RESUMO
The discovery of trace levels of semicarbazide (SEM) in bottled foods (especially baby foods) led to a consideration of the safety of this hydrazine compound by regulatory agencies worldwide. Azodicarbonamide, which is used in the jar-sealing technology known as Press On-Twist Off (or Push-Twist/PT) closures for the formation of a hermetic, plastisol seal, partially degrades with the heat of processing to form trace amounts of SEM. This review has evaluated the potential toxicological risks of resulting exposure to SEM and also the benefit of the PT technology (with azodicarbonamide) in the context of possible microbial contamination. It also considers the potential impact on infant nutrition if parents come to the conclusion that commercial baby foods are unsafe. SEM shows limited genotoxicity in vitro that is largely prevented by the presence of mammalian metabolic enzymes. Negative results were found in vivo in DNA alkaline elution, unscheduled DNA synthesis and micronucleus assays. This pattern is in contrast to the genotoxic hydrazines that also have been shown to cause tumours. Carcinogenicity studies of SEM are of limited quality, show a questionable weak effect in mice at high doses, which are not relevant to human exposure at trace levels, and show no effect in the rat. The IARC has assigned SEM as Group 3, 'Not classifiable as to its carcinogenicity to humans'. Based on estimates of exposure to infants consuming baby foods (with the assumption of SEM levels at the 95th percentile of 20 ng g(-1) in all of the consumed 'ready-to-eat' foods) compared with a no observed adverse effect level (NOAEL) in developmental toxicity studies, the margin of safety is more than 21 000. Since the risk of an adverse effect is negligible, it is clear that any theoretical risk is outweighed by the benefits of continuing use of the PT closure (with azodicarbonamide blowing agent) to ensure both the microbial integrity and availability of commercial baby foods as a valuable source of infant nutrition.
Assuntos
Compostos Azo/farmacocinética , Contaminação de Alimentos/análise , Embalagem de Alimentos/normas , Alimentos Infantis/análise , Semicarbazidas/análise , Carcinógenos/análise , Contaminação de Alimentos/prevenção & controle , Microbiologia de Alimentos , Humanos , Lactente , Alimentos Infantis/microbiologia , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Medição de Risco/métodos , Semicarbazidas/toxicidadeRESUMO
Styrene is a highly reactive monomer widely used in the plastics industry. The potential for styrene to produce genotoxic effects has been studied extensively in experimental systems. Styrene can induce sister chromatid exchanges (SCE) and chromosome aberrations (CA) in vitro under test conditions that enhance metabolism of styrene to styrene 7,8-oxide (SO)or reduce detoxification of 50 by epoxide hydrolase. The in vivo animal data indicate that styrene is not clastogenic at concentrations (doses) likely encountered by humans under ambient or occupational exposure conditions. DNA binding studies with styrene in rats and mice demonstrated no increased adducts in mice compared to rats or in mouse lung compared to liver. As a result, DNA adducts in the lungs are unlikely to be the sole explanation of the development of lung tumors in mice exposed to styrene for 2 yr. Some epidemiological studies reported that DNA and/or protein adducts and DNA strand breaks result from occupational exposure to styrene and/or 50. Results of some of these studies, how-ever, are difficult to interpret, given that the statistical significance of reported effects (SCE, CA, and micronucleus formation) was often near or at p values of .05; dose and/or temporal response relationships often were missing; confounding variables could not be excluded; and, concomitant exposures to other industrial chemicals that are potentially genotoxic may also have occurred. These studies suggest that styrene, through metabolism to SO, could be clastogenic in humans at workplace levels in excess of 125 mg/m3. However, results from controlled animal studies involving in vivo exposure to styrene alone do not show clastogenic effects at exposures of up to 1500 mg/m3/d. In any event, these studies show that there is an apparent threshold for styrene-mediated effects.
Assuntos
Aberrações Cromossômicas/induzido quimicamente , Troca de Cromátide Irmã/efeitos dos fármacos , Estireno/toxicidade , Animais , Adutos de DNA/análise , Monitoramento Ambiental , Humanos , Exposição Ocupacional , Risco , Estireno/metabolismoRESUMO
The IARC Monographs (Vols 1-70) were studied to determine the time of onset of treatment-related tumorigenicity in long-term rodent studies for chemicals classified by IARC as having sufficient evidence of carcinogenicity in animals. The analysis excluded studies on metals and their salts, studies on particulates, studies by parenteral routes of administration that resulted in tumours only at the site of exposure, and studies that did not approximate to the current standard long-term rodent carcinogenicity bioassay, for instance transplacental or multigeneration studies, initiator-promoter studies, lung tumour assays in Strain A mice and studies in newborn animals. Data from a total of 210 chemicals revealed that, overall, evidence of treatment-related tumorigenicity was first apparent within 12 months for 66% of the chemicals and for only 7% were studies of longer than 18 months necessary. All IARC Group 1 chemicals were detected in animals within 18 months, and most within 12 months. Most of the tumour types that required more than 18 months for detection were of dubious relevance to human risk assessment. Termination of rodent carcinogenicity studies at 18 months or earlier would greatly reduce the complications that arise in interpreting the findings in aged animals which often have defective hepatic or renal function and would also markedly reduce the time required for histopathological examination of dozens of tissues taken from the approximately 500 animals routinely employed in these studies.
Assuntos
Testes de Carcinogenicidade/normas , Carcinógenos/toxicidade , Controle de Qualidade , Animais , Humanos , Camundongos , Ratos , Reprodutibilidade dos Testes , Projetos de Pesquisa , Medição de Risco , Fatores de TempoRESUMO
Subchronic/chronic toxicity studies on antimony potassium tartrate (APT) have been reviewed. One of the older studies (H. A. Schroeder et al., 1970, J. Nutr. 100 (1), 59-68), on which are based the EPA reference dose value and a number of state, national, and international drinking water criteria for antimony, has severe inadequacies in study conduct making it uninterpretable and inappropriate for characterization of APT toxicity. In particular, the manner in which control data were generated and utilized in this study is considered invalid. More recent drinking water studies conducted by the NTP (1992, "NTP Technical Report on Toxicity Studies of Antimony Potassium Tartrate in F344/N Rats and B6C3F(1) Mice (Drinking Water and Intraperitoneal Injection Studies)," NTP Toxicity Report Series, No. 11) and Poon et al. (1998, Food Chem. Toxicol. 36, 20-35) showed antimony to be of low toxicity. The NOAEL in the 14-day NTP study was 2500 ppm by the oral route in both rats and mice, while Poon et al. (1998) suggested a NOAEL of 0.5 ppm in their 90-day study. However, upon close examination, it was determined that this value was based on subtle histological changes in the thyroid gland that were physiological, not toxicological, in nature. This conclusion is supported further by an absence of these changes in a well-conducted 13-week intraperitoneal exposure study in rats that utilized APT at much higher doses (NTP, 1992). Thus, the NOAEL by Poon et al. (1998) should more appropriately be 50 ppm. When regulatory criteria for antimony are established and/or reviewed, the findings in the NTP study and this critical reevaluation of the Poon et al. (1998) study should be considered when establishing a NOAEL for subchronic exposure to antimony in the future.
Assuntos
Tartarato de Antimônio e Potássio/toxicidade , Esquistossomicidas/toxicidade , Animais , Humanos , Nível de Efeito Adverso não ObservadoRESUMO
Oral rinse and toothpaste products (Viadent) containing Sanguinaria extract have been shown through extensive clinical trials to be effective against plaque build-up and gingivitis. To establish safety, a comprehensive research program was conducted, including a series of clinical studies and a number of animal studies to evaluate acute, subchronic, and chronic toxicity, and the potential for irritation of mucosal tissues. In 1990 and 1993, an Expert Panel reported on reviews of these data and concluded that Viadent products are safe for their intended use. Despite the large database of information to support the safety of Viadent products, Damm et al. (1999) recently raised the possibility that their usage may be causally associated with development of oral leukoplakia. However, a critique of this recent report shows that it does not fulfil criteria for establishing causation. In particular, the study does not show that exposure to Viadent preceded the onset of leukoplakia, it does not demonstrate dose-response or biological plausibility, and it suffers from selection and information bias and from potential confounding. Furthermore, upon critical evaluation, the Damm et al. (1999) report on a case-series is inconsistent with the weight of available clinical evidence showing that Sanguinaria extract-containing oral health care products cause no cytotoxic or significant irritant effects in the oral mucosa in human studies of up to 6 months duration. The animal data similarly do not support a causal association between Viadent usage and oral leukoplakia in humans. These data demonstrate that Sanguinaria extract and whole Viadent formulations are without significant irritation potential and have no effects on the oral mucosa, even in studies with life-long dietary exposure to Sanguinaria extract. The mutagenicity and genotoxicity data do not indicate that Sanguinaria extract or its components are genotoxic in vivo. The results of 2 GLP-compliant rat oncogenicity studies provide no evidence of any carcinogenic effect of Sanguinaria extract. In conclusion, the available clinical and animal data provide no support for and in fact argue strongly against the hypothesis that the use of Viadent toothpaste and/or oral rinse products may be causally associated with the development of leukoplakia in humans.
Assuntos
Alcaloides/efeitos adversos , Anti-Infecciosos/efeitos adversos , Leucoplasia Oral/induzido quimicamente , Antissépticos Bucais/efeitos adversos , Cremes Dentais/efeitos adversos , Alcaloides/toxicidade , Animais , Anti-Infecciosos/toxicidade , Benzofenantridinas , Humanos , Isoquinolinas , Antissépticos Bucais/toxicidade , Cremes Dentais/toxicidadeRESUMO
p-Dichlorobenzene (PDCB) has been reported to produce tumors in the male and female mouse liver and in the male rat kidney in 2-year gavage studies (NPT, 1987). To elucidate the possible mechanisms of carcinogenicity more fully, UDS and RDS were evaluated in B6C3F1 mouse hepatocytes and F-344 rat kidney cells by autoradiography following in vivo administration of PDCB. All corn oil gavage doses of PDCB (300, 600, and 1,000 mg/kg) and the negative control resulted in < 0 net grains/nucleus (NG) in the mouse liver and rat kidney, indicating that PDCB does not induce UDS in either tissue. Compared to controls with < or = 0.29% hepatocytes in S-phase (%S), treatment of mice induced 0.46, 1.90, and 1.52 %S (males) and 2.61, 1.18, and 4.45 %S (females), which indicates that PDCB acts as an inducer of cell proliferation in the liver. In male rat kidney cells, the same doses produced 0.87, 0.67, and 1.01 %S (0.38% in controls) and in females 0.48, 0.43, and 0.32 %S (0.52% in controls), indicating that PDCB induces cell replication in the male but not the female rat kidney. Therefore, these data demonstrate that PDCB is not genotoxic in the mouse liver or rat kidney at single oral doses comparable to the daily doses given in the National Toxicology Program (NTP) bioassay (NTP, 1987). Furthermore, the increases in RDS support the hypotheses that mouse liver tumor formation occurs via stimulation of hepatocyte proliferation and male rat kidney carcinogenesis via increased renal cell proliferation.
Assuntos
Clorobenzenos/toxicidade , Reparo do DNA , DNA/biossíntese , Inseticidas/toxicidade , Animais , Testes de Carcinogenicidade , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Clorobenzenos/administração & dosagem , Clorobenzenos/metabolismo , Feminino , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Masculino , Camundongos , Testes de Mutagenicidade , Ratos , Ratos Endogâmicos F344RESUMO
A workshop was held to discuss the uses of data on DNA adduct measurement in humans and in experimental systems in vitro and in vivo. The discussions focused principally on the understanding of the toxicological significance of DNA adducts as provided by information from animal models. An Expert Panel concluded that human DNA adduct data have utility in several aspects of risk assessment. The presence and amount of specific adducts that can be correlated with a chemical exposure are relevant for hazard identification and risk evaluation. Data from experimental systems have established dose-response relationships between the level of adducts and exposure, but these remain complex and depend on metabolic fate. Although structure-activity relationships have been useful retrospectively to explain the DNA-reactive nature of some chemicals or classes of chemicals, there are currently no means outside the laboratory to specifically predict the adduct-producing potency of a compound. Analysis of DNA adducts in tissues of laboratory animals and humans has revealed sensitive subpopulations, a finding that has important relevance for human risk assessment. Adduct analysis may be one of the best tools available to characterize exposures to DNA from complex mixtures for purposes of epidemiological investigation. Consensus statements were developed based on presentations by R. Gupta, W. Lutz, R. Nath, and B. Singer [see Regul. Toxicol. Pharmacol. 23(1), 1996] and subsequent discussions. First, rigorous scientific criteria should be met for the detection and characterization of specific DNA adducts in vitro and in target tissues in vivo. Second, the use of adduct data in risk extrapolation has the greatest value when there is characterization of adduct structure, an understanding of the role of repair in DNA adduct removal, and demonstration of biological relevance for each adduct. Third, the detection of DNA adducts in a tissue does not necessarily indicate a specific tumorigenic risk for that tissue. Fourth, the mutagenic potency for specific adducts varies by several orders of magnitude. Fifth, the role of DNA adducts induced by exogenous agents must be placed in perspective of endogenously produced adducts. The biological significance of a type of DNA adduct is related to several factors, including the efficiency of conversion to mutation, the amounts of similar endogenous adducts, and the variety of exogenous DNA adducts found in DNA from humans. The biological relevance of DNA adducts may be deduced from the dose-response relationships for adducts and tumors at physiologically relevant doses as well as from data showing mutations in targets such as oncogenes or tumor suppressor genes. There is convincing evidence in the literature for an association between some specific DNA adducts, mutation, and the carcinogenic process. As a general conclusion, the Panel suggested that the current technological capabilities for detection of DNA adducts exceed our ability to define the biological significance of adducts as it relates to toxicity or health outcome. DNA adducts are likely to play an important role in human risk for cancer induction and progression, but the quantitative aspects of this relationship remain to be determined.
Assuntos
Carcinógenos/efeitos adversos , Adutos de DNA/efeitos adversos , Dano ao DNA/efeitos dos fármacos , Reparo do DNA/genética , Animais , Adutos de DNA/análise , Adutos de DNA/metabolismo , Dieta/normas , Relação Dose-Resposta a Droga , Exposição Ambiental , Humanos , Técnicas In Vitro , Mutação/genética , Medição de Risco , Distribuição TecidualRESUMO
Traditional toxicological procedures have only limited application to the safety assessment of macronutrient substitutes. Experience indicates that spurious effects are often encountered when macronutrients or their replacements are fed to rodents at high dietary levels. These effects may results in nutritional imbalances that lead secondarily to adverse physiological consequences including cancer, renal disease, or reproductive effects. In approaching the safety assessment of macronutrient substitutes, consideration needs to be given to designing and implementing a safety assessment program which acknowledges the physical, chemical, and biological properties of the substance. Factors such as molecular size, physical state, solvent properties, hydrolysis potential, digestibility, absorption potential, and metabolic fate must be well established prior to selection of appropriate test models. Armed with this information, many potential undesirable physiological effects of the substances can be predicted, thus precluding the need for a full spectrum of animal testing. Predicted physiological and metabolic effects, however, should be characterized using in vitro methods and confirmed with in vivo models. Initial short-term toxicity screening tests with rodents should be carried out to identify unanticipated systemic toxicity. Testing in laboratory animals and trials in humans should then proceed with more appropriate models that are specially selected to assess the significance of predicted outcomes, to characterize dose-response relationships, and to identify possible needs to modify the product to mitigate adverse physiological consequences. These might include physical changes to alter particle size, chemical changes to modify digestibility, or nutrient supplementation to overcome impacts on nutrient availability. Thoughtful selection of appropriate and relevant models based on the physical, chemical, and biological properties of test substances will provide more rational approaches to safety assessments and avoid the pitfalls of routine application of traditional tests.
Assuntos
Alimentos Formulados/normas , Qualidade de Produtos para o Consumidor , Alimentos Formulados/efeitos adversosRESUMO
For decades, it has been known that a number of different factors (e.g., species, metabolism, age, animal husbandry, diet) may exhibit a significant modulating effect on the process of carcinogenesis. Often, however, these modulators have been largely uncontrolled and thus have made uncertain the results of many carcinogenicity bioassays. Fortunately, current research into molecular carcinogenesis is beginning to provide methods, not only to understand the molecular basis of known modulators of carcinogenesis, previously described only in empirical terms, but also to allow genetic modulation of carcinogenesis in experimental systems. An expanding body of knowledge regarding the role of oncogenes and tumor suppressor genes in neoplastic events is leading to a better understanding of carcinogenic mechanisms and points to the use of transgenic animal species in carcinogenicity bioassays. The transgenic animal provides methods to examine the molecular basis of carcinogenesis in experimental systems in addition to enhancing the sensitivity of carcinogen identification and the biological specificity of chemical risk extrapolation.
Assuntos
Testes de Carcinogenicidade , Modelos Animais de Doenças , Fatores Etários , Criação de Animais Domésticos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Geneticamente Modificados , Feminino , Genes Supressores de Tumor , Humanos , Masculino , Camundongos , Oncogenes , Ratos , Medição de Risco , Especificidade da EspécieRESUMO
3 chemicals were selected for mutagenicity testing from a priority list, based on production volume and available mutagenicity data. Propargyl alcohol (PA), 2-nitroaniline (2NA), and 5-methyl-1H-benzo-triazole (MBT) were selected for testing using the approach recommended in the Health Protection Branch Genotoxicity Guidelines. The battery of tests included the Salmonella/mammalian microsome mutation assay, the in vitro chromosomal aberration assay, and the bone-marrow micronucleus assay. The results indicate that 2 of the 3 chemicals, PA and 2NA, were clastogenic in vitro. Both PA and 2NA induced chromosomal aberrations in CHO cells in vitro with and without metabolic activation, while none induced reverse mutations detectable with the Salmonella/mammalian microsome assay. Because PA and 2NA were found to be in vitro clastogens, they also were tested in the mouse bone-marrow micronucleus assay. 2NA induced a small increase in micronuclei in males but not females. PA did not induce an increase in micronuclei.
Assuntos
Alcinos/toxicidade , Compostos de Anilina/toxicidade , Medula Óssea/efeitos dos fármacos , Aberrações Cromossômicas , Propanóis , Triazóis/toxicidade , 1-Propanol/toxicidade , Animais , Células CHO/efeitos dos fármacos , Aberrações Cromossômicas/genética , Cricetinae , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Testes para Micronúcleos , Testes de Mutagenicidade , Salmonella/efeitos dos fármacos , Salmonella/genéticaRESUMO
The term chlorinated chemicals is used to describe diverse groups of chemicals of varying chemical structure, including those used in water disinfection, as well as numerous aliphatic, aromatic, and polycyclic chlorinated substances. This report elaborates a number of scientific principles that govern the evaluation of the potential for chlorinated organic chemicals to cause adverse effects on the environment and to human health. The purpose of the report is to demonstrate the importance of applying these scientific principles in the evaluation of potential adverse effects of chlorinated organic chemicals. The four major principles upon which such a scientific analysis must be based are: (1) the fate and biological activity of a compound are determined by the chemical properties of the compound; (2) compounds do not show adverse effects below certain threshold concentrations, and the magnitude of response is related to dose; (3) inherent metabolic processes allow organisms to accommodate low doses of chlorinated organic chemicals; (4) observations associated with the presence of a certain compound must be biologically plausible effects, based on the specificity of the compound's activity in experimental systems. With respect to the first of these principles, there is abundant scientific evidence that the physical and chemical properties of chlorinated organic chemicals govern their bioaccumulative potential, toxicological properties, and thus their potential behavior and effects in the environment. Chemicals that have low solubility in water are highly lipophilic and have low vapor pressure, tend to accumulate in biological systems, and degrade slowly in the environment. Chlorinated organic chemicals that possess these characteristics include those having a carbon ring structure and multiple chlorine substitution. Other chlorinated organic chemicals with lesser degrees of chlorine substitution, such as 2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenol, chloroform, and dichloroethane, do not share the physical and chemical properties of the high molecular weight, cyclic, polychlorinated compounds and, as such, do not have the same potential to bioaccumulate. These differences among chlorinated organic chemicals with respect to their physical and chemical properties and behavior in the environment preclude the generalization that all organic chemicals containing chlorine behave similarly in the environment and act as persistent, bioaccumulative chemicals. The reactivity of chlorinated organic chemicals and hence their potential to produce biological effects depends on their specific molecular features. The substitution of chlorine into an organic molecule may increase or may reduce its biological activity.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Poluentes Ambientais/toxicidade , Hidrocarbonetos Clorados/toxicidade , Animais , Poluentes Ambientais/análise , Humanos , Hidrocarbonetos Clorados/químicaRESUMO
Concentrated organic residues extracted from 5 blended aliquots of commercial beers were evaluated for their ability to induce sister chromatid exchange (SCE), chromosomal aberrations and forward mutation in Chinese hamster ovary (CHO) cells. Each extract was prepared by blending 4 commercial beers of similar ingredients and brewing method, passing the beer pool over XAD-2 resin, extracting the resin and concentrating the extract. Studies were performed both with and without metabolic activation using variable amounts of reconstituted residues from 225-fold concentrates of the blended samples. CHO cultures were treated with 0.75 microliters/ml through 10.0 microliters/ml of the concentrates in the SCE assays, 1.0 microliters/ml through 10.0 microliters/ml of the extracts in the aberration assays and 2.5 microliters/ml up to 20 microliters/ml for forward mutation assays. In preliminary screening for SCE as an indicator of potential DNA damage, a significant increase was observed for 3 of 5 concentrated samples; however, no increase in SCE was induced by any of the 5 samples when S9 was added as a source of exogenous metabolic activation. More definitive tests for induction of genetic events, i.e., chromosome aberrations and forward HGPRT mutations, were negative for all 5 extracts whether or not S9 mix was present. Since SCE were not induced in tests with metabolic activation and since there was no concordant aberration or point mutation induction, the preliminary indication of potential DNA damage shown by elevated SCE under conditions without metabolic activation appears to have little biological significance.
Assuntos
Cerveja/toxicidade , Mutagênicos/toxicidade , Troca de Cromátide Irmã/efeitos dos fármacos , Animais , Biotransformação/genética , Células CHO , Cricetinae , Cricetulus , Dano ao DNA , Testes de MutagenicidadeRESUMO
Toluenesulfonhydrazide (TSH) is a high volume production chemical for which there is relatively little toxicological data. In this study, the mutagenic activity of TSH was determined in the Salmonella/mammalian microsome assay and the in vitro chromosomal aberration assay using Chinese hamster ovary cells. TSH induced gene mutations both with and without metabolic activation in the Salmonella/mammalian microsome assay but that it did not induce chromosomal aberrations in Chinese hamster ovary cells. The results of this study indicate that TSH is an in vitro mutagen and should be assessed for in vivo mutagenicity.
Assuntos
Aberrações Cromossômicas/genética , Mutagênicos/toxicidade , Mutação/efeitos dos fármacos , Compostos de Tosil/toxicidade , Animais , Biotransformação/efeitos dos fármacos , Células CHO , Células Cultivadas , Simulação por Computador , Cricetinae , Microssomos/efeitos dos fármacos , Microssomos/metabolismo , Testes de Mutagenicidade , Mutagênicos/administração & dosagem , Mutação/genética , Ratos , Ratos Sprague-Dawley , Salmonella/efeitos dos fármacos , Compostos de Tosil/administração & dosagemRESUMO
Results from a battery of short-term tests in vitro and in vivo used to assess the genotoxicity of caramel colours are presented and discussed in relation to reports from the literature. No evidence of genotoxicity was found in the Salmonella plate incorporation test using five standard strains or in the Saccharomyces cerevisiae gene conversion assay using strain D4, either with or without S-9 for activation. A weak clastogenic effect for a sample of Caramel Colour III in CHO cells was abolished in the presence of S-9. Two samples of Caramel Colour IV were not clastogenic in CHO cells. Salmonella pre-incubation tests without S-9 also failed to reveal any mutagenic activity for any of the caramel colours tested. The Caramel Colour III sample that showed clastogenic activity in CHO cells in vitro did not induce micronuclei when evaluated in a mouse bone marrow assay. These results are in general agreement with reports in the literature regarding the genotoxicity of caramel colours, and support the conclusion that caramel colours do not pose a genotoxic hazard to humans.
Assuntos
Cromossomos/efeitos dos fármacos , Corantes de Alimentos/toxicidade , Genes/efeitos dos fármacos , Mutagênicos/toxicidade , Animais , Células CHO , Doces , Carboidratos , Cricetinae , Feminino , Masculino , Camundongos , Testes para Micronúcleos , Testes de Mutagenicidade , Compostos Orgânicos , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Salmonella/efeitos dos fármacos , Salmonella/genéticaRESUMO
The most commonly used genotoxicity assays for cultured mammalian cells are mammalian cell mutagenesis, chromosome aberrations/SCE, hepatocyte UDS, and cell transformation. Since their inception, protocols for these assays have been modified in various laboratories. It has been observed that minor but potentially significant method modifications frequently remain unpublished (Swierenga et al., 1983) but should be considered in the development of recommended protocols. The present study was undertaken to determine the current 'state of the art' for these tests. Detailed questionnaires on culture conditions and testing protocols for both stock and test cell populations were designed with the assistance of an international advisory committee and sent to all research and contract laboratories that could be readily identified in Canada, U.S.A. and Europe. Responses from 425 completed questionnaires were analyzed to determine the most commonly used approach and modifications for each procedural step. As expected, the results show a large degree of interlaboratory variation. Detailed protocols for conducting each assay have been prepared and include: stepwise instructions, precautionary measures and practical solutions to common problems associated with each assay; recipes for media and solutions; formulas for quantifying genotoxic responses; reference lists of related assays; guidelines for interpretation; and discussions of the applications, advantages and disadvantages of each test.
Assuntos
Testes de Mutagenicidade/métodos , Projetos de Pesquisa , Animais , Cricetinae , Humanos , Camundongos , Ratos , Inquéritos e QuestionáriosRESUMO
A protocol based primarily on current laboratory practices in the performance of the unscheduled DNA synthesis (UDS) assay with primary rat hepatocyte cultures has been developed. These guidelines were developed using tabulated responses to a detailed questionnaire completed by North American and European governmental, university and contract laboratories involved with the UDS test. This report identifies those modifications to previously described methodologies which are used on a regular basis and also serves to clarify confusing or inconsistent practices. Although this protocol pertains specifically to the use of primary rat hepatocyte cultures, it can be modified to incorporate other types of cells in which certain aspects remain the same.
Assuntos
Replicação do DNA/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Projetos de Pesquisa , Animais , Células Cultivadas , Reparo do DNA , Técnicas In Vitro , Fígado/efeitos dos fármacos , Fígado/metabolismo , RatosRESUMO
Laboratory protocols and guidelines have been developed for the performance of point mutation assays using Chinese hamster ovary (CHO) cells, V79 cells, and L5178Y mouse lymphoma cells. Since only minor differences in the treatment of CHO and V79 cells exist, these two assays could be combined in one procedural guideline. A second protocol was developed for the mouse lymphoma assay in order to incorporate concerns and methods specific to that cell type and genetic locus. The protocols were based primarily on current laboratory practices as determined by responses to a detailed questionnaire completed by North-American and European governmental, university and contract laboratories involved with in vitro mutation testing. This report identifies those modifications to previously described methodologies which are being used on a regular basis, provides recommendations, and also serves to clarify confusing or inconsistent practices.
Assuntos
Testes de Mutagenicidade/métodos , Projetos de Pesquisa , Animais , Células Cultivadas , Cricetinae , Feminino , Técnicas In Vitro , Pulmão/citologia , Pulmão/efeitos dos fármacos , Linfoma/genética , Masculino , Camundongos , Ovário/citologia , Ovário/efeitos dos fármacosRESUMO
A standardized protocol and guidelines for the performance of cell transformation testing in mouse embryo (C3H/10T1/2), mouse fibroblast (BALB/c 3T3) and Syrian hamster embryo (SHE) cells have been developed. The protocol is based primarily on current laboratory practices as determined by responses to a detailed questionnaire completed by North American and European governmental, university and contract laboratories involved with cell transformation experimentation. This report identifies those modifications to previously described methodologies which are being used on a regular basis and also serves to clarify confusing or inconsistent practices.
Assuntos
Transformação Celular Neoplásica/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Projetos de Pesquisa , Animais , Células Cultivadas , Cricetinae , Camundongos , Camundongos Endogâmicos , Testes de Mutagenicidade/normasRESUMO
A recommended protocol has been developed for chromosomal aberration and sister-chromatid exchange assays in CHO, V79 and human lymphocyte cultures. The protocol was based on the responses to a detailed questionnaire completed by North-American and European governmental, university, and contract laboratories using these tests. This report identifies those modifications to previously described methods that are used on a regular basis and clarifies confusing or inconsistent practices. These protocols can be modified for use in other types of cells.
Assuntos
Aberrações Cromossômicas , Testes de Mutagenicidade/métodos , Projetos de Pesquisa , Troca de Cromátide Irmã , Animais , Células Cultivadas , Cricetinae , Feminino , Técnicas In Vitro , Pulmão/citologia , Linfócitos/citologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Testes de Mutagenicidade/normas , Ovário/citologiaRESUMO
The approach used in the survey of current methodologies used in mammalian cell genotoxicity testing (unscheduled DNA synthesis, mutation, cell transformation and cytogenetics testing) is discussed. The recommended protocols, described in the preceding papers, were developed using responses to detailed questionnaires. This summary outlines general observations related to the survey methodology and to the protocols themselves. Also discussed are the qualities of an effective survey; the evolution of a survey and of a protocol; and the contributions and limitations of this study.