Made available in DSpace on 2015-03-26T13:42:19Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1192795 bytes, checksum: 39105ae81e1f922a4d76be541c837833 (MD5) Previous issue date: 2006-09-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The Sw-5 protein codified by the tomato Sw-5 gene confers broad spectrum tospovirus resistance. This protein contains a coil-coiled aminoterminal domain, a central nucleotide binding site domain and a leucine-rich repeat carboxi-terminal domain. These domains suggest that Sw-5 recognizes directly or indirectly virus elicitors and it participates in a signal transduction pathway that leads to the death of infected cells and activation of plant defense responses. The objective of this work was to identifiy and characterize genes encoding proteins that interacts physically with Sw-5 in the yeast two hybrid system. The screening of 5x107 Nicotiana benthamina cDNA clones, using the yeast mating strategy, did not yield any protein capable to interact with Sw-5. Analyzing 1,8 x 105 clones by co-transformation strategy, it was possible identify a cDNA that encodes an chloroplast ferredoxin I (Nb-Fd1) capable to interact with Sw-5. This protein contains a domain and a signature of 2Fe-2S proteins. Virus-induced silencing of the Nb-Fd1 gene leads to a grater number of local necrotic lesions and apical lesions in resistance plants challenged with tospovirus showing that this gene is important for tomato tospovirus resistance. This phenotype indicates that Nb-Fd1 is possibly involved in cellular redox state alteration, reactive oxygen species accumulation and programmed cell death activation processes mediated by Sw-5. / A proteína Sw-5 codificada pelo gene Sw-5 de tomateiro confere resistência de amplo espectro a tospovírus. Essa proteína contém um domínio amino-terminal coil-coiled, um domíno central de ligação a nucleotídeos fosfatados e uma região carboxi-terminal com repetições ricas em leucina. Esses domínios sugerem que a proteína Sw-5 reconhece direta ou indiretamente elicitores produzidos pelo vírus e participa de uma cadeia de transdução de sinais que leva à morte das células inicialmente infectadas pelos tospovírus e, ou, ativação de respostas de defesa da planta. Este trabalho teve como objetivo a identificação e a caracterização molecular de genes que codificam proteínas capazes de interagir fisicamente com a proteína codificada pelo gene de resistência Sw-5 por meio da utilização do sistema duplo-híbrido de leveduras. A triagem 5x107 clones de cDNAs de Nicotiana benthamina por meio da obtenção de diplóides, não revelou nenhum cDNA que codifica proteína capaz de interagir com a proteína Sw-5. A análise de 1,8 x 105 clones pelo processo de co-transformação resultou na identificação de um cDNA que codifica uma proteína ferredoxina I de cloroplasto (Nb-Fd1), capaz de interagir com a proteína Sw-5. Essa proteína contém um domínio e uma assinatura característicos de proteínas 2Fe-2S. Resultados preliminares mostraram que o silenciamento do gene que codifica Nb-Fd1 resulta em uma maior quantidade de lesões necróticas locais e lesões no ápice das plantas resistentes desafiadas com tospovírus evidenciando a não contenção do vírus no sítio de infecção. Esse fenótipo demonstra um papel de Nb-Fd1 na resistência a tospovírus. Assim, é possível que esta proteína participe do processo de alteração do potencial redox da célula, acúmulo de espécies reativas de oxigênio e ativação de morte celular programada que caracterizam a resposta de hipersensibilidade mediada pelo gene Sw-5.

Orientador: Nilson Ivo Tonin Zanchin / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-06T21:20:48Z (GMT). No. of bitstreams: 1 Carneiro_FlaviaRaquelGoncalves_D.pdf: 4565621 bytes, checksum: 26bdfc2a971d3837321625172c6745e8 (MD5) Previous issue date: 2006 / Resumo: O gene que codifica a proteína INT6 corresponde a um dos sítios de inserção do vírus de tumor de glândula mamária em camundongo (MMTV). Esta inserção pode levar à formação de proteínas truncadas sem a porção C-terminal e sem o domínio PCI, descrito como domínio de interação entre proteínas. Neste trabalho, realizamos 3 triagens para a identificação dos ligantes protéicos da proteína humana INT6 (hINT6) pelo método do duplo-híbrido de levedura. Embora interações específicas tenham sido identificadas, não foi possível a confirmação in vitro das novas interações isoladas. Para análises estruturais, usamos a proteína INT6 de Arabidopsis thaliana (AtINT6), visto que a proteína humana expressou em níveis muito baixos na fração solúvel do extrato de Escherichia coli. Análises de dicroísmo circular revelaram que a proteína AtINT6 é rica em estrutura do tipo a-hélice. A região que compreende os aminoácidos 172 a 415, incluindo o domínio PCI, foi identificada por proteólise parcial e espectrometria de massas como um domínio estrutural que após sua clonagem apresentou alto nível de expressão, solubilidade e estabilidade. Este trabalho envolveu também a caracterização da NY-REN-21, que foi isolada pelo duplo-híbrido para a identificação dos ligantes protéicos da proteína hINT6 e representa uma possível ortóloga para o fator de transcrição ZFP38 de camundongo. Ambas as proteínas apresentam um domínio de dimerização (SCAN), 7 domínios dedos de zinco tipo C2H2 na porção C-terminal e uma região central predita como desestruturada. A proteína NY-REN-21 se mostrou parcialmente desenovelada e sua estrutura secundária é afetada pela incubação com EDTA. Ensaios de proteólise limitada, dicroísmo circular e fluorescência confirmaram que sua região central é intrinsicamente desordenada, além de apresentar mobilidade anômala em gel de SDS-PAGE e representa uma fração flexível da proteína. Não foi possível a caracterização da região dos dedos de zinco, pois esta região se mostrou altamente instável. O domínio SCAN da NY-REN-21 é capaz de formar homodímeros e heterodímeros com a proteína SCAND1. Esta interação pode representar uma forma de regulação da atividade da proteína NY-REN-21 / Abstract: The INT6 gene was reported as a frequent genome integration site of Mouse Mammary Tumor Virus (MMTV). This integration may result in truncated forms of INT6 protein lacking its C-terminal region and the PCI domain. It has been reported that this domain is involved in protein-protein interaction. We performed 3 yeast two-hybrid screens in order to identify the human INT6 (hINT6) interaction partners. Although two previously described specific interactions were identified in these screens, it was not possible to confirm the new interactions by in vitro binding assays. The Arabidopis thalina INT6 ortholog (AtINT6) was used for structural analyses, since the hINT6 was insoluble following expression in Escherichia coli. AtINT6 showed CD spectra with high helical content. The region comprising amino acids 172 to 415 forms a compact protease resistant domain as determined by limited proteolysis and mass spectrometry analyses. This domain, comprising the PCI domain sequence, was cloned and expressed in E. coli and showed high solubility and stability. This recombinant protein has the potential to serve as a model protein for three-dimensional structure determination of the PCI domain. We also studied the NY-REN-21 protein, which was isolated as a potential hINT6 interaction partner and represents a putative ortholog of the mouse ZFP38 transcriptional factor. Both proteins are C2H2 type multifinger proteins, containing a conserved oligomerization domain (SCAN) in the N-terminal region and a predicted disordered central region. Our analyses showed that full-length NY-REN-21 is partially unfolded and its secondary structure content is affected by incubation with EDTA. The central region of NY-REN-21 shows an aberrant mobility on SDS-PAGE and is intrinsically unstructured as reveled by circular dichroism, fluorescence and limited proteolysis. The zinc finger region was not characterized because of its unstable nature. The recombinant SCAN domain of NY-REN-21 can form homodimers and heterodimers with the SCAND1 protein. This interaction may represent a novel regulatory mechanism of NY-REN-21 activity / Doutorado / Genetica Animal e Evolução / Doutor em Genetica e Biologia Molecular

La thèse est une contribution à la réduction de la consommation d'énergie primaire et à une meilleure utilisation des sources d'énergie renouvelables dans le cadre des systèmes de refroidissement utilisés dans le bâtiment. Après un état de l'art sur les systèmes de refroidissement, un modèle dynamique d'un système de rafraîchissement solaire à base de machine à absorption est développé et simulé. Ensuite, un facteur d'efficacité pour comparer la pertinence de ce système dans différentes régions du monde est défini. Dans la troisième partie, la notion des systèmes de refroidissement hybride -une méthode efficace contribuant à la réduction de la consommation d'énergie primaire- est présentée. Puis, les systèmes hybrides de refroidissement sont classés en catégories et sont comparés avec les systèmes de refroidissement individuels. Ensuite, un schéma permettant de sélectionner le meilleur système de refroidissement hybride dans des conditions données est proposé. Dans la dernière partie, une méthode de dimensionnement d’un système hybride à base d’énergies renouvelables est établie. Ainsi, le dimensionnement est réalisé en tenant compte de la région spécifique d’utilisation. Pour ce faire, un système de refroidissement hybride, conçu pour une maison standard, est modélisé puis simulé en utilisant le logiciel Trnsys. Finalement, et pour illustrer la méthode proposée, la problématique de dimensionnement est considérée pour deux régions différentes du globe; à savoir Marseille-France et Beyrouth-Liban. Le but est d’évaluer les performances de la méthode, à travers des données effectives, pour diverses conditions climatiques, prix des composants et tarif d'électricité. / This thesis is a contribution towards the reduction of primary energy consumption and a better use of the renewable energy sources within the cooling system for building use. After a state of the art of the cooling machines for building use, a dynamic model for a solar absorption cooling system is developed and simulated. Then, an effectiveness factor (EF) for the comparison of solar absorption chiller suitability in different locations is defined. In the third chapter, the concept of hybrid cooling system -an efficient method contributing to the reduction of primary energy consumption- is presented. Hybrid cooling systems are categorized and reviewed, with the improvement achieved compared to standalone technologies. Then, a scheme for the selection of the best hybrid cooling system for given conditions is proposed. In the last part, an optimal sizing method that defines, in a specific region, a hybrid cooling energy system, economically feasible with maximum renewable energy share is presented. Thereby, the sizing method is performed taking into account the region where it will be used. For this purpose, a hybrid cooling system, used for a standard residential house, is designed. The system is modeled and simulated using a transient system simulation program, called Trnsys. Finally, the problem of sizing is studied for different case studies; namely Marseilles-France and Beirut-Lebanon. The aim is to assess the proposed method according to diverse climatic conditions, component prices and electricity costs.

Cette thèse propose la méthodologie de dimensionnement optimal pour optimiser la configuration de système d'énergie hybride. Pour cela, nous utilisons une approche pour la génération de base de règles floues et une optimisation automatiques au moyen d'algorithme génétique et d'un PSO adaptés avec le floue. Ces algorithmes nous permet d'obtenir le nombre optimal de panneaux photovoltaïques, d'éoliennes et des batteries, minimisant le coût total du système et garantissant la disponibilité permanente de l'électricité pour couvrir les besoins énergétiques. L'historique horaire de vitesse du vent, d’ensoleillement, sont utilisés pour modéliser la production des éoliennes, la production photovoltaïque et de charge. Le coût total est la fonction objective et la taille technique est une contrainte. / This thesis proposes the optimum sizing methodology to optimize the configuration of hybrid energy system. For this, we use an approach for automatic fuzzy rule base generation and optimization by means of Fuzzy-Adaptive Genetic Algorithm and fuzzy adaptive PSO. This Algorithms allows us to obtain the optimal number of photovoltaic panels, wind turbines and storages units, ensuring the minimal global high efficiency system total cost and guaranteeing the permanent availability of energy to cover the load energy requirements. Historical hourly wind speed, solar irradiance and load data are used to stochastically model the wind turbines, photovoltaic generation and load. The total cost is the objective function and the technical size is a constraint.

Le développement des systèmes énergétiques renouvelables et/ou conventionnels pour l’alimentation des zones décentralisées est confronté à de multiples barrières techniques, institutionnelles, socioculturelles et surtout financières. La solution défendue dans cette thèse est de rechercher des approches pour concevoir des systèmes durables et rentables, conditions requises pour le développement de l’électrification rurale par le secteur privé. L’analyse des différents critères économiques permet tout d’abord de définir plusieurs types d’équivalence entre critères économiques : équivalence décisionnelle, équivalence pour le dimensionnement optimal et équivalence pour le classement des solutions techniques. Cette première étude permet de démontrer que les 3 critères économiques habituellement utilisés (le LCOE, le taux de rentabilité interne, et le temps de retour sur investissement) aboutiront à des solutions techniques optimales différentes. Un modèle technico-économique est alors développé pour les systèmes hybrides PV/Diesel sans batteries de stockage en site isolé. Une application au site 2iE-K1 (Ouagadougou) permet de valider la conformité des modèles par comparaison au logiciel HOMER. On montre que les critères de taux de rentabilité interne maximum ou de temps de retour sur investissement minimum limitent l’investissement et le montant de la dette maximale. Plusieurs solutions optimales, dépendant du critère économique considéré, sont alors proposées pour un cas d’application réel : l’électrification de la localité rurale de Bilgo. Le travail réalisé au cours de cette thèse se situe donc dans la branche des outils de dimensionnement optimal et d’optimisation technico-économique des systèmes de production décentralisée d’énergie. / The development of renewable and / or conventional energy systems for power supply to decentralized areas faces many technical, institutional, socio-cultural and financial barriers. The solution recommended in this thesis is to look for approaches to design sustainable and cost-effective systems for the development of rural electrification by the private sector. An analysis of various economic criteria allows first to define several types of equivalence inter-linking them: equivalence for investment decision, equivalence for the optimal sizing and equivalence for ranking technical solutions. This first study shows that the three economic criteria usually used (levelized cost of energy, internal rate of return and discounted payback period) lead to different optimal technical solutions. A techno-economic model is then developed for off-grid batteryless PV/Diesel hybrid systems. An application to 2iE-K1 campus (Ouagadougou) has been done to validate the compliance of models compared to HOMER software. It shows that the maximum internal rate of return criteria or minimum discounted payback period criteria reduces the initial investment and the amount of the maximum debt. Several optimal solutions are then obtained, depending on economic criteria under consideration. The electrification of the rural community of Bilgo has been chosen for a real case study. The work carried out, through this thesis, could be classified in the category of optimal design and techno-economic optimization tools for off-grid energy production systems.

京都大学 / 新制・課程博士 / 博士(情報学) / 甲第23545号 / 情博第775号 / 新制||情||132(附属図書館) / 京都大学大学院情報学研究科数理工学専攻 / (主査)教授 太田 快人, 教授 山下 信雄, 教授 大塚 敏之 / 学位規則第4条第1項該当 / Doctor of Informatics / Kyoto University / DFAM

Quantum sensing, a protocol that takes advantage of the extreme sensitivity of quantum systems to their environment, enables many applications of quantum systems for sensing. Inspired by direct electric field sensing using the Stark effect of a nitrogen-vacancy(NV) center, this work implements an NV-magnet hybrid way to explore the possibilities of overcoming NV’s relatively weak coupling strength to electric fields. The magnetic-noise-induced population relaxation of the NV center serves as the mechanism for sensing. Within this scheme, the magnetic noise spectrum is tuned by modulating the magnetic properties via voltage-controlled magnetic anisotropy (VCMA) or electric-field-induced magnetoelastic effect. In this way, the noise carrying the information of the electric field is taken as a signal - the shift of the noise spectrum leads to a population difference of NV energy levels, which is used for evaluating electric fields. The investigation of the relation between sensitivities and operation points reveals that lower operation frequency is desirable for better performance. The comparison between VCMA and electric-field-induced magnetoelastic effect indicates that the efficiency of converting electric field into magnetic property modulation is a critical parameter for sensitivity enhancement.

Exocyst is a binding protein complex, which is evolutionary conserved in yeast, animal and in plant cells. It has crucial role in regulation of cell morfogenesis and cell polarity. The function of the exocyst complex is binding of secretoric vesicle to the proper side on plasma membrane in penultimate step of exocytosis. This process is essecial for function and survival of cell. Another process crucial for the cell is autophagy. In plants autophagy plays important role in the responses to nutrient starvation, senescence, abiotic and biotic stress. RabG3b are small GTPases, which have positive role in autophagy. In this work I described the interaction between RabG3b and some of subunits of exocyst complex: Exo70B1, Exo70B2 and Exo84b. I also studied changes in morfogenesis of tonoplast by induction and inhibition of authophagy and induction of anthocyans synthesis in Arabidopsis thaliana.

Um Beweise für eine mögliche Mikrokompartimentierung der Glykolyse im pflanzlichen System zu erhalten, sollten in der vorliegenden Arbeit Protein-Protein-Interaktionen der cytosolischen Mais-Aldolase mit anderen Proteinen experimentell nachgewiesen werden. Die in Tieren bekannte Interaktion des glykolytischen Enzyms Aldolase mit Aktin, einem Bestandteil des Cytoskeletts, wurde für Pflanzen in vitro durch Copolymerisationsversuche bestätigt. Die Bindung pflanzlicher Aldolase an Aktinfilamente wurde anders als im tierischen System durch das Substrat Fructose-1,6-bisphosphat auch in hohen Konzentrationen (10 mM) nicht vollständig verhindert, sondern führte lediglich zu einer um 50% verringerten Bindung. Eine ebenfalls hemmende Wirkung auf die Bindung der Aldolase an Aktin wiesen Fructose-6-phosphat und Fructose-2,6-bisphosphat in Konzentrationen von 10 mM auf. Ein eindeutiger Einfluss des Redox-Milieus auf die Aldolase-Aktin-Bindung konnte nicht nachgewiesen werden. Mit Hilfe des im Rahmen dieser Arbeit etablierten Hefe-2-Hybrid-Systems wurden weitere Interaktionspartner der Aldolase identifiziert. Insgesamt wurden neun mögliche Protein-Protein-Interaktionen nachgewiesen, bei denen es sich jedoch zum Teil um falsch-positive Interaktionen handeln kann. Neben einigen noch unbekannten Proteinen konnten Interaktionen mit einem Translations-Initiationsfaktor und dem spannungsabhängigen Anionenkanalprotein VDAC nachgewiesen werden. In Bindeversuchen auf Grundlage der Affinitätschromatographie mit den rekombinanten Proteinen VDAC und Aldolase wurde ein weiterer Hinweis auf eine Interaktion zwischen VDAC und Aldolase erhalten. Aufgrund unspezifischer Bindungen der Aldolase an die Affinitätsmatrix konnte mit dieser Methode jedoch keine eindeutige Verifizierung der Interaktion erzielt werden. Eine eindeutige Bestätigung der Interaktionen zwischen Aldolase und Aktin sowie zwischen Aldolase und VDAC erfolgte durch Far-Western-Blots .

Les activités scientifiques que nous présentons dans ce manuscrit de thèse s’inscrivent dans la thématique de l’intelligence ambiante, axe stratégique ADREAM au sein du LAAS-CNRS. Depuis plusieurs années notre équipe de recherche N2IS fédère l’approche technologique de la SHM avec pour objectif la surveillance de santé structurelle. En effet, la maturité des matériaux innovants tels que les composites suscitent un intérêt certain auprès des constructeurs aéronautiques, ou bien encore l’utilisation des matériaux de type béton pour des ouvrages d’art, sont autant de structures hétérogènes qui nécessitent une surveillance périodique et/ou continue. Ceci, afin de détecter des cracks, des fissures, des corrosions surfaciques ou bien encore des délaminages. Pour ce faire, les solutions existantes s’appuient usuellement sur des technologies de contrôle non destructif (CND) qui intègrent le plus souvent des réseaux de capteurs à faible consommation interfacés avec des systèmes d’analyses des signaux. Ces approches CND présentent des limitations fonctionnelles majeures : elles ne sont pas versatiles et ne permettent pas d’assurer une continuité de service dans un mode « dégradé » lors d’un fonctionnement sur batterie avec un niveau d’énergie minimal. Notre travail de recherche se situe dans une perspective liée à la quantification d’un niveau de robustesse de structure hétérogène. Il a pour ambition le développement et l’intégration de systèmes matériels mixtes (analogiques/numériques) reconfigurables. Au terme d’une investigation sur les principales solutions technologiques matérielles reprogrammables et compte tenu de la problématique liée aux développements d’algorithmes d’analyse embarqués et de la minimisation de la consommation énergétique des capteurs, le choix s’est porté sur des technologies complémentaires FPAA et FPGA. Initialement nos études de recherche se sont focalisées sur l'étude de fonction analogique matérielle reconfigurable analogique. L'objectif est de démontrer une faisabilité conceptuelle en intégrant un système de conditionnement complexe (implémentation d'une technique de détection synchrone), ceci en considérant le compromis entre la prise de décision d’une reconfiguration à la volée vis-à-vis d’une gestion rationnelle de l'énergie du système. Dès lors, se pose la question de comment intégrer et stocker des données nécessaires au développement d’un traitement numérique performant ? Une solution repose sur une approche hybride avec une puce de type Zynq produite par Xilinx et embarquée sur une Zedboard. Cette solution, plus performante qu’une approche PSoC a permis le développement et l’implémentation de techniques de traitement de signal grâce à des outils d'optimisation et de génération de code de haut niveau. Au terme de ce travail de recherche, les résultats obtenus démontrent la validité des concepts mis en œuvre et permettent d'engager dès à présent le développement d’architectures intelligentes de nouvelle génération / Scientific activities described in this PhD thesis are part of the theme of smart environment, strategy axes of ADREAM with the LAAS-CNRS. Since several years, our research team (N2IS) had a field of interest in SHM (Structural Health Monitoring) with the objective of doing a smart diagnostic on different heterogeneous structures. Indeed, the maturity of innovative materials such as composites triggering interest among aircraft manufacturers, or even the use of materials like concrete structures of civil engineering, all those heterogeneous structures that require periodic monitoring and / or continuous one. This is to detect cracks, disbond, surface corrosion or even delamination. To do this, existing solutions usually rely on technologies of nondestructive testing (NDT) that incorporate mostly sensor networks low-power systems interfaced with analysis of signals. These approaches have significant functional limitations: they are not versatile and do not allow for continuity of service in a "degraded" when operating on battery power with a minimum level of energy mode. Our research is a view related to the quantization level of robustness of a heterogeneous structure. Its aim is the development and integration of hardware reconfigurable mixed (A / D ) systems. After an investigation of the main technological solutions reprogrammable hardware and given the problems associated with developments in analytical embedded and minimizing the energy consumption of sensor algorithms. The choice was based on technologies like FPAA and FPGA. Initially our research studies have focused on the study of reconfigurable analog hardware analog. The objective was to show a conceptual feasibility of integrating a complex conditioning system (implementation of a synchronous detection technique), considering the tradeoff between a decision on the fly reconfiguration and a rational energy management system. Therefore, the question of how to integrate and store data necessary for the development of an efficient digital processing. A solution based on a hybrid approach with a chip produced by Xilinx called Zynq and embedded on a Zedboard. This solution is more efficient than a PSoC approach and allowed the development and implementation of signal processing techniques with tools for optimization and provided a solution of self-generation code trough a graphic interface. Following this research, the results obtained demonstrate the validity of the concepts implemented and allow us to imagine the next smart generation architectures