Inflammation is a biological defence mechanism applied by living organisms against foreign invaders. In the response to DAMPs and PAMPs, organisms use inflammatory multi-protein complexes to fight the attackers. The most studied inflammasome proteins are NLRP3, ASC and Caspase-1. This study is aimed at understanding the role of NEK7 protein in the NLRP3 inflammasome’s activation, using CRISPR/Cas9 system. To determine the effect of CRISPR/Cas9 and transfection, mRNA expression was analyzed. The results obtained suggest that neither the transfection nor the NEK7 protein knockout have sufficiently worked. This study could not experimentally establish that NEK7 triggers NLRP3 inflammasome activation because ELISA was not conducted to verify the levels of cytokines emitted, due to there being no statistical differences between the samples. Above all, the research question in this thesis project was not answered because the instability of the ACTB reference gene negatively influenced the results. However, previous related studies conclude that NEK7 plays a crucial role in the activation of the NLRP3 inflammasome.

Orientador: Marco Eustáquio de Sá / Banca: Kuniko Iwamoto Haga / Banca: Stela Maris Kulczynski / Resumo: Nos últimos anos a tecnologia tem sido utilizada visando o aumento da produtividade em cultivos de feijão. O presente trabalho teve como objetivo avaliar o efeito da aplicação de bioestimulante sobre a produtividade e a qualidade fisiológica das sementes de dois cultivares de feijão, cultivados no inverno em condições de Cerrado. A pesquisa foi conduzida no período do outono-inverno de 2007, na Fazenda de Ensino, Pesquisa e Extensão da Faculdade de Engenharia do Campus de Ilha Solteira - UNESP, localizada no Município de Selvíria/MS. No experimento foram utilizadas sementes de feijão do cultivar IAC Apuã e Carioca Precoce. O delineamento experimental foi blocos casualizados, no esquema em faixas com um arranjo fatorial 5 x 2 x 2, com quatro repetições. Os tratamentos foram constituídos pela combinação de cinco doses (0; 0,5; 1,0; 1,5 e 2,0 L p.c. ha-1) de Stimulate, em duas épocas de aplicação em dois cultivares: no estádio vegetativo (V4) e no reprodutivo (R5). Avaliou-se a população final de plantas, altura média das plantas e inserção da primeira vagem, nº de vagens/planta, nº de sementes/vagem, nº de sementes/planta, massa de 100 sementes e a produtividade. O Stimulate proporcionou aumento no número de vagens/planta (Cultivar IAC Apuã), número de sementes/planta e na produtividade, e para o número de sementes/planta e a produtividade a melhor época de aplicação foi em R5. As doses e as épocas de aplicação do Stimulate não influenciaram significativamente na altura das plantas e inserção da primeira vagem, número de sementes/vagem, massa de 100 sementes e nos teores foliares dos nutrientes em nenhum dos cultivares. E na qualidade fisiológica das semente as doses do Stimulate influenciaram significativamente na porcentagem de plântulas fortes, porém não afetaram a taxa de germinação, primeira contagem, comprimento de plântulas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Lately, technology has been used to increase bean productivity. Therefore, this research work was aimed at evaluating the application effect of bio stimulant on the seeds productivity and physiological quality of two common bean seeds cultivars, grown winter in Cerrado conditions. The research was carried out during autumn-winter of 2007, on the Fazenda de Ensino, Pesquisa e Extensão da Faculdade de Engenhaira do Campus de Ilha Solteira - UNESP, in Selvíria-MS. Seeds of IAC Apuã and Carioca Precoce cultivars were used in the experiment. The experimental design was the randomized blocks in lanes with a 5 x 2 x 2 factorial outline and four replications. The treatments consisted of the combination of five Stimulate doses (0; 0.5; 1.0; 1.5 and 2.0 L p.c. ha-1), in two application periods: at vegetative stage (V4) and at reproductive stage (R5) in two common bean cultivars. Final plants population, plants average height and first pod insertion, number of pods/plant, number of seeds/pod, number of seeds/plant, 100-seed mass and productivity were evaluated. Stimulate provided an increase in the number of pods/plant, number of seeds/plant and also in the productivity and as for the number of seeds/plant and productivity, the most appropriate application period was at R5. The plants height and the first pod insertion, number of seeds/pod, 100-seed mass and leaf contents were not influenced by the Stimulate doses nor the application times. Concerning the seeds physiological quality of the cultivars, Stimulate doses significantly influenced the percentage of healthy seedlings; however, they did not influence the germination rate, the first counting, the seedlings length, emergence, speed emergence, seedlings height, fresh and dry matter of the seedlings, fast aging, cold test and electrical conductivity. / Mestre