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La régulation de la photosynthèse est cruciale pour les organismes photoautotrophes et est habituellement opérée par la modulation de l'absorption de la lumière ou par la réorientation des électrons vers des puits alternatifs afin de redistribuer l'énergie entre plusieurs voies métaboliques. Parmi les différents mécanismes décrits, le remodelage de l'appareil photosynthétique est crucial dans des conditions de carences nutritives ou de fluctuations de la lumière. Il est bien connu que l'oxyde nitrique (NO) joue un rôle de signalisation dans de nombreuses réponses au stress abiotique, agissant comme second messager et / ou modifiant les protéines cibles par des modifications post-traductionnelles redox. Sa participation a été récemment décrite au cours de la carence en azote chez Chlamydomonas reinhardtii. Ce travail se concentre sur le remodelage de l'appareil photosynthétique lors de la carence en soufre et lors des fluctuations de lumineuses chez Chlamydomonas reinhardtii, avec un intérêt particulier pour la voie de signalisation impliquée dans ces réponses. Tout d'abord, nous avons caractérisé la carence en soufre en conditions d’hétérotrophie ou de photo-autotrophie. En faible lumière ou à l’obscurité, l'inactivation photosynthétique est obtenue grâce à la dégradation spécifique de la Rubisco et du cytochrome b6f et ne se produit qu'en présence de carbone réduit dans le milieu. Nous avons également montré une forte production de NO après le début de la carence, avec des sondes fluorescentes sensibles au NO visualisées par microscopie confocale. Nous fournissons des preuves pharmacologiques que la production de NO intracellulaire régit cette voie de dégradation. En outre, ici, nous fournissons des preuves claires de l’existence d’un circuit régulateur qui contrôle la traduction cytosolique du LHCII en réponse à des changements de quantité de lumière. Ce circuit nécessite la protéine de liaison à l'ARN cytosolique NAB1 pour réprimer la traduction de certains ARNm de LHCII. La nitrosylation spécifique de la Cys-226 diminue l'activité de NAB1 et a été démontrée in vitro et in vivo. La forme moins active et nitrosylée de NAB1 se trouve dans les cellules acclimatées à un apport de lumière limité, ce qui permet l'accumulation de protéines des antennes et la capture efficace de la lumière. En revanche, une intensité lumineuse plus élevée provoque la dénitrosylation de NAB1, activant ainsi la répression de la synthèse des protéines LHCII et diminuant ainsi la pression de la lumière au niveau du PSII. La dénitrosylation de NAB1 est efficacement réalisée par le système thiorédoxine cytosolique in vitro. À notre connaissance, NAB1 est le premier exemple de dénitrosylation induite par un stimulus dans le contexte de l'acclimatation photosynthétique. Dans l’ensemble, nos données suggèrent un rôle pivot pour la signalisation NO dans le contrôle des réponses au stress environnemental. / The regulation of photosynthesis is crucial for photoautotrophic organisms and is usually operated by the modulation of light absorption or by redirection of electrons towards alternative sinks, in order to redistribute energy among several metabolic pathways. Between different mechanisms described, the remodeling of the photosynthetic apparatus is crucial under conditions of nutrient starvation or light fluctuations. It is well known that nitric oxide (NO) plays a signaling role in many abiotic stress responses, acting as a second messenger and/or modifying target proteins through redox post translational modifications. Its involvement has been recently described during nitrogen starvation in Chlamydomonas reinhardtii. This work focuses on the remodeling of the photosynthetic apparatus upon sulfur starvation and light fluctuations in Chlamydomonas reinhardtii, with particular interest for the signaling pathway involved in the responses. First we characterized sulfur starvation under heterotrophy and photo-autotrophy. Photosynthetic inactivation under low light and darkness is achieved through specific degradation of Rubisco and cytochrome b₆f and occurs only in the presence of reduced carbon in the medium. We have also shown a strong NO production after the onset of starvation, with NO-sensitive fluorescence probes visualized by confocal microscopy. We provide pharmacological evidence that intracellular NO production governs this degradation pathway using NO scavengers, NO synthesis inhibitors and NO donors. Furthermore, here, we provide clear evidence for a regulatory circuit that controls cytosolic LHCII translation in response to light quantity changes. This circuit requires the cytosolic RNA-binding protein NAB1 to repress translation of certain LHCII mRNAs. Specific nitrosylation of Cys-226 decreases NAB1 activity and could be demonstrated in vitro and in vivo. The less active, nitrosylated form of NAB1 is found in cells acclimated to limiting light supply, which permits accumulation of light harvesting proteins and efficient light capture. In contrast, elevated light supply causes NAB1 denitrosylation, thereby activating the repression of light-harvesting protein synthesis and decreasing the light pressure at the level of PSII. Denitrosylation of NAB1 is efficiently performed by the cytosolic thioredoxin system in vitro. To our knowledge, NAB1 is the first example of stimulus-induced denitrosylation in the context of photosynthetic acclimation. Taken together, our data suggest a pivotal role for NO-signaling in the control of environmental stress responses.

The dissertation thesis deals with an experimental study of an elastohydrodynamic contact under insufficient lubricant supply. Theoretical studies published in this research area focus mainly on the development of theoretical models and there is an insufficient number of experimental studies which validate these models. There are two basic methods of starvation severity control in published theoretical models: by positioning of an inlet meniscus and by setting the thickness of an oil layer in the vicinity of contact inlet. The second method is more suitable when considering a wide range of operating conditions, however it is difficult to ensure experimentally. This thesis presents original experimental results and aims to describe the starvation severity level as a function of the inlet film thickness. Deeper understanding of fundamental processes in starved elastohydrodynamic contacts enables the development of more precise models, better ability of prediction of the starvation and improves the ability to avoid starvation in the early design process of key tribological components.

This master’s thesis deals with the experimental study of an elastohydrodynamic contact under conditions of insufficient lubricant supply. Starvation level of this type of the contact may be experimentally determined based on the input position of the meniscus, but this way can’t determine all its levels of starvation. Consequent development in the field of tribology achieved theoretical model that can determine at all levels of starvation depending on the thickness of the lubricant film entering the contact, but it is relatively difficult for experimental verification. The main goal of this work is an experimental study and describes the behavior of elastohydrodynamic contact at an exactly defined thickness of the lubricant film at the contact input. The benefit of this work is a validation of that mentioned theoretical model for base oils and greases made from them, and mutual comparison of their behavior.

The liver is the central metabolic hub in organisms and a complex, intertwining regulatory network guarantees efficient liver processes. The morphogenic Hedgehog pathway was recently shown to play a role in regulating the underlying genetic program. Transgenic mouse models with hepatocyte-specific inactivation of Hedgehog signaling showed alterations in insulin-like growth factor homeostasis and in energy metabolism associated with increased lipid accumulation in the liver. In this thesis, it was possible to connect the observed infertility of female knockout mice with an unexpected activation of sex steroid synthesis in the liver. Associated with increased steroidogenic gene expression exclusively in hepatocytes, the plasma testosterone level was significantly elevated, which led to androgenization and an anovulatory phenotype. With these characteristics, the mouse model mimicked the human polycystic ovarian syndrome and suggested an influence of liver and hepatic Hedgehog signaling on reproduction under disease conditions. Further, murine liver metabolism was challenged with starvation starting at different times of day. The transcriptomic results were analyzed with a self-organizing map approach, allowing an intuitive interpretation of data and a thus far unknown diurnally different response of hepatic regulatory mechanisms due to starvation was revealed. In contrast to the manifoldly published and observed switch from energy-consuming to energy-providing processes due to starvation started in the morning, evening starvation led to a novel hepatic expression signature with decreased gluconeogenic gene expression and increased levels of lipid and steroid metabolism-related genes. These differences can be explained by the equally diurnally regulated expression of the corresponding regulatory transcription factors and hormones. Additionally, lipidome analysis confirmed the diurnal differences after starvation. Thus, this study emphasized the immense impact of circadian regulation on liver metabolism and suggests high accuracy when starvation is the focus of research to avoid varying results.:BIBLIOGRAPHISCHE DARSTELLUNG ................................................................................ II LIST OF ABBREVIATIONS .................................................................................................. III TABLE OF CONTENTS ....................................................................................................... IV SUMMARY ............................................................................................................................ 1 ZUSAMMENFASSUNG ......................................................................................................... 5 INTRODUCTION ................................................................................................................... 9 Liver architecture and metabolism ..................................................................................... 9 Diverse possibilities of liver metabolism regulation .......................................................... 10 Connection of Hedgehog signaling to hepatic metabolism ............................................... 10 Impact of feeding schemes on hepatic metabolism .......................................................... 13 Aims of the thesis ............................................................................................................ 14 References ...................................................................................................................... 15 CHAPTER 1 ........................................................................................................................ 18 CHAPTER 2 ........................................................................................................................ 39 PERSPECTIVE ................................................................................................................... 64 CURRICULUM VITAE ........................................................................................................... V PUBLICATIONS AND PRESENTATIONS ............................................................................ VI Publications ...................................................................................................................... VI Oral presentations ............................................................................................................ VI Poster presentations ........................................................................................................ VII AUTHOR CONTRIBUTION STATEMENT .......................................................................... VIII SELBSTSTÄNDIGKEITSERKLÄRUNG .............................................................................. XII DANKSAGUNG .................................................................................................................. XIII

In order to meet their energy budget animals must often increase their risk of predation, either through their choice of foraging location or by decreasing anti-predation behaviours, which are incompatible with foraging. I investigated the starvation-predation risk trade-off in redshanks overwintering in the area of the Firth of Forth in Scotland over different spatial scales. On a small spatial scale, where redshanks foraged in an area where risk of attack was high I investigated the role of competition for food and decreased individual vigilance within groups and how this related to predation risk, additionally I looked at the relative roles of individual variation in time spent exposed to risk and variation in anti-predation behaviours on individual survival time. On larger spatial scales of 100s of meters and over several kilometres, I considered how choice of overwintering site was affected by predation risk, profitability and population density. Time available to feed increased with increased group size, allowing redshanks to compensate for increased competition and allowing large groups to form, thus decreasing individual predation risk. Individuals that spent less time exposed to attacking predators survived for longer, however individuals constrained by cold weather to spend long periods exposed to risk could increase their survival through increased intake rates and vigilance. On an intermediate spatial scale redshanks selected overwintering sites based on profitability rather than risk, and only used less profitable site when population density was high. On a large spatial scale redshanks increased their use of less profitable sites in warmer weather, but did this without increasing their risk of predation. Overall this suggests that across most spatial scales redshanks can minimise their predation risk by their choice of foraging location, but when forced by weather conditions or competition to be exposed to attack, capture reducing behaviours also reduce predation risk.

Obesity results from an excess of adipose tissue and is a major risk factor for type 2 diabetes, cardiovascular disease, and cancer. Adipose tissue exists in two main forms: white adipose tissue (WAT), which stores energy as triglycerides, and brown adipose tissue (BAT), which dissipates stored energy as heat. White adipose tissue is composed of several subcutaneous and visceral depots, each possessing distinct molecular and functional characteristics. Brown-like adipocytes can emerge in WAT depots in response to cold or beta-adrenergic stimulation. These cells have been called “beige” or “brite” (brown-in-white) cells. The reduction of obesity in mice treated with beta-adrenergic agonists is correlated with the emergence of beige cells. Beige cell development occurs most readily in subcutaneous depots, and to the least extent in visceral depots. Understanding the molecular mechanisms underlying beige cell development in different WAT depots may be important in discovering new therapies against obesity and related diseases. Our lab has previously discovered that Tyrosine Kinase 2 (Tyk2), an important mediator of cytokine signaling, promotes the development of classical brown adipose tissue. Due to the lack of functional BAT, Tyk2-knockout (Tyk2-/-) mice become grossly obese with age and develop several symptoms of the metabolic syndrome. In the present study, we have found a potential role of Tyk2 in the development of beige cells. Here, we show that mRNA expression of BAT-selective genes (UCP1, Cidea, Cox8b, and Elovl3) is significantly reduced in subcutaneous WAT of Tyk2-knockout (Tyk2-/-) mice compared to wild-type mice. Surprisingly, BAT-selective genes are induced in Tyk2-/- subcutaneous WAT by acute starvation. These findings suggest that Tyk2 is required for the development of beige cells under ambient conditions, and that the need for Tyk2 in beige cell development is bypassed during nutritional stress, a stimulus of the sympathetic response.

La photosynthèse oxygénique peut être le lieu de formation des Formes Réactives de l’Oxygéne (FROs). Les FROs altèrent toutes les macromolécules de la cellule, générant ainsi un stress oxydant. Toute perturbation du métabolisme cellulaire peut conduire à ce type de stress. Les cyanobactérie sétant les premiers organismes à avoir émis de l’oxygène sur terre, elles ont du développer très tôt au cours de l’évolution des mécanismes de perception et de défense pour lutter contre ce stress. Nous nous sommes intéressés à l’étude des mécanismes qui permettent à la cyanobactérie filamentuse et diazotrophe Anabaena PCC 7120 de s’adapter à diverses conditions de stress et de carence : stress oxydant, carence en fer et en azote combiné. En réponse à une carence en azote combiné, elle différencie en 24 h des hétérocystes : cellules spécialisées dans la fixation de l’azote atmosphérique. Nous avons étudié la réponse transcriptomique globale de cette bactérie à la fois au stress oxydant et à la carence en fer et nous avons établit la connection existant entre ces deux stress. Nous avons pu identifier le régulateur transcriptionnel pleiotrope impliqué dans la perception et la signalisation du stress peroxyde et nous en avons élucidé le mécanisme d’action. Nous avons également étudié une Ser/Thr kinase qui joue un rôle important à la fois dans la réponse au stress oxydant et à la carence en azote combiné. Notre étude a montré que cette kinase pourrait être le lien moléculaire entre ces deux conditions, puisque une cible potentielle de cette kinase semble être la protéine HetR qui est le régulateur clé du processus de différenciation cellulaire. / Oxygenic photosynthesis may generate Reactive forms of Oxygéne (ROS). These reactive oxygen species can damage all the macromolecules of the cell, inducing oxidative stress. Any disruption of cellular metabolism can lead to oxidant damage. Cyanobacteria were the first organisms producing oxygen on Earth, they therefore had to develop very early during evolution the mechanisms of perception and defence to cope with this tstress. We are interested in studying the mechanisms that allow the diazotrophic filamentous cyanobacterium Anabaena PCC 7120 to adapt to various conditions of stress and stravations: oxidative stress, iron and combined nitrogen starvations. Anabaena PCC 7120 is a simple model for the study of cell differentiation. In response to combined nitrogen stravation it can differentiate heterocysts, cell specialized in molecular nitrogen fixation. We studied the global transcriptomic response of this bacterium to both oxidative stress and iron deficiency and we establish the crosstalk between these two stresses. We were able to identify the global transcriptional regulator involved in the perception and in the signaling of a peroxide stress. Its mechanism of action was elucidated. We also studied a Ser / Thr kinase that plays an important role both in the response to oxidative stress and combined nitrogen stravation. Our study showed that this kinase may be the molecular link between these two conditions, as a potential target of this kinase appears to be the HetR protein which is the key regulator of cellular differentiation process.

La grande diversité des applications, dans tous les domaines industriels, des plus simples aux plus complexes, que ce soit pour un usage quotidienne ou pour des applications high-tech, nécessite la conception continue de systèmes d'étanchéité de plus en plus performants. Les vérins pneumatiques sont des éléments fondamentaux de l'automatisation dans des domaines industriels très variés : les dispositifs de levage, la robotique, l'aérospatial, les systèmes respiratoire, etc. Cependant, un dysfonctionnement d'étanchéité conduit à des performances plus faibles et même à des disfonctionnements. Inversement, une meilleure connaissance des phénomènes aux interfaces, permettra l’amélioration du rendement, la prolongation de la durée de vie et de la fiabilité des différentes applications pneumatiques.Ce travail de thèse est focalisé sur l'étude expérimentale et numérique des systèmes d'étanchéité par joints pneumatiques. Un banc d'essais modulable a été conçu et réalisé pour permettre l'investigation expérimentale de différents types de joints pneumatiques, pour une large gamme de conditions de fonctionnement, en termes de vitesse linéaire, pression pneumatique et solutions constructives. Il permet principalement de caractériser le frottement des joints mais aussi la visualisation des phénomènes physiques au niveau de la zone d'étanchéité et de son environnement.En parallèle, un modèle théorique ElastoHydroDynamique, incluant la prise en compte des conditions de lubrification mixte et le comportement rhéologique non-newtonien de lubrifiant (graisse) a été développé. Il est basé sur le couplage d'un logiciel de calcul non-linéaire (permettant la prise en compte du comportement mécanique hyperélastique des joints) et un modèle d'écoulement en film mince. Les comparaisons de mesures expérimentales avec les prédictions numériques ont permis de valider le modèle théorique ainsi que d'améliorer la compréhension des conditions de fonctionnement et d'alimentation de la zone d'étanchéité. / The great diversity of applications in all industrial fields, from the simplest to the most complex, whether for everyday use or for high-tech applications, requires the continuous design of increasingly efficient sealing systems. Pneumatic cylinders are fundamental elements of automation in a wide variety of industrial fields: lifting devices, robotics, aerospace, respiratory systems, etc. However, a sealing failure leads to lower performance and even malfunctions. Conversely, a better knowledge of the phenomena at the interfaces will allow the improvement of efficiency, the extension of the service life and of the reliability of the various pneumatic applications.This thesis work focuses on the experimental and numerical study of pneumatic seal systems. A modular test bench has been designed and built to allow the experimental investigation of different types of pneumatic seals, for a wide range of operating conditions, in terms of linear velocity, pneumatic pressure and constructive solutions. It mainly allows to characterize the friction of the joints but also the visualization of physical phenomena at the level of the sealing zone and its environment.In parallel, an ElastoHydroDynamic theoretical model, including the consideration of mixed lubrication conditions and the non-Newtonian rheological behaviour of lubricant (grease) has been developed. It is based on the coupling of a non-linear comercial software (allowing the modelling of the hyperelastic mechanical behaviour of the joints) and a thin film flow model. Comparisons of experimental measurements with numerical predictions validated the theoretical model and improved the understanding of the operating and supply conditions of the sealing area.

Fatores sigma de carência de ferro, representados por FecI de E. coli, direcionam a transcrição de genes de transporte de sideróforos (quelantes de ferro), e são geralmente regulados por fatores antissigma (FecR), que liberam o fator sigma após ligação do sideróforo no receptor de membrana externa (FecA). Caulobacter crescentus possui quatro genes para fatores sigma desta família. Ensaios de expressão gênica e crescimento indicaram que estes genes não respondem à disponibilidade de ferro. Em microarranjos de cDNA, apenas o gene fecA2 foi induzido em DfecR2 comparado à linhagem parental, sugerindo que este é o único gene alvo do fator sigma FecI2. Já DfecR4 mostrou indução em mais de 50 genes, alguns envolvidos na utilização de fontes alternativas de carbono. Ensaios fenotípicos com DfecI4 sugeriram que este gene é importante para o crescimento em g-ciclodextrina ou ácido caproico. Os resultados sugerem que o fator sigma FecI2 é bem específico, enquanto FecI4 parece regular uma resposta geral relacionada a compostos carbônicos, e não à homeostase de ferro. / Iron starvation sigma factors, whose prototype is E. coli FecI, direct transcription of genes involved in siderophore (iron chelators) transport, being usually regulated by anti-sigma factors (FecR), which release the sigma factor after siderophore binding to the outer membrane receptor (FecA). Caulobacter crescentus possesses four genes encoding FecI-like sigma factors. Gene expression and growth assays indicated that these genes do not respond to iron availability. In cDNA microarrays, only the fecA2 gene was induced in DfecR2 relative to the wild-type strain, suggesting that this is the only target gene of the FecI2 sigma factor. However, in DfecR4 there was induction of over 50 genes, some of them involved in utilization of alternative carbon sources. Phenotypic microarrays with the DfecI4 strain showed that this gene is important for growth in g-cyclodextrin or caproic acid. The results suggest that the FecI2 sigma factor is very specific, whereas FecI4 seems to regulate a more general response, related to carbon compounds rather than iron homeostasis.