車両の電動化(ハイブリッド化)と内燃機関システムの電動化を最適に組み合わせた技術と、それらを効果的に稼働させるハイブリッド制御ロジックを適用した新方式大型ハイブリッドトラックを提案した。長距離貨物輸送時の燃料消費率の改善に対し効果があることを明らかとした。同時に、従来大型トラックに対し、荷室搭載性の確保や車両コストの抑制が可能となることが分かり、実用性の面でも優位性があることを示した。 / 博士(工学) / Doctor of Philosophy in Engineering / 同志社大学 / Doshisha University

The potential of Solid Oxide Fuel Cell Gas Turbine (SOFC/GT) hybrid systems for fuel flexibility makes this technology greatly attractive for system hybridization with various fuel processing units in advanced power generation systems and/or polygeneration plants. Such hybrid technologies open up the possibility and opportunities for improvement of system reliabilities and operabilities. However, SOFC/GT hybrid systems have not yet reached their full potential in term of capitalizing on the synergistic benefits of fuel cell and gas turbine cycles. Integrating fuel cells with gas turbine and other components for transient operations increases the risk for exposure to rapid and significant changes in process dynamics and performance, which are primarily associated with fuel cell thermal management and compressor surge. This can lead to severe fuel cell failure, shaft overspeed, and gas turbine damage. Sufficient dynamic control architectures should be made to mitigate undesirable dynamic behaviours and/or system constraint violations before this technology can be commercialized. But, adequate understanding about dynamic coupling interactions between system components in the hybrid configuration is essential. Considering this critical need for system identification of SOFC/GT hybrid in fuel flexible systems, this thesis investigates the dynamic performance of SOFC/GT hybrid technology in response to fuel composition changes. Hardware-based simulations, which combined actual equipment of direct-fired recuperated gas turbine system and simulated fuel cell subsystem, are used to experimentally investigate the impacts of fuel composition changes on the SOFC/GT hybrid system, reducing potentially large inaccuracies in the dynamic study. The impacts of fuel composition in a closed loop operation using turbine speed control were first studied for the purpose of simplicity. Quantification of safe operating conditions for dynamic operations associated with carbon deposition and compressor stall and surge was done prior to the execution of experimentation. With closed loop tests, the dynamic performance of SOFC/GT hybrid technology due to a transition in gas composition could be uniquely characterized, eliminating the interactive effects of other process variables and disturbances. However, for an extensive system analysis, open loop tests (without turbine speed control) were also conducted such that potential coupling impacts exhibited by the SOFC/GT hybrid during fuel transients could be explored. Detailed characterization of SOFC/GT dynamic performance was performed to identify the interrelationship of each fuel cell variable in response to fuel composition dynamics and their contributions to operability of the system. As a result of lowering LHV content in the fuel feed, which involved a transition from coal-derived syngas to humidified methane composition in the SOFC anode, the system demonstrated a dramatic transient increase in fuel cell thermal effluent with a time scale of seconds, resulting from the conversion of fuel cell thermal energy storage into chemical energy. This transient was highly associated with the dynamics of solid and gas temperatures, heat flux, heat generation in the fuel cell due to perturbations in methane reforming, water-gas shifting, and electrochemical hydrogen oxidation. In turn, the dramatic changes in fuel cell thermal effluent resulting from the anode composition changes drove the turbine transients that caused significant cathode airflow fluctuations. This study revealed that the cathode air mass flow change was a major linking event during fuel composition changes in the SOFC/GT hybrid system. Both transients in cathode air mass flow and anode composition significantly affected the hybrid system performance. Due to significant coupling between fuel composition transitions and cathode air mass flow changes, thermal management of SOFC/GT hybrid systems might be challenging. Yet, it was suggested that modulating cathode air flow offered promise for effective dynamic control of SOFC/GT hybrid systems with fuel flexibility. / Thesis / Doctor of Philosophy (PhD)

Toxoplasmose ist weltweit eine der am häufigsten auftretenden parasitären Zoonosen mit einer geschätzten Infektionsrate von über 30%. Toxoplasma gondii (Phylum: Apicomplexa) besitzt ein Plastid ähnliches Organell, den Apicoplasten. In diesem befindet sich das einzig bekannte Redoxsystem, welches aus der Ferredoxin-NADP+-Reduktase und Ferredoxin (Fd) besteht. Fd als Elektonendonator liefert Elektronen an verschiedene essentielle Stoffwechselwege, wie der Isoprenoidvorstufen- und Liponsäuresynthese. Um die bei einem Elektronentransfer benötigte direkte Protein-Protein-Interaktion eingehend zu analysieren, wurde ein bakterielles Reverse Two Hybrid System verwendet, womit die Interaktion von TgFd und TgLipA gezeigt werden konnte. Da angenommen wird, dass Fd eine zentrale Rolle in verschiedenen Stoffwechselwegen übernimmt, ist für einen Fd Knockout ein komplexer biochemischer Phänotyp zu erwarten, der möglicherweise zum Absterben der Parasiten führt. Zur Untersuchung dessen wurden zwei komplementäre Wege verfolgt. Eine der Strategien basierte auf dem grundsätzlichen Nachweis, dass Fd unerlässlich für das Überleben von T. gondii ist. Mit Hilfe des DiCre Systems sollte ein definierter genetischer Fd Knockout hergestellt werden, welcher jedoch nicht zweifelsfrei generiert werden konnte. Bei der zweiten Strategie kam ein konditionales Knockdown System zur Anwendung, bei welchem die Expression des Fd Gens nach Induktion herabreguliert wird. Mit Hilfe dessen konnten weitreichende Auswirkungen der Fd Defizienz auf T. gondii gezeigt werden: die Fettsäuresynthese der im Apicoplasten synthetisierten Fettsäuren ist reduziert sowie die Motilität durch eine beeinträchtigte Isoprenoidsynthese verringert, wodurch insgesamt drastische Auswirkungen auf das Parasitenwachstum gezeigt werden konnten. Beide Stoffwechsel sind vom Elektronendonator Fd abhängig und durch die Fd Herabregulation betroffen. Die Ergebnisse unterstreichen die essentielle Rolle des Fd-Redoxsystems von T. gondii. / Toxoplasmosis is one of the most common parasitic zoonoses world-wide, around 30% of human beings are infected. Toxoplasma gondii (phylum: Apicomplexa) contains a unique intracellular organelle derived from plastids, called apicoplast. The only known redox system in the apicoplast consists of the ferredoxin NADP+-reductase and its redox partner, ferredoxin (Fd). The latter donates electrons to different essential metabolic pathways in the apicoplast like the last two enzymes of the isoprenoid precursor biosynthesis and the lipoic acid synthesis. To dissect protein protein interactions for an electron transfer a bacterial reverse two hybrid system was used. The physical interaction of both proteins TgFd and TgLipA could be shown. Fd is supposed to play an important role in diverse metabolic pathways, hence a knock-out of the Fd gene is expected to generate a complex biochemical phenotype and be lethal to the parasite. Therefore two complementary approaches were used to analyze the role of TgFd in this context. The first strategy shall verify the essentiality of TgFd for the survival of T. gondii. It is based on the DiCre system whereby a defined genetic knock out of TgFd is produced. Respectives parasites have been generated, but at the end no genetic Fd knock out could be produced. In the second approach a conditional knock-down was generated, where the expression of the TgFd gene is repressed after induction. The Fd deficiency has wide ranging effects on T. gondii: The fatty acid synthesis of the apicoplast-synthesized fatty acids is reduced as well as the motility is decreased due to an affected isoprenoid synthesis. In total this leads to a dramatic inhibition of parasite growth. Both metabolic pathways depend upon the electron carrier Fd and thus are affected by Fd deficiency. The results underline the essential role of the ferredoxin redoxsystem of T. gondii.

Proteinmethylierung spielt eine immer größere Rolle in der Regulierung zellulärer Prozesse. Die Entwicklung effizienter proteomweiter Methoden zur Detektion von Methylierung auf Proteinen ist limitiert und technisch schwierig. In dieser Arbeit haben wir einen neuen Hefe-Zwei-Hybrid-Ansatz (Y2H) entwickelt, der Proteine, die miteinander wechselwirken, mit Hilfe von Sequenzierungen der zweiten Generation identifiziert (Y2H-Seq). Der neue Y2H-Seq-Ansatz wurde systematisch mit dem Y2H-Seq-Ansatz verglichen. Dafür wurde ein Bait-Set von 8 Protein-Arginin-Methyltransferasen, 17 Protein-Lysin-Methyltransferasen und 10 Demethylasen gegen 14,268 Prey-Proteine getestet. Der Y2H-Seq-Ansatz ist weniger arbeitsintensiv, hat eine höhere Sensitivität als der Standard Y2H-Matrix-Ansatz und ist deshalb besonders geeignet, um schwache Interaktionen zwischen Substraten und Protein-Methyltransferasen zu detektieren. Insgesamt wurden 523 Wechselwirkungen zwischen 22 Bait-Proteinen und 324 Prey-Pr oteinen etabliert, darunter 11 bekannte Methyltransferasen-Substrate. Netzwerkanalysen zeigen, dass Methyltransferasen bevorzugt mit Transkriptionsregulatoren, DNA- und RNA-Bindeproteinen wechselwirken. Diese Daten repräsentieren das erste proteomweite Wechselwirkungsnetzwerk über Protein-Methyltransferasen und dienen als Ressource für neue potentielle Methylierungssubstrate. In einem in vitro Methylierungsassay wurden exemplarisch mit Hilfe massenspektrometrischer Analysen die methylierten Aminosäurereste einiger Kandidatenproteine bestimmt. Von neun getesteten Proteinen waren sieben methyliert, zu denen gehören SPIN2B, DNAJA3, QKI, SAMD3, OFCC1, SYNCRIP und WDR42A. Wahrscheinlich sind viele Methylierungssubstrate im Netzwerk vorhanden. Das vorgestellte Protein-Protein-Wechselwirkungsnetzwerk zeigt, dass Proteinmethylierung sehr unterschiedliche zelluläre Prozesse beeinflusst und ermöglicht die Aufstellung neuer Hypothesen über die Regulierung Molekularer Mechanismen durch Methylierung. / Protein methylation on arginine and lysine residues is a largely unexplored posttranslational modification which regulates diverse cellular processes. The development of efficient proteome-wide approaches for detecting protein methylation is limited and technically challenging. We developed a novel workload reduced yeast-two hybrid (Y2H) approach to detect protein-protein interactions utilizing second generation sequencing. The novel Y2H-seq approach was systematically evaluated against our state of the art Y2H-matrix screening approach and used to screen 8 protein arginine methyltransferases, 17 protein lysine methyltransferases and 10 demethylases against a set of 14,268 proteins. Comparison of the two approaches revealed a higher sensitivity of the new Y2H-seq approach. The increased sampling rate of the Y2H-seq approach is advantageous when assaying transient interactions between substrates and methyltransferases. Overall 523 interactions between 22 bait proteins and 324 prey proteins were identified including 11 proteins known to be methylated. Network analysis revealed enrichment of transcription regulator activity, DNA- and RNA-binding function of proteins interacting with protein methyltransferases. The dataset represents the first proteome-wide interaction network of enzymes involved in methylation and provides a comprehensively annotated resource of potential new methylation substrates. An in vitro methylation assay coupled to mass spectrometry revealed amino acid methylation of candidate proteins. Seven of nine proteins tested were methylated including SPIN2B, DNAJA3, QKI, SAMD3, OFCC1, SYNCRIP and WDR42A indicating that the interaction network is likely to contain many putative methyltransferase substrate pairs. The presented protein-protein interaction network demonstrates that protein methylation is involved in diverse cellular processes and can inform hypothesis driven investigation into molecular mechanisms regulated through methylation.

O sistema celulolítico do fungo filamentoso Trichoderma reesei é induzido transcricionalmente em pelo menos 1000 vezes pelo crescimento do fungo na presença de celulose e fortemente reprimido por glicose. Usando a abordagem de deleção no promotor, determinou-se que a região localizada entre -241 e -72 bp, em relação ao TATA box, denominada UARcb1, é responsável pela transcrição estimulada por celulose da enzima celobiohidrolase I (cbhl). Neste trabalho mostramos que essa região controla a transcrição de um gene repórter, sofrendo repressão por glicose, em Saccharomyces cerevisiae, um microrganismo que não possui os genes necessários para a utilização de celulose. A transcrição mediada por UARcbl, que é controlada por glicose, requer o produto do gene SNFl, uma proteína quinase, e dois repressores: SSN6 e TUP1, cujos papéis no controle de genes reprimidos por glicose, na levedura, são bem estabelecidos. Nossos resultados indicam um mecanismo conservado de controle por glicose em microrganismos eucarióticos. / The cellulotic system of the filamentous fungus Trichoderma reesei is transcriptionally induced 1000 -fold in presence of cellulose and is strongly repressed by glucose. Using the promoter deletion approach, the upstream activating region (UARcbl) responsible for cellulose-stimulated transcription of the major member of the cellulase system, cellobiohydrolase I, was localized between -241 and -72 relative to the TATA box. In this work we show that this region controls transcription and mediates glucose repression of a reporter gene in Saccharomyces cerevisiae, a unicellular microorganism that lacks the genes required for the utilization of cellulose. Glucose-controlled transcription mediated by the UARcbl requires the product of SNF1 gene, a protein kinase, and two repressors SSN6 and TUP1, which are well estalished in controlling glucose-represible yeast genes. Our results indicate a conserved mechanism of glucose control in eukariotic microorganisms.

O sistema celulolítico do fungo filamentoso Trichoderma reesei é induzido transcricionalmente em pelo menos 1000 vezes pelo crescimento do fungo na presença de celulose e fortemente reprimido por glicose. Usando a abordagem de deleção no promotor, determinou-se que a região localizada entre -241 e -72 bp, em relação ao TATA box, denominada UARcb1, é responsável pela transcrição estimulada por celulose da enzima celobiohidrolase I (cbhl). Neste trabalho mostramos que essa região controla a transcrição de um gene repórter, sofrendo repressão por glicose, em Saccharomyces cerevisiae, um microrganismo que não possui os genes necessários para a utilização de celulose. A transcrição mediada por UARcbl, que é controlada por glicose, requer o produto do gene SNFl, uma proteína quinase, e dois repressores: SSN6 e TUP1, cujos papéis no controle de genes reprimidos por glicose, na levedura, são bem estabelecidos. Nossos resultados indicam um mecanismo conservado de controle por glicose em microrganismos eucarióticos. / The cellulotic system of the filamentous fungus Trichoderma reesei is transcriptionally induced 1000 -fold in presence of cellulose and is strongly repressed by glucose. Using the promoter deletion approach, the upstream activating region (UARcbl) responsible for cellulose-stimulated transcription of the major member of the cellulase system, cellobiohydrolase I, was localized between -241 and -72 relative to the TATA box. In this work we show that this region controls transcription and mediates glucose repression of a reporter gene in Saccharomyces cerevisiae, a unicellular microorganism that lacks the genes required for the utilization of cellulose. Glucose-controlled transcription mediated by the UARcbl requires the product of SNF1 gene, a protein kinase, and two repressors SSN6 and TUP1, which are well estalished in controlling glucose-represible yeast genes. Our results indicate a conserved mechanism of glucose control in eukariotic microorganisms.

Die vorliegende Arbeit beschäftigt sich mit Cbs1p und Cbs2p, zwei spezifische Translationsaktivatoren der COB-mRNA. Im Mittelpunkt standen sowohl die weitere molekularbiologische und biochemische Charakterisierung von Cbs1p als auch ein Screening von Interaktionskandidaten, die mit Cbs1p und/oder Cbs2p physikalisch wechselwirken könnten. Cbs1p liegt als peripheres Membranprotein fest mit der inneren Mitochondrienmembran matrixseitig assoziiert vor. Dabei spielen möglicherweise hydrophobe und/oder Protein-Protein-Wechselwirkungen mit integralen Membranproteinen eine essentielle Rolle bei der Membranverankerung von Cbs1p. Durch die Identifizierug von atmungsdefekten Cbs1p-Mutanten, deren Mutationen in Bereichen mit Homologie zu RNA-bindenden Proteinen liegt, verstärken sich die Hinweise zur Beteiligung von Cbs1p an der direkten physikalischen Wechselwirkung mit dem 5´-leader der COB-mRNA. Darüber hinaus konnte gezeigt werden,, dass die abspaltbare Präsequenz nicht notwendig für einen mitochondrialen Import ist. Die Ergebnisse präzisieren und erweitern das vorliegende Modell der Wirkungsweise der Translationsaktivatoren Cbs1p und Cbs2p (Michaelis, 1991). Aufgrund der Membranverankerung von Cbs1p wird auch die gebundene COB-mRNA in räumlicher Nähe zur Membran gebracht. Darüber hinaus definiert Cbs1p damit möglicherweise auch den Ort der Insertion des nascierenden Apocytochrom b in die Membran. Cbs2p vermittelt die Bindung zur kleinen Untereinheit der mitochondrialen Ribosomen und könnte seinerseits ebenfalls in Interaktionen mit Untereinheiten des bc1-Komplexes involviert sein.

La capacité des systèmes conventionnels de production d'énergie éolienne et marémotrice à fournir au réseau une énergie fiable et stable à tout moment est un nouveau défi en raison des fluctuations météorologiques, qui ont un impact significatif et direct sur la production d'énergie. C'est pourquoi l'hybridation des systèmes de production d'énergie éolienne et hydrolienne ont été étudiées pour améliorer l'intégration des énergies éolienne et marémotrice sur le réseau électrique.Cette étude nous a amené à développer des contributions liées à deux axes principaux :Le premier axe est focalisé sur un nouveau concept d'hybridation de deux sources énergétiques différentes en termes de propriétés physiques, l’éolien et l’hydrolienne à axe horizontal, basé sur un couplage électromécanique de ces deux systèmes. Les deux ressources sont l’énergie éolienne et l’énergie des courants marins. Le concept est développé en utilisant les similitudes fonctionnelles des turbines et les similarités en conversion d’énergie de leurs chaînes énergétiques. Pour appliquer ce concept en premier lieu, les paramètres de la génératrice asynchrone à double alimentation installée dans l’émulateur du GREAH sont identifiés. Ensuite, la chaîne de conversion de puissance est modélisée mathématiquement et simulée dans un environnement MATLAB / SIMULINK. Nous avons développé deux stratégies de commande.Une stratégie à vitesse fixe appelé "Contrôle direct de la vitesse", et une stratégie à vitesse variable basée sur la recherche de puissance maximale, dénommée "Contrôle indirecte de vitesse". Enfin, ce concept a été implémenté pratiquement sur l’émulateur en temps réel du laboratoire. Les résultats obtenus ont été analysés et discutés suite à ces travaux.Le deuxième axe est consacré à un concept que l’on appelle «temps accélérée» de simulation ou « temps virtuel ». Par la suite, ce concept a été mis en œuvre sur l’émulateur multi physique disponible au laboratoire GREAH. Ce concept (temps accélérée) est basé sur la réduction des échantillons de profil de vent afin de diminuer le temps de simulation et faciliter la commande en temps réel.Les résultats principaux sont obtenus d’abord dans MATLAB / SIMULINK, puis ont été vérifiés sur l’émulateur en temps réel. L’objectif principal de cette thèse est d’étudier le concept d’hybridation éolienne offshore / éolienne basée sur la flexibilité d’un émulateur multifonctions permettant diverses architectures d’émulation : éoliennes, éoliennes, et systèmes hybrides éoliennes - éoliennes. Nous analysons son impact sur la puissance de sortie du système. Les résultats obtenus sont corrélés aux profils de vitesse du vent et des marées, dans lesquels les propriétés statistiques ayant un impact sur les chaînes énergétiques mondiales pourraient être complémentaires et en particulier en fonction des sites donnés.Contributions principales et perspectives- Développement du concept de couplage électromécanique. Lorsque deux sources d’énergie renouvelables sont « intégrées », on stabilise la fluctuation rapide de la puissance générée, mais sous certaines conditions telles que la présence d’unités de stockage ou d’un système d’embrayage automatique.- Le concept temps accéléréeCette méthode est utilisée pour réduire la taille des données enregistrées du vent ou des courant marins, afin d’accélérer le temps de simulation des unités de production d'énergie avec des résultats raisonnables qui se rapprochent pertinemment des situations réelles.- Etudier et développer le concept de régime d’arbre électrique :Si le couplage électromécanique est difficile à réaliser du point de vue mécanique et que les découplages à arbre unique sont trop fréquents et que les contraintes mécaniques sont élevées, on peut étudier le régime de l'arbre électrique avec deux machines à induction DFIG. Le système peut fonctionner en mode synchrone avec des structures et configurations spécifiques. / The ability of conventional wind and tidal generation systems to provide the grid with reliable and stable power at all times is a new challenge due to weather fluctuations, which have a significant and direct impact on energy production. This is why the hybridization of wind and tidal power generation systems has been studied to improve the integration of wind and tidal power into the electricity grid.This study led us to develop contributions related to two main axes:The first axis is focused on a new concept of hybridization of two different energy sources in terms of physical properties, wind and horizontal axis turbines, based on an electromechanical coupling of these two systems. The two resources are wind energy and marine energy. The concept is developed using the functional similarities of turbines and similarities in energy conversion of their energy chains. To apply this concept first, the parameters of the double fed asynchronous generator installed in the GREAH emulator are identified. Then, the power conversion chain is modeled mathematically and simulated in a MATLAB / SIMULINK environment. We have developed two control strategies.A fixed speed strategy called "Direct Speed Control", and a variable speed strategy based on the search for maximum power, called "Indirect Speed Control". Finally, this concept has been implemented practically on the real-time emulator of the laboratory. The results obtained were analyzed and discussed following this work.The second axis is devoted to a concept called "accelerated time" simulation or "virtual time". Subsequently, this concept was implemented on the multi-physics emulator available at the GREAH laboratory. This concept (accelerated time) is based on reducing wind profile samples in order to decrease simulation time and facilitate real-time control.The main results are obtained first in MATLAB / SIMULINK, then verified on the emulator in real time.The main objective of this thesis is to study the concept of offshore wind / tidal turbine hybridization based on the flexibility of a multi-function emulator that allows various emulation architectures: wind turbines, tidal turbines, and hybrid wind - tidal turbines systems. We analyze its impact on the output power of the system; the obtained results are correlated with wind and tidal speed profiles, in which statistical properties impacting global power chains could be complementary and in particular in function of the given sites. Main contributions and perspectives- Development of the concept of electromechanical coupling.When two renewable energy sources are "integrated", the rapid fluctuation of the power generated is stabilized, but under certain conditions such as the presence of storage units or an automatic clutch system.- The accelerated time conceptThis method is used to reduce the size of the recorded wind or sea current data, to speed up the simulation time of the power generation units with reasonable results that are close to actual situations.- Study and develop the concept of electric shaft regime: If the electromechanical coupling is difficult to achieve from the mechanical point of view and the single shaft decouples are too frequent so high mechanical stress, one can study the electric shaft regime with two DFIG induction machines.There is a regime in which the ratios between the speeds of the different machines are rigorously constant. The system can operate in synchronous mode with specific structures and configurations.

Les systèmes à énergies renouvelables couplés à un stockage hydrogène apportent des solutions nouvelles et innovantes à l'alimentation électrique des milieux peu ou non électrifiés. Le concept de batterie H2 qui équipe ce type de système est une forme de stockage originale qui apporte l'autonomie et l'indépendance électrique pour des longues durées (typiquement stockage saisonnier). Le fonctionnement de cette batterie H2 est le suivant : un électrolyseur produit des gaz (H2 et O2) avec les surplus d'énergie de la source renouvelable ; l'hydrogène, voire l'oxygène, est ensuite stocké dans des réservoirs pour être utilisé ultérieurement grâce à une pile à combustible lorsque la source renouvelable est insuffisante. Dans cette étude, nous nous intéresserons spécifiquement au couplage entre des générateurs photovoltaïques avec une batterie H2/O2 pour l'alimentation d'un site isolé sans interruption. Ces travaux de recherche s'inscrivent dans le projet ANR PEPITE (ANR-PanH 2007-2012) et ont été menés en partenariat avec HELION Hydrogen Power, le CEA Liten et l'Université de Corse. Le projet est également labellisé par les pôles de compétitivité CAPENERGIES et TENERRDIS. Tout d'abord, une réflexion générale s'appuyant sur les propriétés d'une batterie H2/O2 démontre la nécessité d'introduire une batterie (ici au plomb) pour garantir un fonctionnement instantané et sans interruption. Puis, une étude qualitative sur les architectures électriques possibles (bus de tension DC, AC…) a été menée pour s'achever sur une étude quantitative réalisée spécifiquement pour le projet PEPITE. Parallèlement à cela, différentes stratégies de gestions énergétiques ont été proposées afin d'utiliser les deux stockages dans les meilleures conditions, de limiter leur vieillissement ainsi que les pertes. Deux bancs d'essais à échelle réduite (un premier à bus DC et un second à bus AC) ont été réalisés au sein du laboratoire LAPLACE afin de valider les études et de préparer le prototype final qui sera testé sur le site de HELION Hydrogen Power au cours de l'été 2011. / Renewable energy systems coupled to a hydrogen storage bring new and innovative solutions to supply power to environments with little or no electricity. The concept of H2 battery which is a part of such system is a form of storage that gives autonomy and electric independence for long periods (typically seasonal storage). The operation of this H2 battery is this: an electrolyser produces gases (H2 and O2) with the extra energy from the renewable source. Hydrogen or oxygen is then stored in tanks for later use with a fuel cell when the renewable source becomes insufficient. In this study, we focus specifically on the coupling between photovoltaic arrays with a H2/O2 battery to supply power to a remote site without interruption. This work is part of the PEPITE Project, partially funded by the french National Research Agency (ANR-Panh 2007-2011) and was conducted in partnership with HELION Hydrogen Power, CEA-Liten and the University of Corsica. The project is also accredited by the CAPENERGIES and TENERRDIS clusters. First, a general discussion based on the properties of a H2/O2 battery demonstrates the need to introduce a secondary battery (lead in our case) to ensure an instant and uninterrupted operation. Then, a qualitative study on the possible electrical architectures (DC bus or AC bus) was conducted and resulted in a quantitative study conducted specifically for the PEPITE project. At the same time, various energy management strategies have been proposed to use both storage in the best conditions, limiting their losses and aging. Two small scale bench tests (one with a DC bus and a second with an AC bus) were performed in the LAPLACE laboratory to validate our strategies and prepare the final prototype which will be tested on the site of HELION Hydrogen Power during the summer of 2011.

Proteínas PUF regulam a estabilidade e a tradução através da ligação a seqüências específicas nas regiões 3\' não traduzidas (3\' UTR) dos mensageiros. A ligação é mediada por um domínio de ligação conservado constituído por 8 repetições de aproximadamente 36 aminoácidos cada. Experimentos realizados no sistema triplo-híbrido de levedura mostraram que os homólogos PUF de Arabidopsis APUM-1, APUM-2 e APUM-3 são capazes de ligar especificamente à seqüência chamada de Elemento de Resposta a NANOS (NRE) reconhecida pelo homólogo PUF de Drosophila. A utilização de bibliotecas de expressão de RNA em ensaios no sistema triplo-híbrido permitiu a identificação de seqüências de ligação consenso para as três proteínas APUM. Análises computacionais identificaram elementos de ligação a APUM em regiões 3\' UTR de importantes transcritos relacionados ao controle do meristema do caule e à manutenção das células totipotentes. Nós mostramos que os homólogos APUM-l, APUM-2 e APUM-3 reconhecem elementos de ligação a APUM nas regiões 3\' UTR dos transcritos WUSCHEL, CLAVATA-1, ZWILLE e FASCIATA-2. Ensaios de RT-PCR e Western blot semiquantitativos mostraram que a quantidade dos transcritos WUSHEL e CLAVATA-1 é alterada em plantas antisenso induzíveis para APUM-l, APUM-2 e APUM-3. A relevância biológica dessas interações foi observada através de ensaios de coimunoprecipitação, confirmando, portanto, o primeiro caso de regulação traducional descrito para os mensageiros WUSCHEL e CLAVATA-1. Análises computacionais adicionais para a identificação de outros homólogos PUF em Arabidopsis encontraram vinte e cinco proteínas possuindo repetições PUF. Entre elas, os homólogos APUM-4, APUM-S e APUM-6 apresentam alta similaridade com as proteínas APUM-l, APUM-2 e APUM-3, sendo capazes de ligar especificamente à seqüência NRE e aos elementos de ligação a APUM presentes nas regiões 3\' UTR dos transcritos WUSCHEL, CLAVATA-1, ZWILLE e FASCIATA-ts resultados indicam que vários homólogos PUF podem agir como reguladores traducionais em Arabidopsis através de um mecanismo molecular conservado entre as espécies, podendo abrir uma nova área de investigação da regulação de mRNA em plantas. / PUF proteins regulate stability and translation through sequence-specific binding to 3\' untranslated regions (UTR) oftarget mRNA transcripts. Binding is mediated by a conserved PUF domain which contains 8 repeats of approximately 36 amino acids each. Through three-hybrid assays, we have found that APUM-1, APUM-2 and APUM-3 Arabidopsis PUF homologs can bind specifically to the NANOS Response Element sequence (NRE) recognized by Drosophila PUF homologo Using Arabidopsis RNA libraries in three-hybrid screenings, we were able to identify APUM binding consensus sequences. A computational analysis allowed us to identify the APUM binding element within the 3\' UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot and stem cell maintenance. We show that APUM-1, APUM-2 and APUM-3 are able to bind specifically to APUM binding elements in the 3\' UTR of WUSCHEL, CLAVATA-1, ZWILLE and FASCIATA-2 transcripts. RT-PCR and Western blot semiquantitatives assays showed altered WUSHEL and CLAVATA-1 amounts in APUM-1, APUM-2 and APUM-3 antisense plants. The biologic relevance of these interactions was observed with co-immunoprecipitation assay, which confirmed the first example of translational regulation to WUSCHEL and CLA VATA-1 transcripts. Computational analysis to identify others PUF homologs in Arabidopsis found twenty five proteins presenting PUF repeats. Among them, we found that APUM-4, APUM-S and APUM-6 homologs are very similar to APUM-1, APUM2 and APUM-3 and also are able to bind specifically to the NRE sequence and to APUM binding elements in the 3\' UTR of WUSCHEL, CLAVATA-1, ZWILLE and FASCIATA-2 transcripts. Our results indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach to investigate mRNA regulation in plants.