Para se avaliar a influência da adubação orgânica, a que foi submetida cana-de-açúcar, na fisiologia de bactérias diazotróficas endofíticas foram identificados e estudados 36 isolados de plantas submetidas a tratamentos orgânico e convencional ou sem tratamento (controle). Foi evidenciado predomínio da família Enterobacteriaceae (75%). Essas bactérias foram avaliadas quanto à excreção de substâncias promotoras de crescimento vegetal (aminoácidos, poliaminas, AIA e etileno), escolhidas como indicadoras da ação do tratamento. Os resultados mostraram que foram significativamente maiores: a excreção de aminoácidos por bactérias provenientes do tratamento orgânico assim como a produção de etileno por bactérias isoladas de plantas controle. Além das 36 linhagens acima citadas, foram identificadas sete linhagens de Rhizobium por sequenciamento dos genes 16S rRNA, recA e gapA. Três dessas linhagens foram identificadas como Rhizobium trifolii, Rhizobium etli e Rhizobium hainanense. O gênero Rhizobium ainda não havia sido descrito como endofítico de cana-de-açúcar. / Aiming to evaluate the influence of the organic fertilization, applied to sugarcane, on the physiology of endophytic nitrogen-fixing bacteria, 36 isolates of plants submitted to organic and conventional fertilization or not fertilized (control) were identified and studied. Results showed that the major part of the strains belong to the Enterobacteriaceae family (75%). The excretions of amino acids, polyamines, IAA and ethylene substances chosen as indicators of the action of the treatment were evaluated in the 36 strains. The release of amino acids was significantly higher in cultures of bacteria isolated from organic treated plants and the production of ethylene was produced in greater amounts by strains isolated from control plants. Seven isolates, different from the 36 tested for the release of plant growth promoters, were identified as Rhizobium strains through 16S rRNA, recA and gapA sequences. Three strains were identified as Rhizobium trifolii, Rhizobium etli and Rhizobium hainanense. This genus was not described as endophytic of sugarcane previously.

A doença Huanglongbing (HLB) associada a bactéria Candidatus Liberibacter spp., que coloniza os vasos do floema, é considerada uma das mais graves doenças de citros. Uma importante estratégia para o controle desta doença consiste na produção de plantas transgênicas, expressando genes que codificam peptídeos antibacterianos especificamente no local de colonização do patógeno. O objetivo deste trabalho foi obter plantas transgênicas de laranja doce, expressando o gene que codifica o peptídeo antibacteriano atacina A (attA) dirigido por promotores específicos para a expressão gênica no floema. O trabalho foi iniciado com a elaboração de construções gênicas contendo o gene attA (associado ou não ao peptídeo sinal), sob o controle dos promotores AtSuc2 (transportador de sacarose), AtPP2 (proteína de floema 2), clonados de Arabidopsis thaliana, ou CsPP2 (proteína de floema 2), clonado de Citrus sinensis. Os experimentos de transformação genética foram realizados com C. sinensis cv. \'Hamlin\', \'Valência\', \'Pêra\' e \'Natal\', via Agrobacterium tumefaciens, utilizando-se segmento de epicótilo como explante. A identificação de plantas transgênicas foi realizada por meio da análise de PCR. Plantas PCR+ foram aclimatizadas e transferidas para casa-de-vegetação específica para o cultivo de plantas transgênicas. Análises de Southern e Northern blot foram realizadas em plantas aclimatizadas, confirmando-se a integração e transcrição do gene attA, respectivamente. A expressão do gene attA também foi confirmada pela análise de RT-qPCR. Plantas de laranja \'Hamlin\' contendo o gene attA (associado ou não ao peptídeo sinal), sob o controle dos promotores AtSuc2 ou AtPP2 foram propagadas por enxertia, para futura avaliação da resistência a Candidatus Liberibacter asiaticus / Huanglongbing (HLB) associated to Candidatus Liberibacter spp., which colonizes the phloem, is considered one of the most serious diseases of citrus. One important strategy to control this disease consists of producing transgenic plants expressing, in the bacteria colonization tissue, genes encoding antibacterial peptides. The objective of this work was to produce transgenic sweet orange plants expressing genes encoding the antibacterial peptide attacin A (attA) driven by phoem-specific promoters. The work started with the development of the gene constructs, containing the attacin A gene (with or without signal peptide) controlled by either sucrose transporter gene (AtSuc2) or phloem protein 2 gene promoters (AtPP2) from Arabidopsis thaliana, or phloem protein 2 gene promotor (CsPP2) from Citrus sinensis. The genetic transformation of C. sinensis \'Hamlin\', \'Valencia\', \'Pera\' and \'Natal\' cultivars was done via Agrobacterium tumefaciens. Epicotyls segments collected from in vitro germinated seedlings were used as explants. Transgenic plants were identified by PCR analyses. PCR positive plants were acclimatized and transferred to specific greenhouse. Integration and transcription of the attA gene was confirmed in acclimatized transgenic plants by Southern and Northern blot analysis, respectively. The attA gene expression was validated by RT-qPCR analysis. \'Hamlin\' transgenic cultivars containing the AtSuc2 or AtPP2 promoters controlling the expression of attA (with or without signal peptide) were propagated by grafting, for future evaluation of Candidatus Liberibacter asiaticus resistance

Preocupações recentes a respeito da resistência de microrganismos a antimicrobianos têm aumentado o interesse por estudos de novos aditivos alimentares para ruminantes, alternativos ao uso dos ionóforos. Assim, objetivou-se com este trabalho avaliar o efeito da suplementação da cultura viva de levedura (Saccharomyces cerevisiae, cepa 1026, 5 x 106 ufc/g, Beef-Sacc®, Alltech, Inc.) e da monensina (Rumensin®, 10% de monensina sódica, Elanco, Inc.) e da combinação dos dois aditivos, sobre as características de desempenho e carcaça de novilhos Nelore confinados. Foram utilizados 72 novilhos da raça Nelore, com peso vivo inicial de 339,5 kg e 20 meses de idade, submetidos a um dos quatro tratamentos: dieta controle (sem aditivos), dieta com levedura (0,6g de Beef Sacc®/kg de matéria seca), dieta com monensina (0,3g de Rumensin®/kg de matéria seca) e dietas com monensina e levedura. Os animais foram alimentados com uma ração contendo bagaço de cana in natura (21%), casca de soja (29%), milho grão moído grosso (39%), farelo de soja (7%) e uréia (1%), por um período médio de 84 dias. Medições individuais de consumo de alimentos foram possibilitadas pelo uso de baias individuais e sistemas de Calan Gates. O pH das carcaças foi medido a 1 e 24 horas após o abate. A meia carcaça esquerda foi dividida em músculos, ossos e aparas para a determinação da porcentagem de porção comestível. A área de olho de lombo e a espessura de gordura subcutânea foram medidas no músculo Longissimus dorsi na região entre as 12ª e 13ª costelas. Bifes daquele músculo foram tirados e maturados por 0, 7 ou 14 dias para mensurações de força de cisalhamento e perdas totais por cozimento. Os tratamentos não influenciaram o ganho de peso, consumo de alimentos ou conversão alimentar dos animais. A suplementação com levedura aumentou o rendimento de carcaça, porém não houve efeitos importantes sobre outras características de carcaça e percentagem de porção comestível. A qualidade da carne também não foi influenciada pelos aditivos. Os aditivos estudados não apresentaram efeitos significativos nas características de desempenho e carcaça de novilhos Nelore alimentados com dietas de alto concentrado em confinamento. / A recent concern about anti-microbial resistance by microorganisms has increased the interest for studies of new feed additives for ruminants, alternatively to ionophores utilization. In this way, It was aimed with this work to evaluate the effects of live yeast culture (Saccharomyces cerevisiae, strain 1026, 5 x 106 cfu/g, Beef Sacc®, Alltech, Inc.), monensin (Rumensin, 10% of sodium monensin, Elanco®, Inc.) and the combination of both additives, on the performance and carcass traits of Nellore steers fed a high concentrate diet in feedlot. Seventy-two animals, with 330 kg of live weight and 20 months of age were allotted to one of four treatments: control (no additives), diet with yeast (0,6g of Beef Sacc®/ kg of dry matter), diet with monensin (0,3g of Rumensin®/ kg of dry matter) and diet with yeast and monensin. Animals were fed ration with sugarcane bagass (21%), soybean hulls (29%), corn ground (39%), soybean meal (7%) and urea (1%), for an 84-day period. Individual feed intake measurements were possible through individual pens and Calan Gates system. Carcass pH was measured at 1 and 24 hours after slaughter. Left half-carcass was split in muscle, bone and fat for retail cuts percentage determination. Rib eye area and subcutaneous fat thickness were measured on Longissimus dorsi muscle in the region between 12th and 13th ribs. Steakes of Longissimus dorsi muscle were taken and aged 0, 7 and 14 days to shear force and cooking total losses measurements. Treatments didn\'t affect weight gain, feed intake or feed efficiency. Yeast increased carcass dressing percentage, but there weren\'t important effects of additives on other carcass traits and on retail cuts percentage. Meat quality wasn\'t influenced by additives. Additives studied showed no significant effects on the performance and carcass traits of Nellore steers fed high concentrate diets in feedlot.

A síntese de Fischer-Tropsch é um processo de conversão do gás de síntese (CO + H2) em hidrocarbonetos de cadeias longas. Os catalisadores clássicos para a hidrogenação do CO são, principalmente, o Fe e o Co suportados em diferentes óxidos. O desempenho catalítico do catalisador é influenciado pelo tamanho, dispersão e grau de redução das partículas metálicas. Estudos recentes mostram uma promissora aplicação de materiais à base de nanofibras de carbono na catálise heterogênea. Estes materiais apresentam algumas vantagens em relação aos suportes catalíticos tradicionais, tais como: uma baixa interação metal-suporte, elevada área superficial, ausência de poros fechados, alta condutividade térmica, elevada inércia química e hidrofobicidade. Neste trabalho foram fabricados suportes catalíticos macroscópicos à base de nanofibras de carbono, empregando o método de vapor deposição, a partir da decomposição do etano. Os catalisadores foram preparados pela impregnação incipiente do Co e de promotores na superfície do suporte. Foram empregados os metais nobres, Ir, Pt e Ru, como promotores catalíticos, com o objetivo de incrementar a redutibilidade e a atividade do catalisador. Todos os catalisadores foram caracterizados por Quimissorção de CO, Fisissorção de N2, Redução a Temperatura Programada (TPR), Espectroscopia Fotoeletrônica de Raios X (XPS) e Microscopia Eletrônica de Transmissão (MET). Os catalisadores foram, então, testados na síntese de Fischer-Tropsch, utilizando um reator de leito fixo e fluxo contínuo, com análise simultânea dos produtos gasosos e controle sistemático da temperatura, pressão e vazão dos reagentes. Finalmente, foram analisados os produtos líquidos obtidos na reação com objetivo de conhecer a influência dos promotores na seletividade dos hidrocarbonetos formados. / Fischer-Tropsch synthesis is a process of converting the syngas (CO + H2) to long-chain hydrocarbons. The traditional catalysts for the CO hydrogenation are Fe and Co supported on different oxides. Catalytic performance of the catalyst is influenced by size, dispersion and degree of reduction of metal particles. Recent studies show a promising application of materials based on carbon nanofibers in heterogeneous catalysis. These materials have some advantages compared to traditional catalyst supports, such as a low metal support interaction, high surface area, no closed pores, high thermal conductivity, high chemical resistance, and hydrophobicity. In this work, based on macroscopic carbon nanofiber catalyst supports have been manufactured by employing the method of chemical vapor deposition from ethane decomposition. Catalysts were prepared by incipient wetness impregnation of Co and promoters on the support surface. Noble metals, Ir, Pt and Ru were used as catalytic promoters, with the aim of increasing the reductibility and catalyst activity. All catalysts were characterized by CO Chemisorption, N2 Physisorption, Temperature Programmed Reduction (TPR), X-ray Photoelectron Spectroscopy (XPS), and Transmission Electron Microscopy (TEM). The catalysts were then tested in the Fischer-Tropsch synthesis using a fixed bed reactor, continuous flow, with simultaneous analysis of gaseous products and systematic temperature control, pressure, and flow rate of the reactants. Finally, the liquid products obtained in the reaction were analyzed in order to determine the influence of promoters on the selectivity of hydrocarbons formed.

Chan Ka Ping Sophie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 140-144). / Abstracts in English and Chinese. / Thesis/Assessment Committee --- p.i / Acknowledgements --- p.ii / Abstract --- p.viii / 摘要 --- p.x / Abbreviations --- p.xi / List of Figures --- p.xii / List of Tables --- p.xiv / Chapter 1 --- Introduction / Chapter 1.1 --- Pathogenesis of HBV Infection --- p.1 / Chapter 1.2 --- Classification and Structure --- p.2 / Chapter 1.3 --- HBV Genome --- p.4 / Chapter 1.4 --- Replication Cycle --- p.7 / Chapter 1.5 --- HBV Genotypes and Nomenclature --- p.9 / Chapter 1.5.1 --- Asian prevalent genotypes --- p.9 / Chapter 1.5.2 --- Numbering system --- p.9 / Chapter 1.6 --- Identification of Markers in HBV Genome for HCC Development --- p.11 / Chapter 1.7 --- Project Objective --- p.13 / Chapter 1.8 --- Promoters of HBV --- p.14 / Chapter 1.8.1 --- Pre-S1 promoter --- p.14 / Chapter 1.8.2 --- X promoter and enhancer I --- p.14 / Chapter 1.8.3 --- Core promoter and enhancer II --- p.15 / Chapter 1.8.4 --- Pair of mutations at BCP --- p.17 / Chapter 2 --- Materials and Methods / Chapter 2.1 --- Construction of pGL3-promoter Plasmids --- p.18 / Chapter 2.1.1 --- Templates selection --- p.18 / Chapter 2.1.2 --- Amplification of promoters --- p.19 / Chapter 2.1.3 --- Cloning into pGL3-basic vector --- p.21 / Chapter 2.1.4 --- Screening and plasmid preparation --- p.21 / Chapter 2.2 --- Construction of Mutant Promoter Clones --- p.23 / Chapter 2.2.1 --- Site-directed mutagenesis --- p.23 / Chapter 2.2.2 --- pPreS 1 /2712C mutant clone --- p.24 / Chapter 2.3 --- Cloning of Full-length HBV Genomes --- p.26 / Chapter 2.3.1 --- Replication-competent HBV clone --- p.26 / Chapter 2.3.2 --- Amplification of full-length HBV genome --- p.28 / Chapter 2.3.3 --- Cloning into pUC19 vector --- p.28 / Chapter 2.3.4 --- Screening for insert and sequence confirmation --- p.29 / Chapter 2.3.5 --- Excision of full-length HBV from plasmid --- p.29 / Chapter 2.4 --- Re-construction into a 1.3-fold HBV Clone --- p.32 / Chapter 2.4.1 --- Cloning of HBV fragment nucleotide 979-2617 (nt 979-2617) --- p.32 / Chapter 2.4.2 --- Screening for insert and sequence confirmation --- p.33 / Chapter 2.4.3 --- Cloning of HBV fragment (nt 905-2000) --- p.33 / Chapter 2.4.4 --- Construction of a 1.3-fold HBV genotype Cs clone --- p.34 / Chapter 2.5 --- Cell Culture --- p.37 / Chapter 2.5.1 --- Cell culture maintenance --- p.37 / Chapter 2.5.2 --- Transient transfection of promoter clones --- p.37 / Chapter 2.5.3 --- Transient transfection of HBV genomes --- p.38 / Chapter 2.6 --- Dual-Luciferase® Reporter Assay System --- p.40 / Chapter 2.6.1 --- Principle of the assay --- p.40 / Chapter 2.6.2 --- Cell harvest --- p.43 / Chapter 2.6.3 --- Luciferase assay --- p.43 / Chapter 2.7 --- Data Analysis --- p.44 / Chapter 2.8 --- Extraction of HBV DNA from Intracellular Cores --- p.45 / Chapter 2.8.1 --- Harvest of intracellular cores --- p.45 / Chapter 2.8.2 --- Phenol/chloroform extraction --- p.45 / Chapter 2.9 --- Southern Blotting --- p.47 / Chapter 2.9.1 --- Transfer of DNA to membrane --- p.47 / Chapter 2.9.2 --- Preparation of probes --- p.47 / Chapter 2.9.3 --- Hybridization with radiolabeled probes --- p.48 / Chapter 2.10 --- Detection of HBeAg and HBsAg --- p.50 / Chapter 2.10.1 --- HBsAg assays --- p.50 / Chapter 2.10.2 --- HBeAg assays --- p.51 / Chapter 2.11 --- SEAP Reporter Gene Assay --- p.52 / Chapter 3 --- Results / Chapter 3.1 --- Templates Selected --- p.53 / Chapter 3.2 --- Results of Luciferase Assays --- p.58 / Chapter 3.2.1. --- BCP mutation of genotype A as control --- p.58 / Chapter 3.2.2. --- Effect of C1165T mutation on Xpro/enhI activity of HBV genotype B --- p.60 / Chapter 3.2.3. --- Effect ofT2712C mutation on pre-S1 promoter activity of HBV Genotype B --- p.60 / Chapter 3.2.4. --- Effect of G1613A mutation on core pro/enhII activity of HBV Genotype Cs --- p.64 / Chapter 3.2.5. --- G1613A and BCP mutation --- p.67 / Chapter 3.3 --- Full-length HBV Genome Clones --- p.70 / Chapter 3.3.1. --- Construction of replication-competent full-length HBV genome clones --- p.70 / Chapter 3.3.2. --- Drawbacks of the system --- p.78 / Chapter 3.4 --- Construction of a Replication-competent 1.3-fold HBV Clone --- p.82 / Chapter 3.4.1. --- Construction of the HBV (nt 979-2617) clone --- p.82 / Chapter 3.4.2. --- Construction of the HBV (nt 905-2000) clone --- p.86 / Chapter 3.4.3. --- Construction of 1.3-fold genotype Cs HB V clone --- p.89 / Chapter 3.4.4. --- Test for replication competency --- p.92 / Chapter 4 --- Discussion / Chapter 4.1 --- BCP Mutation as Control of the Luciferase Assay --- p.94 / Chapter 4.2 --- Promoter Activities Not Altered by T2712C and C1165T --- p.96 / Chapter 4.3 --- Mutation G1613A of Core pro/enhll --- p.98 / Chapter 4.3.1 --- Mutation resides in negative regulatory element of core promoter --- p.98 / Chapter 4.3.2 --- NRE and NRE-binding protein --- p.98 / Chapter 4.3.3 --- Relationship with BCP mutation --- p.101 / Chapter 4.4 --- HBV Constructs --- p.103 / Chapter 4.4.1 --- Rationale in re-construction of 1.3-fold HB V clone --- p.103 / Chapter 4.4.2 --- Replication competency --- p.104 / Chapter 4.5 --- Conclusion --- p.106 / Chapter 4.6 --- Future Work --- p.107 / Appendix --- p.108 / References --- p.140

Made available in DSpace on 2015-03-05T13:57:00Z (GMT). No. of bitstreams: 0 Previous issue date: 11 / Nenhuma / A região promotora é uma seqüência de DNA que localiza-se anteriormente a uma determinada região gênica. Ela é responsável pelo início do processo de transcrição de um gene ou conjunto de genes. Assim, ela também atua como um elemento regulador da expressão gênica. O estudo da regulação da expressão gênica é relevante porque é essencial para a compreensão da maquinária vital dos seres vivos, já que a diferença entre duas espécies está mais relacionada em como e quando seus genes estão “ativos” ou “inativos” do que com a estrutura destes em si. Embora exista métodos computacionais para a predição de genes com boa acurácia, o mesmo não é conseguido para os promotores. Esta dificuldade deve-se ao pequeno e pouco conservado padrão das seqüências, gerando assim resultados com alto número de falsos positivos. Além dos motivos consensuais, os promotores possuem características físicas que os diferem de seqüências não-promotoras. No entanto, estas ainda não são amplamente utilizadas no problema de predição in silic / The promoter region is localized few base pairs before the gene region. It is responsible by initiate the gene expression process, thus, it plays a regulatory role. The study about the gene expression regulation is a great area, since it can assist in the comprehension of complex metabolic network presented by several organisms and, because the difference between two different species is how and when your genes are turn off and turn on than your structure. The computational methods to gene prediction have a good accuracy, but this is not achieved in the promoter prediction. This difficulty occurs because the length of promoter and the degenerate pattern presented, thus the results presented a great number of false positives. This work aims employed Neural Networks to promoter prediction and recognition of Escherichia coli by two approaches: whit the orthogonal codification and stability values of the promoter sequence. For characterization, realize the extraction rules of type if … then. The results in this

Made available in DSpace on 2015-03-05T13:57:00Z (GMT). No. of bitstreams: 0 Previous issue date: 28 / Nenhuma / Microarranjos têm sido fortemente usados para monitorar, somultaneamente, o padrão de expressão de milhares de genes. Assim, uma grande quantidade de dados tem sido gerada e o desafio atual é descobrir como extrair informações úteis destes conjuntos de dados. Dados de Microarranjos são fortemente especializados, envolvendo diversas variáveis de forma não linear e temporal, necessitando de modelos recorrentes não lineares, os quais são complexos para formular e analisar. Este trabalho propõe a utilização de Redes Nunes Recorrentes (RNR) como modelo para os dados devido às suas habilidades de aprendizado de sistemas nâo-lineares e complexos. Uma vez obtido um modelo para os dados utilizando uma RNR, é possível extrair regras que representam as características aprendidas. Analisando as regras em conjunto com a base de dados, propõe-se a representação do conhecimento utilizando Cadeias de Markov. Tais Cadeias são facilmente visualizadas, na forma de grafos de estados, apresentando as interações entre os níveis de / Array technologies have made it strainghtforward to simultaneously monitor the expression pattern of thousands of genes. Thus, a lot fot data is being generated and the challenge now is to discover how to extract useful information from these data sets. Microarray data is highly specialized. It involves several variables in a nonlinear and temporal way, demanding nonlinear recurrent free models, which are complex to formulate and to analyse. So, this work proposes the use of Rucurrent Neural Networks(RNN) for data modeling, due to their learning hability of nonlinear and complex systems. Once a model is obtained with a RNN for the data, it is possible to extract rules to represent the knowledge acquired by them. From rule analisys, this work proposes the representation of the knowledge by Markov Chains model, which is easily visualized in the form of a graph of states, which show the interactions among the gene expression levels and their changes in time. In this work, we propose a new approach to microarra

Submitted by Filipe dos Santos (fsantos@pucsp.br) on 2017-03-15T13:33:41Z No. of bitstreams: 1 André Luís Alves de Melo.pdf: 2741804 bytes, checksum: 1369fd561e394e8df2a6f9b550bb1a1b (MD5) / Made available in DSpace on 2017-03-15T13:33:41Z (GMT). No. of bitstreams: 1 André Luís Alves de Melo.pdf: 2741804 bytes, checksum: 1369fd561e394e8df2a6f9b550bb1a1b (MD5) Previous issue date: 2017-02-21 / The present work proposed to investigate and discuss the unconstitutionality, or not, that is, the non-acceptance of the principle of mandatory public prosecution, in the face of the constitutional principle of functional independence, acting the Public Prosecutor as a political agent in criminal policies And its limits. Sedimented in the innovative proposal of a Doctoral Thesis, the work focused on unconstitutionality (not reception) and not on the convenience or inconvenience of mandatory criminal action. However, the research sought to analyze empirical data and not just arguments of rhetoric, and established comparisons between the routine and results obtained by Public Prosecutors who adopt the finalist theory (more traditional) and the functionalist (more innovative), verifying that the functionalism has better Results with fewer processes, because it focuses on the system as a whole. It also addressed foreign laws and practices, verifying that functionalism currently prevails in practically all countries, except in Brazil. In addition, even in countries that do not expressly guarantee functional independence to members of the Public Prosecutor's Office, they have the authority to establish criminal public policy priorities and results. Thus, the Code of Criminal Procedure must conform to the Federal Constitution and not the contrary, which implies a revision of articles 24 and 28 of the CPP, and for this the work elaborated a whole research on the ends of criminal law, On criminology, on criminal policy, on criminal action, including historical aspects of criminal law and criminal procedure in Brazil, as well as pointed out that several writers defend the opportunity of criminal action, which is little discussed in the most used works in undergraduate courses Of Law, and finally, stressed the need that the works of Criminal Procedure also address and confront Constitutional Law with the Criminal Procedure of 1941 / O presente trabalho propôs-se a pesquisar e discutir a inconstitucionalidade, ou não, ou seja, a não recepção do princípio da obrigatoriedade da ação penal pública, em face do princípio constitucional da independência funcional, atuando o Ministério Público como agente político nas políticas criminais e os seus limites. Sedimentando na proposta inovadora de uma Tese de Doutorado o trabalho focou na inconstitucionalidade (não recepção) e não na conveniência ou inconveniência da obrigatoriedade da ação penal. No entanto, a pesquisa buscou analisar dados empíricos e não apenas argumentos de retórica, e estabeleceu comparações entre a rotina e resultados obtidos por Promotorias que adotam a teoria finalista (mais tradicional) e a funcionalista (mais inovadora), verificando que o funcionalismo tem melhores resultados com menor número de processos, pois foca no sistema como um todo. Também abordou legislações e práticas estrangeiras, verificando que o funcionalismo prevalece atualmente em praticamente todos os países, menos no Brasil. Além disso, mesmo em países que não asseguram expressamente a independência funcional aos Membros do Ministério Público, estes acabam tendo autoridade para estabelecer política pública criminal de prioridades e resultados. Dessa forma, o Código de Processo Penal é que deve ser amoldado à Constituição Federal e não o contrário, o que implica uma revisão dos artigos 24 e 28 do CPP, e para isto o trabalho elaborou toda uma pesquisa sobre os fins do direito penal, sobre a criminologia, política criminal, sobre ação penal, inclusive com aspectos históricos das leis penais e processuais penais no Brasil, bem como apontou vários doutrinadores que defendem a oportunidade da ação penal, o que é pouco discutido nas obras mais usadas nos cursos de graduação de Direito, e por fim, ressaltou a necessidade de que as obras de Processo Penal também abordem e confrontem o Direito Constitucional com o Processo Penal de 1941

Phakopsora pachyrhizi Syd.& P.Syd. est le plus important fléau du soja (Glycine max (L.) Merrill). Introduit au Brésil dans les années 2000, ce champignon s’est rapidement répandu sur les deux continents Américains. Seule l’utilisation de fongicides associée à des pratiques culturales strictes permet de maintenir le niveau de production. L’utilisation répétitive de ces fongicides ainsi que la plasticité génétique de ce champignon ont rapidement entraîné une diminution d’efficacité de certaines molécules. Par ailleurs, la plupart des résistances verticales identifiées dans les ressources naturelles du soja restent inefficaces contre certains isolats du champignon. La compréhension des mécanismes de l’immunité des plantes permet de proposer des solutions biotechnologiques pour le contrôle des maladies. L’utilisation antérieure du système barnase/barstar induisant une mort cellulaire artificielle, a permis de générer des pommes de terre résistantes à Phytophtora infestans. Cette technologie est basée sur l’expression de la barnase une ribonucléase toxique pour les cellules, et la barstar un inhibiteur de la barnase. Il a été proposé d’évaluer ce système pour le contrôle de P. pachyrhizi. Le point critique de cette approche est de trouver le bon rapport de l’expression des gènes barnase/barstar. Pour ce faire la barnase sera placée sous le contrôle d’un promoteur induit par le pathogène, permettant une régulation spatiotemporelle. La recherche de tels promoteurs a été effectuée en utilisant des données transcriptomiques et bibliographiques. Des sojas stables exprimant les différentes fusions promoteur:GFP ont été créées afin d’étudier l’ expression spatiotemporelle de ces promoteurs en présence du champignon. Les promoteurs pGmCHIT1 (de G. max) et pgst1 (de Solanum tuberosum) contrôlant respectivement l’expression d’une chitinase et d’une glutathione-S-transférase ont été identifiés comme induits par le pathogène. L’impact de différents stress sur ces deux promoteurs a été évalué. Les constructions génétiques « barnase/barstar » comprenant les différentes combinaisons des promoteurs ont été générées. Nicotiana benthamiana a été utilisé pour exprimer transitoirement les construits et évaluer leur phytotoxicité en absence du pathogène. Un seul construit contenant le promoteur gst1 s’est avéré non phytotoxique. Il a été transféré avec succès dans le soja. Ces sojas n’ont pas montré de gain de tolérance à la rouille. Une proposition d’amélioration du système barnase/barstar est discutée afin de mieux cerner les possibilités et les limites de ce système pour le contrôle de la rouille du soja / Phakopsora pachyrhizi Syd.& P.Syd, the fungus responsible for Asian soybean rust, is the most devastating soybean (Glycine max (L.) Merrill) pathogen. First observed in the 2000s in Latin America, the pathogen has spread throughout the Americas. The control of this pathogen depends on the use of fungicides and strict agricultural practices. The repetitive use of the 3 classes of fungicides and the genome plasticity of the pathogen have led to a decreased efficacy of certain molecules. Although vertical resistance genes have been mapped in the soybean germplasm, most of them are not effective against all Asian soybean rust isolates. A deeper understanding of plant immunity facilitates the development of biotechnological approaches for plant disease control. Artificial cell death was previously developed to control Phytophthora infestans development in potato. The technology was based on a barnase ribonuclease that is highly toxic to the plant cell and that consequently needed to be expressed only in the presence of the pathogen. The lethal expression of barnase was counterbalanced by barstar, a highly specific inhibitor of barnase. We propose to evaluate this technology in soybean to control P. pachyrhizi. The key objective is the modulation of the ratio of barnase/barstar based on the identification of an adequate inducible promoter to control the expression of barnase. The previous literature and transcriptomic data were used to identify candidate promoters for barnase expression. Stable transgenic soybean expressing the different promoter:GFP fusions were generated to test the spatiotemporal activity of the promoters in the presence of the pathogen. pGmCHIT1 (from G. max) and pgst1 (from Solanum tuberosum) promoters controlling a chitinase and a glutathione-Stransferase, respectively, were identified as induced by soybean rust. The impacts of different stresses on these promoters were evaluated. Molecular constructs with different promoters driving the barnase and barstar gene combination were generated. Nicotiana benthamiana was used to evaluate construct toxicity in the absence of the pathogen. One single construct containing the promoter pgst1 was shown to be non-phytotoxic. This construct was successfully introduced in soybean plants. The generated soybeans were challenged with rust, but no protection was observed. Based on these results, we discuss how to improve the barnase/barstar system to control soybean rust

Plant promoters are the principle cis-acting regulatory sequences responsible for the temporal and spatial expression of genes. One method for isolating plant promoters is based on the ability of a common soil bacterium, <i> Agrobacterium tumefaciens </i>, to transfer a specific segment of DNA (T-DNA) into plant cells. This specific T-DNA has been shown to integrate stably into the recipient plant genome. If the T-DNA contains a promoterless marker gene, then T-DNA integration events occurring adjacent and downstream to a promoter region can be detected by the activation of the marker gene. These T-DNA-mediated gene fusions, consisting of an unknown plant promoter sequence and the coding sequence of a marker gene, can be isolated using the marker gene as a promoter tag. The key objective of this work was to develop a novel, bifunctional selectable marker gene and assess its use as: a selectable marker gene in bacterial and plant transformation systems, and as a promoter tag for T-DNA promoter-tagging studies in dicots. A bifunctional fusion gene was produced between phosphinothricin acetyltransferase and neomycin phosphotransferase (PAT::NPT II), by fusing an NPT II coding sequence to the 3' terminus of the PAT gene. The PAT gene product confers tolerance to a non-selective herbicide L-phosphinothricin (Ignite, Hoechst AG). The neomycin phosphotransferase ('npt II') gene allows for direct selection of transformed cells with the antibiotic, kanamycin. Using an <i>in vivo Escherichia coli </i> selection system, a translational fusion gene between these two reporter genes was achieved. The resulting protein had activities of both parent enzymes. This was demonstrated both in transformed <i>Escherichia coli</i> and in transformed <i>Nicotiana tabacum</i> and <i>Brassica napus</i> plants. Using this bifunctional selectable marker gene, a T-DNA promoter tagging vector, pBAU2, was constructed and its utility was demonstrated in <i>Nicotiana tabacum</i>. One of the <i>N. tabacum</i> promoter tagged events was selected for subsequent promoter isolation studies. The promoter from this regenerant was isolated by screening a Lambda subgenomic library and also by thermal asymmetric interlaced (TAIL-)PCR. The isolated upstream regulatory sequence was fused to a reporter gene, â-glucuronidase ('gus'), and subjected to a preliminary evaluation in <i> Nicotiana tabacum</i> and in <i>Brassica napus</i>.