Quantum Chemical Topology (QCT) descriptors, calculated from ab initio wave functions, have been utilised to model pKa and mutagenicity for data sets of pharmaceutically relevant compounds. The pKa of a compound is a pivotal property in both life science and chemistry since the propensity of a compound to donate or accept a proton is fundamental to understanding chemical and biological processes. The prediction of mutagenicity, specifically as determined by the Ames test, is important to aid medicinal chemists select compounds avoiding this potential pitfall in drug design. Carbocyclic and heterocyclic aromatic amines were chosen because this compounds class is synthetically very useful but also prone to positive outcomes in the battery of genotoxicity assays.The importance of pKa and genotoxic characteristics cannot be overestimated in drug design, where the multivariate optimisations of properties that influence the Absorption-Distribution-Metabolism-Excretion-Toxicity (ADMET) profiles now features very early on in the drug discovery process.Models were constructed using carboxylic acids in conjunction with the Quantum Topological Molecular Similarity (QTMS) method. The models produced Root Mean Square Error of Prediction (RMSEP) values of less than 0.5 pKa units and compared favourably to other pKa prediction methods. The ortho-substituted benzoic acids had the largest RMSEP which was significantly improved by splitting the compounds into high-correlation subsets. For these subsets, single-term equations containing one ab initio bond length were able to accurately predict pKa. The pKa prediction equations were extended to phenols and anilines.Quantitative Structure Activity Relationship (QSAR) models of acceptable quality were built based on literature data to predict the mutagenic potency (LogMP) of carbo- and heterocyclic aromatic amines using QTMS. However, these models failed to predict Ames test values for compounds screened at GSK. Contradictory internal and external data for several compounds motivated us to determine the fidelity of the Ames test for this compound class. The systematic investigation involved recrystallisation to purify compounds, analytical methods to measure the purity and finally comparative Ames testing. Unexpectedly, the Ames test results were very reproducible when 14 representative repurified molecules were tested as the freebase and the hydrochloride salt in two different solvents (water and DMSO). This work formed the basis for the analysis of Ames data at GSK and a systematic Ames testing programme for aromatic amines. So far, an unprecedentedly large list of 400 compounds has been made available to guide medicinal chemists. We constructed a model for the subset of 100 meta-/para-substituted anilines that could predict 70% of the Ames classifications. The experimental values of several of the model outliers appeared questionable after closer inspection and three of these have been retested so far. The retests lead to the reclassification of two of them and thereby to improved model accuracy of 78%. This demonstrates the power of the iterative process of model building, critical analysis of experimental data, retesting outliers and rebuilding the model.

Dietary factors have been implicated in the etiology of colon cancer. The salient components of high-risk diets are thought to be high intakes of meat, especially beef, and fat, especially animal fat, and low intakes of fiber. Low-risk diets are thought to be high in fiber, and low in meat and animal fat. The present study examines the effects of short-term consumption of diets hypothesized to increase or decrease the risk for colon cancer on mutagenic activity of feces. Whether the fecal mutagens responsible for the mutagenic activity observed in the study are directly involved in the etiology of colon cancer is not known. However, most known mutagens are potentially carcinogenic, and fecal mutagenic activity may be an indicator of risk for colon cancer. Six healthy adult subjects consumed the following diets in sequence a baseline diet for one week, a low-risk lacto-ovo vegetarian, high fiber diet for two weeks, and a high-risk, high meat, low fiber diet for two weeks. Quantitative daily food intake records were kept, and daily bowel habits were recorded. Fecal samples were collected at the end of each diet period. Analyses were performed of the diets for food and nutrient intake, and of feces for percent dry weight and pH. Mutagenic activity of the fecal samples was assayed using the fluctuation test for mutagens. The subjects' habitual diets, although omnivorous, were found to closely resemble a low-risk diet pattern. Analysis of the vegetarian and high meat diets confirmed that the subjects had consumed foods which respectively represented the components of high-risk and low-risk diets. The overall fecal mutagenic activity obtained with samples on the high meat diet was higher than with the vegetarian or baseline diets using Salmonella typhimurium TA 98 and TA 100. The trend towards higher mutagenicity on the high meat diet over the vegetarian diet was consistent for all six subjects using TA 100, and for five of the six using TA 98. The vegetarian and baseline diets resulted in similar overall mutagenic activity. Analysis of the fecal sample parameters using the Kruskal-Wallis one-way analysis of variance showed no significant differences among fecal samples from the three diet periods with respect to wet weight, dry weight, percent dry weight, pH or number of daily bowel movements. However, a sign-test analysis showed a significant trend (p<0.05) towards fewer bowel movements on the high meat diet than on the vegetarian diet. There were significant differences among subjects for all of the fecal sample parameters (p<0.01 or p<0.001). Spearman rank correlations were significantly positive between mutagenic activities using bacterial strains Salmonella typhimurium TA 98 and TA 100 for the baseline diet (p<0.01) and the vegetarian diet (p<0.05). There were also significant positive correlations (p<0.001) between pH and fecal mutagenicity on the high meat' diet using tester strain TA 100, and between wet weight and dry weight. The results of this study indicate that the overall mutagenic activity of human feces can be increased over a period of two weeks by the consumption of a diet high in meat and low in fiber, which is considered to be a high-risk diet for colon cancer. / Land and Food Systems, Faculty of / Graduate

Mutagens in feces have been suggested to be an indicator for risk of colon cancer. Groups consuming vegetarian diets are known to have lower mortality from colon cancer. The purpose of this study was to assess mutagenic activity in feces of persons habitually consuming vegetarian or non-vegetarian diets and to try to identify dietary factors or other health habits which contributed to fecal mutagenicity. Eleven strict vegetarians, six ovo-lacto vegetarians and twelve non-vegetarians, all from the Greater Vancouver area, participated in this study. Data on certain demographic variables and health habits, as well as dietary intake (food frequency and food records), were taken. One fecal sample was collected from each subject for the study. Aqueous extracts of the feces were prepared and analyzed for mutagens using the fluctuation test with Salmonella typhimurium TA100 and TA98. Levels of mutagenicity on each organism were then statistically correlated with frequency of consumption of food groups, nutrient intake, demographic data and health habits. Ovo-lacto vegetarians and strict vegetarians, as groups, had significantly lower levels of fecal mutagens than non-vegetarians in the TA100 assay. With TA98, only the strict vegetarians had lower levels of mutagens compared to the non-vegetarians. The presence of several different mutagenic compounds was indicated. Significant negative correlations were found with mutagenicity on TA98 for all subjects with the following dietary variables: fruits and juices, fiber and iron. Similar negative correlations were found for total carbohydrate and Southgate fiber intakes and mutagenicity on TA100. Within the group of non-vegetarians, there were negative correlations with mutagenicity on TA98 and total protein and with mutagenicity on TA100 and calcium. With the demographic variables and health habits, no clear pattern emerged to indicate factors which would predict lowered mutagenicity for all subjects. It is concluded that vegetarians have lower levels of fecal mutagenicity and that several dietary factors are likely to contribute to this phenomenon. / Land and Food Systems, Faculty of / Graduate

Orientadores: Nelma de Mello Silva Oliveira, Gisela de Aragão Umbuzeiro, Simone Valente Campos / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Tecnologia / Made available in DSpace on 2018-08-27T02:06:39Z (GMT). No. of bitstreams: 1 Resende_MariellyReis_M.pdf: 1924615 bytes, checksum: 1591718b0b86c23e64a29629b4589246 (MD5) Previous issue date: 2015 / Resumo: Embora tenha grande utilidade e diversas aplicações nos setores industriais, há anos a discussão sobre o potencial genotóxico do corante tartrazina vem sendo abordada, uma vez que há vários resultados controversos descritos na literatura. É provável que a presença de impurezas nas amostras possa ser uma das causas do possível potencial mutagênico. Dessa forma, esse estudo visa avaliar a atividade mutagênica do corante tartrazina com diferentes graus de pureza e possíveis interferentes presentes nas amostras, utilizando o ensaio Salmonella/microssoma a partir das linhagens recomendadas pela OECD 471. Os resultados obtidos demonstraram que o corante tartrazina ?99 % e o corante tartrazina comercial 90%, não apresentaram atividade mutagênica para as linhagens TA97a, TA98, TA100, TA1535 e TA102 demonstrando ausência de impurezas mutagênicas ou que as mesmas estejam em baixas concentrações nas amostras avaliadas / Abstract: Although very useful and diverse applications in industry, for years the discussion on the genotoxic potential of the dye tartrazine has been addressed, since there are several controversial results in the literature. It is likely that the presence of impurities in the samples may be a cause of the possible mutagenic potential. Thus, this study aims to evaluate the mutagenic activity of the dye tartrazine with different degrees of purity and possible interferences present in the samples, using the Salmonella / microsome test from the lines recommended by the OECD 471. The results showed that the dye tartrazine ?99 % and the dye tartrazine commercial 90% showed no mutagenic activity for the strains TA97, TA98, TA100, TA1535 and TA102 showing absence of mutagenic impurities or that they are in low concentrations in the analyzed samples / Mestrado / Tecnologia e Inovação / Mestra em Tecnologia

Made available in DSpace on 2016-01-26T18:55:37Z (GMT). No. of bitstreams: 1 Mariliza.pdf: 910731 bytes, checksum: 21aa8bdaf81f9c020e8cdee92ae30dcb (MD5) Previous issue date: 2013-03-26 / Studies with some local anesthetics showed that these can cause genetic damage. But local anesthetics has not been tested against the genotoxicity of repeated use. Objective: The aim of this study was to evaluate the genotoxic potential of local anesthetics with single dose and repeated, through the micronucleus test. Methods: We used 80 Wistar rats, male, 12 weeks old, divided into 6 groups: A - 16 rats received lidocaine hydrochloride intraperitoneally (4.4 mg / kg), B - 16 rats received 3% mepivacaine intraperitoneally (4.4 mg / kg), C - 16 rats received 4% articaine intraperitoneally (7.0 mg / kg) D - 16 rats received prilocaine 3% intraperitoneally (6.0 mg / kg) E - 08 rats received cyclophosphamide in single subcutaneous dose (50mg/kg) (positive control group) F - 08 rats received 0.5 ml of saline intraperitoneally (negative control group). Eight rats from Group A, B, C and D received the dose of anesthetic only once at the first day of the experiment and the remainder were dosed daily for five days. The Kruskal-Wallis test followed by a multiple comparisons Student-Newman-Keuls test was used for statistical analysis. Results: The median of micronuclei in the lidocaine group exposed for 1 day was 1.00 and by 5 days was 0.50, mepivacaine for 1 day and for 5 days was 1.00, articaine for 1 day was 1.00 and for 5 days 0.00, prilocaine for 1 day and 5 days was 0.00, cyclophosphamide was 10.00 and the negative control group exposed for 1 day was 1.00 and exposed for 5 days was 0.00 (p<0.0001). There was a statistical difference between the number of micronuclei in relation to the positive control group and all local anesthetics studied (related to the anesthetic type and duration of exposure) (p=0.0001) but not for the negative control group (p>0.05). Conclusion: There was no increase in micronuclei frequency in relation to exposure to 1 or 5 days in all local anesthetics evaluated. / Estudos com alguns anestésicos locais mostraram que estes podem causar dano genético. Porém ainda não foi testada a genotoxicidade frente ao uso repetido destes. Objetivo: O objetivo deste estudo foi avaliar o potencial genotóxico de anestésicos locais em dose única e repetida, por meio do teste de micronúcleo em ratos Wistar. Material e métodos: Foram utilizados 80 ratos Wistar, machos, com 12 semanas, divididos em 6 grupos: A 16 ratos receberam cloridrato de lidocaína via intraperitoneal (4,4 mg/kg); B 16 ratos receberam mepivacaína a 3% via intraperitoneal (4,4 mg/kg); C 16 ratos receberam articaína a 4% via intraperitoneal (7,0 mg/kg); D 16 ratos receberam prilocaína a 3% via intraperitoneal (6,0 mg/kg); E 08 ratos receberam ciclofosfamida em dose única subcutânea (50mg/kg) (grupo controle positivo); F 08 ratos receberam 0,5ml de soro fisiológico via intraperitoneal (grupo controle negativo). Oito ratos do grupo A, B, C e D receberam a dose do anestésico apenas uma vez no primeiro dia do experimento e os demais receberam doses diárias durante cinco dias. O teste de Kruskal-Wallis, seguido das comparações múltiplas de Student-Newman-Keuls, foi utilizado para análise estatística. Resultados: A mediana de micronúcleos no grupo exposto a Lidocaína por 1 dia foi de 1.00 e por 5 dias foi de 0.50, a Mepivacaína por 1 dia e por 5 dias foi de 1.00, a Articaína por 1 dia 1.00 e por 5 dias 0.00, a Prilocaína por 1 dia e por 5 dias 0.00, a ciclofosfamida foi 10.00 e no grupo de controle negativo exposto por 1 dia foi 1,00 e no exposto por 5 dias foi de 0,00 (p<0,0001). Houve diferença estatística entre o número de micronúcleos em relação ao grupo controle positivo e todos os anestésicos locais estudados (quanto ao tipo e tempo de exposição) (p=0,0001), porém não em relação ao grupo controle negativo (p>0,05). Conclusão: Não foi observado aumento da frequência de micronúcleos com relação à exposição de 1 ou 5 dias em todos os anestésicos locais avaliados.

Made available in DSpace on 2016-01-26T18:55:40Z (GMT). No. of bitstreams: 1 Gustavo Logar.pdf: 371330 bytes, checksum: 461b03ba76909955dcf6db3c8b004b60 (MD5) Previous issue date: 2014-05-23 / The Biodentine is a new material suitable for various clinical situations in dentistry. Despite being a promising material, there are few studies evaluating the characteristics of this material, especially its genotoxicity and mutagenicity. Objective: This study aims to evaluate the genotoxic and mutagenic potential of Biodentine "in vivo". Methods: We used 24 Wistar albino rats, males were divided into 3 groups: A - 8 rats where specimens of Biodentine measuring 2 mm in diameter x 10mm length on the dorsum were placed, B - 8 rats, which received cyclophosphamide in single subcutaneous dose (50mg/kg) on the first day of the experiment (positive control group), C - 8 rats where specimens measuring 2 mm in diameter x 10mm long without Biodentine on the dorsum were placed (negative control group). After 24 hours, all animals were euthanized and material from bone marrow of femurs was collected to perform the Comet assay and micronucleus test. Results: Biodentine showed levels of DNA damage (tail intensity %) in bone marrow cells of 23.57 ± 7.70, cyclophosphamide of 27,43 ± 7,40 and negative control of 24.75 ± 5 55 (p < 0.05). The average number of micronuclei in the exposed Biodentine group was 6.25 (standard deviation - SD = 3.53), in the group exposed to cyclophosphamide was 9.75 (SD = 2.49) and negative control group was 0.75 (SD = 1.03) (p < 0.0001). Conclusion: The Biodentine showed an increase in the frequency of micronuclei, but no genotoxicity effect by the Comet assay. / O Biodentine é um novo material indicado para diversas situações clínicas na odontologia. Apesar de ser um material promissor, ainda há poucos trabalhos avaliando as características deste material, em especial sua genotoxicidade e mutagenicidade. Objetivo: Este estudo visa avaliar o potencial genotóxico e mutagênico do Biodentine in vivo . Material e métodos: Foram utilizados 24 ratos Wistar albinos, machos, distribuídos em 3 grupos: A 8 ratos onde foram colocados corpos de prova medindo 2mm de diâmetro x 10mm de comprimento com Biodentine no subcutâneo do dorso; B 8 ratos, que receberam ciclofosfamida em dose única subcutânea (50mg/kg) no primeiro dia do experimento (grupo controle positivo); C 8 ratos onde foram colocados corpos de prova medindo 2mm de diâmetro x 10mm de comprimento sem Biodentine no subcutâneo do dorso (grupo controle negativo). Após 24 horas, todos os animais foram eutanasiados e material da medula óssea dos fêmures foi coletado para realização do Teste do Cometa e do Teste do micronúcleo. Resultados: O Biodentine apresentou níveis de danos no DNA (tail intensity %) em células da medula óssea de 23,57 ± 7,70, a ciclofosfamida de 27,43 ± 7,40 e o controle negativo de 24,75 ± 5,55 (p<0,05). A média de micronúcleos no grupo exposto ao Biodentine foi de 6,25 (desvio padrão DP= 3,53), no grupo exposto a ciclofosfamida foi de 9,75 (DP= 2,49) e no grupo de controle negativo foi 0,75 (DP= 1,03) (p<0,0001). Conclusão: O Biodentine apresentou aumento na frequência de micronúcleos, mas não apresentou efeito genotóxico observado pelo teste do Cometa.

Made available in DSpace on 2016-05-02T13:54:44Z (GMT). No. of bitstreams: 1 Dissertacao completa Patricia Scotini Freitas.pdf: 380463 bytes, checksum: 593d35f67c31f8a091c16952ef946b32 (MD5) Previous issue date: 2007-04-27 / Coordenacao de Aperfeicoamento de Pessoal de Nïvel Superior / Blueberry Vaccinium corymbosum is a vegetable very rich in anthocyanins which have strong antioxidant capacity and other potential health benefits and because of that is widely consumed in the world as medicinal plant In this work the mutagenic potential of the crude extract from this plant was studied in mice after acute treatment using the comet and micronucleus assay Animals were treated orally with three different concentrations of the extract (1000 1500 and 2000 mg/kg) Peripheral blood cells of Swiss mice were collected 4 and 24 hours after the treatment for the comet assay and 48 and 72 hours for the micronucleus test The results have shown that the extract of Vaccinium corymbosum did not induce statistically significant increases in the average number of damages to desoxyribonucleic acid in peripheral blood cells However a significant increase in the mean of the micronucleated polychromatic erythrocytes was observed at three tested doses It is suggested that its consumption could be moderate until a definitive risk for humans is established / Vaccinium corymbosum é um vegetal muito rico em antocianinas que tem uma grande capacidade antioxidante e outros potenciais benefícios à saúde e por este motivo é altamente utilizado no mundo inteiro como planta medicinal Neste trabalho foi analisado o potencial mutagênico da administração aguda do extrato bruto de frutos desta planta em células de camundongos utilizando o ensaio cometa e o teste do micronúcleo Os animais foram tratados oralmente com três diferentes concentrações do extrato (1000, 1500 e 2000 mg/kg de peso corporal) Células do sangue periférico de camundongos foram coletados 4 e 24 horas após o tratamento para a realização do ensaio cometa e 48 e 72 horas para o teste do micronúcleo Os resultados mostraram que o extrato de Vaccinium corymbosum não induziu aumentos estatisticamente significativos de danos ao ácido desoxirribonucléico nas células de sangue periférico Entretanto o teste do micronúcleo evidenciou um aumento significativo na média de eritrócitos policromáticos micronucleados nas três concentrações testadas Sugere-se que o consumo deste extrato seja moderado até que seu risco definitivo para os seres humanos seja melhor estabelecido

Made available in DSpace on 2016-07-18T17:53:11Z (GMT). No. of bitstreams: 1 Mariliza.pdf: 910731 bytes, checksum: 21aa8bdaf81f9c020e8cdee92ae30dcb (MD5) Previous issue date: 2013-03-26 / Studies with some local anesthetics showed that these can cause genetic damage. But local anesthetics has not been tested against the genotoxicity of repeated use. Objective: The aim of this study was to evaluate the genotoxic potential of local anesthetics with single dose and repeated, through the micronucleus test. Methods: We used 80 Wistar rats, male, 12 weeks old, divided into 6 groups: A - 16 rats received lidocaine hydrochloride intraperitoneally (4.4 mg / kg), B - 16 rats received 3% mepivacaine intraperitoneally (4.4 mg / kg), C - 16 rats received 4% articaine intraperitoneally (7.0 mg / kg) D - 16 rats received prilocaine 3% intraperitoneally (6.0 mg / kg) E - 08 rats received cyclophosphamide in single subcutaneous dose (50mg/kg) (positive control group) F - 08 rats received 0.5 ml of saline intraperitoneally (negative control group). Eight rats from Group A, B, C and D received the dose of anesthetic only once at the first day of the experiment and the remainder were dosed daily for five days. The Kruskal-Wallis test followed by a multiple comparisons Student-Newman-Keuls test was used for statistical analysis. Results: The median of micronuclei in the lidocaine group exposed for 1 day was 1.00 and by 5 days was 0.50, mepivacaine for 1 day and for 5 days was 1.00, articaine for 1 day was 1.00 and for 5 days 0.00, prilocaine for 1 day and 5 days was 0.00, cyclophosphamide was 10.00 and the negative control group exposed for 1 day was 1.00 and exposed for 5 days was 0.00 (p<0.0001). There was a statistical difference between the number of micronuclei in relation to the positive control group and all local anesthetics studied (related to the anesthetic type and duration of exposure) (p=0.0001) but not for the negative control group (p>0.05). Conclusion: There was no increase in micronuclei frequency in relation to exposure to 1 or 5 days in all local anesthetics evaluated. / Estudos com alguns anestésicos locais mostraram que estes podem causar dano genético. Porém ainda não foi testada a genotoxicidade frente ao uso repetido destes. Objetivo: O objetivo deste estudo foi avaliar o potencial genotóxico de anestésicos locais em dose única e repetida, por meio do teste de micronúcleo em ratos Wistar. Material e métodos: Foram utilizados 80 ratos Wistar, machos, com 12 semanas, divididos em 6 grupos: A 16 ratos receberam cloridrato de lidocaína via intraperitoneal (4,4 mg/kg); B 16 ratos receberam mepivacaína a 3% via intraperitoneal (4,4 mg/kg); C 16 ratos receberam articaína a 4% via intraperitoneal (7,0 mg/kg); D 16 ratos receberam prilocaína a 3% via intraperitoneal (6,0 mg/kg); E 08 ratos receberam ciclofosfamida em dose única subcutânea (50mg/kg) (grupo controle positivo); F 08 ratos receberam 0,5ml de soro fisiológico via intraperitoneal (grupo controle negativo). Oito ratos do grupo A, B, C e D receberam a dose do anestésico apenas uma vez no primeiro dia do experimento e os demais receberam doses diárias durante cinco dias. O teste de Kruskal-Wallis, seguido das comparações múltiplas de Student-Newman-Keuls, foi utilizado para análise estatística. Resultados: A mediana de micronúcleos no grupo exposto a Lidocaína por 1 dia foi de 1.00 e por 5 dias foi de 0.50, a Mepivacaína por 1 dia e por 5 dias foi de 1.00, a Articaína por 1 dia 1.00 e por 5 dias 0.00, a Prilocaína por 1 dia e por 5 dias 0.00, a ciclofosfamida foi 10.00 e no grupo de controle negativo exposto por 1 dia foi 1,00 e no exposto por 5 dias foi de 0,00 (p<0,0001). Houve diferença estatística entre o número de micronúcleos em relação ao grupo controle positivo e todos os anestésicos locais estudados (quanto ao tipo e tempo de exposição) (p=0,0001), porém não em relação ao grupo controle negativo (p>0,05). Conclusão: Não foi observado aumento da frequência de micronúcleos com relação à exposição de 1 ou 5 dias em todos os anestésicos locais avaliados.

Made available in DSpace on 2016-07-18T17:53:13Z (GMT). No. of bitstreams: 1 Gustavo Logar.pdf: 371330 bytes, checksum: 461b03ba76909955dcf6db3c8b004b60 (MD5) Previous issue date: 2014-05-23 / The Biodentine is a new material suitable for various clinical situations in dentistry. Despite being a promising material, there are few studies evaluating the characteristics of this material, especially its genotoxicity and mutagenicity. Objective: This study aims to evaluate the genotoxic and mutagenic potential of Biodentine "in vivo". Methods: We used 24 Wistar albino rats, males were divided into 3 groups: A - 8 rats where specimens of Biodentine measuring 2 mm in diameter x 10mm length on the dorsum were placed, B - 8 rats, which received cyclophosphamide in single subcutaneous dose (50mg/kg) on the first day of the experiment (positive control group), C - 8 rats where specimens measuring 2 mm in diameter x 10mm long without Biodentine on the dorsum were placed (negative control group). After 24 hours, all animals were euthanized and material from bone marrow of femurs was collected to perform the Comet assay and micronucleus test. Results: Biodentine showed levels of DNA damage (tail intensity %) in bone marrow cells of 23.57 ± 7.70, cyclophosphamide of 27,43 ± 7,40 and negative control of 24.75 ± 5 55 (p < 0.05). The average number of micronuclei in the exposed Biodentine group was 6.25 (standard deviation - SD = 3.53), in the group exposed to cyclophosphamide was 9.75 (SD = 2.49) and negative control group was 0.75 (SD = 1.03) (p < 0.0001). Conclusion: The Biodentine showed an increase in the frequency of micronuclei, but no genotoxicity effect by the Comet assay. / O Biodentine é um novo material indicado para diversas situações clínicas na odontologia. Apesar de ser um material promissor, ainda há poucos trabalhos avaliando as características deste material, em especial sua genotoxicidade e mutagenicidade. Objetivo: Este estudo visa avaliar o potencial genotóxico e mutagênico do Biodentine in vivo . Material e métodos: Foram utilizados 24 ratos Wistar albinos, machos, distribuídos em 3 grupos: A 8 ratos onde foram colocados corpos de prova medindo 2mm de diâmetro x 10mm de comprimento com Biodentine no subcutâneo do dorso; B 8 ratos, que receberam ciclofosfamida em dose única subcutânea (50mg/kg) no primeiro dia do experimento (grupo controle positivo); C 8 ratos onde foram colocados corpos de prova medindo 2mm de diâmetro x 10mm de comprimento sem Biodentine no subcutâneo do dorso (grupo controle negativo). Após 24 horas, todos os animais foram eutanasiados e material da medula óssea dos fêmures foi coletado para realização do Teste do Cometa e do Teste do micronúcleo. Resultados: O Biodentine apresentou níveis de danos no DNA (tail intensity %) em células da medula óssea de 23,57 ± 7,70, a ciclofosfamida de 27,43 ± 7,40 e o controle negativo de 24,75 ± 5,55 (p<0,05). A média de micronúcleos no grupo exposto ao Biodentine foi de 6,25 (desvio padrão DP= 3,53), no grupo exposto a ciclofosfamida foi de 9,75 (DP= 2,49) e no grupo de controle negativo foi 0,75 (DP= 1,03) (p<0,0001). Conclusão: O Biodentine apresentou aumento na frequência de micronúcleos, mas não apresentou efeito genotóxico observado pelo teste do Cometa.

Made available in DSpace on 2016-07-18T17:53:15Z (GMT). No. of bitstreams: 1 Dayane.pdf: 321360 bytes, checksum: c2b06ee1c47438b8e086585e5a6fc8c5 (MD5) Previous issue date: 2015-09-16 / Background: Pesticides are substances used for pest control. Because of their persistence in the environment, they can induce toxicity in humans and animals. Indoxacarb is an oxadiazine insecticide that acts against insects of the order Lepidoptera. Furthermore, indoxacarb has demonstrated strong insecticide activity against fleas; therefore, it is used in ectoparasite treatment. Choice of species used in this study (rats and cats) is based on several factors such as these animals have a high count of micronucleated polychromatic erythrocytes, thereby facilitating the execution of the chosen assay; rats are the standard animal model for mutagenicity studies; and the product is intended for use in cats. Limited data in literature pertaining to the mutagenic effects of this product motivated this study. The aim of this study was to evaluate the mutagenicity of indoxacarb when administered in one and tenfold therapeutic doses in rats and cats. This evaluation was performed using the micronucleus test. Materials, Methods & Results: Forty male Wistar rats aged 70 days and weighing 280 ± 10 g and 20 mixed breed male and female adult cats weighing 4 ± 0.2 kg were selected. These animals were obtained from the central animal facility and cattery, respectively, of the university of origin. Rats were reared in individual cages with a controlled temperature of 22°C ± 2°C, humidity of 55% ± 5% and 12-h light dark cycle. Cats were reared in individual stalls with water and food ad libitum. Animals were randomly distributed into four groups comprising 10 rats and 5 cats in respective groups: negative control group, which received 0.9% sodium chloride solution as a single topical administration; positive control group, which received 50 mg/kg cyclophosphamide as a single intra-peritoneal (rats) or intravenous injection (cats); indoxacarb group, which received indoxacarb as a single topical dose according to the manufacturer s recommendations; and high dose indoxacarb group, which also received indoxacarb in a single topical dose, but at a tenfold concentration. Rats were evaluated 24 h after indoxacarb administration. After euthanasia, rat femurs were obtained and their medullary canals were washed using fetal bovine serum. The content was centrifuged and used for smear preparations. Cats were evaluated using the micronucleus test before and 24 h after indoxacarb administration. For smear preparations, a drop of peripheral blood samples was obtained from the tail tip. Smear fixation, staining, and microscopic analysis were similar for cats and rats. Statistical analyses were performed using analysis of variance with contrast through Tukey s method and paired t test to compare time points. The significance level was 5%. Indoxacarb showed no mutagenic activity in the two species and doses evaluated. Even in animals subjected to high doses, the micronuclei count remained within the normal range (p > 0.05). Discussion: The results of this study corroborate those of other studies, where in the mutagenic capacity of indoxacarb was similar in different animal species and product delivery routes. In addition, it is of note the speed and ease of micronucleus testing, which makes it an important mutagenicity bioassay. / Introdução: Os pesticidas são venenos intencionalmente dispersos no ambiente para o controle de pragas e, com sua persistência nesse meio podem induzir a toxicidade em seres humanos e animais. Indoxacarbe é um inseticida, oxidiazínico, que apresenta atividade sobre insetos da ordem lepidóptera. No tratamento contra pulgas em pequenos animais, o indoxacarbe tem demonstrado uma alta atividade inseticida associada a esses ectoparasitas. A escolha da utilização dessas duas espécies animais neste estudo se deu pelo fato de que ratos e gatos apresentam uma grande quantidade de eritrócitos policromáticos micronucleados, facilitando desta forma a realização do ensaio escolhido e, pelo rato ser o modelo biológico padrão para avaliações de mutagenicidade, e por fim pelo fato do produto ser indicado para o uso em gatos. A escassez na literatura sobre o efeito mutagênico causado pelo uso desse produto levou a realização dessa pesquisa, cujo objetivo foi avaliar por meio do teste de micronúcleo a mutagenicidade do indoxacarbe em dose terapêutica e em dez vezes maior em ratos e gatos. Materiais, métodos e resultados: Foram selecionados 40 ratos, com 70 dias de idade, da linhagem Wistar, machos, peso de 280g±10 e 20 gatos adultos, sem raça definida, machos e fêmeas, com peso de 4kg±200gr, ambos do biotério central e gatil da universidade de origem, respectivamente. Os ratos foram mantidos em caixas individuais em ambiente com controle de temperatura 22ºC±2, umidade 55%±5, fotoperíodo 12 horas claro e escuro, os gatos foram mantidos em baias individuais, e ambos com água e ração ad libitum. Os animais foram distribuídos aleatoriamente em quatro grupos, com 10 animais em cada grupo de ratos e 5 animais nos grupos compostos por gatos, e foram tratados com: Grupo controle negativo (GCN)-solução de cloreto de sódio 0,9% por via tópica, em dose única. Grupo controle positivo (GCP)-ciclofosfamida em única dose de 50mg/kg por via intraperitoneal nos ratos e intravenosa nos gatos. Grupo indoxacarbe (GI)- indoxacarbe por via tópica, em dose única recomendado pelo fabricante. Grupo indoxacarbe dose alta (GIDA)-indoxacarbe por via tópica, em dose única, porém dez vezes maior que a dose recomendada pelo fabricante. Os ratos foram avaliados 24 horas após o uso do indoxacarbe, após eutanásia e retirada do fêmur para a realização da lavagem do canal medular com soro fetal bovino, posterior centrifugação, retirada do sobrenadante e realização do esfregaço. Os gatos foram avaliados por meio do teste de micronúcleo antes e 24 horas após a aplicação do indoxacarbe, através da coleta de uma gota de sangue periférico da extremidade do rabo para a realização do esfregaço. A fixação, coloração e leitura das lâminas foram semelhantes para os ratos e gatos. Os testes estatísticos utilizados foram de análise de variância com contraste pelo método de Tukey e, para a comparação entre momentos utilizou-se teste t pareado. O nível de significância adotado foi de 5%. Indoxacarbe não apresentou atividade mutagênica em nenhuma das espécies e doses estudadas, mesmo nos animais submetidos a altas doses, a quantidade de micronúcleos permaneceu dentro da normalidade (p>0,05).Discussão: Os resultados obtidos neste estudo corroboraram com os poucos relatos encontrados na literatura, afirmando a ausência da capacidade mutagênica do indoxacarbe em diferentes espécies animais e vias de administração do produto. Observou-se também a rapidez, e facilidade na realização do teste para micronúcleo, caracterizando-o como um importante bioensaio para avaliações de mutagenicidade.