Comparação do perfil de mutagenicidade e da composição quí­mica do material particulado atmosférico de Limeira, Estocolmo e Quioto [thesis]

Bianca de Souza Maselli
Dedico este trabalho aos meus pais, José Maselli e Maria Beatriz, aos meus irmãos Alessandra, Bruno e Larissa, pelo amor, carinho e apoio incondicional que sempre me deram, pois sem vocês não teria chegado até aqui. Agradecimentos A Deus, por eu estar aqui, por estar ao meu lado em todos os desafios e por todas as pessoas e oportunidades colocadas em meu caminho. Aos meus pais José Maselli e Maria Beatriz, pela estrutura familiar e educação que me deram, as quais foram de extrema importância
more » ... rema importância para a minha formação pessoal, pela compreensão nos momentos que estive ausente, e pelo amor e apoio incondicionais em todos os momentos da minha vida. Aos meus irmãos Alessandra (em especial a você, irmã) e Bruno, e minha irmã/prima/afilhada Larissa, pelo amor, carinho e apoio que sempre me deram, pelas longas conversas de otimismo, sempre que ficava aflita, pela admiração no meu trabalho e pelo exemplo de pessoas batalhadoras que são para mim. Ao meu eterno amigo Fabrício Miranda Follador, pelo apoio nos momentos mais difíceis, pela compreensão nos momentos que estive ausente, por me acalmar nos momentos de aflição, e por estar sempre na torcida em todas as minhas conquistas. À família Souza, Gilda, Waldemar, Juliana, Sidney, Arminda, Jairo e Jaime, pelo apoio, principalmente nos momentos que estive ausente, e pelos momentos de descontração. Ao meu orientador Prof. Dr. Fábio Kummrow, pela orientação, pelas oportunidades que me proporcionou, pelos conhecimentos transmitidos e, acima de tudo, pela amizade e cumplicidade em todos esses anos trabalhando juntos. À Profa. Dra. Gisela de Aragão Umbuzeiro, pela orientação, por todo conhecimento compartilhado, mas principalmente, pelo acolhimento excepcional e por ter cedido e cuidado deste projeto com tanto carinho. e Vivien por todos os momentos compartilhados, dentro e fora do laboratório, e por todos os alunos que me ajudaram de alguma forma. A todos os amigos de Limeira, pelos momentos de diversão e descontração, e pelo apoio e compreensão nos momentos que estive ausente. Às instituições, Faculdade de Ciências Farmacêuticas -USP e Faculdade de Tecnologia -UNICAMP, pelo apoio e estrutura fornecida para o desenvolvimento deste trabalho. À CAPES e FAPESP pelo apoio financeiro e bolsas concedidas, sem os quais não seria possível realizar este trabalho. Muito obrigada! Palavras-chaves: Partículas totais suspensas; Ensaio Salmonella/microssoma em microssuspensão; Razões diagnósticas moleculares; Processo de envelhecimento do PTS, Miniaturização do teste Salmonella/microssoma. ABSTRACT MASELLI, B. S. Comparison of mutagenicity and chemical profile of the atmospheric particulate matter from Limeira, Stockholm and Kyoto. 2018. 134f. Tese (Doutorado) -Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, 2018. Atmospheric particulate matter (PM) is associated with various diseases and has recently been classified as carcinogenic to humans (Group 1) by the International Agency for Research on Cancer (IARC). Although the PM toxicity, including genotoxicity, is known to be related to particle size, the contribution of its chemical composition to the observed effects has not yet been fully elucidated. The size and composition of the particles are influenced by meteorological and climatic characteristics, especially the temperature and the period of solar radiation. The Salmonella/microsome assay is the most used for the evaluate of the mutagenicity of PM samples, however a small number of strains are used. The objective of this study was to investigate and compare the influence of the different atmospheric and climatic conditions of the cities of Limeira (Brazil), Stockholm (Sweden) and Kyoto (Japan) on mutagenicity profiles, using 11 strains with different selectivity, and chemical composition profiles of total particles suspended (TPS) composed samples collected during the winter of these cities. For the results to be directly compared, the same methodology was adopted, including the sampling procedure, the sample preparation protocol, the mutagenicity test protocol, and the chemical analysis techniques for the identification of polycyclic aromatic hydrocarbons (PAH) and their alkylated derivatives. Limeira presented the highest concentration of TPS (99.0 μg/m 3 ), followed by Kyoto (28.0 μg/m 3 ) and Stockholm (6.2 μg/m 3 ). Although the TPS concentration in Limeira was 16-fold higher than in Stockholm and 3.5-fold higher than in Kyoto, the percentage of extracted organic material (EOM) obtained was 9, 15 and 5%, respectively. The extracts from the TPS samples collected in the three cities presented mutagenic activity for all strains, both in the absence and presence of S9, except for TA102 that did not detect the mutagenic activity in any of the extracts, and for YG7108 in the presence of S9, only for the Limeira sample. Despite the differences in meteorological and climatic conditions of these cities, their mutagenicity profiles were similar. The lower mutagenic potency for YG7108 in the absence of S9 and the nonmutagenicity in the presence of S9 reveals the lower contribution of alkylating agents to the mutagenicity of the Limeira sample in relation to the other cities. Considering the selectivity of the strains used, we can also observe a greater contribution of compounds that cause DNA oxidative damage and of alkylating agents for the mutagenicity of the Kyoto sample. However, according to the responses of the strains that have increased activity of the nitroreductase (NR) and/or O-acetyltransferase (OAT) enzymes, YG1021, YG1024 and YG1041, show the highest contribution of nitroarenes and aromatic amines for the mutagenicity of the three cities, with emphasis on those mutagens that depend on OAT activation. The mutagenic potencies expressed as a function of EOM mass (rev./μg EOM) were similar for all samples. When expressed by air volume (rev./m 3 ), the mutagenic potencies were proportional to TPS concentrations of the three cities (Limeira> Kyoto> Stockholm). When applying the diagnostic ratios, it was possible to verify a mixture of sources of pollution in Limeira and Stockholm. The diagnostic ratios of the PAH indicate that Limeira sample is composed of fresh particles with a slight predominance of combustion sources. The Stockholm sample present aged particles with a predominance of petrogenic sources. This result was unexpected, since Limeira's atmospheric conditions are more favorable to particle aging than Stockholm's. The study of the transport of air masses explains, at least in part, the presence of aged particles in the Stockholm sample. It also suggests that the reduction of PAH concentrations in Limeira depends on the control of local sources, while in Stockholm control of local sources would not be efficient in reducing these pollutants. In parallel to the evaluation of the TPS samples, a new miniaturized protocol of the Salmonella/microsome assay in microsuspension was also developed. The protocol using 12-well microplates was validated by employing thirteen mutagenic compounds tested with three selected Salmonella strains based on their different spontaneous reversion frequencies (low, medium and high). Both the miniaturized microplate agar (MPA) and the microsuspension assay presented similar sensitivities, concluding that MPA is a promising tool and can be particularly suitable for environmental studies such as effectdirected analysis (EDA) or monitoring programs.
doi:10.11606/t.9.2018.tde-18062018-113553 fatcat:keotxn36yre6rkfssmg4ggjl2y