Desenvolvimento e validação de uma técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata

The comet assay is a method of DNA damage analysis widely used to quantify oxidative damage, crosslinks of DNA, apoptosis and genotoxicity of chemicals substances as chemical, pharmaceuticals, agrochemicals products, among others. This technique is suitable to detect DNA strand breaks, alkali-labile sites and incomplete excision repair sites and is based on the migration of DNA fragments by microeletroforesis, DNA migrates for the anode forming a “tail”, and the formed image has the appearance of a comet. The slides can be stained with fluorescence or silver, having differences in the microscopy type used for the analysis and the possibility of storage of the slides, moreover, the first one is a stained-method with more difficulties of accomplishment. The image analysis can be performed by a visual way, however, there is a disadvantage as the subjectivity on the results, that can be minimized by an automated method of digital analysis. This process was studied in this report with the aim to perceive the validation of the digital analysis turning it a quantitative method with larger reproductibility, minimizing the variability and imprecision due to the subjective analysis. For this validation we selected 50 comets photographed in a standardized way and printed, afterwards, pictures were submitted to three experienced appraisers, who quantified them manually. Later, the images were processed by free software ImageJ 1.38x, printed and quantified manually by the same appraisers. The intraclass correlation was higher to comet measures after image processing. Following, an algorithm of automated digital analysis from the measures of the comet was developed; the values obtained were compared with those 12 estimated manually after the processing resulting high correlation among the measures. The use of image analysis systems increases ...(Complete abstract click electronic access below)

Avaliação dos efeitos dos inseticidas malation e fipronil sobre a viabilidade e regeneração de girardia tigrina (Platyhelminthe, tricladida)

In this study were carried out toxicity tests and the evaluation of sub lethal effects of the insecticides malathion and fipronil, over the test system of Girardia tigrina, using for these analyses a great variety of methods. For evaluation of the lethal concentrations of the products, adult planarians were exposed to several different concentrations. All the tests were carried out in duplicate, maintained on the shelter of the light, on a temperature 24-26ºC and they had the duration of 96h, with observations at each 24h. The results were statically valued by the tests U of Mann- Whitney, Spearman-Karber, ANOVA and Pearson correlation. The effects over the system-test, in the different tested concentrations, varied in accordance with the time of exposition. The EC50 given by the test of Spearman-Karber was of 12,5mg/L for malathion and 13,4mg/L for fipronil. The evaluation of sub lethal effects was composed by analyses of the potential of regeneration; histological, for tissue damage evaluation; d; and the “comet assay”, for DNA damage detection. The specimens split up and exposed to the insecticides demonstrated, between other effects, retardation in development of the regenerative blastema. The histological evaluation has shown that exposed planarians to lower concentrations of the insecticides cause more significant damage against the tissues and some specific cell types. Through the comet assay, significant damages were noticed in the DNA in all the treatments, when compared with the control group. Such damages may result in reproduction and immunity alterations, which reduce the competitive efficiency and, consequently, the survival chances of these organisms.

Indução de morte celular por apoptose decorrente da irradiação com Raios-X: um ensaio com células mamárias humanas tumorais

Apoptosis is a form of programmed cell death selectively removes abnormal cells, and thus contributes to maintaining the balance of the dynamics of cell reproduction. Therefore the verification of the occurrence of apoptotic cell death after a pathological stimulus is crucial for the analysis of the maintenance of normal cell cycle of a given tissue or organ. In this experiment were used cells lines human mammary tumor MDAMB231, T47, MCF7, which were irradiated with X-rays at a dose of 5 Gy in a time interval of 15 seconds, and filtration of 1mm aluminum. Samples containing the cells were grown in a specific culture medium, containing fetal bovine serum and growth factor, and two samples were prepared with each of the cell lines, one to be irradiated, and another that has not been irradiated, which denoted by negative control of the irradiation. The primary goal of the experiment was to verify and compare the rates of apoptosis in each cell lines, in which were irradiated and that were not irradiated, using flow cytometry as a method for detecting apoptotic cell death in together with specific markers annexin V and propidium iodide. Data from the readings made by flow cytometry were analyzed and interpreted using the software WinMDI statistical graph. By comparing the indices relating to the readings of positive and negative for specific markers of apoptosis, based on differences in the statistical data presented lectures regarding the cellular irradiated and not irradiated, collude cells in question once... (Complete abstract click electronic access below)

Avaliação da toxicidade da vinhaça utilizando Oreochromis niloticus (Perciformes: Cichlidae) como organismo teste

Contamination of rivers and lakes, waste disposed by infiltration into the soil as fertilizer, for example, has been highlighted around the world, since such compounds can contain toxic elements, causing changes in aquatic and/or terrestrial ecosystem. Due to the possible presence of these elements in these wastes, it is possible assess damage to organisms exposed through breaks in genetic material and mutations. In this sense, this work aimed at exploring the cytotoxic, mutagenic and genotoxic vinasse pontential, residue used as fertilizer, from the processing of sugar cane into alcohol, using fish (Oreochromis niloticus - Cichlidae) as a test organism. The individuals were exposed to different dilutions of vinasse in the water and later carried out analysis of their erythrocytes using the micronucleus test and nuclear abnormalities associated and the comet assay. In the first bioassay, tilapia exhibited 5% and 10% vinasse concentrations died after 48h. The number of micronucleated erythrocytes was statistically significant for the 1% vinasse concentration and also for nuclear abnormalities such as broken-egg and nuclear bud, when compared with negative and positive controls; were also significant lobed type erythrocytes, when compared to the negative control. The test results of the comet assay were not significant for 1% vinasse when compared to the negative control, due to the high rate of comets with damage control, which is unusual. The same was observed for changes as notched and blebbed type. In the second chemical analyzes conducted, the detection of heavy metals in the vinasse sample, charged only the presence of copper and chromium, the latter with a concentration 89 times higher than in the first test. Differentially the first bioassay, fish exposed to dilution of 5% did not die... (Complete abstract click electronic access below)

Comparação de dois métodos de profilaxia dental: avaliação da pressão arterial e do conforto do paciente. Ensaio clínico controlado e randomizado

GOALS: This research evaluated the change in arterial pressure before and after two procedures of dental prophylaxis: Jet system baking soda and conventional prophylaxis and patient's opinion regarding the comfort of each one. MATERIAL AND METHOD: Were selected 32 patients with age between 18 to 30 years old, who need prophylaxis to remove biofilm and were subjected to three different types of treatment: sodium bicarbonate jet (G1), prophylaxis conventional (G2) and placebo (G3) at intervals of one month between them. Patients were divided randomly. Arterial pressure was measured by wrist digital Omron HEM – 6111. The measurements were realized in four times: before the prophylaxis, immediately the end of procedure, 15 and 30 minutes after finished of treatment. Patient comfort was measured by a Visual Analog Scale (VAS) after the end of the treatment. The data were analyzed using the variance test. The results showed that there was statistically significant difference to the comfort of the procedures. RESULTS: There was a statistically significant difference to the comfort of the procedures, and G2 and G3 better than G1. Regarding the variation of arterial pressure there was no statistically significant difference between groups. CONCLUSIONS: The methods of prophylaxis no effect on arterial pressure, but conventional prophylaxis is more comfortable than others treatments

Helibacter pylori: preaparação de Lapdesf-IUNI e ensaio de inibição de urease (IC50) de derivados de tiouréia e guanidínicos

O presente trabalho de conclusão de curso visou à otimização e preparação da síntese de Lapdesf-IUNI e avaliação do potencial de inibição de urease de três compostos previamente sintetizados, direcionados especificamente para o tratamento de infecções por Helicobacter pylori, bactéria mundialmente reconhecida como uma das principais causadoras de infecções no mundo, responsável pelos casos de infecções gástricas, como gastrites crônicas, úlceras (gástricas e duodenais) e carcinomas. Lapdesf-IUNI é um composto derivado de metronidazol, com atividade potencialmente inibidora de urease, enzima presente neste microrganismo e essencial a sua sobrevivência no ambiente gástrico. O produto foi obtido, porém com baixo rendimento, por isso sua metodologia ainda necessita ser aperfeiçoada, para que, em seguida este composto possa ser testado. Outros três compostos, um derivado de tiouréia e dois derivados guanidínicos, com potencial atividade inibidora de urease, T-6, R-17 e R-21, sintetizados pela equipe do Prof. Eduardo Perez, foram avaliados com relação a sua atividade inibidora frente à enzima urease e IC50, apresentando bons resultados, que permitem a continuidade dos estudos, comparando inclusive com fármacos utilizados nos protocolos de tratamento padrões para o tratamento das infecções causadas por H. pylori

Ensaio de ligações de estruturas de madeira por chapas com dentes estampados em quatro espécies da família leguminosas

In the last years, the use of industrialized systems of lattice structures of wood as an alternative in the construction processes, is becoming more popular in Brazil, mainly for their commitment to the environment.The industrialized system of wooden lattice structures consists in a production of lattice structures, composed of pieces of wood where their mechanical connections are made with the sheet multi-toothed connectors (CDE).Among the many challenges to make this system competitive, the whole system of the connections between the pieces of wood not only must show functionality, but also speed, strength, versatility and economy. Referenced at Brazilian Standard for Wood Structures (NBR 7190/1997 - Project of timber structures) the sheet multi-toothed connectors, are analyzed using three test methods: tensile strength parallel to grain, tensile strength normal to the fibers and shear strength, all of them in two positions, αCH0=0o e αCH0=90o to four types of wood: Angelim (Vatairea heteroptera Ducke); Red-Angico (Parapiptadenia rigida (Benth) Brenae); Garapa (Apuleia leiocarpa (Vog.) Macbr) and Jatoba (Hymenaea stilbocarpa Hayne), belonging to the Leguminosae family and founded in several regions of Brazil.The purpose of this manuscript consists to analyze the mechanical connections with the sheet multi-toothed connectors through tests from NBR7190/1997

Um contexto histórico e sócio-ambiental dos povos da floresta do Acre: ensaio sobre história recente desses povos e as políticas públicas estaduais do Governo da Floresta

O Estado do Acre, território mais recentemente incorporado ao Brasil, até meados do século XIX apresentava em seu território apenas povos tribais ameríndios, que lá habitavam desde milênios. O histórico de povoamento mais recente do estado (a partir da segunda metade do século XIX) está atrelado com o mercado internacional da borracha, a região foi o destino de milhares de pessoas em busca de fazer a vida explorando as seringueiras. A partir de então uma série de disputas fundiárias ocorreram na região, o que foi um dos fatores que influenciaram na conformação da sociedade acreana. O movimento ambientalista surgiu no estado como resultado dessas disputas fundiárias na segunda metade do século XX. No final da década 90 o governo estadual, assimilando o discurso ambientalista, assume o governo com a Frente Popular do Acre, composta por uma série de partidos, com o autodenominado “Governo da Floresta” e usando o neologismo “florestania” como símbolo do projeto do governo. Apesar de utilizar amplamente em seu discurso temáticas ambientais e dos povos da floresta, acabou ao longo dos anos tendo atitudes com um cunho mais doutrinador do que emancipador. A discrepância entre a prática e o discurso do “Governo da Floresta” é então analisada a partir de conceitos como “Ecologia dos Saberes” e “Biopoder”, conceitos que auxiliam na identificação das atitudes de cunho doutrinador postas em prática pelo governo. Considerando como “ecologia dos saberes” aquela forma de conhecimento que tem a premissa um reconhecimento da pluralidade de conhecimentos heterogêneos que em interações sustentáveis e dinâmicas entre eles não comprometem suas autonomias e “biopoder” como aquela forma de poder que exerce a sua dominação sobre a própria vida da população, estabelecendo como é que esses devem viver

Estudo da deformação heterogênea no ensaio do anel de atrito

The plastic deformation is widely used in the metallurgical market due to its positive factors such as low prices and high speed production. Forming process products are obtained in high quality, both surface quality and mechanical properties. Friction is an importante factor in metal forming. Friction study in metal forming can be accomplished indirectly, such as the ring test of friction. Two samples of different materials being mild steel and copper alloy were used. The results showed the influence of friction in the flow behavior of the deformation of the second phase, as evidenced by standard metallography. It is observed that in the outer regions of the ring, plastic deformation occured in the radial direction. In the central region of the disc deformation occured in the direction of compression and the inner region of the ring flux lines showed a significant deformation in the radial direction towards the center of the ring

Avaliação da atividade quimiopreventiva da substância Morelloflavona isolada de Garcinia xanthochymus

Câncer é o nome dado a um conjunto de mais de 100 doenças que têm em comum o crescimento desordenado (maligno) de células que invadem tecidos e órgãos, podendo espalhar-se (metástase) para outras regiões do corpo. Dividindo-se rapidamente, estas células tendem a ser muito agressivas e incontroláveis, determinando a formação de tumores (acúmulo de células cancerosas) ou neoplasias malignas. As causas de câncer são variadas, podendo ser externas ou internas ao organismo, estando inter-relacionadas. As causas externas referem-se ao meio ambiente e aos hábitos ou costumes próprios de uma sociedade. As causas internas são, na maioria das vezes, geneticamente pré-determinadas, e estão ligadas à capacidade do organismo de se defender das agressões externas (Sistema Imunológico ou acúmulo de mutações no DNA). A compreensão dos exatos mecanismos e a tentativa de inativar genes ou impedir as ações tóxicas de substâncias contribuirá para a elucidação de diversos pontos importantes para o entendimento da carcinogênese e, portanto, para novas estratégias de tratamento e de prevenção do câncer. O presente estudo tem como objetivo, avaliar o efeito da substância pura, morelloflavona, isolada da folha de Garcinia xanthochymus, planta típica do Cerrado no Estado de São Paulo, conhecida por sua variada atividade biológica (analgésica, antibacteriana, antioxidante, antiviral e antitumoral) devido aos seus grupos de metabólitos: xantonas, flavonóides, fenóis e acridonas. Assim, foram realizados: o ensaio da Quinona Redutase em paralelo com o ensaio do Violeta Cristal para análise da viabilidade celular em células de hepatocarcinoma murino (Hepa 1c1c7); o ensaio de Sulforrodamina B para análise da viabilidade celular em células de hepatocarcinoma humano (HepG2) e o ensaio do Cometa nos protocolos de genotoxicidade e antigenotoxicidade em células... (Resumo completo, clicar acesso eletrônico abaixo)

Reprodução de papagaio-verdadeiro (Amazona aestiva) em cativeiro: perfil anual de esteróides sexuais e ensaio de estímulo hormonal exógeno

Pós-graduação em Medicina Veterinária - FCAV

Influência das substâncias húmicas aquáticas na genotoxicidade dos metais em tilápias do Nilo (Oreochromis niloticus)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biodiesel etílico de babaçu x diesel B S1800: ensaio de opacidade da fumaça do trator agrícola variando os horários do dia

Pós-graduação em Agronomia (Ciência do Solo) - FCAV

Bloqueio pericoal guiado por ultrassom: ensaio clínico randonizado

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito quimiopreventivo e modulador de HLA-G (Human Leukocyte Antigen - G) de Morelloflavona

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)