Amazonian Tambaqui (Colossoma macropomum) is a commercial important fish species that is intensively cultivated in rice fish-cultures in a variety of water paddies in the area of the São Francisco river in the northeast of Brazil. Commercial formulations of herbicides containing Trifluralin (α,α,α-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine) are commonly used to control weed on the rice fields of the area. The present study aimed to investigate the sensitivity of Colossoma macropomum to trifluralin using biomarkers to understand the effects of an acute exposure to this substance. On the in vivo toxicity tests, C. macropomum juvenile were exposed to four concentrations of trifluralin: 0.74, 1.5, 2.23 and 3.0 μmols / L for 96 hours, and compared with negative, positive controls and solvent, respectively. The herbicide trifluralin shows a dose-dependent inhibition profile on the brain C. macropomum cholinesterase activity. On the highest trifluralin concentration was observed a drop in the cholinesterase activity around 62 % and the inhibition of the muscle was 80 % at the highest concentration tested. The estimated value of 96 h LC50 for trifluralin was 1.25 μmols / L. In vitro assays had shown that brain cholinesterase-ChE C.macropomum has a moderate sensitivity toward trifluralin (2.34 μmols / L) whereas the sensitivity to the carbamate eserine is much higher (0.158 μmols / L). In order to get the apparent kinetic constants for C.macropomum brain cholinesterase, kinetic assays were performed on brain homogenate in the presence of acetylthiocholine as substrate. The kinetic results in the presence of acetylthiocholine as substrate, the brain homogenate containing activity were 0.043 ± 0.015 mmol / L and 0.301 ± 0.014 for mmol.min-1.mg-1 protein Kapp m and Vapp max apparent kinetic constants, respectively. We also evaluated the mutagenic, genotoxic and apoptotic exposure of C. macropomum juvenile trifluralin. In the Comet assay, erythrocytes analyzed, compared to the negative control showed induction of DNA damage at all concentrations tested. The higher frequency of micronuclei and high numbers of apoptotic cells were observed in fish treated with concentrations of 1.5 μmols / L with a significant effect (p < 0.01). The results of this study indicate that brain ChE C. macropomum is a reliable biomarker for use in environmental monitoring of pesticides in the delta of the São Francisco river. It was also confirmed the genotoxic and apoptotic trifluralin at concentrations from 1.5 μmols / L for juvenile C. macropomum. (Note: check some nomeclaturas in the original summary because lost formatting in this field) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Conselho Nacional de Desenvolvimento Científico e Tecnológico / O Colossoma macropomum (Tambaqui Amazônico) é uma espécie de peixe comercial importante e intensamente cultivado em consórcio com as culturas de Oryza sativa (arroz) em uma variedade de corpos de água na área do rio São Francisco, no Nordeste do Brasil. Formulações comerciais de herbicida contendo Trifluralina (α, α, α-trifluoro-2,6-dinitro-N, N-dipropil-p-toluidina) são comumente utilizadas para controlar ervas daninha em campos de produção de arroz. O presente estudo teve como objetivo avaliar a sensibilidade de C. macropomum à trifluralina utilizando biomarcadores para analisar os efeitos de uma exposição aguda a esta substância. Nos testes de toxicidade in vivo, juvenis de C. macropomum foram expostos a quatro concentrações de trifluralina: 0,74; 1,5; 2,23 e 3,0 μmols / L durante 96 horas com os controles negativos, positivos e solvente, respectivamente. O herbicida trifluralina apresentou um perfil de inibição dose-dependente na atividade da colinesterase-ChE do cérebro de C. macropomum. Na maior concentração de trifluralina foi observada uma diminuição na atividade da ChE em torno de 62%, enquanto que no músculo a inibição foi de 80% na maior concentração testada. O valor estimado da CL50 em 96 h para trifluralina foi de 1,25 μmols / L. Os ensaios in vitro mostraram que a ChE cerebral de C. macropomum tem uma moderada sensibilidade à trifluralina (2,34 μmols / L), porém, com alta sensibilidade ao controle positivo - carbamato de eserina (0,158 μmols / L). Os resultados cinéticos, em presença de acetiltiocolina como substrato, com o homogenato de cérebro contendo atividade foram de 0,043 ± 0,015 mmol / L e 0,301 ± 0,014 mmol / min-1.mg proteína para as constantes cinéticas aparentes Kapp m e Vapp max, respectivamente. Também foram avaliados os efeitos mutagênico, genotóxico e apoptótico da exposição de C. macropomum juvenil à trifluralina. No ensaio do cometa, os eritrócitos analisados, quando comparados ao controle negativo, apresentaram indução de danos ao DNA em todas as concentrações testadas. A maior frequência de micronúcleos e o elevado número de células em apoptose foram observados nos peixes tratados com concentrações de 1,5 μmols / L com efeito significativo (p<0,01). Os resultados desse estudo indicam que a ChE do cérebro C. macropomum é um biomarcador confiável para ser utilizado no monitoramento ambiental de agrotóxicos no delta do rio São Francisco e outros rios do Brasil. Também ficou confirmado o potencial genotóxico e apoptótico da trifluralina em concentrações a partir de 1,5 μmols/L para juvenis de C. macropomum. (OBS.: verificar algumas nomeclaturas no resumo original pois perderam a formatação neste campo)

Aquatic environments are ecosystems that most affected impacts caused by anthropogenic action, since it is the final compartments of various products produced by human activity. This study evaluated the fish specie Serrasalmus brandtii (pirambeba), collected at two sites in the Itaparica reservoir (Brazil) during March/12 to January/2013 through biochemical and genotoxic biomarkers. Around the reservoir resettlement areas exist with agricultural practices using agrochemicals. Thus, the study aimed to use S. brandtii as specie sentinel for this region of the Sao Francisco river. There was no significant difference in the physicochemical parameters measured such as temperature and dissolved oxygen over the sample period. Among the morphological parameters, the masses of the specimens varied between sampling period. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities were quantify on the brain and muscle specimens collected from the two sites. Brain AChE from pirambeba collected at site 1 showed a significant decay in specific activity over the study period, however protein content did not show the same profile. Brain BChE specific activity had significant variation during the sampling period. On the contrary, pirambeba collected at site 2 showed no significant difference in the brain AChE specific activity and protein content. Brain BChE of these specimens showed significant variation on the specific activity values. On muscle tissue, pirambeba collected at site 1 showed significant variation in the values of AChE specific activity as well as on the protein content during the sample period. BChE specific activity on the muscle also had significant variation. Pirambeba collected at site 2 showed significant variation on muscle AChE specific activity and protein content values over the analyzed period. For muscle BChE activity,values, the results showed pattern similar to the one observed on site 1. Genotoxicity assays were performed on pirambeba erythrocyte cells collected at two sites. The results showed frequency of micronuclei (MN) and significant variation between the mean frequencies of MN over the study period was observed. Nuclear abnormalities (NA) were also analyzed and the results indicated that pirambeba the site 1 had higher nuclear abnormalities frequencies than the pirambeba from site 2. The Comet assay indicated that majority of erythrocyte nuclei analyzed from both sites shown class 1damage indicating slight DNA damage. In vitro studies were also performed for the characterization of brain AChE from pirambeba and kinetic assays showed values of 0.324 ± 0.04 mmol /L and 0.709 ± 0.014 mmol / min-1.mg protein for the apparent kinetic constants Kmapp and Vmáxapp respectively. Brain AChE inhibitory assays showed that muscle AChE (0.362±0.2 μM) was more sensitive to carbamate eserine that brain AChE (0.887 ± 0.1 μM) AChE and when compared brain AChE between eserine (0.887 ± 0,1μM) and glyphosate (2,276 ± 0,02μM), there was a greater sensitivity for brain AChE with eserine. We conclude that among the biomarkers genotoxic tested, the results showed that the MN test was considered the most sensitive; S. brandtii specie can be a specie sentinel for the submedium San Francisco region, since was present in all samples of the river, even during the most severe period of drought and low water level, but also by having a direct relationship with the AChE activity and forming MN. / Ambientes aquáticos são ecossistemas que mais sofrem impactos causados pela ação antropogênica, uma vez que constitui compartimentos finais de vários produtos gerados pela atividade humana. O presente trabalho avaliou a espécie de peixe Serrasalmus brandtii (pirambeba), coletada em dois sítios no reservatório Itaparica (Brasil) durante o período de março/12 a janeiro de 2013 através de biomarcadores bioquímicos e genotóxicos. No entorno do reservatório existem reassentamentos com práticas agrícolas empregando agroquímicos. Desta forma, o trabalho visou a utilização de S. brandtii como sentinela no rio Sao Francisco. Não houve diferença marcante nos parâmetros físico-químicos temperatura e oxigênio dissolvido mensurados ao longo do período amostral. Entre os parâmetros morfológicos, as massas dos espécimes variaram entre os meses de amostragem. Foi realizada quantificação de atividade acetilcolinesterase (AChE) e butirilcolinesterase (BChE) cerebral e muscular de espécimes coletados nos dois sítios. AChE cerebral de pirambeba coletada no sítio 1 apresentou decaimento significativo na atividade específica ao longo do período estudado, no entanto o conteúdo protéico não mostrou o mesmo perfil. A atividade específica BChE cerebral mostrou variação significativa durante o período de amostragem. Todavia, pirambeba coletada no sítio 2 não mostrou diferença significativa na atividade específica de AChE cerebral e conteúdo protéico. BChE cerebral nesses espécimes apresentou variação significativa nos valores de atividade específica. No tecido muscular, pirambeba coletada no sítio 1 apresentou variação significativa nos valores de atividade específica AChE, bem como conteúdo protéico durante período amostrado. A atividade específica de BChE muscular também teve variação significativa. Espécimes coletadas no sítio 2 apresentaram variação significativa na atividade específica da AChE muscular e no conteúdo protéico ao longo do período estudado. Para valores de atividade de BChE muscular, os resultados apresentaram um padrão semelhante ao observado no sítio 1. Os ensaios genotóxicos foram realizados em eritrócitos de pirambeba coletadas nos dois sítios. Os resultados mostraram frequência de micronúcleos (MN) e variação significativa entre as freqüências de MN foi observada ao longo do período estudado. Anormalidades nucleares (AN) também foram analisadas e resultados indicaram que espécimes do sítio 1 tiveram maiores freqüências de AN do que espécimes do sítio 2. O ensaio Cometa indicou que a maioria dos núcleos de eritrócitos analisados nos dois sítios apresentou dano na classe1, indicando danos leves no DNA das células. Estudos in vitro foram também realizados para caracterização de AChE cerebral de pirambeba e ensaios cinéticos mostraram valores de 0,324±0,04 mmol/L e 0,709±0,014 mmol/min-1.mg de proteína para constantes cinéticas aparentes Kmapp e Vmáxapp, respectivamente. Ensaios inibitórios de AChE mostraram que AChE muscular (0,362±0,2) foi mais sensível ao carbamato de eserina que a AChE cerebral (0,887±0,1) e quando comparados AChE cerebral entre a eserina (0.887±0,1μM) e o glifosato (2,276±0,02μM), verificou-se uma maior sensibilidade para a AChE cerebral com eserina. Concluímos que dentre os biomarcadores genotóxicos testados, os resultados mostraram que teste de MN foi considerado o mais sensível; a espécie S. brandtii pode ser a espécie sentinela para a região do submédio São Francisco, visto que esteve presente no rio em todas as coletas, mesmo durante o período mais rigoroso de estiagem e baixo nível da água, como também por apresentar relação direta com a atividade de AChE e a formação de MN.

Made available in DSpace on 2014-10-09T12:35:46Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:54Z (GMT). No. of bitstreams: 0 / O câncer é uma doença de alta incidência e é considerado um problema de saúde pública no mundo todo. O resveratrol é um polifenol de defesa que tem a habilidade de inibir a carcinogênese em múltiplos estágios. Este polifenol é sintetizado por uma grande variedade de plantas em resposta a exposição à radiação ultravioleta (UV) ou também, pelo estresse mecânico produzido pela ação de patógenos, agentes químicos e físicos. As videiras são consideradas as plantas que têm uma capacidade elevada de produzir o resveratrol, portanto suco de uva e vinho, principalmente o vinho tinto são considerados uma boa fonte de resveratrol. Os efeitos protetores exercidos pelo resveratrol promovem efeitos como indução da resposta antiinflamatória, atividade antitumoral, prevenção ou inibição de doenças degenerativas, diminuição da incidência de doenças cardiovasculares, inibição da agregação plaquetária, entre outros. Assim, o resveratrol é considerado um protetor celular. Porém, em concentrações mais elevadas, o resveratrol promove o efeito contrário, sensibilizando as células a algum tipo de efeito, como por exemplo, o efeito da radiação ionizante. O objetivo desse trabalho foi estudar o efeito radiomodificador do resveratrol em cultura de células RD (rabdomiossarcoma humano) aplicando o teste do cometa para avaliação do dano e capacidade de reparo celular. Foi obtida a DL50 das células RD em 403 Gy e o índice de citotoxicidade do resveratrol (IC50%) nas células RD foi de 150 &mu;M. A partir destes resultados foram definidas as doses de radiação gama (50 Gy e 100 Gy) e as concentrações de resveratrol (15 &mu;M 30 &mu;M 60 &mu;M) que foram analisadas no trabalho. Foram evidenciados os efeitos do resveratrol como protetor celular na concentração de 15 &mu;M e seu efeito citotóxico em 60 &mu;M. Com a interação da radiação gama, a concentração de 60 &mu;M não mostrou efeito radiossensibilizador estatisticamente significante. / Dissertação (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2017-09-21T19:58:03Z No. of bitstreams: 2 Tese - Fernanda Craveiro Franco - 2017.pdf: 4803005 bytes, checksum: 35b5c5c99f5a0689a4c30ebe8ea513cd (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-09-22T11:38:57Z (GMT) No. of bitstreams: 2 Tese - Fernanda Craveiro Franco - 2017.pdf: 4803005 bytes, checksum: 35b5c5c99f5a0689a4c30ebe8ea513cd (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-09-22T11:38:57Z (GMT). No. of bitstreams: 2 Tese - Fernanda Craveiro Franco - 2017.pdf: 4803005 bytes, checksum: 35b5c5c99f5a0689a4c30ebe8ea513cd (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-09-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The interaction of some compounds and DNA molecule can cause genotoxic changes. The tests used to evaluate genotoxicity have evolved over the years, and they have great efficiency. Currently, several traditional methodologies are used, such as the comet assay, and some more modern, based on molecular biology. Considering the importance of detecting the effects caused by exposure to potentially genotoxic agents, this thesis presents three chapters with different approaches to this topic. The first chapter aimed at evaluating published studies on the mutagenic, genotoxic and oxidative effects caused by occupational exposure to pesticides. For this, a systematic review of articles published between the years of 2011 and 2015 was carried out, focusing on these exposure conditions. The results showed that 44% of the studies addressed only men and 44% approached men and women, and 78% used rural workers as an exposed population. The most evaluated class of pesticide was organophosphorus. The studies were separated by methodologies of analysis used and all showed some type of alteration in the populations occupationally exposed to pesticides. The second chapter, published in the journal Environmental Science and Pollution Research, considered the dengue scene in the state of Goias and techniques of mosquitoes control, and sought to understand the genotoxic effects caused by exposure of endemic agents to pesticides used against Aedes aegypti, being the first study published with this group of workers. The main pesticides used were insecticides and larvicides and an increase in DNA damage was observed in the exposed population, without influence of the lifestyle factors. In the whole transcriptome assay the differential expression of genes related to various chronic diseases was observed. Finally, the third chapter was carried out in collaboration with the Institute of Experimental Medicine of the Czech Academy of Sciences, aiming the determination of cytotoxic and genotoxic effects caused by in vitro exposure to different types of metallic nanomaterials. In nanomaterials that showed cell viability reduction (cytotoxic), an increase in genotoxicity was also identified. In addition, three of the four non-cytotoxic nanomaterials evaluated showed genotoxic effects. Studies aimed at detecting the compounds genotoxic potential are extremely important for the regulatory definition and identification of risks caused by exposure. In this study, the toxicological and genotoxic effects of different compounds and mixtures were evaluated using traditional and modern techniques, in vitro or by human biomonitoring, seeking to understand and determine the risks involved in exposure to these substances. / Compostos que interagem com a molécula de DNA podem provocar alterações genotóxicas. Os testes utilizados para avaliação de genotoxicidade evoluíram ao longo dos anos, apresentando grande eficiência. Atualmente, utilizam-se várias metodologias tradicionais, como o ensaio cometa, e também algumas mais modernas, baseadas na biologia molecular. Considerando a importância de detectar os efeitos causados pela exposição a agentes potencialmente genotóxicos, essa tese apresenta três capítulos com formas de abordagem distintas, referente a essa temática. O primeiro capítulo visou a avaliação de estudos publicados acerca dos efeitos mutagênicos, genotóxicos e oxidativos causados pela exposição ocupacional a pesticidas pela realização uma revisão sistemática de artigos publicados entre os anos de 2011 a 2015. Observou-se que 44% dos estudos abordaram exclusivamente indivíduos do sexo masculino e 44% abordaram ambos os sexos, e 78% utilizaram trabalhadores rurais como população exposta. A classe de pesticida mais avaliada foi a de organofosforado. Os estudos foram separados por metodologias de análise utilizada e todos apresentaram algum tipo de alteração nas populações expostas a pesticidas no ambiente de trabalho. O segundo capítulo, publicado na revista Environmental Science and Pollution Research, considerou o cenário da dengue no estado de Goiás e as formas de combate ao mosquito, e buscou compreender os efeitos genotóxicos causados pela exposição de Agentes de combate a endemias aos pesticidas utilizados contra o mosquito, sendo o primeiro estudo publicado com esse grupo de trabalhadores na região centro-oeste. Os principais pesticidas utilizados foram inseticidas e larvicidas e foi observado um aumento no dano ao DNA na população exposta, sem influência dos fatores relacionados ao estilo de vida. No ensaio de transcritoma foi observada a expressão diferencial de genes relacionados a diversas doenças crônicas. Finalmente, o terceiro capítulo foi realizado em colaboração com o Instituto de Medicina Experimental, da Academia Tcheca de Ciências, visando a determinação de efeitos citotóxicos e genotóxicos, causados pela exposição in vitro a diferentes tipos de nanomateriais metálicos. Nos nanomateriais que apresentaram redução na viabilidade celular (citotóxicos) também foi identificado aumento na genotoxicidade. Além disso, três dos quatro nanomateriais não citotóxicos avaliados apresentaram efeitos genotóxicos. Estudos que visam detectar o potencial gentoxicológico de compostos são de extrema importância para definição de normas e regulamentações de aplicação e identificação dos riscos causados pela exposição. Nesse estudo, foram avaliados os efeitos toxicológicos e genotóxicos de diferentes compostos e misturas, com a utilização de técnicas tradicionais e modernas, in vitro ou por biomonitoramento humano, buscando compreender e determinar os riscos envolvidos na exposição a essas substâncias.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-02-06T09:39:55Z No. of bitstreams: 2 Dissertação - Rangel Moreira Silva - 2016.pdf: 3905320 bytes, checksum: 6b2881f7623815dc402d04f906727e47 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-02-06T09:40:57Z (GMT) No. of bitstreams: 2 Dissertação - Rangel Moreira Silva - 2016.pdf: 3905320 bytes, checksum: 6b2881f7623815dc402d04f906727e47 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-02-06T09:40:57Z (GMT). No. of bitstreams: 2 Dissertação - Rangel Moreira Silva - 2016.pdf: 3905320 bytes, checksum: 6b2881f7623815dc402d04f906727e47 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-06-30 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Myrciaria cauliflora (Mart), commonly known as “Jabuticaba” (Brazilian Grape) occurs in all of South America, especially in Brazil. This species presents medicinal properties and it is popularly used to treat diarrhea and respiratory diseases. It has already been proven several biological activities such as antioxidant, antimutagenic and anti-proliferative. The phytochemical analysis of M. cauliflora detected the presence of several compounds, including the ellagitannin pedunculagin. Due to the widespread use of this species for therapeutic and food purposes, this study aimed to evaluate the genotoxic, antigenotoxic, cytotoxic and anticytotoxic actions of the aqueous fraction (AF) and pedunculagin from M. cauliflora seed, by micronucleus (MN) test and the comet assay in mice bone marrow cells. In order to analyze AF and pedunculagin activities prior to CP’s toxic effects, the animals were co-, pre- and post-treated with these substances. The results showed that AF at 24 h treatment did not show genotoxic or cytotoxic effects in the MN test and comet assay. However, AF showed cytotoxic action, but did not show genotoxic effect at 120 h treatment in the MN test. In the comet assay it was able to significantly reduce the frequency of DNA breaks when compared to the negative control. In the evaluation of the antigenotoxic and anticytotoxic effects, it was observed that in all treatments, in both assays, AF and pedunculagin showed antigenotoxic and anticytotoxic activities, but in the post-treatment pedunculagin increased the cytotoxic effect of CP. In general, our results indicated that AF and pedunculagin were able to protect the mice bone marrow cells against DNA damage induced by CP in the MN test and comet assay. These findings indicated that AF and pedunculagin could be probable candidates for the development of new drugs. / Myrciaria cauliflora (Mart), comumente conhecida como jabuticaba, ocorre em toda América do Sul e especialmente no Brasil. Esta espécie apresenta propriedades medicinais e é popularmente usada para tratar diarreia e doenças respiratórias. A essa espécie já foram atribuídas várias atividades biológicas, tais como: antioxidante, antimutagênica e antiproliferativa. A análise fitoquímica de M. cauliflora detectou a presença de vários compostos, incluindo o elagitanino pedunculagina. Devido ao grande uso desta espécie para fins terapêuticos e alimentícios, este estudo teve como objetivo avaliar as ações genotóxica, antigenotóxica, citotóxica e anticitotóxica da fração aquosa (FA) e do composto pedunculagina da semente de M. cauliflora pelo teste do micronúcleo e ensaio do cometa em células da medula óssea de camundongos. A fim de avaliar a atividade da FA e da pedunculagina frente aos efeitos genotóxicos da CP, os animais foram co, pré e pós-tratados com essas substâncias. Os resultados mostraram que a FA no tempo de 24 horas não apresentou efeitos genotóxico e citotóxico no teste do micronúcleo e no ensaio do cometa. No tempo de 120 horas, pelo teste do micronúcleo, a FA não mostrou ação genotóxica, porém exibiu efeito citotóxico e pelo ensaio do cometa, a frequência de quebras de DNA foi menor, quando comparada ao controle negativo. Na avaliação dos efeitos da antigenotoxicidade e anticitotoxicidade, em ambos os ensaios, a FA e a pedunculagina mostraram atividades antigenotóxica e anticitotóxica, porém, no pós-tratamento, o composto pedunculagina potencializou o efeito citotóxico da CP. De modo geral, nossos resultados indicaram que a FA e a pedunculagina foram capazes de proteger as células da medula óssea de camundongos contra os danos no DNA induzidos pela CP no teste do micronúcleo e no ensaio do cometa. Esses resultados indicaram que a FA e a pedunculagina podem ser prováveis candidatos para o desenvolvimento de novas drogas.

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

La majeure partie de mon travail de thèse a porté sur l’étude de la composition chimique de la surface d’Europe. Afin d’apporter une plus-value par rapport aux résultats de l’instrument NIMS à bord de la sonde Galileo qui orbita dans le système jovien de 1995 à 2003, une campagne d’observations depuis le sol a été menée avec le spectromètre imageur infra-rouge SINFONI au VLT. Cinq observations en optique adaptative possédant une résolution spatiale d’environ 160 km et une résolution spectrale R=1500 dans le proche infra-rouge furent acquises. Une procédure complexe de réduction des données a été mise en place afin de construire un cube hyperspectral global en réflectance de la surface. La modélisation linéaire de chacun des spectres du cube a permis d’aboutir aux premières cartes d’abondances absolues jamais obtenues pour la surface d’Europe. Ces cartes confirment la présence des deux espèces majoritaires, la glace d’eau et l’acide sulfurique hydraté. La distribution de l’acide sulfurique est centrée sur l’hémisphère orbital arrière qui est préférentiellement impacté par un flux d’ions de soufre originaires du tore de plasma produit par l’activité volcanique d’Io. Cependant, deux résultats inattendus ont été obtenus. Le premier concerne la glace d’eau, dont la forme cristalline est près de deux fois plus abondante que la forme amorphe selon les résultats de la modélisation. Ce résultat, surprenant compte tenu du taux d’irradiation très élevé auquel la surface est soumise, pourrait s’expliquer par l’existence d’un fort gradient de cristallinité au sein de la couche de glace, mais aussi par l’existence d’une activité endogène relativement soutenue qui se traduit visuellement par une surface peu cratérisée, donc jeune. La corrélation entre la distribution des grains cristallins et la géomorphologie semble accréditer la seconde hypothèse. Le second résultat concerne la détection de sels chlorés à partir de la modélisation des spectres hautement résolus de SINFONI, et non de sulfates, remettant en cause les détections marginales annoncées par les observations de l’instrument NIMS/Galileo. La distribution des sels chlorés, tout comme celle de la glace d’eau cristalline, est corrélée à la géomorphologie, ce qui confirme le rôle important des apports endogènes. Des processus tectoniques et cryovolcaniques mis en évidence récemment pourraient être à l’origine de cette distribution. L’autre versant de ma thèse a été consacré aux caractéristiques physiques des grains cométaires de 67P/C-G. L’instrument COSIMA embarqué sur l’orbiter de la sonde Rosetta a permis la collecte, l'imagerie et l'analyse chimique élémentaire des grains présents dans l’environnement proche de 67P/C-G. Une approche automatisée de la détection des grains à partir des images prises par la caméra « COSISCOPE » a été mise en place et a permis de détecter environ 35000 grains ayant une surface de plus de 100 µm2 entre août 2014 et mai 2016. La résolution de 13.7 µm/pixel a rendu possible la caractérisation en détail de la forme et de la structure des grains, et le nombre important de détections a permis d’obtenir des statistiques robustes concernant la distribution en taille et l’évolution de celle-ci au cours du temps. Deux grandes familles de grains ont été identifiées : les grains compacts, qui ne représentent qu’une faible minorité des grains et qui ont été majoritairement collectés en début de mission, et les agrégats, qui ont une structure très poreuse similaire à celle des IDPs et des micrométéorites collectées en Antarctique. La distribution en taille obtenue suit une loi de puissance intégrale en r-2.66. La comparaison avec des lois obtenues à des échelles différentes par d’autres instruments met en évidence des différences qui peuvent être interprétées par des mécanismes d’éjection dépendant de la taille associés à un biais du processus de collecte en orbite. / The major part of my work focused on the study of the chemical composition of Europa’s surface. In order to provide additional insights in comparison to the results of the NIMS instrument onboard the Galileo spacecraft that orbited in the Jovian system from 1995 to 2003, a ground-based observations campaign was conducted with the infrared imaging spectrometer SINFONI on the VLT. Several observations using adaptive optics with a spatial resolution of about 160 km and a spectral resolution R = 1500 in the near-infrared were acquired and then combined. A specific data reduction pipeline was developed to build a global hyperspectral cube in surface reflectance. The linear modeling of each spectra of this cube leads to the first global abundance maps ever obtained for the surface of Europa. These maps confirm the presence of the two major species, namely water ice and hydrated sulfuric acid. The distribution of the hydrated sulfuric acid is centered on the trailing orbital hemisphere preferentially affected by a sulfur ion flux coming from the plasma torus produced by Io volcanic activity. Two surprising results were obtained. The first one concerns water ice, which crystalline form is about twice more abundant than the amorphous form according to the modeling results. This result, unexpected given the very high radiation rate on the surface, could be explained by a strong crystallinity gradient through the ice slab. However, it could also point out an endogenous activity possibly strong as first suggested by its poorly well-known cratered surface. The correlation between the crystalline grains distribution and the geomorphology seems in favor to the second hypothesis. The second result is related to the detection of chlorinated salts from the modeling of highly resolved spectra from SINFONI. Sulfates, first reported by several analyses of NIMS observations are marginal in the modeled composition, hence challenging their presence on the surface of Europa. The chloride distribution, as well as the one of the crystalline water ice, is correlated to geomorphology, potentially confirming significant endogenous contributions as the result of tectonic and cryovolcanic processes recently highlighted. The second part of my PhD was dedicated to the physical characteristics of the 67P/C-G’s cometary grains. The COSIMA instrument onboard the Rosetta orbiter allowed the collection, imaging and elemental chemical analysis of grains present in the immediate environment of 67P/C-G. An automated approach of the grains detection based on images taken by the camera "COSISCOPE" was set up and able to detect about 35.000 grains having an area of more than 100 µm² between August 2014 and May 2016. The resolution of 13.7 µm/pixel allowed to perform a detailed characterization of the shape and the structure of the grains, and the large number of detections permitted to obtain significant statistics on the size distribution and its evolution over the time. Two large families of grains have been identified: the compact grains, which represent only a small minority of grains mostly collected at the beginning of the mission, and aggregates, which have a very porous structure similar to those of IDPs and micrometeorites collected in Antarctica. The size distribution obtained follows an integrated power law in r-2.66. The comparison with the laws obtained at different scales by other instruments highlights differences that can be interpreted by ejection mechanisms depending on the size associated with a selection bias during the collection process in orbit.

Recent missions to solar system comets, such as ESA's Rosetta mission, raise interest for models and descriptions of their plasma environment. The interaction with various space phenomena such as stellar wind make the construction of an analytical description difficult. Instead, a simplified view of the comet environment is considered where the effects of magnetism and departures from radial symmetry are neglected. This is done in an effort to construct an approximation of the comet plasma behaviour later to be compared against observational accounts to find which plasma features are dependent on more complex phenomena and which plasma features arise as a result of the simpler comet view. Several attempts are made to construct an analytical description of comet plasma as based on the description within another branch of plasma physics: fusion. Previous work regarding the vacuum expansion of plasma after a stationary target is rapidly ablated via high-intensity lasers appears promising for adaptation to the comet environment. Before the comet environment can be considered the different natures of the two problems have to be considered. For example, the comet case is a stationary expansion problem as opposed to fast-ignition fusion where the expansion is treated as an initial value problem. Having accounted for the problems' inherent differences, a few methods are proposed to convert solutions of lab fusion distribution functions to the comet case. Additionally, a numerical approach to calculate the distribution function of comet electrons is presented employing ergodic invariance. Lastly, a toy-model simulation of the timescale for variations in the potential show that the error in the ergodic invariance may in practice have a faster convergent timescale dependence than theoretical bounds suggest. Optimistically, this suggest the possibility of future use in numerical attempts at modelling comet plasma.

The first cases of patients with pneumonia which grew into an acute respiratory distress syndrome and caused breathing problems began to appear in December 2019. Coronavirus disease 2019 (COVID-19) is the cause of a global pandemic and it is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A complex interplay of factors is responsible for the progression of the disease. Some studies suggest that it promotes oxidative stress and thus may lead to oxidative damage to cells and DNA. The purpose of this study was to observe the relationship between oxidative DNA damage and a critical condition caused by COVID-19 using a comet assay technique. The basic principle of the used method consists in fixation of lymphocytes in an agarose gel, removal of the membrane and cytoplasm of cells, incubation with specific enzymes and electrophoresis. In the process of electrophoresis, negatively charged DNA fragments migrates towards the anode and the cell thus acquires the typical shape of a comet. Comets are visualized using the DNA intercalation dye ethidium bromide. We quantified single - strand breaks and oxidized pyrimidines and purines by using specific enzymes (modification of the method for detecting specific lesions). Results are reported as % tail DNA, thus the percentage of DNA in the...

The genotoxicological effects of five intra-peritoneal administered oestrogens (17β- oestradiol, daidzein, diethylstilboestrol, genistein, and equol), were examined. Male hooded- Lister rats were used to examine to what extent DNA damage occurred. The alkaline Comet assay was the chosen method used to assess double-strand DNA breakage by examining the Olive tail moment and %age tail DNA. Tissues from the testis, bone marrow, liver and blood were analysed after an 8-day duration of exposure. Statistically significant increases in DNA damage were observed in the testis with daidzein and in the blood with diethylstilboestrol. In addition, a further study was carried out to examine the effects of bulk and nanotised forms of non-steroidal anti-inflammatory drugs (NSAIDs), aspirin and ibuprofen, in the Comet and micronucleus assays, on whole blood taken from prostate cancer patients or volunteers. These were used because it is known that the sensitivity of DNA to genotoxins can be heightened in patients with cancer. Patients’ and volunteers’ blood was cultured with either the bulk or nano-forms for 44 hours at 37°C, 5% CO2. Data were obtained for the Comet assay as above and the number of binucleated cells scored for the micronucelus assay. The results show the nanotised forms of the NSAIDs decreased the levels of strand breakage and lowered the numbers of micronuclei generated compared with their bulk forms. There was no clear difference between the sensitivity of the healthy controls and the prostate cancer patients, with only one individual showing evidence of heightened sensitivity.