análise de genotoxicidade: otimização de método laboratorial e avaliação em recém-nascidos de mães com restrição de crescimento intrauterino exercitadas durante a prenhezOliveira Netto, Aline de [UNESP] 22 February 2013 (has links) (PDF)
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000758181.pdf: 1360078 bytes, checksum: 426974081d9d61117a57cddef7eb84c6 (MD5) / Para ampliar os conhecimentos dos efeitos do diabete na gestação e danos no DNA, foi realizada uma revisão de literatura utilizando dois conjuntos de palavras-chave: 1) diabetes, DNA damage, pregnancy e 2) rat, diabetes, DNA damage, pregnancy, no site da base de dados do National Center of Biotechnology Information (NCBI-PUBMED). No primeiro conjunto, foram encontrados 28 artigos e, no segundo, 13 artigos. Destes, 11 eram iguais aos do primeiro conjunto e dois não tinham coerência com as palavras-chave. Dos 28 artigos analisados, 28,58% foram considerados adequados para a redação da revisão e 71,42% inadequados, pois tinham títulos e resumos fora de contexto. Dos adequados, 37,5% eram relacionados à pesquisa em humanos, 50% em animais de experimentação e 12,5% em humanos e animais. De forma geral, os artigos demonstraram que o diabete aumenta espécies reativas de oxigênio gerando danos em ácidos nucleicos e isto exarceba o estresse oxidativo materno, embrionário e fetal / To contribute the knowledge of the diabetes effects during pregnancy in the DNA damage, we performed a literature review using two groups of keywords: 1) diabetes, DNA damage, pregnancy and 2) rat, diabetes, DNA damage, pregnancy, in site of the database of the National Center for Biotechnology Information (NCBI-PUBMED). In the first group of keywords were found 28 articles. In the second group, there were 13 articles, which 11 were equal to the first group. Then, 28 articles were analyzed. Of these articles, 28,28% were considered adequate for writing the review, while 71,42% were considered inappropriate because they presented no coherent titles and abstracts. Of these articles, 37.5% were related to human research, 50% in experimental animals and 12.5% in humans and animals. Thus, in general, the articles showed that diabetes increases the reactive oxygen species that causes nucleic acid damage and this exacerbates maternal, embryonic and fetal oxidative stress
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Avaliação da biocompatibilidade e atividade antimicrobiana in vitro da matriz de Hidrogel associada ao extrato glicólico do Punica granatum L. (romã) /Gomes, Livia Aparecida Procopio. January 2017 (has links)
Orientador: Samira Esteves Afonso Camargo / Banca: Luana Marotta Reis de Vasconcelos / Banca: Jonatas Rafael de Oliveira / Resumo: A matriz de hidrogel é um biomaterial de nanofibra peptídica tridimensional que induz o crescimento, migração e proliferação celular, assim favorecendo a regeneração tecidual. O objetivo deste trabalho foi avaliar a biocompatibilidade e a atividade antimicrobiana da matriz de hidrogel associado ao extrato romã e ao antimicrobiano ciprofloxacino. Para isso, foram realizadas análises microbiológicas sobre Enterococcus faecalis (ATCC 4083) em cultura planctônica, por meio d ensaio de microdiluição em caldo e em biofilme pelo teste de MTT. Para os ensaios da biocompatibilidade, foi utilizada cultura de macrófagos (RAW 264.7). A citotoxicidade foi avaliada pelo ensaio de MTT e a genotoxicidade foi realizado pelo teste de micronúcleo. A análise estatística foi realizada pelos testes ANOVA e Tukey, adotando o nível de significância 5%. Os resultados demonstraram que a matriz de hidrogel associado ao extrato de romã não apresentou atividade antimicrobiana para a cultura planctônica como para o biofilme de E. faecalis. Porém, não foi citotóxico e genotóxico para RAW 264.7. Por outro lado, o antimicrobiano ciprofloxacino apresentou atividade antimicrobiana sobre E. faecalis,além de ter apresentado efeitos citotóxico e genotóxico para os macrófagos. Com isso, foi possível concluir que o extrato de romã associado ou não a matriz de hidrogel apresenta ausência de citotóxico, genotóxico e efeito antimicrobiano / Abstract: The hydrogel matrix is a three-dimensional peptide nanofiber biomaterial that induces cell growth, migration and proliferation, thus favoring tissue regeneration. The objective of this work was to evaluate the biocompatibility and antimicrobial activity of the hydrogel matrix associated with the pomegranate extract and the antimicrobial ciprofloxacin. For this, microbiological analyzes on Enterococcus faecalis (ATCC 4083) were carried out in planktonic culture, by means of the microdilution test in broth and in biofilm by the MTT test. For the biocompatibility assays, macrophage culture (RAW 264.7) was used. Cytotoxicity was assessed by the MTT assay and genotoxicity was performed by the micronucleus test. Statistical analysis was performed by the ANOVA and Tukey tests, adopting the significance level 5%. The results showed that the hydrogel matrix associated with the pomegranate extract did not show antimicrobial activity for the planktonic culture as for the E. faecalis biofilm. However, it was not cytotoxic and genotoxic for RAW 264.7. On the other hand, the antimicrobial ciprofloxacin showed antimicrobial activity on E. faecalis, besides having cytotoxic and genotoxic effects for the macrophages. With this, it was possible to conclude that the pomegranate extract associated or not with the hydrogel matrix shows absence of cytotoxic, genotoxic and antimicrobial effect / Mestre
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Avaliação da genotoxicidade e estresse oxidativo em planárias aquáticas (Dugesia schubarti) tratadas com formulações do herbicida glifosatoBordin, Diana Lilian 19 November 2007 (has links)
Atualmente, diversas formulações de herbicidas a base de glifosato estão disponíveis no mercado. No entanto, diferentes surfactantes são também adicionados às formulações a fim de aumentar a eficácia do produto, o que pode contribuir significativamente para o aumento da toxicidade do herbicida aos organismos não alvos. Este estudo avaliou a genotoxicidade e o estresse oxidativo em planárias aquáticas (Dugesia schubarti) expostas às formulações de glifosato Roundup Original, Roundup Transorb e Roundup Ready. As planárias foram expostas à soluções de 0.5 mg/L de equivalente ácido de glifosato das três formulações durante 4, 8 e 16h. A genotoxicidade foi avaliada através do ensaio cometa e o estresse oxidativo foi medido através da atividade das enzimas superóxido dismutase (Sod) e catalase (Cat) e pelas substâncias reativas com o ácido tiobarbitúrico (TBARS). Todas as formulações foram capazes de induzir danos ao DNA e estresse oxidativo. A atividade de Sod aumentou entre 4 e 16h, enquanto os níveis de TBARS diminuíram entre 4 e 16h de tratamento. A atividade de Cat permaneceu diminuída em todos os tempos de exposição para as três formulações testadas. Esses resultados demonstram que formulações de herbicida contendo glifosato são capazes de causar dano ao DNA e alterar o balanço oxidativo em planárias indicando que a toxicidade deste herbicida não está restrita às plantas. / Submitted by Marcelo Teixeira (mvteixeira@ucs.br) on 2014-05-30T16:17:08Z
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Dissertacao Diana Lilian Bordin.pdf: 300600 bytes, checksum: 95ba8b94d9994115a4d1aaa7a0f08bc0 (MD5) / Currently, several glyphosate-based herbicide formulations are available in the market. However, different surfactants are also added to the formulation in order to increase the effectiveness of the product, what it can contribute significantly to the toxic effect exerted by the herbicide to the non target organisms. This study evaluated the genotoxicity and the oxidative stress in freshwater planarians exposed to the glyphosate formulations Roundup Original, Roundup Transorb and Roundup Ready. The planarians were submitted to 0.5 mg/L of glyphosate formulations during 4, 8, and 16h. The genotoxicity was evaluated through comet assay, and the oxidative stress was measured through the superoxide dismutase (Sod) and catalase (Cat) activity and by thiobarbituric acid reactive substances (TBARS). All three formulations were able to induce genotoxicity and to disturb the antioxidants activity in different ways. While Sod increased between 4 and 16h, TBARS decreased between 4 and 16h, and Cat stayed depressed during all time-points tested. These results demonstrate that herbicide formulations which contend glyphosate can induce damage to the DNA and to modify the oxidative balance in planarians, indicating that the toxicity of this herbicide is not restricted to the plants.
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Avaliação da genotoxicidade espermática em pacientes HIV/AIDS usuários de terapia antirretroviral de alta potênciaBuffon, Viviane Raquel 15 December 2015 (has links)
A Síndrome da Imunodeficiência Adquirida (AIDS) é doença causada pelo vírus da imunodeficiência humana. Apesar dos benefícios proporcionados pela terapia de supressão viral, algumas doenças como lipodistrofia, doenças cardiovasculares e infertilidade aumentaram a sua prevalência. No paciente com AIDS, considera-se a carga viral e o uso de terapia antirretroviral como possíveis agentes de genotoxicidade. Apesar do espermograma não poder detectar a causa precisa da infertilidade, o mesmo ainda é o exame mais realizado para averiguar a qualidade seminal. Por outro lado, a integridade do DNA do espermatozóide tem sido proposta como um parâmetro adicional de qualidade do sêmen. O ensaio cometa alcalino detecta a genotoxicidade de células germinativas humanas e pode ser usado para demonstrar a capacidade de uma substância interagir com o material genético das células gonadais. O presente estudo incluiu 50 pacientes com AIDS, atendidos no Ambulatório de Infectologia do Município de Caxias do Sul e no Ambulatório da Universidade de Caxias do Sul, sendo analisados dois grupos: usuário de terapia antirretroviral e naive. Realizou-se espermograma e o ensaio cometa alcalino, comparando com o número de linfócitos T CD4+, a carga viral, a idade, etc. Os dados demonstraram que o uso de antirretrovirais reduziu a genotoxicidade espermática, mas não se encontrou correlação com o espermograma. O presente trabalho demonstra que o ensaio cometa alcalino é um método eficiente para mensurar a genotoxicidade espermática dos pacientes portadores do vírus da imunodeficiência adquirida. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2017-02-10T16:43:52Z
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Previous issue date: 2017-02-10 / The Acquired Immune Deficiency Syndrome (AIDS) is a disease caused by the human immunodeficiency virus. Despite all the benefits provided by the viral suppression therapy, some diseases such as lipodystrophy, heart diseases and infertility increased their prevalence. In patients with AIDS, viral load and anti-retroviral therapy are possible agents for genotoxicity. The World Health Organization guidelines are reference for semen analysis and sperm DNA integrity has been proposed as an additionalparameter for semen quality and a potential predictor of fertility. The alkaline comet assay has the potential to detect genotoxic human germ cells and can be used to demonstrate the ability of a substance or metabolite to interact directly with the genetic material of gonadal cells. The study has included 50 patients with AIDS treated at the Outpatient Center for Infectious Diseases and at the University of Caxias do Sul Clinical Center, evaluated as two groups: one treated with anti-retroviral therapy and the other was a naive group. Semen and sperm genotoxicity were analysed by alkaline comet assay, comparing CD4 cell count, viral load, age and other factors of potential genotoxicity between the two groups. Data showed that the use of any anti-retroviral class has reduced sperm genotoxicity. This study shows that the alkaline comet assay is a eficcientmethod to measure sperm genotoxicity in patients infected with the human immunodeficiency virus.
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Análise de genotoxicidade : otimização de método laboratorial e avaliação em recém-nascidos de mães com restrição de crescimento intrauterino exercitadas durante a prenhez /Oliveira Netto, Aline de. January 2013 (has links)
Orientador: Débora Cristina Damasceno / Banca: Mariana Gobbo Braz / Banca: Maricelma da Silva Soares de Souza / Resumo: Para ampliar os conhecimentos dos efeitos do diabete na gestação e danos no DNA, foi realizada uma revisão de literatura utilizando dois conjuntos de palavras-chave: 1) diabetes, DNA damage, pregnancy e 2) rat, diabetes, DNA damage, pregnancy, no site da base de dados do National Center of Biotechnology Information (NCBI-PUBMED). No primeiro conjunto, foram encontrados 28 artigos e, no segundo, 13 artigos. Destes, 11 eram iguais aos do primeiro conjunto e dois não tinham coerência com as palavras-chave. Dos 28 artigos analisados, 28,58% foram considerados adequados para a redação da revisão e 71,42% inadequados, pois tinham títulos e resumos fora de contexto. Dos adequados, 37,5% eram relacionados à pesquisa em humanos, 50% em animais de experimentação e 12,5% em humanos e animais. De forma geral, os artigos demonstraram que o diabete aumenta espécies reativas de oxigênio gerando danos em ácidos nucleicos e isto exarceba o estresse oxidativo materno, embrionário e fetal / Abstract: To contribute the knowledge of the diabetes effects during pregnancy in the DNA damage, we performed a literature review using two groups of keywords: 1) diabetes, DNA damage, pregnancy and 2) rat, diabetes, DNA damage, pregnancy, in site of the database of the National Center for Biotechnology Information (NCBI-PUBMED). In the first group of keywords were found 28 articles. In the second group, there were 13 articles, which 11 were equal to the first group. Then, 28 articles were analyzed. Of these articles, 28,28% were considered adequate for writing the review, while 71,42% were considered inappropriate because they presented no coherent titles and abstracts. Of these articles, 37.5% were related to human research, 50% in experimental animals and 12.5% in humans and animals. Thus, in general, the articles showed that diabetes increases the reactive oxygen species that causes nucleic acid damage and this exacerbates maternal, embryonic and fetal oxidative stress / Mestre
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Avaliação da genotoxicidade e estresse oxidativo em planárias aquáticas (Dugesia schubarti) tratadas com formulações do herbicida glifosatoBordin, Diana Lilian 19 November 2007 (has links)
Atualmente, diversas formulações de herbicidas a base de glifosato estão disponíveis no mercado. No entanto, diferentes surfactantes são também adicionados às formulações a fim de aumentar a eficácia do produto, o que pode contribuir significativamente para o aumento da toxicidade do herbicida aos organismos não alvos. Este estudo avaliou a genotoxicidade e o estresse oxidativo em planárias aquáticas (Dugesia schubarti) expostas às formulações de glifosato Roundup Original, Roundup Transorb e Roundup Ready. As planárias foram expostas à soluções de 0.5 mg/L de equivalente ácido de glifosato das três formulações durante 4, 8 e 16h. A genotoxicidade foi avaliada através do ensaio cometa e o estresse oxidativo foi medido através da atividade das enzimas superóxido dismutase (Sod) e catalase (Cat) e pelas substâncias reativas com o ácido tiobarbitúrico (TBARS). Todas as formulações foram capazes de induzir danos ao DNA e estresse oxidativo. A atividade de Sod aumentou entre 4 e 16h, enquanto os níveis de TBARS diminuíram entre 4 e 16h de tratamento. A atividade de Cat permaneceu diminuída em todos os tempos de exposição para as três formulações testadas. Esses resultados demonstram que formulações de herbicida contendo glifosato são capazes de causar dano ao DNA e alterar o balanço oxidativo em planárias indicando que a toxicidade deste herbicida não está restrita às plantas. / Currently, several glyphosate-based herbicide formulations are available in the market. However, different surfactants are also added to the formulation in order to increase the effectiveness of the product, what it can contribute significantly to the toxic effect exerted by the herbicide to the non target organisms. This study evaluated the genotoxicity and the oxidative stress in freshwater planarians exposed to the glyphosate formulations Roundup Original, Roundup Transorb and Roundup Ready. The planarians were submitted to 0.5 mg/L of glyphosate formulations during 4, 8, and 16h. The genotoxicity was evaluated through comet assay, and the oxidative stress was measured through the superoxide dismutase (Sod) and catalase (Cat) activity and by thiobarbituric acid reactive substances (TBARS). All three formulations were able to induce genotoxicity and to disturb the antioxidants activity in different ways. While Sod increased between 4 and 16h, TBARS decreased between 4 and 16h, and Cat stayed depressed during all time-points tested. These results demonstrate that herbicide formulations which contend glyphosate can induce damage to the DNA and to modify the oxidative balance in planarians, indicating that the toxicity of this herbicide is not restricted to the plants.
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The use of genotoxic and stress proteins in the active biomonitoring of the Rietvlei system, South AfricaMbizi, Zviregei 14 October 2008 (has links)
The use of biomarkers in assessing the health of aquatic ecosystems is widely being used. Biomarkers have been proposed as sensitive early warning tools for biological measurement in environmental quality assessment. In this study, DNA damage, HSP 70 expression, Lactate dehydrogenase and alkaline phosphatase were evaluated for their usefulness and applicability in the active biomonitoring of the Rietvlei system using Melanoides tuberculata and Oreochromis mossambicuss as test organisms. Levels of DNA strand breakage were measured in the exposed test organisms and the references that were kept in the laboratory under unpolluted environment. Different levels of DNA damage were recorded in M tuberculata that were exposed during high-flow and those from the low-flow exposure period. There was no difference in DNA between the control and high-flow exposure M. tuberculata but the low-flow exposure duration had significantly higher DNA damage for all the three sites when compared to the high-flow and the controls. There was no difference in the amount of DNA damage in both exposed and control O. mossambicus. Fish samples were only available for the high-flow four weeks exposure because of the test organisms dying during the low-flow exposure. Heat shock protein 70 expression was determined in O. mossambious only and significant differences in expression between sites were demonstrated with highest expression at site 1 decreasing down stream at sites 3 and 5. The activity of LDH in M. tuberculata was not different in both control and exposed fish at all the sites. There was no difference in the activity of LDH between the high-flow and low-flow exposure groups in M. tuberculata. In O. mossambicus, the activity of LDH was significantly inhibited at site 1 with the effects of the contaminants decreasing downstream at sites 3 and 5 where LDH activity did not differ from the controls. There was no alkaline phosphatase activity in both exposed and control M. tuberculata but in O. mossambicus alkaline phosphatase was significantly inhibited at the three sites although inhibition did not differ from site to site. This suggests that LDH and alkaline phosphate are not sensitive biomarkers in M. tuberculata in the Rietvlei system but are sensitive in O. mossambicus. DNA damage was shown to be a sensitive biomarker using M. tuberculata but it was difficult to assess this in O. mossambicus since fish were only available during the 4 weeks high-flow exposure, which did not show any differences from the control. The lack of DNA damage in O. mossambicus maybe a result of low concentration of contaminants during high-flow or because this biomarker is not sensitive in fish. / Prof. J.H.J. Van Vuren
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Analysis of the possible confounding effects of cytotoxicity and exogenous metabolism on in vitro genotoxicity assessmentsTate, Matthew January 2011 (has links)
Recent evaluations of the regulatory in vitro genotoxicity tests have revealed that the mammalian cell tests are sensitive for the prediction of rodent carcinogenicity, but not very specific; they produce positive results for many non-carcinogens. Studies to investigate the potential causes of this poor specificity have focused on the high test concentrations of chemicals and the often associated high cytotoxicity. Others have sought to develop alternative in vitro assays that retain high sensitivity but also have higher specificity. One such test is the GADD45a-GFP “GreenScreen HC” genotoxicity assay, which uses elements of the cellular DNA damage response in a green fluorescent protein (GFP) reporter system to detect genotoxic hazard. Published validation studies for this assay have shown both high sensitivity and high specificity. Suggestions that the high specificity of the assay is attributable to the method of cytotoxicity estimation, relative cell density (RCD), and further that this method leads to overestimation of cytotoxicity and hence lower maximum test concentrations, are investigated in this study.In order to investigate whether high specificity of the GADD45a-GFP assay is a result of inaccurate estimation of cytotoxicity, RCD was compared with other cytotoxicity methods used in screening assays, as well as the cytotoxicity methods used in the regulatory in vitro mammalian genotoxicity tests. Results showed that RCD actually underestimates cytotoxicity in the GADD45-GFP assay and as a consequence, test data are collected up to and beyond the cytotoxicity-limited test concentrations reached in the regulatory tests. The use of RCD was found not to contribute to the high specificity of the GADD45a-GFP assay and indeed, its high specificity was retained regardless of the method used to estimate cytotoxicity.A largely overlooked element in the consideration of the poor specificity of in vitro mammalian genotoxicity assays is the supplementation of assays with exogenous metabolism systems, the most common of which is ‘S9’, a liver-derived preparation. S9 is used to enable the detection of compounds (pro-genotoxins) that undergo metabolic transformation in vivo resulting in the formation of genotoxic carcinogens. However, both the preparation and usage of S9 create a bias towards phase I metabolism, specifically the cytochrome P450 monooxygenases that can catalyse the formation of DNA-reactive species. The neglect of phase II metabolism (detoxification/conjugation) could potentially contribute to the poor specificity of the in vitro mammalian tests and this was explored in the study presented.A review of published data revealed that the prevalence of positive results amongst in vitro mammalian genotoxicity data generated in the presence of S9 was similar in collections of both rodent carcinogens and non-carcinogens. Thus, any increased sensitivity from using S9 is offset by decreased specificity for these tests. However, as inclusion of S9 affected sensitivity and specificity in equal measure it was concluded that S9 is not a confounding factor on the accuracy of the in vitro mammalian genotoxicity tests.
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