REGIANE MARTINS
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Artigo IPEN-doc 27057 Evaluation of cytotoxicity through MTS test of 2-ACBs (2-dDCB AND tDCB) after exposure to hepatic cells2019 - BARBEZAN, A.B.; CARVALHO, L.R.; VIEIRA, D.P.; MACHADO-SANTELLI, G.M.; MARTINS, R.; VILLAVICENCIO, A.L.C.H.Food irradiation has been approved in more than 60 countries for many applications in a wide variety of foods. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products formed when foods containing fatty acids are irradiated. Despite the importance of food irradiation, the toxicological potential of 2-ACBs in irradiated food is still no fully understood. In this study we investigated the cytotoxic effects of irradiated palmitic and stearic fatty acids byproducts, 2-dDCB and 2-tDCB, in hepatic cell lines (HepG2, BRL3A and HTC). The cytotoxic effects of 2-dDCB and 2-tDCB were evaluated at 100, 300 and 500 μM for 24 and 48 hours and the cell viability was measured using the MTS assay. While no toxicity was observed for 2-tDCB in all cells for all tested conditions, 2-dDCB was found to be toxic to BRL3A cells (at 100 μM after 48 hours) and HTC cells (at 24 hours in all tested concentrations). HepG2 cells on the other hand, were found to be resistant to 2-dDCB-induced toxicity. Overall our data shows that the byproduct 2-tDCB is not toxic for hepatic cells while 2-dDCB can be used has a potential marker for food irradiation-induced toxicity.Artigo IPEN-doc 27906 In vivo genotoxicity of 2-alkylcyclobutanones in liver cells from rats fed with irradiated cocoa butter using flow cytometry2021 - MARTINS, R.; VIEIRA, D.P.; CARVALHO, L.R.; BARBEZAN, A.B.; VILLAVICENCIO, A.L.C.H.Food irradiation proves to be an effective technique of eliminating some pathogens from food and this has gained significant attention to its potential for food safety. Since 1990, studies on the toxicological safety of 2-Alkylcyclobutanones have been conducted extensively. 2-Alkylcyclobutanones are unique radiolytic products generated by the radiation-induced breakage of triglycerides in food, 2-dodecylcyclobutanone (2-dDCB) and 2-tetradecylcyclobutanone (2-tDCB) are the predominant compounds detected . Despite studies showing non-genotoxicity of 2-ACBs (2-Alkylcyclobutanones), the results are conflicting and therefore we continue the studies in order to confirm the compounds are safety for human health. In vivo micronucleus test were performed to verify the 2-ACBs genotoxic effects in hepatic cells using flow cytometry. We used cocoa butter irradiated with 20 kGy. A group with animals was treated with daily intake of irradiated cocoa butter, synthesized 2-dDCB and 2-tDCB for one month. Hepatic cells were selected for genotoxicity analysis due to the liver importance in the compounds metabolization. The improvement of the analytical techniques is important for the research future since the irradiation process is already consolidated. The results confirmed the safety of the food irradiation process, as they did not indicate the genotoxic potential of the samples.Artigo IPEN-doc 26253 In vivo genotoxicity of 2-Alkylcyclobutanones in liver cells from rats fed with irradiated cocoa butter using flow citometry2019 - MARTINS, REGIANE; VIEIRA, DANIEL P.; CARVALHO, LUMA R. de; BARBEZAN, ANGELICA B.; VILLAVICENCIO, ANNA L.C.H.Food irradiation proves to be an effective technique of eliminating some pathogens from food and this has gained significant attention to its potential for food safety. Since 1990, studies on the toxicological safety of 2-Alkylcyclobutanones have been conducted extensively. 2- Alkylcyclobutanones are unique radiolytic products generated by the radiation-induced breakage of triglycerides in food, are exclusively found in irradiated lipid containing foods. 2-dodecylcyclobutanone (2-dDCB) and 2-tetradecylcyclobutanone (2-tDCB) are the predominant compounds detected in irradiated food. Despite studies showing non- genotoxicity of 2-ACBs (2-Alkylcyclobutanones), the results are conflicting and therefore we continue the studies in order to confirm the compounds safety for human health. In vivo micronucleus test were performed to verify the 2-ACBs genotoxic effects in hepatic cells using flow citometry. We used cocoa butter irradiated with 20 kGy at IPEN GAMACELL. A group with animals (IPEN Ethical Animal Experimentation Committee, process number 148/14) was treated with daily intake of irradiated cocoa butter, synthesized 2- Dodecylcyclobutanone and 2-Tetradecylcyclobutanone for two months. Hepatic cells were selected for genotoxicity analysis due to the liver importance in the compounds metabolization. Analyzes were made by micronucleus test with specific cells extracted from hepatic tissue using flow cytometry, which is an alternative to conventional techniques, allowing faster analysis and reduction in the animals number that is a subject much approached in research today. The improvement of the analytical techniques is important for the research future since the irradiation process is already consolidated. The results confirmed the safety of the food irradiation process, as they did not indicate the genotoxic potential of the samples.Artigo IPEN-doc 24270 In vivo evaluation of genotoxic potential of liver cells from rats fed with irradiated diet using flow cytometry2017 - MARTINS, R.; CARVALHO, L.R. de; VIEIRA, D.P.; BARBEZAN, A.B.; VILLAVICENCIO, A.L.C.H.Radiation sources became widely available since 1960’s, and between its main uses are the applications in food irradiation and research of effects of ionizing radiation on food products. Despite some public concern, the process is safe, free from chemical residues and presents advantages for preservation and storage. Nevertheless, safety dose parameters must be adopted in irradiation procedures to inhibit formation of undesirable and/or toxic products, for example, 2-ACB’s (2-alkylcyclobutanones) that are cyclic compounds containing four carbon rings that can be formed in food when its fat content is irradiated through breakdown of fatty acids. 2-ACBs are considered a unique class of compounds due to divergences between results of its mutagenicity potential collected from different studies. In this study, a cell population collected from rat livers were chosen for in vivo genotoxicity analysis because the importance of the liver in the metabolization of compounds. Analysis was performed using the micronuclei test using flow cytometry, allowing faster analysis, use of few materials and reduction in the number of animals, what is a subject much addressed currently in research. Irradiated rat diet did not show any genotoxic effect on liver cell populations. The improvement of the techniques is important for the future of the research since the irradiation process is already consolidated.