Submitted by Silvana Teresinha Dornelles Studzinski (sstudzinski) on 2016-02-11T11:31:09Z No. of bitstreams: 1 Liriana Zanon Stefanello_.pdf: 11645045 bytes, checksum: e9a1d6a5118168de4fe492a58c1bcee7 (MD5) / Made available in DSpace on 2016-02-11T11:31:09Z (GMT). No. of bitstreams: 1 Liriana Zanon Stefanello_.pdf: 11645045 bytes, checksum: e9a1d6a5118168de4fe492a58c1bcee7 (MD5) Previous issue date: 2015-11-05 / Milton Valente / A tese é um estudo sobre a construção das memórias familiares de imigrantes na região da Quarta Colônia Imperial de Imigração Italiana do Rio Grande do Sul – Brasil. Ela fala da trajetória das famílias Pippi e Pigatto, da Itália ao Brasil, buscando analisar quais os mecanismos e dinâmicas usados para a inserção dessas famílias na sociedade sul-rio-grandense e como estas vêm mantendo as suas memórias. Para tanto, estudamos não só as estratégias usadas por estes imigrantes para se inserirem na sociedade receptora, mas também, como a memória de si e sobre si foi e vem sendo construída por e para seus descendentes. A elaboração da memória é uma construção social, então, selecionar o que deve ser lembrado e o que se relega ao esquecimento é parte de todo e qualquer processo de construção de memória e de identidade. E mais que isso, compreendendo o indivíduo como ator social responsável pela sua história, os documentos familiares tornam-se patrimônio de uma comunidade quando a representam, uma vez que carregam consigo uma carga simbólica de representar no presente o passado de cada família. Para estas duas famílias, como para centenas de outras, a construção de uma memória que chamamos de oficial ocorreu através da criação do Centro de Pesquisas Genealógicas (CPG) de Nova Palma, organizado pelo Padre Luiz Sponchiado (01/06/1984) com o objetivo de preservação da memória deste grupo imigrante. Uma das famílias estudadas, entretanto, reelaborou suas memórias também de forma particular, a família Pippi. Ao construir uma outra memória de si, esta criou um acervo privado que deu a ler uma outra história/memória familiar, que em última instância tencionou a memória oficial organizada no CPG. Já os documentos da família Pigatto, guardados no CPG, representam e são aceitos como a sua memória na Quarta Colônia de Imigração Italiana do Rio Grande do Sul. Além desse caráter memorial, todos os documentos são parte do patrimônio cultural desta comunidade quer os guardados no acervo do CPG, quer no acervo particular da Sra. Maria Neli Donatto Pippi ou de outros não identificados. Juntos, eles fazem parte desta busca dos indivíduos pela identificação e pela afirmação de uma memória familiar. Cada documento “escolhido” torna-se, então, um símbolo de rememoração que objetiva reconstruir a trajetória, a história e a memória familiar. / The thesis is a study on the construction of family memories of immigrants in the region of the Fourth Imperial Colony of Italian Immigration of Rio Grande do Sul - Brazil. It addresses the trajectory of Pippi and Pigatto families, from Italy to Brazil seeking to analyze the mechanisms and dynamics used for insertion in the south of Rio Grande society and how they have maintained their memories. For that we study not only the strategies used by these immigrants to integrate into the host society but also how the memory itself and on itself was and is being built by and for their descendants. The development of memory is a social construct, therefore selecting what should be remembered and what is relegated to oblivion is part of any construction process of memory and identity. And more than that, understanding the individual as a social actor responsible for its history, family documents become heritage of a community when they represent it, because they carry the symbolic burden of representing in the present the past of each family. For these two families as well as hundreds of others, the construction of a memory - we call official - occurred through the creation of the Center for genealogy Research (CPG) of Nova Palma organized by Father Luiz Sponchiado (01/06/1984) in order to preserve the memory of this immigrant group. However one of the studied families has also reworked their memories in a particular way. The Pippi family, on building another memory itself, has created a private collection that has given to read another story / family memory, which ultimately tensed the official memory organized at the CPG. Nonetheless Pigatto family documents stored in CPG represent and are accepted as its memory at the Fourth Colony of Italian Immigration of the Rio Grande do Sul. In addition to this memorial character all documents are part of the cultural heritage of this community which want them stored in the CPG's collection, either in the private collection of Mrs. Maria Neli Donatto Pippi or unidentified others. Together they are part of this individuals search for identification and affirmation of a family memory. Each "chosen" document becomes then a symbol of remembrance that aims to reconstruct the trajectory, history and family memory.

Dans cette thèse nous proposons un outil d’aide à la décision multicritère qui permet aux décideurs de sélectionner un scénario optimal dans un graphe de projet qui contient toutes les alternatives de choix de conception et de réalisation d’un nouveau système, tout en tenant compte des risques inhérents aux choix réalisés. Le modèle du graphe est construit en considérant toutes les décisions collaboratives des différents acteurs impliqués dans le projet. Cet outil d’aide à la décision est basé principalement sur les techniques de l’optimisation combinatoire. En effet, nous avons choisi de travailler avec la métaheuristique ACO (algorithme d’optimisation par colonies de fourmis) vu sa capacité à fournir des solutions optimales dans un temps raisonnable. Les objectifs à minimiser sont le coût global du projet, sa durée totale de réalisation et l’incertitude sur ces critères (coût, durée). La modélisation des incertitudes a été abordée suivant deux approches différentes. La première approche consiste à modéliser l’incertitude en utilisant des intervalles simples et en la considérant comme un objectif à part entière à optimiser avec le coût et la durée. Quant à la deuxième approche, elle permet de modéliser l’incertitude sur les objectifs du projet (coût, durée) sous formes de distributions de probabilités. L’outil d’optimisation proposé dans la thèse fait partie d’un processus intégré et plus global qui se base sur les standards industriels (processus d’ingénierie système et de management de projet) qui sont largement connus et utilisés dans les entreprises. Ainsi, le travail développé dans cette thèse constitue un vrai guide pour les industriels dans leurs processus de conception et de réalisation des systèmes complexes innovants dans le domaine d’ingénierie système. / In this thesis, we propose a multi-criteria decision making tool that allows decision-makers to select an optimal scenario in a project graph that includes all the alternative choices of a new system’s conception and realization, while taking into account the risks inherent to these choices. The model of the graph is constructed by considering all the collaborative decisions of the different actors involved in the project. This decision making tool is based mainly on the techniques of combinatorial optimization. Indeed, we have decided to work with the metaheuristic ACO (Ant Colony Optimization algorithm) for its ability to provide optimal solutions in a reasonable amount of time. The objectives to be minimized are the total cost of the project, its global duration and the uncertainties about these criteria (cost, duration). The uncertainties modeling is performed according to two different approaches. The first approach consists in using single intervals to model the uncertainty and it is considered as a third objective to optimize besides cost and duration. As for the second approach, the uncertainty about project objectives (cost, duration) is performed by using probabilities distributions. The optimization tool proposed in this thesis is a part of an integrated and more global process, based on industrial standards (the systems engineering process and the project management one) that are widely known and used in companies. Thus, the work developed in this thesis is a real guide for companies in their process of design and realization of innovative complex systems in the systems engineering field.

The growing demand for strawberries has imposed challenges, especially regarding the control of pests. Many farmers report problems with reduced chemical control efficiency, probably due to selection of resistant populations of insects and mites. An alternative is the use of biological control using pathogenic fungi as a tool in integrated pest management. Metarhizium spp. (Hypocreales: Clavicipitaceae) are generalist entomopathogenic fungi with worldwide distribution and can cause diseases in a large number of hosts. Many studies on the development of Metarhizium as a biological control agent were performed, but this bulk of knowledge is in remarkable contrast to the lack of research on the fundamental ecology of Metarhizium in agroecosystems. This thesis aimed to evaluate the establishment, persistence and dispersal of these entomopathogenic fungi in strawberry crop soil in Inconfidentes, Minas Gerais, Brazil; and to study the diversity and abundance of species of Metarhizium isolated from organic and conventional strawberry crop soils, and the field margins in Brazil and Denmark. The effectiveness of new species of Metarhizium recently found in Brazil, was evaluated against two spotted spider mite, Tetranychus urticae. Applied isolates of M. anisopliae (ESALQ1037) and M. robertsii (ESALQ1426) were able to persist for up to 12 months after the application within the soil, and disperse to other plots and colonize the rhizosphere of strawberry plants. In the plots where ESALQ1037 and ESALQ1426 were applied, 25% and 87.5% of the isolates recovered after 12 months consisted of the same isolates inoculated. A new taxonomically unassigned lineage, referred to as Metarhizium sp. Indet. 5 in this study, was found in strawberry crop margins. The dominant species of Metarhizium in Brazil and Denmark was Metarhizium robertsii and M. brunneum respectively. Further, Metarhizium pemphigi was first detected in Denmark in this study. Soil in organically grown strawberries harbored a more diverse population of Metarhizium spp. compared with conventionally grown strawberries. These studies showed for the first time the potential of new species of Metarhizium as spider mite biological control agents, the lowest median lethal time (LT50 = 4 ± 0.17 days) was observed in mites treated with the isolate ESALQ1638 of Metarhizium sp. indet. 1. The best isolates were ESALQPL63 of B. bassiana, ESALQ1608 and ESALQ1638 of Metarhizium sp. indet. 1 and ESALQ3069 and ESALQ3222 of M. pingshaense based on the survival curve, total mortality, percentage of sporulated cadavers and LT50. Knowledge of the diversity of Metarhizium spp. and persistence in strawberry soil generated in this study may be useful in developing conservation strategies and maximize the natural biological pest control. / A crescente demanda por morangos vem impondo desafios, especialmente quanto ao controle das pragas. Muitos agricultores relatam problemas com a redução da eficiência do controle químico, provavelmente devido à seleção de populações resistentes de insetos e ácaros. Uma alternativa é o uso de controle biológico com fungos entomopatogênicos como ferramenta dentro do manejo integrado de pragas. Metarhizium spp. (Hypocreales: Clavicipitaceae), são fungos entomopatogênicos generalistas com distribuição cosmopolita e que podem causar doenças em um grande número de hospedeiros. Muitos estudos sobre o desenvolvimento de Metarhizium como agente de controle biológico foram realizados, mas este leque de conhecimento está em contraste com a notável falta de investigação sobre a ecologia de Metarhizium nos agroecossistemas. Esta tese teve como objetivo avaliar o estabelecimento, persistência e dispersão destes fungos entomopatogênicos em solo de morangueiro em Inconfidentes, Minas Gerais, Brasil; bem como estudar a diversidade e abundância de espécies de Metarhizium isolados do solo de cultivos orgânico e convencional de morangueiro, e das margens das plantações no Brasil e Dinamarca. A eficácia de novas espécies de Metarhizium, encontradas recentemente no Brasil, foi avaliada contra o ácaro rajado, Tetranychus urticae. Os isolados inoculados de M. anisopliae (ESALQ1037) e M. robertsii (ESALQ1426) foram capazes de persistir por até 12 meses após a aplicação no solo, além de dispersar para outras parcelas e colonizar a rizosfera dos morangueiros. Nas parcelas onde ESALQ1037 e ESALQ1426 foram aplicados, 25% e 87,5% dos isolados recuperados após 12 meses consistiam dos mesmos isolados inoculados. Uma nova linhagem não taxonomicamente identificada, referida neste trabalho como Metarhizium sp. Indet. 5, foi encontrada nas margens de morangueiros cultivados. A espécie dominante de Metarhizium no Brasil e Dinamarca foi Metarhizium robertsii e M. brunneum, respectivamente. Além disso, Metarhizium pemphigi foi detectado pela primeira vez na Dinamarca neste estudo. Solos de cultivo orgânico de morangueiro em geral apresentaram uma diversidade maior de Metarhizium do que solos de cultivos convencionais. Estes estudos revelaram pela primeira vez o potencial de novas espécies de Metarhizium como agentes de controle biológico do ácaro rajado, sendo o menor tempo letal mediano (TL50= 4 ± 0.17 dias) observado em ácaros tratados com o isolado ESALQ1638 de Metarhizium sp. indet. 1. Os melhores isolados foram ESALQPL63 de B. bassiana, ESALQ1608 e ESALQ1638 de Metarhizium sp. indet. 1 e ESALQ3069 e ESALQ3222 de M. pingshaense baseado na curva de sobrevivência, mortalidade total, porcentagem de cadáveres esporulados e TL50. O conhecimento da diversidade de Metarhizium spp. e persistência em solos de morango, gerados neste estudo, poderão ser úteis no desenvolvimento de estratégias de conservação e maximizar o controle biológico natural de pragas.

A aplicação de sistemas de múltiplos robôs é desejável em várias tarefas. Algumas delas são: exploração de ambientes, mineração, detecção de minas terrestres, segurança e operações de resgate. Uma estratégia eficiente de coordenação é decisiva para alcançar melhoras no desempenho. Neste projeto, duas novas estratégias são propostas para a coordenação de sistemas de múltiplos robôs, aplicadas para as tarefas de exploração, vigilância e formação. Elas são distribuídas, descentralizadas e ocorrem em tempo de execução. A inspiração para ambas advém de mecanismos biológicos que definem uma organização social de sistemas coletivos. Especificamente, considerou-se nesta tese uma versão modificada do sistema de colônia de formigas. As estratégias são adaptáveis para cenários em que o número de robôs e a estrutura do ambiente mudam. Em relação à primeira estratégia, os experimentos consideram dois critérios de desempenho: a média de ciclos de vigilância e a média de iterações em cada intervalo de segurança. Os resultados de simulação confirmam que a exploração e vigilância emergem da sinergia dos comportamentos individuais dos robôs. Os dados obtidos mostram que a estratégia de coordenação é eficiente e satisfatória para realizar as tarefas de exploração e vigilância. Quanto à segunda estratégia, o sistema apresenta as características desejáveis para que a formação seja mantida: separação, alinhamento e coesão. Evidências empíricas mostraram que o sistema possui boa habilidade dispersiva, o que promoveu o aumento da cobertura, e que o mesmo foi capaz de se adaptar a novas topologias de grupo e configurações de ambiente / The application of systems of multiple robots is desirable in various tasks. Some of them include: exploration, mining, land mine detection, security and rescue operations. An effective strategy for coordination is crucial to achieve performance improvements. In this project, two new strategies are proposed for the coordination of multiple robot systems, applied to the tasks of exploration, surveillance and formation. They are distributed, decentralized and performed in real time. The inspiration for both of them comes from the biological mechanisms that define a social organization of collective systems. Specifically, it was considered in this thesis a modified version of the Ant Colony System. The strategies are adaptable for scenarios where the number of robots and structure of the environment change. Regarding the first strategy, the experiments consider two performance criteria: average surveillance cycles and average iterations for each patrolling interval. Simulation results confirm that the exploration and surveillance emerge from the synergy of individual behaviors of robots. The data obtained show that the coordination strategy is efficient and and suitable to perform the tasks of exploration and surveillance. Regarding the second strategy, the system presents the characteristics desirable to maintain the formation: separation, alignment and cohesion. Empirical evidence showed that the system has good dispersive ability, which promoted an increase in coverage, and that it was able to adapt to new group topologies and environment settings

A manutenção do estado progenitor durante o cultivo de células mesenquimais progenitoras derivadas do tecido adiposo (MSCs-TA), caracterizado pelo potencial de diferenciação e da capacidade de autorrenovação, é atualmente um dos maiores desafios da terapia celular. Sabendo da influência da angiogênese no desenvolvimento de tecidos de origem mesenquimal, avaliamos se um ambiente pro-angiogênico mimetizado em cultura forneceria condições para manutenção de um estado progenitor durante o processo de expansão celular. Utilizando como modelo de um ambiente pró-angiogênico o cultivo no meio EGM-2, o qual é suplementado pelos fatores de crescimento EGF, FGF-2, IGF e VEGF, nós demonstramos que a presença de tais fatores pró-angiogênicos é fundamental para a manutenção do estado progenitor de MSCs-TA em cultura. Verificamos que a presença de tais fatores de crescimento possibilitaram às MSCs-TA apresentarem um alto potencial de diferenciação adipogênico e osteogênico em comparação ao meio convencional DMEM/F12 e ao meio EBM, ausente de fatores. Além disso, o cultivo na presença de fatores pró-angiogênicos aumentou o potencial clonogênico das MSCs-TA, ao mesmo tempo em que aumentou a capacidade proliferativa destas células. Dentre os fatores de crescimento, EGF e FGF-2 foram responsáveis pelos efeitos mais robustos. Ao mesmo tempo, células cultivadas nas presença destas citocinas foram capazes de manter a morfologia fibroblastóide e apresentaram alta expressão do fator de pluripotência Klf-4. Em concordância com estes achados, o transplante subcutâneo de MSCs-TA cultivadas nestas condições mostrou que aquelas mantidas em EGM-2 geram um tecido semelhante ao tecido formado pela fração estromal vascular não cultivada. Estes resultados reforçam o papel do ambiente pró-angiogênico na manutenção do estado progenitor de MSCs-TA, e que tal estado foi proporcionado pela ação dos fatores de crescimento pró-angiogênicos EGF, FGF-2, IGF e VEGF nas células em cultivo, com destaque para as citocinas EGF e FGF-2. Em conclusão, o uso do ambiente pró-angiogênico no cultivo de MSCs-TA mostrou-se como uma abordagem promissora para a manutenção do estado progenitor destas células in vitro. / The maintenance of the progenitor state in the culture of adipose tissue derived- mesenchymal progenitor cell (TA-MSCs), characterized by the differentiation potential and self-renewal capability, is currently one of the major challenges of cell therapy. The information that the angiogenesis influences the development of mesenchymal tissues, has led us to evaluate how a pro-angiogenic environment mimicked in culture would provide conditions for maintaining a progenitor state during the cell expansion process. We designe a model for a pro-angiogenic environment in which cells grown in EGM-2 supplemented with the following growth factors: EGF, FGF-2, IGF and VEGF, and demonstrated that the presence of such pro-angiogenic growth factors was crucial for maintenance of the progenitor of AT-MSCs in culture. We observed that the presence of such growth factors allowed to AT-MSCs a high potential of adipogenic and osteogenic differentiation compared to conventional DMEM/F12 medium and the EBM medium, in the absence of the factors. Furthermore, the culture in presence of pro-angiogenic growth factors increased the clonogenic potential of AT-MSCs and increased the proliferative capability of these cells. Among the growth factors, EGF and FGF-2 were responsible for most robust effects. At the same time, cells cultured in the presence of these cytokines were able to maintaining the fibroblastoid morphology and presented high expression levels of Klf-4 pluripotency factor. In agreement with these observations, the subcutaneous transplantation of AT-MSCs cultured under these conditions showed that those cells kept in EGM-2 generated a tissue-like to tissue formed by the stromal vascular fraction uncultivated. These results reinforce the role of the pro-angiogenic environment in the maintenance of the progenitor state of AT-MSCs, and that such a state was provided by the action of the pro-angiogenic growth factors EGF, FGF-2, IGF and VEGF in cultured cells, highlighting EGF and FGF-2 cytokines. In conclusion, we showed that the use of a pro-angiogenic environment in AT-MSCs culture is a promising approach to the maintain the progenitor state of these cells in vitro.

by Ng Wing Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 156-174). / Abstracts in English and Chinese. / Acknowledgements --- p.iii / Abstract --- p.v / 摘要 --- p.vii / Table of Contents --- p.ix / List of Figures --- p.xiii / List of Tables --- p.xvi / List of Graphs --- p.xvii / List of Abbreviations --- p.xviii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter Chapter 2 --- Literature Review --- p.3 / Chapter 2.1 --- Human granulocyte colony-stimulating factor (hG-CSF) --- p.3 / Chapter 2.1.1 --- Historical background --- p.3 / Chapter 2.1.2 --- Physiological Roles --- p.5 / Chapter 2.1.3 --- Molecular properties --- p.8 / Chapter 2.1.4 --- Biochemical properties --- p.9 / Chapter 2.1.5 --- Comparison to G-CSF of other species --- p.11 / Chapter 2.1.6 --- Biological Activities --- p.12 / Chapter 2.1.7 --- Clinical Applications --- p.14 / Chapter 2.1.7.1 --- Clinical use in myelosuppressive chemotherapy and neutropenic fever --- p.14 / Chapter 2.1.7.2 --- Clinical use in bone marrow transplantation (BMT) and peripheral blood progenitor cell (PBPC) transplantation --- p.14 / Chapter 2.1.7.3 --- Clinical use in HIV infection --- p.16 / Chapter 2.1.7.4 --- Clinical use in diabetes mellitus --- p.17 / Chapter 2.1.7.5 --- Clinical use in severe chronic neutropenia --- p.18 / Chapter 2.1.7.6 --- Future prospects --- p.18 / Chapter 2.1.7.7 --- Dosages and adverse effects --- p.19 / Chapter 2.1.8 --- Economic value --- p.20 / Chapter 2.2 --- Plant as bioractor --- p.20 / Chapter 2.2.1 --- Medical molecular farming --- p.20 / Chapter 2.2.2 --- Commercial biopharmaceutical proteins --- p.25 / Chapter 2.2.3 --- Transgenic plants producing hematopoietic growth factors --- p.25 / Chapter 2.2.3.1 --- Granulocyte-macrophage colony-stimulating factor (GM-CSF) --- p.26 / Chapter 2.2.3.2 --- Interleukin-2 (IL-2) --- p.28 / Chapter 2.3 --- Rice as expression system --- p.29 / Chapter 2.3.1 --- Characteristics --- p.29 / Chapter 2.3.2 --- Advantages of using rice as bioreactor --- p.30 / Chapter 2.3.3 --- Previous studies --- p.31 / Chapter 2.3.4 --- Transformation method --- p.33 / Chapter 2.3.5 --- Super-binary vector --- p.34 / Chapter 2.4 --- Strategies for enhancing protein expression level --- p.36 / Chapter 2.4.1 --- Vacuolar targeting --- p.36 / Chapter 2.4.1.1 --- Protein targeting signals --- p.38 / Chapter 2.4.1.2 --- Binding protein of 80kDa (BP-80) --- p.39 / Chapter 2.4.1.3 --- a-Tonoplast intrinsic protein (α-TIP) --- p.39 / Chapter 2.4.1.4 --- Receptor homology region-transmembrane domain-Ring H2 motif (RMR) --- p.40 / Chapter 2.4.2 --- Fusion with glutelin in rice --- p.41 / Chapter 2.5 --- Hypotheses and aims of this study --- p.43 / Chapter Chapter 3 --- Materials and Methods --- p.45 / Chapter 3.1 --- Introduction --- p.45 / Chapter 3.2 --- Chemicals --- p.45 / Chapter 3.3 --- Bacterial strains --- p.46 / Chapter 3.4 --- Chimeric genes construction --- p.46 / Chapter 3.4.1 --- Protein targeting constructs --- p.51 / Chapter 3.4.2 --- Enterokinase site constructs --- p.60 / Chapter 3.4.3 --- Glutein signal peptide constructs --- p.65 / Chapter 3.4.4 --- Glutelin fusion constructs --- p.70 / Chapter 3.4.5 --- Sequence fidelity of chimeric genes --- p.77 / Chapter 3.4.6 --- Cloning of chimeric genes into rice super-binary vector --- p.77 / Chapter 3.5 --- Rice transformation --- p.79 / Chapter 3.5.1 --- Plant materials --- p.79 / Chapter 3.5.2 --- Agrobacterium transformation --- p.79 / Chapter 3.5.3 --- A grobacterium-mediated transformation of rice --- p.79 / Chapter 3.6 --- Transgenic expression --- p.81 / Chapter 3.6.1 --- Extraction of leaf genomic DNA --- p.81 / Chapter 3.6.2 --- Synthesis of DIG-labeled double-stranded DNA probe --- p.82 / Chapter 3.6.3 --- Southern blot analysis --- p.83 / Chapter 3.6.4 --- Extraction of total RNA from immature rice seeds --- p.84 / Chapter 3.6.5 --- Northern blot analysis --- p.85 / Chapter 3.6.6 --- Protein extraction --- p.86 / Chapter 3.6.7 --- Tricine SDS-PAGE --- p.86 / Chapter 3.6.8 --- Western blot analysis --- p.87 / Chapter 3.6.9 --- Enterokinase digestion of EK fusion proteins --- p.88 / Chapter 3.7 --- Confocal immunoflorescence studies of rhG-CSF in rice grain --- p.89 / Chapter 3.7.1 --- Preparation of sample sections --- p.89 / Chapter 3.7.2 --- Double-labeling of fluorescence probes --- p.89 / Chapter 3.7.3 --- Image collection --- p.90 / Chapter 3.8 --- Functional analysis of rhG-CSF --- p.91 / Chapter 3.8.1 --- Culture of NFS-60 cells --- p.91 / Chapter 3.8.2 --- MTT cell proliferation assay --- p.92 / Chapter 3.9 --- Bacterial expression of anti-hG-CSF --- p.93 / Chapter 3.9.1 --- pET expression in E. coli --- p.93 / Chapter 3.9.2 --- Purification of His-hG-CSF --- p.97 / Chapter 3.9.3 --- Immunization of rabbits --- p.97 / Chapter Chapter 4 --- Results --- p.99 / Chapter 4.1 --- Construction of chimeric genes for rice transformation --- p.99 / Chapter 4.2 --- "Rice transformation, selection and regeneration" --- p.103 / Chapter 4.3 --- Southern blot analysis --- p.105 / Chapter 4.4 --- Northern blot analysis --- p.109 / Chapter 4.5 --- Western blot analysis --- p.114 / Chapter 4.6 --- Enterokinase digestion of EK fusion proteins --- p.125 / Chapter 4.7 --- Confocal immunofluorescence studies of rhG-CSF in transgenic rice grain --- p.128 / Chapter 4.8 --- Functional analysis of rhG-CSF --- p.132 / Chapter 4.9 --- Bacterial expression of anti-hG-CSF --- p.135 / Chapter 4.9.1 --- Expression and purification of recombinant His-hG-CSF in E. coli --- p.135 / Chapter 4.9.2 --- Titer and specificity of the anti-serum --- p.137 / Chapter Chapter 5 --- Discussion --- p.139 / Chapter 5.1 --- Introduction --- p.139 / Chapter 5.2 --- Fusion of hG-CSF with protein sorting determinants --- p.141 / Chapter 5.3 --- Fusion of hG-CSF with rice glutelin --- p.145 / Chapter 5.4 --- Glutelin signal peptide --- p.146 / Chapter 5.5 --- O-glycosylation --- p.148 / Chapter 5.6 --- Enterokinase digestion --- p.148 / Chapter 5.7 --- Expression level of rhG-CSF --- p.149 / Chapter 5.8 --- Functional analysis of rhG-CSF --- p.151 / Chapter 5.9 --- Future perspectives --- p.151 / Chapter Chapter 6 --- Conclusion --- p.155 / References --- p.156

Perçues jusqu’au XIXe siècle comme des masses gélatineuses aux propriétés urticantes, les méduses ont été classées dans le groupe des zoophytes, organismes intermédiaires entre les animaux et les végétaux. Les savants s’en sont alors servis pour démontrer la continuité entre les êtres vivants végétaux et animaux et elles sont devenues des arguments essentiels dans l’établissement de l’échelle des êtres. A partir du XIXe siècle, on découvre, en particulier grâce à la microscopie, la complexité de leur anatomie, ce qui permet enfin de les caractériser de manière positive. Au milieu du XIXe siècle, on met en évidence les différentes phases de leur cycle de vie et l’organisation en colonie de certaines méduses. Les méduses deviennent alors l’un des principaux centres d’intérêt des zoologistes et des embryologistes qui cherchent à trouver la signification évolutive des différentes étapes de leur développement. Le terme de méduse apparaît alors pour désigner un de leurs stades de vie. En parallèle de cette histoire conceptuelle, les différentes étapes de cette classification ont été marquées par des difficultés d’étude qui ont fait que les méduses constituaient un des rares groupes non définis par des caractères positifs à la fin du XVIIIe siècle. En effet, si les expéditions scientifiques ont permis de récupérer de nombreux échantillons, bien vite s’est posé le problème de leur conservation. Face aux difficultés de maintenir leur morphologie et leurs couleurs après leur prélèvement, les images naturalistes de méduses sont donc devenues des objets d’étude essentiels dans la connaissance et la classification de ces organismes. Ces images donnent un reflet de l’évolution conceptuelle et technique, qui a accompagnée la classification des méduses / Perceived until the 19th century as gelatinous masses with stinging properties, jellyfish were classified in the group of zoophytes, intermediate organisms between animals and plants. Scientists then used it to demonstrate continuity between plant and animal living beings and became essential arguments in establishing the scale of beings. From the nineteenth century, we discover, particularly through microscopy, the complexity of their anatomy, which finally allows to characterize them positively. In the mid-nineteenth century, we highlight the different phases of their life cycle and the colony organization of some jellyfish. Jellyfish become one of the main interests of zoologists and embryologists who seek to find the evolutionary significance of the different stages of their development. The term jellyfish then appears to designate one of their life stages. In parallel with this conceptual history, the various stages of this classification were marked by study difficulties which made jellyfish one of the few groups not defined by positive characters at the end of the eighteenth century. Indeed, if the scientific expeditions allowed to recover many samples, quickly the problem of their conservation was posed. Faced with the difficulties of maintaining their morphology and their colors after their collection, the naturalistic images of jellyfish have thus become essential objects of study in the knowledge and classification of these organisms. These images give a reflection of the conceptual and technical evolution, which accompanied the classification of jellyfish

Introdução: O leite pode ser considerado um dos alimentos mais completos por apresentar alto teor de proteínas e sais minerais, porém, também é considerado excelente meio de cultura para microrganismos. Objetivos: Identificar o número de UFC de Staphylococcus spp. e Klebsiella pneumoniae, cultivados isoladamente ou em associação, em leite integral estéril, a 6 oC, 27 oC e 37 oC e descrever a curva de morte térmica, quando submetidas ao processo de pasteurização lenta isoladamente ou em associação. Material e Métodos: Avaliação através da contagem das UFC, do comportamento de Staphylococcus spp. e K. pneumoniae, isoladas de tanque de refrigeração de leite, submetidas a temperaturas que simulam o leite em condições de refrigeração (6 °C), condições ambientais (25 °C) e na temperatura ideal de crescimento de patógenos mesófilos (36°C). Avaliação da sensibilidade de Staphylococcus spp. e K. pneumoniae isoladas e em associações ao processo de pasteurização lenta. Resultados: Na escala 1 de Mac Farland a média de UFC de K. pneumoniae foi maior que a de Staphylococcus spp. A 6 °C as bactérias no leite, isoladas ou em associação crescem na mesma velocidade. A 25 °C a K. pneumoniae cresce mais que o Staphylococcus spp. A 25 °C K. pneumoniae associada ao Staphylococcus spp. cresce mais do que quando encontra-se isolada. A 36 °C K. pneumoniae associada ao Staphylococcus spp. cresce mais do que quando encontra-se isolada e ainda mais que a 25 °C. A pasteurização lenta foi efetiva, pois reduziu em no mínimo 90% as UFC no leite após 30 minutos a 65 °C. Conclusão: a medida que aumenta a temperatura, até 36 °C, a Klebsiella pneumoniae apresenta crescimento superior ao Staphylococcus spp., a contaminação de leite por Klebsiella pneumoniae (contaminação ambiental) irá influenciar mais na qualidade do produto final, comparando-se à contaminação por Staphylococcus spp., oriundo de mastite e a interação de microrganismos altera a morte dos mesmos, recomendando-se que novos estudos sejam realizados para que se entenda melhor esse processo. / Introduction: The milk can be considered one of the most complete food for its high contents of protein and minerals, but also it is considered an excellent medium of culture for microorganisms. Objective: To identify the number of the Colony Forming Units of Staphylococcus spp. and Klebsiella pneumoniae, in integral sterile milk, at different temperatures (6ºC, 27ºC e 37ºC) and to describe a thermal curve death, when submitted to the process of slow pasteurization isolated or in association. Materials and methods: Evaluation by counting of the colony forming units and the behavior of the Staphylococcus spp. and Klebsiella pneumoniae, isolated and in association occurred in the cooling tank, submitted to temperatures that simulate the conditions of milk in cooling (6ºC), environmental condition (25ºC) and the optimal temperature for the growth of microorganisms (36ºC). Evaluation of the sensibility of Staphylococcus spp. and Klebsiella pneumoniae isolated and in association during the pasteurization process. Results: In Scale 1 of Mac Farland the average of the colony form units of the K. pneumoniae was greater than the colony of Staphylococcus spp. At 6ºC the microorganisms in the milk isolated or in association grew at the same speed. At 25ºC K. pneumoniae associated with Staphylococcus spp. grew more than when they were isolated. At 36ºC K. pneumoniae associated with Staphylococcus spp. grew more then when it was isolated and more than at 25ºC. So we can conclude that the slow pasteurization was effective, because it has reduced at least 90% of the colony form unity in the milk after 30 minutes at 65ºC. Conclusion: By raising the temperature up to 36ºC, a Klebsiella pneumoniae had a superior growth when compared with Staphylococcus spp.; the contamination of milk by Klebsiella pneumoniae (environmental contamination) influences the most the quality of the final product when compared to the contamination by Staphylococcus spp. from mastitis, we can also conclude that the interaction of microorganisms modify the death rate of them. New studies and researches are recommended to a deeper and better comprehension of this process.

O objetivo deste trabalho é apresentar uma nova abordagem para obtenção de configurações para sistemas de distribuição de energia elétrica com o intuito de minimizar o valor de perdas ativas sem violar as restrições operacionais. Para isso, considera-se que os sistemas de distribuição estão operando em regime permanente e que suas fases estão equilibradas e simétricas, podendo o sistema ser representado por um diagrama unifilar. A reconfiguração é feita de forma a redistribuir os fluxos de corrente nas linhas, transferindo cargas entre os alimentadores e melhorando o perfil de tensão ao longo do sistema. O problema de reconfiguração do sistema pode ser formulado como um problema de programação não-linear inteiro misto. Devido à explosão combinatorial inerente a este tipo de problema, a resolução do mesmo por técnicas de otimização clássicas torna-se pouco atraente, dando espaço para técnicas heurísticas e metaheurísticas. Essas outras, mesmo não garantindo o ótimo global, são capazes de encontrar boas soluções em um espaço de tempo relativamente curto. Para a resolução do problema de reconfiguração, utilizou-se uma nova metodologia baseada no comportamento de colônias de formigas em busca de alimento na natureza. Nesta, formigas artificiais (agentes) exploram o meio ambiente (sistema de distribuição) e trocam informações para tentar encontrar a topologia que apresente os menores valores de perdas ativas. Para o cálculo das perdas, este trabalho também apresenta uma nova abordagem para resolução do problema de fluxo de potência (FP) em sistemas de distribuição radial. O fluxo de potência é uma ferramenta básica utilizada pelos centros de controle para determinar os estados e condições operacionais desses sistemas de potência. Basicamente, as metodologias empregadas para o cálculo do fluxo de potência são baseadas nos métodos clássicos de Newton ou Gauss. Mas em sistemas de distribuição de energia, devido a particularidades inerentes a estes, como a alta relação entre resistência e reatância das linhas (r/x) e a operação radial, estes métodos apresentam problemas de convergência e se tornam ineficientes na maioria das vezes. A abordagem consiste na associação dos métodos da função penalidade e de Newton. O mal-condicionamento da matriz Jacobiana de Newton é resolvido pela associação com o método da função penalidade. São apresentados testes realizados em sistemas de 5 barras, 16 barras, 33 barras, 69 barras e 136 barras para avaliar a potencialidade das técnicas propostas. Os resultados são considerados bons ou muito bons quando comparado com as técnicas existentes atualmente. / The objective of this work is to present a novel methodology for obtaining new configurations of the distribution system in order to minimize the active power losses without violating operational constraints. For this, it is considered that any distribution system is operating in a steady state and that it is balanced, therefore it can be represented by a one-line diagram. The reconfiguration is done in order to redistribute de current flows on the distribution power lines, transferring loads among the feeders and improving the voltage profile along the system. Such problem can be formulated as a mixed integer nonlinear programming problem. Due to its inherent combinatorial characteristic and since its solution by classic optimization techniques is not appealing, heuristic and metaheuristic techniques are thus better suited for its solution. Although these latter do not guarantee a global optimum, they are able to find good solutions in a relatively short time. The solution of the reconfiguration problem in this approach makes use of a novel methodology based on ant colony behavior, when these search for victuals in nature. In this technique, the artificial ants (agents) explore the environment (distribution system) and exchange information among them in order to find the topology that provides the smallest active losses. For the active losses calculation, this work also presents a novel approach for the solution of the power flow problem for radial distribution systems. The solution of the power flow problem is used by system operators in order to determine the state and operational conditions of power systems. Basically, the most common techniques used in the power flow solution are based on either Newton\'s or Gauss\' approaches. However, due to particular characteristics of distribution systems such as the high ratio of r/x and the radial topology, these methods present convergence problems and are not efficient in most of the cases. Thus, this novel technique consists in associating Newton\'s and the penalty function approaches. The matter of the ill-conditioned Jacobian matrix in Newton\'s method is overcome with the penalty function method. Some tests performed in different systems are then presented in order to assess the effectiveness of both proposed techniques.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / A pesquisa de campo foi conduzida no NÃcleo de ProduÃÃo ComunitÃria Santa Clara, pertencente a empresa Brasil Ecodiesel, no municÃpio de Canto do Buriti â PI, entre maio e julho de 2006. As anÃlises laboratoriais foram realizadas nos LaboratÃrios da Universidade Federal do CearÃ, Universidade de Fortaleza, Grupo Edson Queiroz e Instituto Centro de Ensino TecnolÃgico (CENTEC) do Cariri. Os experimentos foram realizados com o objetivo de investigar a possibilidade de agregar valor a cultura da mamona (Ricinus communis) por meio da apicultura. Os seguintes aspectos foram investigados: desenvolvimento de colÃnias de Apis mellifera em meio a plantio de mamoneira; potencial da mamoneira para a produÃÃo de mel por abelhas A. mellifera; possÃvel toxidez do pÃlen e/ou nÃctar da mamoneira para abelhas melÃferas e humanos; caracterÃsticas fÃsico-quÃmicas e organolÃpticas do mel de mamona e potencial comercial deste mel. Os resultados mostraram que a cultura da mamoneira oferece boas condiÃÃes de desenvolvimento para colÃnias de A. mellifera uma vez que oferece pÃlen e nÃctar, e que estes sÃo prontamente utilizados pelas abelhas; a mamoneira mostrou-se uma importante fonte de nÃctar para a exploraÃÃo apÃcola, tendo sido produzido em apenas 49 dias medias entre 18,82 kg (apiÃrio em Ãrea de cultivo limpo de mamona) e 23,52 kg (apiÃrio em Ãrea de mamona com ervas silvestres) de mel, semelhantes à mÃdia anual do paÃs. O pÃlen e o mel da mamoneira nÃo sÃo tÃxicos para abelhas melÃferas e para roedores, indicando que, provavelmente, seja prÃprio para o consumo humano. O mel da mamoneira apresenta caracterÃsticas fÃsico-quÃmicas dentro da legislaÃÃo brasileira e caracterÃsticas organolÃpticas bem aceitas pela populaÃÃo, demonstrando que o mesmo possui potencial comercial. Conclui-se que a exploraÃÃo apÃcola da cultura da mamona pode aumentar a rentabilidade das Ãreas de cultivo e minimizar os impactos ecolÃgicos da agricultura, alÃm de gerar um produto natural, nutritivo e bem aceito pela populaÃÃo em geral. / The research was carried out in the NÃcleo de ProduÃÃo ComunitÃria Santa Clara, belonging to the company Brasil Ecodiesel, in the county of Canto do Buriti, state of PiauÃ, Brazil, from May to July 2006. Laboratorial analyses were carried out in the laboratories of Universidade Federal do CearÃ, Universidade de Fortaleza, Grupo Edson Queiroz and Instituto Centro de Ensino TecnolÃgico (CENTEC). The experiments aimed to assess the possibility of aggregating value to castor bean (Ricinus communis) plantations placing honey bee (Apis mellifera) colonies in the area. The following aspects were investigated: colony development in castor bean plantations; potential of castor bean plantations for honey production; possible toxicity of castor bean pollen and/or nectar to honey bees and humans; physical-chemical and organoleptic characteristics of castor bean honey and its commercial potential. Results showed that castor bean plantations offer good conditions for development of A. mellifera colonies because it provides pollen and nectar which are readily used by honey bees; castor bean showed to be an important nectar source for beekeeping leading in only 49 days to an average production between 18.82 (apiary in area of castor bean clean cultivation) and 23.52 kg (apiary in area of castor bean with wild weeds), similar to the annual national mean production; castor bean pollen and nectar are not toxic to honey bees or mammals, suggesting that, probably, are also adequate for human consumption. Castor bean honey presents physical-chemical characteristics within the Brazilian regulations for honey, and its organoleptic characteristics were well rated by population in test trials suggesting commercial potential. It was concluded that beekeeping in castor bean plantations can improve the area profitability, contribute to mitigate ecological impacts caused by agriculture, besides generating a natural, nutritive and well accepted product by the population.