Orientador: Mariana Gobbo Braz / Resumo: A carência e a divergência de dados sobre o potencial genotóxico e mutagênico da exposição ocupacional aos anestésicos inalatórios mais modernos e mais utilizados atualmente estimularam a realização da presente pesquisa, que teve como objetivos avaliar os danos sistêmicos no DNA e os diferentes tipos celulares e biomarcadores de instabilidade genômica em células esfoliadas da mucosa oral em anestesiologistas. O estudo foi realizado em 60 médicos que atuavam no Hospital das Clínicas da Faculdade de Medicina de Botucatu – UNESP, os quais foram alocados em dois grupos: exposto, constituído por anestesiologistas expostos aos resíduos de gases anestésicos (isoflurano, sevoflurano, desflurano e óxido nitroso – N2O) e controle, constituído por médicos não expostos ocupacionalmente, os quais foram pareados por idade, sexo e estilo de vida com o grupo exposto. Amostras de sangue e de células esfoliadas orais foram coletadas de todos os participantes; os danos basais no DNA foram avaliados em linfócitos periféricos pelo teste do cometa e o teste do micronúcleo (MN) bucal foi utilizado para avaliação de diferentes parâmetros em células esfoliadas orais. As concentrações de N2O e dos halogenados foram mensuradas nas salas de operação com a utilização de espectrofotômetro infravermelho portátil. As concentrações médias de resíduos de gases anestésicos para todos os halogenados foi maior que 5 partes por milhão (ppm) e superior a 150 ppm para o N2O. Não houve diferença significativa entre ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Due to the lack and controversial data on the genotoxic and mutagenic potential of occupational exposure to the modern and most used inhaled anesthetics, the aim of the current study was to evaluate the systemic DNA damage, and the different cell types and genomic instability biomarkers in buccal exfoliated cells in anesthesiologists. The study was conducted in 60 physicians who worked in the Hospital of Botucatu Medical School - UNESP, and were allocated in two groups: the exposed group, consisted of anesthesiologists exposed to waste anesthetic gases (isoflurane, sevoflurane, desflurane and nitrous oxide - N2O) at least two years, and the control group consisted of non-exposed physicians, who were matched by age, sex and lifestyle with the exposed group. Venous blood and buccal exfoliated cells samples were collected from all participants; basal DNA damage was evaluated in peripheral lymphocytes by the comet assay, and the buccal micronucleus (MN) cytome (BMCyt) assay was used to evaluate different parameters in oral cells. The concentrations of N2O and halogenated anesthetics were measured in the operating rooms, by using of portable infrared spectrophotometer. The average concentrations of waste gases were greater than 5 parts per million (ppm) for all the halogenated anesthetics and more than 150 ppm for N2O. There was no significant difference between the groups in relation to DNA damage and there was no influence of the variables age, sex and time of exposure in the co... (Complete abstract click electronic access below) / Mestre

Submitted by LORENA MENDES DE CARVALHO LUCIO null (limcarvalho@yahoo.com.br) on 2016-09-12T13:56:26Z No. of bitstreams: 1 TESE FINAL LORENA POS BANCA Repositorio UNESP 12.09.16.pdf: 1673690 bytes, checksum: 578ad77e846c1bc6a83776f9b4d1b8bf (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-09-14T18:44:12Z (GMT) No. of bitstreams: 1 lucio_lmc_dr_bot.pdf: 1673690 bytes, checksum: 578ad77e846c1bc6a83776f9b4d1b8bf (MD5) / Made available in DSpace on 2016-09-14T18:44:12Z (GMT). No. of bitstreams: 1 lucio_lmc_dr_bot.pdf: 1673690 bytes, checksum: 578ad77e846c1bc6a83776f9b4d1b8bf (MD5) Previous issue date: 2016-08-25 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O presente estudo objetivou determinar as concentrações dos resíduos de gases anestésicos (RGA) em salas de operação (SO) e o impacto dessa exposição ocupacional em relação aos danos genômicos e estresse oxidativo em profissionais recém-expostos. O estudo foi conduzido no Hospital das Clínicas, Faculdade de Medicina de Botucatu-UNESP. As concentrações de isoflurano, sevoflurano e óxido nitroso (N2O) foram medidas nas SO por espectrofotometria infravermelha, com equipamento portátil. Sessenta e três médicos residentes, ao final de três anos do Programa de Residência Médica, foram alocados em dois grupos: exposto (Anestesiologia e Cirurgia, n=32) e controle (Clínica Médica, n=31). Amostras de sangue periférico e células bucais foram coletadas e protegidas da luz. Avaliaram-se danos no material genético (teste do cometa - danos basais, purinas e pirimidinas oxidadas em linfócitos; 8-hidroxi-2’-desoxiguanosina no plasma), micronúcleo (MN) em células bucais, proteínas carboniladas, marcadores de lipoperoxidação (malonaldeído e 4-hidroxinonenal) e capacidade antioxidante plasmática (ferric reducing antioxidant power, oxygen radical absorbance capacity e total antioxidant performance). As concentrações médias dos RGA foram superiores aos limites internacionalmente recomendados (2,7 vezes: isoflurano; 4,9 vezes: sevoflurano; 7,2 vezes: N2O). Os grupos não diferiram quanto aos dados demográficos (p>0,05). Detectou-se aumento significativo de danos basais no DNA (p=0,01) e maior frequência de MN em células bucais (2,3 vezes; p=0,07) no grupo exposto em relação ao grupo controle, mas não houve diferença (p>0,05) entre os grupos em relação a todos os marcadores de estresse oxidativo. Em conclusão, o estudo mostra que médicos residentes expostos a altas concentrações de RGA apresentam aumento significativo de danos sistêmicos no DNA e frequência aumentada de instabilidade genômica (MN bucais), mas não de estresse oxidativo. Portanto, este estudo mostra que jovens profissionais já apresentam alterações genômicas, o que reforça a importância do biomonitoramento e da diminuição da exposição aos RGA. / This study determined the waste anesthetic gases (WAG) in operating rooms (OR) and evaluated the impact of the occupational exposure in genetic damage and oxidative stress in medical residents. The study was performed at “Hospital das Clínicas, Faculdade de Medicina de Botucatu-UNESP”. The concentrations of isoflurane, sevoflurane and nitrous oxide (N2O) were measured in ORs. Sixty-three medical residents completing their three-year Medical Residency Program were recruited for the study and were assigned to two groups: exposed group (n=32) of Anesthesiology and Surgery areas and control group (n=31) of Internal Medicine area. Blood and buccal cells were concomitantly collected from both groups and protected from light to measure genetic instability by buccal micronucleus (MN), basal and oxidized DNA damage (comet assay and 8-hydroxy-2′-deoxyguanosine), biomarkers of protein and lipid oxidation, and three different assays for plasma antioxidant activity. Mean WAG concentrations were above international thresholds (2.7-fold: isoflurane; 4.9-fold: sevoflurane; 7.2-fold: N2O). There was no significant difference between groups regarding demographic data. Basal DNA damage (p=0.01) and buccal MN frequency (by 2.3-fold; p=0.07) were increased in the exposed group compared to the control group. Results showed no significant difference for oxidative stress biomarkers between groups. In conclusion, this study shows that medical residents exposed to high WAG concentrations have increased systemic DNA damage and genomic instability (buccal MN), but not oxidative stress. Thus, these young professionals already have genetic damage in the beginning of their career. Our results reinforce the importance of the biomonitoring and also the adequate measures to decrease ambient air pollution in the OR. / FAPESP: 2013/21130-0 / CNPq: 472453/2013-0 / CAPES/PGCI: 14527-13-8

Orientador: Leandro Gobbo Braz / Resumo: O presente estudo objetivou determinar as concentrações dos resíduos de gases anestésicos (RGA) em salas de operação (SO) e o impacto dessa exposição ocupacional em relação aos danos genômicos e estresse oxidativo em profissionais recém-expostos. O estudo foi conduzido no Hospital das Clínicas, Faculdade de Medicina de Botucatu-UNESP. As concentrações de isoflurano, sevoflurano e óxido nitroso (N2O) foram medidas nas SO por espectrofotometria infravermelha, com equipamento portátil. Sessenta e três médicos residentes, ao final de três anos do Programa de Residência Médica, foram alocados em dois grupos: exposto (Anestesiologia e Cirurgia, n=32) e controle (Clínica Médica, n=31). Amostras de sangue periférico e células bucais foram coletadas e protegidas da luz. Avaliaram-se danos no material genético (teste do cometa - danos basais, purinas e pirimidinas oxidadas em linfócitos; 8-hidroxi-2’-desoxiguanosina no plasma), micronúcleo (MN) em células bucais, proteínas carboniladas, marcadores de lipoperoxidação (malonaldeído e 4-hidroxinonenal) e capacidade antioxidante plasmática (ferric reducing antioxidant power, oxygen radical absorbance capacity e total antioxidant performance). As concentrações médias dos RGA foram superiores aos limites internacionalmente recomendados (2,7 vezes: isoflurano; 4,9 vezes: sevoflurano; 7,2 vezes: N2O). Os grupos não diferiram quanto aos dados demográficos (p>0,05). Detectou-se aumento significativo de danos basais no DNA (p=0,01) e maior frequê... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study determined the waste anesthetic gases (WAG) in operating rooms (OR) and evaluated the impact of the occupational exposure in genetic damage and oxidative stress in medical residents. The study was performed at “Hospital das Clínicas, Faculdade de Medicina de Botucatu-UNESP”. The concentrations of isoflurane, sevoflurane and nitrous oxide (N2O) were measured in ORs. Sixty-three medical residents completing their three-year Medical Residency Program were recruited for the study and were assigned to two groups: exposed group (n=32) of Anesthesiology and Surgery areas and control group (n=31) of Internal Medicine area. Blood and buccal cells were concomitantly collected from both groups and protected from light to measure genetic instability by buccal micronucleus (MN), basal and oxidized DNA damage (comet assay and 8-hydroxy-2′-deoxyguanosine), biomarkers of protein and lipid oxidation, and three different assays for plasma antioxidant activity. Mean WAG concentrations were above international thresholds (2.7-fold: isoflurane; 4.9-fold: sevoflurane; 7.2-fold: N2O). There was no significant difference between groups regarding demographic data. Basal DNA damage (p=0.01) and buccal MN frequency (by 2.3-fold; p=0.07) were increased in the exposed group compared to the control group. Results showed no significant difference for oxidative stress biomarkers between groups. In conclusion, this study shows that medical residents exposed to high WAG concentrations have increase... (Complete abstract click electronic access below) / Doutor

Made available in DSpace on 2014-08-20T14:30:17Z (GMT). No. of bitstreams: 1 Dissertacao_Maria_Raquel_Fernandez_Morinigo.pdf: 1321282 bytes, checksum: 91f2eb8e65e4b67c642c6df035df822a (MD5) Previous issue date: 2009-04-28 / Aim: To evaluate the cytotoxicity and genotoxicity of sodium percarbonate in comparison with bleaching agents used in discolored pulpless teeth from a mouse fibroblast cell culture. Methodology: The bleaching agents cytotoxicity and genotoxity were evaluated both in their pure form of products as well as in concentration commonly used in clinical practice. Hydrogen peroxide (HP), carbamide peroxide (CP), sodium perborate (SP) and sodium percarbonate (SPC) were diluted in DMEM in series. To evaluate the cytotoxicity, the survival of 3T3 mouse fibroblasts were measured photometrically by using MTT assay after a 24h exposure period. Genotoxicity was indicated by the micronuclei (MN) formation and the modification of the normal cell was analyzed by light microscopy (400x). The statistical analysis was performed by one-way ANOVA, followed by a multiplecomparison Tukey post hoc test (p < 0.05). Results: All tested groups exhibited a dose-dependent cytotoxicity. However, CP has showed a similar cytotoxic effect when compared with DMEM untreated control (UC) group. PH and SPC were significantly more cytotoxic than SP. Genotoxicity test has showed that SPC and SP have presented an intermediate rate of MN frequency over the UC group. The mean rate of MN frequency to HP was higher and statistically more significant than other groups tested. No difference was observed when CP and UC groups were compared. Conclusions: The results suggest that the cytotoxicity of SPC was similar to HP and significantly more cytotoxic than SP and CP. On the other hand, SPC and SP has shown an intermediate genotoxic effect, lower than HP and higher than CP, which was non-genotoxic / Objetivo: Avaliar a citoxicidade e genotoxidade do percarbonato de sódio e comparar com os agentes clareadores comumente utilizados no tratamento de dentes despolpados em cultura celular. Metodologia: A citotoxidade e genotoxicidade dos agentes clareadores foram avaliadas tanto na forma pura de cada produto quanto nas concentrações utilizadas na prática clínica. Peróxido de Hidrogênio (PH), peróxido de carbamida (PC), perborato de sódio (PS) e percarbonato de sódio (PCS) foram diluídos em DMEM. A viabilidade de fibroblastos da linhagem 3T3/NIH após 24h de exposição aos diferentes agentes foi medida fotometricamente por meio do teste colorimétrico com MTT. A genotoxicidade foi indicada pela formação e contagem de micronúcleos (MN) analisados em microscópio óptico comum (400x). A análise estatística dos dados foi feita utilizando o teste ANOVA de uma via complementado com o teste de comparações múltiplas Tukey post hoc (p < 0, 005). Resultados: Todos os grupos testados apresentaram um efeito citotóxico dosedependente. Entretanto, o PC mostrou citotoxicidade semelhante ao grupo controle (GC). PH e PCS foram significativamente mais citotóxicos que PS. O teste de genotoxicidade mostrou que PCS e PS apresentaram uma frequência intermediária de MN quando comparados ao GC. Por outro lado, a frequência média de MN usando PH foi significantemente superior aos outros grupos testados (p < 0, 005). Não houve diferença estatística entre PC e o GC. Conclusão: A citotoxicidade do PCS foi similar ao PH e significantemente maior do que o PS. Por outro lado, PCS e PS apresentaram um efeito genotóxico intermediário, inferior ao PH, porém superior ao PC que não teve efeito genotóxico

Gewässer sind Lebensgrundlage, jedoch gleichzeitig Schadstoffsenken für eine Vielzahl von Kontaminanten. Biologische Wirkungstests und das Biomonitoring aquatischer Proben sind daher besonders wichtig, um Umwelt-Gefahrenpotenziale erkennen zu können. Der "Comet Assay" (Einzelzell-Gelelektrophorese) ist ein Indikator von DNA-Strangbrüchen und wurde hier als Test auf gentoxische Wirkungen erprobt und angewandt. Mit bekannten, gentoxischen Substanzen wurden Nachweisgrenzen und Dosis-Wirkungs-Beziehungen für die Zelllinien RTG-2 und RTL-W1 (aus der Regenbogenforelle, Oncorhynchus mykiss) in vitro ermittelt und methodische Parameter an die Zellen angepasst. Der Test reagierte sehr sensitiv auf 4-Nitrochinolin-1-oxid. Die Substanz war daher geeignet, um in weiteren Versuchen als Positivkontrolle zu dienen. Zur Bewertung der Messdaten wurde ein geeignetes statistisches Verfahren gefunden, das auch historische Kontrollen mit einbezog. Der zeitliche Verlauf der DNA-Schädigung des Testsystems mit RTG-2-Zellen wurde ermittelt, und durch Inhibition der DNA-Reparatur mit Aphidicolin wurden Zusammenhänge zwischen der Entstehung von DNA-Strangbrüchen, der DNA-Reparaturkapazität sowie der Metabolisierungskapazität untersucht. In einer zweiten Phase wurden unbehandelte Wasserproben aus Rhein, Elbe sowie weitere Oberflächenwasserproben mit dem Comet Assay an RTG-2-Zellen getestet. Bei 15 von 49 Proben zeigten sich gentoxische Effekte. In einer dritten Phase wurden Erythrozyten von freilebenden Döbeln, Leuciscus cephalus, aus der Mosel mit dem Comet Assay untersucht. Die Fische von drei Messstellen zeigten erhöhte Werte von DNA-Schädigungen, gegenüber einer vierten, stromabwärts gelegenen Messstation. Korrelationen mit den Ergebnissen zusätzlicher Biomarker ergaben sich nur teilweise. Chemische Analysen von Wasser- oder Gewebeproben ließen keine Rückschlüsse auf verursachende Kontaminanten zu - gerade dies unterstreicht jedoch die Wichtigkeit biologischer Tests bei komplexen Proben. / Bodies of Water are both vital resources and pollutant sinks for a multitude of contaminants. Therefore, biological effect tests and biomonitoring of aquatic samples are of particular importance to detect potential environmental hazards. The "comet assay" (single cell gel electrophoresis) is an indicator for DNA strand breaks and was explored and applied as a genotoxicity test in the present study. Known genotoxic substances were used to determine the detection limits and dose-response relationships for the cell lines RTG-2 and RTL-W1 (from rainbow trout, Oncorhynchus mykiss) in vitro, and to adapt methodological parameters to the cells. The test was very sensitive to 4-Nitroquinoline-1-oxide. This substance was therefore well-suited to serve as positive control in further experiments. In order to evaluate the measurement data, an appropriate statistical procedure was developed, which also took "historical" controls into account. The time course of DNA damage in the test system using RTG-2 cells was determined, and relationships between the origin of DNA strand breaks, DNA repair capacity and the metabolizing capacity of the cells was investigated by means of inhibition of DNA repair with Aphidicoline. In the second stage, native water samples from the rivers Rhine and Elbe and further surface waters were tested with the comet assay, using RTG-2 cells. 15 out of 49 samples showed genotoxic effects. In a third stage, erythrocytes of feral chub, Leuciscus cephalus, from the Moselle river were examined with the comet assay. The fish from three measuring stations showed elevated values of DNA damage compared to fish sampled from a downstream station. There were only partly correlations with the results from additional biomarkers. Chemical analyses of water and tissue samples did not permit conclusions on effect-causing substances.However, this emphasizes the importance of biological tests in dealing with complex environmental samples.