Visando o aumento da resistência a fungos patogênicos, o presente trabalho foi realizado com o objetivo de introduzir o gene SnOLP, que codifica uma osmotina de Solanum nigrum var. americanum, em cultivares de soja. A estratégia escolhida foi a transformação por biobalística, utilizando o plasmídeo pCL1390-UBQ3-SnOLP, que contém o gene SnOLP e o gene hpt II, que confere resistência ao antibiótico higromicina. Conjuntos de embriões somáticos globulares das cultivares IAS-5, Bragg e BRSMG 68 Vencedora foram utilizados como alvo. Os conjuntos bombardeados foram transferidos para meio seletivo visando obter material estavelmente transformado. Os conjuntos higromicinaresistentes correspondendo a cinco, 12 e 13 eventos de transformação independentes nas cultivares Bragg, IAS-5 e BRSMG 68 Vencedora, respectivamente, foram sequencialmente transferidos para meio de proliferação D20 (sem higromicina), maturação (MSM6) e regeneração (MSO). Um total de 114, 70 e 211 embriões histodiferenciados das cultivares Bragg, IAS-5 e BRSMG 68 Vencedora, respectivamente, foram obtidos. A partir destes, foram regeneradas oito plantas da cultivar IAS-5, correspondentes a três eventos de transformação independentes e 30 plantas da cultivar Bragg, de um evento de transformação. Nenhuma planta da cultivar BRSMG 68 Vencedora foi regenerada. Em conseqüência de um acidente, foram recuperadas apenas duas plantas adultas de IAS-5, cada uma proveniente de um evento de transformação independente e 12 plantas de Bragg, todas do mesmo evento de transformação. A presença do transgene nas plantas foi detectada por PCR e a expressão da proteína recombinante através de Western blot. A herança do transgene seguiu o padrão Mendeliano, para um gene dominante, na linhagem I4 de IAS-5. As progênies das plantas transgênicas de Bragg apresentaram uma segregação excepcional, com deficiência de plantas transformadas. Resultados preliminares dos bioensaios utilizando extratos protéicos totais não mostraram atividade antifúngica dessas plantas transgênicas. / Aiming to enhance resistance to fungal pathogens, the present work was carried out with the objective of introducing a gene (SnOLP) coding an osmotinlike protein from Solanum nigrum var. americanum in soybean cultivars [Glycine max (L.) Merrill]. Biolistic transformation was the strategy elected, using the plasmid pCL1390-UBQ3-SnOLP, which contain the SnOLP gene and the selectable marker hpt II gene. Somatic globular embryo clusters of IAS-5, Bragg and BRSMG 68 Vencedora cultivars were used as target tissues. Bombarded embryo clusters were transferred to selective medium containg hygromycin, aiming to obtain stable transformed material. Hygromycin-resistant embryogenic clusters corresponding to five, 12 and 13 independent transformation events of Bragg, IAS-5 and BRSMG 68 Vencedora cultivars, respectively, were sequentially transferred to proliferation D20 (without hygromycin), maturation and regeneration media. A total of 70, 114 and 211 histodifferentiated embryos were obtained from IAS-5, Bragg and BRSMG 68 Vencedora cultivars, respectively. Eight plants corresponding to three independent transformation events were recovered for cultivar IAS-5 and 30 plants from one transformation event for Bragg cultivar. No one plant for BRSMG 68 Vencedora cultivar was regenerated. As a consequence of an accident, only two adult plants for IAS-5 cultivar, each one proceeding from an independent transformation event and 12 plants for Bragg, all them from the same transformation event, were obtained. The integration and expression of the SnOLP transgene into the genomes of transformed plants were confirmed by PCR and Western blot. The I4 progeny from IAS-5 cultivar segregated as a single dominant locus as predicted by Mendelian principles. The transgenic Bragg progenies segregated in an exceptional manner, with fewer SnOLP-positive plants. Preliminary bioassays using total protein extracts of transgenic plants did not show antifungal activity.

Ureases (EC 3.5.1.5) são amplamente distribuídas em bactérias, fungos e plantas onde sua função biológica não é completamente conhecida. Acredita-se que ureases estejam envolvidas na biodisponibilidade de nitrogênio e mecanismos de defesa contra predadores e patógenos. Plantas de soja Glycine max (L.) Merril contêm duas isoformas de urease. Neste trabalho, clonamos e seqüenciamos um fragmento de 300 pb que corresponde a uma região interna do gene de urease. Relatamos, também, a utilização de um gene de soja como modo de gerar resistência a doenças fúngicas em plantas. Plantas transgênicas de tabaco (Nicotiana tobacum var. Turkish) contendo o cDNA codificador completo da urease ubíqua de soja sob a regulação do promotor 35S do vírus do mosaico da couveflor (CaMV) e do terminador do gene da nopalina sintase, foram geradas a partir da transformação de discos foliares por Agrobacterium tumefaciens. Extratos proteicos obtidos a partir das folhas das plantas transgênicas foram analisados quanto à atividade ureásica e à imunorreatividade contra anticorpos da urease do feijão-de-porco. A habilidade dos extratos proteicos em inibir o crescimento de fungos fitopatogênicos foi comparada com a atividade fungicida da urease embrião-específica isolada de sementes tipo-selvagem. Nossos resultados demonstraram a atividade antifúngica de ambas as isoformas de urease e apresentaram uma correlação positiva entre a inibição do crescimento de fungos e o conteúdo/atividade da urease ubíqua de soja recombinante. Os dados sugerem que a superexpressão da urease, em plantas transgências, pode auxiliar na resistência das plantas contra fungos fitopatogênicos, além de seus efeitos conhecidos sobre insetos. / Ureases (EC 3.5.1.5) are largely distributed in bacterial, fungi and plants, where theis physiological role is not completely understood. It is thought that ureases are involved in nitrogen bioavailability and defense mechanisms against predators and pathogens. Soybean [Glycine max (L.) Merril] plants contain two isoforms of urease. Here we describe the cloning and sequencing of a fragment of 300 bp corresponding to an internal region of one of the soybean urease genes. Here we also reported the employment of a gene from soybean as a tool to confer resistance to fungal diseases in plants. Transformed tobacco (Nicotiana tobacum var. Turkish) plants harbouring the full length cDNA encoding the soybean ubiquitous urease under the control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and the nopaline synthase gene (nos) terminator were obtained after leaf disc transformation by Agrobacterium tumefaciens. Leaf protein extracts of transgenic plants were analyzed for urease activity and immunoreactivity against antibodies to the jackbean urease. The ability of leaf protein extracts to impair growth of selected phytopathogens was compared to the fungicidal activity of the embryo-specific urease isolated from wild-type seeds. Our results demonstrated the antifungal activity of both soybean ureases and showed a positive correlation between the inhibiton of fungal growth and content/activity of the recombinant soybean ubiquitous urease in leaves of transgenic tobacco. The data suggest that urease overexpression in transgenic plants may help to improve plant resistance against phytopathogenic fungi, besides its known effect on insects.

Ureases (EC 3.5.1.5.) catalisam a hidrólise de ureia em NH3 e CO2, sendo sintetizadas por plantas, fungos e bactérias. No solo, a urease é encontrada em microrganismos, raízes de plantas e como uma enzima extracelular ligada a compostos orgânicos e inorgânicos. Em plantas e fungos, as ureases consistem em trímeros ou hexâmeros formados por uma subunidade de 90 kDa, enquanto que enzimas bacterianas são complexos com duas ou três subunidades. A inserção de dois átomos de níquel no sítio ativo requer pelo menos três proteínas acessórias, UreD, UreF e UreG em bactérias, ou seus ortólogos em plantas e fungos. Bradyrhizobium japonicum é uma bactéria do solo que forma nódulos fixadores de nitrogênio em plantas de soja. Esse microrganismo produz uma urease, e seu papel na sinalização, tanto para a planta de soja quanto para outros organismos no complexo ambiente da rizosfera, ainda não foi investigado. Desta forma, o presente estudo objetivou purificar e caracterizar a urease de B. japonicum (BJU), bem como avaliar o papel desta enzima, tanto a de origem vegetal quanto a de origem bacteriana, no processo de nodulação da soja. A capacidade da enzima em induzir exocitose/secreção foi avaliada no teste de agregação plaquetária, utilizando-se plasma rico em plaquetas obtido de sangue de coelho e monitorando-se a agregação por turbidimetria. Observamos que a urease de B. japonicum possui a propriedade de agregar plaquetas, implicando em uma provável atividade indutora de exocitose. Ensaios de quimiotaxia demonstraram a atração exercida pela urease ubíqua recombinante de soja sobre células de B. japonicum. Para os ensaios de nodulação, sementes pré-germinadas de soja tiposelvagem (Williams 82) e de mutantes deficientes na proteína urease (eu1-sun/eu4) foram expostas a culturas de B. japonicum USDA110 (tipo selvagem), B. japonicum ΔureG (ausência de atividade ureásica) ou B. japonicum ΔureABC (ausência de urease), e semeadas em vasos de Leonard modificados. Os nódulos foram contados e pesados em diferentes tempos após a inoculação. Além disso, foi determinado o conteúdo de leghemoglobina destes nódulos e o conteúdo de nitrogênio na parte aérea das plantas, como uma maneira de estimar a eficiência da fixação biológica de nitrogênio. Plantas deficientes em urease formam nódulos maiores e em menor número que as selvagens, independente do fenótipo da bactéria. O pico de produção de leghemoglobina em plantas tipo-selvagem é maior e anterior ao pico observado nas plantas mutantes. Inibição de toda a atividade enzimática de urease nas plantas selvagens pelo inibidor fenilfosforodiamidato não causou as alterações observadas pela ausência da proteína urease nas plantas mutantes. Esses resultados sugerem que o desenvolvimento do nódulo em plantas requer a proteína urease, de maneira independente de sua atividade enzimática. Em contraste, a urease da bactéria parece não influenciar a nodulação ou a fixação biológica de N2 na planta. Concluímos que a urease da soja apresenta um papel relevante na simbiose planta - B. japonicum, independente de sua atividade ureolítica, e não compartilhado com a urease bacteriana. / Ureases (EC 3.5.1.5.) catalyze the hydrolysis of urea in NH3 and CO2, and are synthesized by plants, fungi and bacteria. In the soil, urease occurs in microorganisms and plant roots, and as an extracellular enzyme bound to organic and inorganic compounds. In plants and fungi, ureases consist of trimers or hexamers formed by a subunit of 90 kDa, whereas bacterial enzymes are complexes with two or three subunits. The insertion of two nickel atoms into the active site requires at least three accessory proteins, ureD, ureF, and ureG in bacteria, or their orthologs in plants and fungi. Bradyrhizobium japonicum is a soil bacterium that forms nitrogen fixing nodules on soybean plants. This bacterium produces a urease, and its role in signaling for both the soybean plant and other organisms in the complex environment of the rhizosphere, has not yet been investigated. Thus, the present study aimed to purify and characterize B. japonicum urease (BJU), and to evaluate the role of this enzyme, from both plant and bacteria, in the process of soybean nodulation. The induction of secretion was assessed by the ability of the enzyme to induce platelet aggregation in rabbit platelet-rich plasma monitored by turbidimetry. We found that the urease of B. japonicum possesses the property of aggregating platelets, implying a secretion inducing activity. Chemotaxis assays demonstrated the attraction of recombinant soybean ubiquitous urease upon B. japonicum cells. For nodulation assays, pre-germinated seeds of wild-type soybeans (Williams 82) and of mutants deficient in the urease protein (eu1-sun/eu4) were exposed to cultures of B. japonicum USDA110 (wild-type), B. japonicum ΔureG (lack of urease activity) or B. japonicum ΔureABC (no urease), and planted in modified Leonard jars. The nodules were counted and weighed at different times after inoculation. Additionally, we determined the leghemoglobin content of nodules and the nitrogen content in the shoots, as a way to estimate the efficiency of biological nitrogen fixation. Plants deficient in urease (eu1-sun/eu4) form fewer but larger nodules than wildtype plants, regardless of the phenotype of the bacteria. The peak of leghemoglobin production in wild-type plants is higher and earlier than the peak observed in mutant plants. Inhibition of all the enzymatic activity of urease in wild-type plants by phenylphosphorodiamidate did not result in the alterations seen in mutant plants lacking urease. These results suggest that the development of nodule requires the protein urease, but not its enzyme activity. In contrast, the bacterial urease seems to play no roles in the nodulation and biological N2 fixation in the plant. We conclude that the soybean urease plays an important role in the soybean - B. japonicum symbiosis, which is independent of its ureolytic activity and is not shared by the bacterial urease.

Orientador: Renato Pinto Ortiz / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Filosofia e Ciencias Humanas / Made available in DSpace on 2018-07-27T18:35:43Z (GMT). No. of bitstreams: 1 Rodrigues_JuliaMariadeSouza_M.pdf: 8871250 bytes, checksum: 1cdf4639e8a7d7f2ab71e20f3326ee0a (MD5) Previous issue date: 2001 / Resumo: A religião é, para Weber, a chave de interpretação para o entendimento de processos culturais mais amplos, como o desencantamento do mundo e a secularização. Além disso, ele a define como estilo de vida próprio fomentado pelo indivíduo, que, conseqüentemente, interfere na conduta de um grupo ou de uma coletividade historicamente determinados. A análise da religião possibilita também a compreensão interpretativa de "individualidades históricas", ou seja, das seis religiões ou sistemas como o budismo, confucionismo, judaísmo, o islamismo, protestantismo e hinduísmo, cuja descrição histórica e tipológica possibilitou a formulação mais desenvolvida da relação entre religião e economia, ou seja, da conexão de sentido entre protestantismo e capitalismo. Mediante a análise comparativa e descritiva das religiões mundiais (éticas religiosas), Weber aponta a religião (o protestantismo) como um dos determinantes causais da ética econômica do capitalismo europeu / Abstract: For Weber, religion is the key to the interpretation and understanding of wider cultural processes, such as disenchantment of the world and secularization. Moreover, he defines it as a personal style of life promoted by the individual, which consequently interferes in the conduct of a historically detennined group or collective. Analysis of religion pennits the interpretive comprehension of various "historical individualities", that is, innumerous religions such as Buddhism, Confucianism, Judaism, Protestantism, and Hinduism, among others, whose historic and typological description permitted the most complete formulation of the relationship between religion and economy, that is, of the meaningful connection between Protestantism and Capitalism. Based on a comparative and descriptive analysis of world religions (religious ethics), Weber considers religion (Protestantism) to be one of the causal detenninants of European capitalism / Mestrado / Mestre em Sociologia

Orientador: Luiz Roberto Monzani / Tese (doutorado) - Universidade Estadual de Campinas,Instituto de Filosofia e Ciencias Humanas / Made available in DSpace on 2018-08-08T21:51:09Z (GMT). No. of bitstreams: 1 Araujo_SaulodeFreitas_D.pdf: 1513410 bytes, checksum: 53dd6729adaeb6f5cdf31f74e9b6fd3f (MD5) Previous issue date: 2007 / Resumo: O objetivo do presente trabalho é analisar a constituição do projeto de uma psicologia científica na obra de Wilhelm Wundt. Tendo em vista as inúmeras falhas e lacunas na literatura secundária, propõe-se uma nova interpretação de seu pensamento, amparada principalmente no exame de fontes primárias, envolvendo tanto suas obras oficiais quanto materiais inéditos do ¿Espólio-Wundt¿ da Universidade de Leipzig. A tese central é a de que o projeto wundtiano de uma psicologia científica só pode ser adequadamente compreendido se levarmos em consideração os interesses e os pressupostos filosóficos que o fundamentam. Em outras palavras, demonstra-se que a psicologia de Wundt não pode ser separada de seu projeto filosófico, do qual ela é uma parte essencial. Argumenta-se que somente dentro desta perspectiva é possível compreender a ruptura teórica operada por Wundt em relação ao seu projeto psicológico inicial ¿ ruptura esta que permanece mal explicada, quando não ignorada na literatura secundária ¿, assim como a evolução e o amadurecimento posterior de seu sistema de psicologia. Finalmente, discutem-se a questão da unidade teórico-conceitual de seu projeto final e os limites do mesmo / Abstract: The aim of this work is to analize the formation of the project of a scientific psychology in the work of Wilhelm Wundt. Considering the many flaws in contemporary literature a new interpretation of his thought is proposed, which is based essentialy on the examination of primary sources, including both his published works and the unpublished material from the ¿Wundt-Estate¿ of the Leipzig University. The central thesis claims that the Wundtian project of a scientific psychology can only be properly understood if we take into account the philosophical interests and premisses on which it is based. In other words, it is demonstrated that Wundt¿s psychology cannot be separated from his philosophical project, to which it belongs as an essential part. It is argued that only this perspective enable us to comprehend Wundt¿s theoretical rupture with his early psychological project ¿ a rupture which still remains unexplained, when not ignored in the literature ¿ as well as the evolution and the later maturation of his psychology system. Finally, the question of the conceptual unity of his final project and its very limits is discussed. / Doutorado / Doutor em Filosofia

Submitted by BKAB Setor Proc. Técnicos FGV-SP (biblioteca.sp.cat@fgv.br) on 2013-04-12T15:30:17Z No. of bitstreams: 1 1198301247.pdf: 5160976 bytes, checksum: 48939e61ebcde68db395bcd3a82e2709 (MD5) / O objeto original da dissertação era a análise crítica da teoria das organizações, através da 'sociologia compreensiva' de Max Weber. Em resumo, essa dissertação é uma tentativa de penetrar uma pequena parte do universo intelectual de Max Weber.

La production de semences très vigoureuses est un levier clépour augmenter les rendements agricoles. Chez le soja, lesrisques de production de semences de faible vigueur germinativesont élevés sous les climats tropicaux, ce qui conduit àune mauvaise installation de la culture, voire un déclassementdes lots en grain. La longévité (c-a-d la capacité de survivre àl’état sec pendant de longues périodes) représente une caractéristiqueimportante de la vigueur germinative. Cependant,la façon dont elle est acquise au cours de la maturation dela graine n’est pas connue. L’objectif de la thèse était d’obtenirun aperçu des mécanismes de régulation de l’acquisitionde la longévité chez le soja. Celle-ci est acquise peu aprèsla phase de remplissage et l’acquisition de la tolérance à ladessiccation (stade R7.2) et continue d’augmenter jusqu’enfi n de maturation. Des différences de longévité ont été observéesentre les récoltes 2013 et 2014.Elles seraient liées auxtempératures élevées au cours de la formation des graines,un facteur connu pour impacter la qualité des semences. Leséquençage Illumina de graines récoltées à 7 stades de développementa révélé des changements complexes se produisantau niveau transcriptionnel dès le stade R7.2, pourtantconsidéré comme correspondant à la maturité physiologique.Les analyses d’enrichissement GO ont révélé une associationpositive entre longévité et les processus de maturation desARN et régulateurs de la post-transcription et une associationnégative avec les processus liés à la photosynthèse. L’acquisi / Soybean is one of the most important oil crop species usedfor food and feed as well as a range of industrial applications.However, producing highly vigorous seeds is a key leverto increase crop production effi ciency. Low physiologicalseed quality, which is more prone to occur under tropicalenvironment, leads to poor stand establishment and decreasedin yields. Seed longevity is the ability to survive the dry statefor prolonged periods of time and represents an importanttrait for seed quality. Here, the objective was to obtain insightsinto the mechanisms regulating the progressive acquisitionof longevity. Using Illumina high-throughput sequencing,RNA was sequenced from seven different stages duringthe acquisition of longevity, generating between 14 and 38million of reads.These reads were aligned to the Glycine maxWm82.a2.v1 gene model. Differentially expressed transcripts(DET) were correlated with the increase in seed longevity.Transcriptome and GO enrichment analyses of these DETrevealed a signifi cant over-representation of terms associatedwith response to stress and RNA processing and modifi cation.Photosynthesis biological process was related to low seedlongevity. HSF and several TF associated with biotic defense(WRKY3 and NLFX1) are candidate genes whose putative rolein seed longevity deserve further characterization. We alsoperformed the determination of the content of non-reducingsoluble sugars, and we observed that the accumulation ofnon-reducing soluble sugars are related to acquisition oflongevity but only the a

This thesis aims to better represent a framework for asynchrony. Traditional asynchronous models, particularly those used to simulate cellular automata, have used stochasticity or randomness to generate update times. We claimthat, while they may make good representations of their application, such asynchronousmethods rid themodel of the essence of interesting asynchronous processes. Thus, we attempt to better harness the aspects internal to the decision process of such discretely dynamic cells as those in cellular automata.We propose the maxmin-m model as a suitable model for the asynchronous computation of cellular automata. The model uses maxmin-plus algebra, a special case of which is max-plus algebra. This algebra arises naturally from the cellular automaton requirement that a cell receives the state of its neighbours before updating. The maxmin-m model allows each cell to update after it receives m out of a possible n neighbours' states.The max-plus model shows that, while update times may be asynchronous in real time, there is no loss of information, since the corresponding asynchronous process is bijectively related to the synchronous model. In turn, the cellular automaton output, measured by the Shannon and word entropies, is shown to vary little from the synchronous model. Moreover, this type of asynchrony is simple, i.e. it is deterministically obtained due to the linearity of max-plus algebra.Indeed, the maxmin-m model is also shown to be deterministic and always reaches periodic behaviour. In the long time limit, this model is shown to be represented by a max-plus model, supporting its determinism further. Consequently, the complexity of such a model may be thought to be limited. However, we show through large scale experiments that the case where m is approximately n/2 generates most complex behaviour in terms of large periods and transients to the aforementioned periodic orbits. In particular, the complexity is empirically shown to obey a bell form as a function of m (where m ranges from 1 to n). The resulting cellular automaton simulations indicate a correspondence from the complexity of the update times. Therefore, cellular automaton behaviour may be predictable with the type of asynchrony employed in this thesis.

Cette thèse porte sur la résolution du problème d'optimisation Maximum Satisfiability (Max-SAT). Nous y étudions en particulier les mécanismes liés à la détection et à la transformation des sous-ensembles inconsistants par la règle de la max-résolution. Dans le contexte des solveurs de type séparation et évaluation, nous présentons plusieurs contributions liées au calcul de la borne inférieure. Cela va du schéma d'application de la propagation unitaire utilisé pour détecter les sous-ensembles inconsistants à l'extension des critères d'apprentissage et à l'évaluation de l'impact des transformations par max-résolution sur l'efficacité des solveurs. Nos contributions ont permis l'élaboration d'un nouvel outil de résolution compétitif avec les meilleurs solveurs de l'état de l'art. Elles permettent également de mieux comprendre le fonctionnement des méthodes de type séparation et évaluation et apportent des éléments théoriques pouvant expliquer l'efficacité et les limites des solveurs existants. Cela ouvre de nouvelles perspectives d'amélioration, en particulier sur l'augmentation de l'apprentissage et la prise en compte de la structure interne des instances. Nous présentons également un exemple d'utilisation de la règle de la max-résolution dans un algorithme de recherche local. / This PhD thesis is about solving the Maximum Satisfiability (Max-SAT) problem. We study the mechanisms related to the detection and transformations of the inconsistent subsets by the max-resolution rule. In the context of the branch and bound (BnB) algorithms, we present several contributions related to the lower bound computation. They range from the study of the unit propagation scheme used to detect inconsistent subsets to the extension of the learning criteria and to the evaluation of the impact of the max-resolution transformations on the BnB solvers efficiency. Thanks to our contributions, we have implemented a new solver which is competitive with the state of art ones. We give insights allowing a better understanding of the behavior of BnB solvers as well as theoretical elements which contribute to explain the efficiency of these solvers and their limits. It opens new development perspectives on the learning mechanisms used by BnB solvers which may lead to a better consideration of the instances structural properties. We also present an example of integration of the max-resolution inference rule in a local search algorithm.

Orientador : João Carlos Kfouri Quantim de Morais / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Filosofia e Ciencias Humanas / Made available in DSpace on 2018-07-16T17:05:20Z (GMT). No. of bitstreams: 1 Pierre_HectorLuisSaint_M.pdf: 34171943 bytes, checksum: 6f71de5ec6f43e51c9e96dd1203853a6 (MD5) Previous issue date: 1988 / Resumo: Não informado / Abstract: Not informed. / Mestrado / Mestre em Filosofia