Myxococcus xanthus, a Gram-negative bacterium with a developmental cycle, displays a type IV pili (TFP) mediated surface motility known as social (S) gliding. Beside the polarly localized TFP, the fibril or extracellular polysaccharide (EPS) is also required for S-motility to function. It is proposed that S-motility, along with the related bacterial twitching motility in other species, is powered by TFP retraction. EPS is proposed to anchor and trigger such retractions in M. xanthus. EPS production is known to be regulated by TFP and the Dif signal transduction pathway. Two genetic screens were performed previously to identify additional genes important for EPS production. The first was for the isolation of pilA suppressors, the second for the identification of mutants underproducing EPS in a difA suppressor background. Both screens identified transposon insertions at the stk locus. In particular, StkA, a DnaK homolog, was identified as a possible negative regulator of EPS production by a stkA transposon insertion that suppressed a pilA mutation. A stkB transposon insertion was found to have diminished EPS production in a difA suppressor background. In this study, in-frame deletion mutants of the five genes at the stk locus, stkY, stkZ, stkA, stkB and stkC, were constructed and examined. In addition, mutations of rbp and bskL, two genes downstream of the stk locus, were constructed. Like transposon insertions, the stkA in-frame deletion resulted in overproduction of EPS. The stkB and to a less extent the stkC mutants underproduced EPS. Mutations in the other genes had no obvious effects on EPS production. Genetic epistasis suggests that StkA functions downstream of TFP and upstream of the Dif sensory proteins in EPS regulation in M. xanthus. Epistasis analysis involving stkB was inconclusive. It is unresolved whether StkB plays a role in the biosynthesis or the regulation of EPS production in M. xanthus. / Master of Science

Dans l'étude des éléments combustibles des réacteurs à eau pressurisée, cette thèse s'inscrit dans la compréhension et la modélisation du comportement viscoplastique du dioxyde d'uranium (UO2) à l'échelle du polycristal. Lors de fonctionnement de type incidentel du réacteur, le combustible subit une forte élévation de la température avec un gradient thermique de la pastille engendrant des déformations viscoplastiques contrôlées par des mouvements de dislocations. D'abord, un modèle de plasticité cristalline a été développé de manière à décrire l’anisotropie viscoplastique du matériau en fonction de la température et de la vitesse de sollicitation. Des simulations par éléments finis (EF) sur monocristaux ont permis d’identifier que les trois modes de glissement généralement observés dans l'UO2 sont importants pour décrire le comportement anisotrope du matériau. Dans un second temps, les coefficients de la matrice d'interactions entre dislocations ont été déterminés spécifiquement pour l’UO2 afin d’améliorer la modélisation des polycristaux. En effet, en calculant par EF les dislocations géométriquement nécessaires, qui sont responsables d’une forte augmentation de la densité de dislocations stockées dans les polycristaux, les interactions entre dislocations permettent de simuler l’effet dé taille de grain et l’écrouissage des pastilles. Finalement, le modèle, adapté pour les polycristaux, a été validé par comparaison avec les essais expérimentaux sur pastille et par comparaison du comportement intra-granulaire simulé avec des mesures EBSD. Grâce à cette dernière comparaison, il est possible de remonter indirectement aux hétérogénéités de déformation dans les grains / This thesis is part of the study of fuel elements of pressurized water reactors and, more specifically, focus on the understanding and modelling of the viscoplastic behavior of uranium dioxide (UO$_2$) at polycrystalline scale. During the incidental operation of the reactor, the fuel undergoes a strong increase of temperature and thermal gradient between the center and the periphery of the pellet leading to viscoplastic strains due to dislocation movement mechanisms. First, a crystal plasticity model was developed in order to describe the viscoplastic anisotropy of the material considering the temperature and the loading rate. Finite element (FE) simulations on single crystals enabled to highlight that the three slip modes generally observed in UO$_2$ are crucial to describe the anisotropic behavior of the material. Secondly, coefficients of the interaction matrix have been identified specifically for UO$_2$ in order to improve the polycrystal modelling. Indeed, by calculating geometrically necessary dislocations (GNDs), which are responsible of the great increase of the stored dislocation density in polycrystals, the interactions between dislocations enable to simulate de grain size sensitivity and hardening of the fuel pellet. Finally, the model adapted for polycrystals, have been validated by comparing FE simulations with pellet compression tests and by comparing the simulated intra-granular behavior with EBSD measurements. Thanks to the latter comparison, it is possible to indirectly compare the strain heterogeneities in the grains

Baigiamajame magistro darbe apžvelgiamos sklandytuvų motorizavimo tendencijos, būdai bei reikiama teorinė medžiaga. Nagrinėjamas papildomas pasipriešinimo padidėjimas dėl motorizavimo elektros varikliu, tiriamos būsimos motorizuoto sklandytuvo LAK-12 tiesiaeigio skridimo bei kilimo charakteristikos. Darbo gale pateikiami gauti rezultatai bei suformuluojamos išvados. / The theses provides an overview of the engine types used for motorizing gliders, tendency and theoretical material. Additional drag due to engine and propeller blades were studied as well as straight and level flight and climb performance. The results and conclusions are given at the end.

In this thesis I deal with the project of a robotic apparatus that could be used for material treatment by using the gliding arc discharge as well as the appropriate design and choice of materials for the atmospheric pressure discharge jet with regards to a possibility of an industrial use. The input and output parameters were studied, such as the air flow regulation with regards to cooling, the design and choice of materials. Then I observed the temperatures near electrodes depending on the air flow. The applicability of the created atmospheric discharge was taken into account.

Lors de la prospection de bactéries d’intérêt dans le milieu marin, nous avons isolé des colonies aux couleurs brillantes sous illumination directe, ressemblant à l’iridescence de certains papillons. L’iridescence est une couleur physique induite par des structures nanoscopiques périodiques. Ce phénomène est peu détaillé chez les procaryotes. Dans cette thèse nous avons souhaité caractériser le phénomène par des analyses physico-optiques,microbiologiques et écologiques. La souche fut identifiée comme appartenant à l’espèce Cellulophaga lytica.Les premières étapes ont permis d’illustrer le phénomène et de prouver une coloration structurale des colonies par microspectrophotométrie. Une comparaison de diverses souches a démontré l’iridescence singulière de C. lytica,dénommée iridescence « glitter-like ». Nous avons ainsi proposé la première classification des iridescences bactériennes. Certaines souches du même phylum Bacteroidetes exprimaient le phénomène. Un lien entre cette iridescence et la motilité « gliding » fut mis en évidence. De façon intéressante, l’iridescence était conservée en conditions mimant le biotope C. lytica. De plus, diverses souches marines iridescentes ont pu être ré-isolées. Elles étaient particulièrement abondantes sur les macro algues (rouges, brunes), mollusques, cnidaires et dans l’eau de mer. Nous supposons alors que ce phénomène pourrait avoir des rôles bio-écologiques. Grâce à la microscopie électronique à transmission et une modélisation, nous avons pour la première fois élucidé des structures reponsables d’une iridescence bactérienne : une organisation populationnelle de cellules alignées équidistantes.Les bases fondamentales et méthodologiques de l’iridescence bactérienne sont données dans cette thèse. Les applications correspondraient aux domaines des nanotechnologies et de la biomimétique. / During collections in the marine environment, colony with bright coloration under direct illumination was isolated. Colors were similar to butterly iridescence. Iridescence is a property of structural color where periodic nanoscopic structures are responsible of intense coloration. Bacterial iridescence is poorly described in the literature. In this work, we aimed to characterize the phenomenon by physico-optical, microbiological and ecological analysis. The marine bacterium was identified as Cellulophaga lytica.In the first experiment we illustrated and proved the physical color by micro spectrophotometry. Comparison with diverse bacteria permitted to demonstrate the uniqueness of C. lytica iridescence, named “glitter-like”. We proposed the first classification of iridescences within the prokaryotes. Others Bacteroidetes strains showed “glitterlike”iridescence effects. A link between iridescence and gliding motility was additionally identified. A biotic factors demonstrated conservation of the phenomenon under conditions that mimic C. lytica stressing biotope. Moreover,other iridescent marine strains were isolated. They were particularly associated to macroalgae (red, brown),mollusks, cnidarians and seawater. Biological and ecological functions of iridescence are then supposed. By using transmission electron microscopy and modeling, structures responsible for iridescence in the colony were first identified: alligned and equidistant cells at a population level.Bacterial iridescence’s fundamental and technical skills are described in this thesis. Applications could lead to biomimetic and nanotechnology of iridescent structures.

The diploma thesis is focused on the study of chemical reaction products in prebiotic atmospheres. The theoretical part deals with the main theories of composing the atmosphere of the early Earth, hypotheses of the origin of life and genetic code in the form of RNA, the atmosphere of the moon Titan and the basic characteristics of the gliding arc discharge, and the analytical techniques used in the diploma thesis. In the experimental part, study of products was carried out by experimental atmospheric simulations in a special stainless steel reactor in various mixtures of gases N2, CH4, CO2 and O2. The electrical discharge for atmospheric simulation was generated by an electrode configuration for the gliding arc discharge and glow discharge. Detection and identification of chemical reaction products was performed by Gas Chromatography with Mass Spectrometry (GC-MS) and Flame Ionization Detector (GC-FID), Testo 350 M/XL and Proton Transfer Reaction Mass Spectrometry (PTR-MS). A theoretical kinetic model was created to predict and confirm the detected products. During the experiments, the dependence of the product intensity on changes in the composition of the gaseous mixture and the applied value of the electric current was also observed.

Diploma thesis deals with gliding arc discharge in non-equilibrium plasma, its properties and usage for dissociation of volatile organic compounds. Plasma techniques create a perspective alternative to classical methods such as adsorption, biofilters, thermal processes, freezing and condensation. The method used for analysis of dissociation products is described in the theoretical part as well. Method is gas chromatography and it is combined with mass spectroscopy. The experimental part contains the overall description of GlidArc reactor where volatile organic compounds were dissociated. The volatile organic compound is brought into the reactor from the reservoir by carrier gas (nitrogen); synthetic air was used as working gas. The reactor is connected to the device for sampling. This device has several openings for sampling by SPME fibre method, sorption tubes method and a special opening for probe entry from the device Testo 350 M/XL, which enables instant analysis of low-molecular compounds. GC-MS was used for determination of high-molecular products of dissociation. Products sorbed on SPME fibres were analysed directly, compounds sorbed by active carbon were extracted by carbon disulphide. Samples obtained using SPME fibres were analysed. The major products of the dissociaton were found due to this analysis, those major products are in the case of hexane: pentanal, 4 methyl-3-pentanal, 2-butoxy-ethanol, pentane a 2-hexene. Major products detected when dissociating cyclohexane were: l,3-dimethyl-butane, propanal, cyklohexanone, 5-hexenal a 2-pentyn-1-ol. Major products of xylene dissociation were methyl-benzene, benzaldehyde, 4-methyl-benzaldehyde, 1-nitroethyl-benzene a benzenmethanol. After analysis obtained using sorption tubes showed that mainly the dissociated compound was sorbed, but the products of the dissociation were presented in too low concentration for an adequate analysis. For that reason the sorption tubes were used only for quantitative determination of the compounds depending on the power supply. The dependence of the dissociation of the compounds on the power supply was observed using SPME fibres which were used for quantitative determination. The Results obtained by both obtained techniques were almost in accordance. Speaking of low-molecular compounds, dissociation of all compounds gave the same products, i.e. carbon dioxide, hydrogen, nitrogen oxide, nitrogen dioxide and water. The results show that the dissociation of VOC gives - besides many potentially dangerous dissociation products - significant amounts of nitrogen oxides which is harmful to the environment. Hence it is necessary to be concerned with problems studied in the future mainly with respect to limitation of NOx generation. It will be possible to optimalize the conditions of the VOC dissociation on the basis of future kinetic analysis.

The surface gliding motility of Myxococcus xanthus is required for a multicellular developmental process initiated by unfavorable growth conditions. One form of the M. xanthus surface motility, social (S) gliding, is mediated by the extension and retraction of polarly localized type IV pili (Tfp). Besides Tfp, exopolysaccharides (EPS), another cell surface associated component, are also required for M. xanthus S motility. Previous studies demonstrated that the Dif chemotaxis-like signal transduction pathway is central to the regulation of EPS production in M. xanthus. Specifically, difA, difC and difE mutants were found to be defective in EPS production and S motility. DifA, DifC and DifE, homologous to methyl-accepting chemotaxis proteins (MCPs), CheW and CheA, respectively, are therefore positive regulators of EPS. This study, undertaken to better understand the regulation of EPS production, led to a few major findings. First, DifD and DifG, homologous to CheY and CheC, respectively, were found to be negative regulators of EPS production. Both DifD and DifG likely function upstream of the DifE kinase in EPS regulation. DifB, which has no homology to known chemotaxis proteins, was found not to be involved in EPS production. Secondly, this study led to the recognition that Tfp likely function upstream of the Dif pathway in the regulation of EPS production. Extracellular complementation experiments suggest that Tfp may act as sensors instead of signals for the Dif chemotaxis-like pathway. We propose a regulatory feedback loop that couples EPS production with Tfp function through the Dif signaling proteins. Lastly, we sought to identify additional genes involved in EPS production. Our efforts identified a mutation in a separate chemotaxis gene cluster as a suppressor of difA mutations, suggesting potential cross-talks among the multiple chemotaxis-like pathways in M. xanthus. In addition, we identified twenty-five previously uncharacterized genes that are predicted to be involved in M. xanthus EPS production. These genes appear to encode additional EPS regulators and proteins with biosynthetic function. / Ph. D.

The Myxococcus xanthus Dif and Frz chemosensory pathways play important roles in the regulation of gliding motility. The Dif system regulates the production of exopolysaccheride (EPS), which is essential for social motility and fruiting body formation. The Frz pathway controls reversal frequency, which is fundamental for directed movement by this surface-gliding bacterium. In addition, both pathways are involved in the chemotactic response towards several phosphatidylethanolamine (PE) species such that the Dif pathway is required for excitation while the Frz pathway is essential for adaptation. In this study we addressed three crucial questions regarding the signal processing of these two chemosensory pathways by focusing on DifA and FrzCD, the MCP homologs from their respective pathways. First, the receptor protein in the Dif pathway, DifA, lacks a perisplasmic domain, the typical signal-sensing structure. To examine whether DifA shares similar transmembrane signaling mechanism with typical transmembrane sensor proteins (MCPs and sensor kinases), we constructed a chimeric protein that is composed of the N-terminus of NarX (nitrate sensor kinase) and the C-terminus of DifA. This NarX-DifA chimera restores the DifA functionality (EPS production, agglutination, S-motility and development) to a "difA mutant in a nitrate-dependent manner, suggesting DifA shares a similar transmembrane signaling mechanism with typical MCPs and sensor kinases despite its unorthodox structure. Second, the M. xanthus chemotaxis is still controversial. It has been argued that the taxis-like response in this slowly gliding bacterium could result from physiological effects of certain chemicals. To study motility regulation by the Frz pathway, we constructed two chimeras between the N-terminus of NarX and C-terminus of FrzCD, which is the receptor protein of the Frz pathway. The two chimeras, NazDF and NazDR, are identical except that NazDR contains a G51R mutation in the otherwise wild-type NarX sensory module. This G51R mutation was shown to reverse the signaling output of a NarX-Tar chimera to nitrate. We discovered that nitrate specifically decreased the reversal frequency of NazDF-expressing cells and increased that of NazDR-expressing cells. These results show that directional motility in M. xanthus can be regulated independently of cellular metabolism and physiology. Surprisingly, the NazDR strain failed to adapt to nitrate in temporal assays, as did the wild type to known repellents. Therefore, the lack of temporal adaptation to negative stimuli is an intrinsic property in M. xanthus motility regulation. Third, the Dif and Frz pathways are both involved in the chemotactic response towards certain PE molecules such that the Dif pathway is required for excitation and while the Frz system is essential for adaptation. In addition, 12:0 PE, known to be sensed by DifA, results in increased FrzCD methylation. These findings suggested that in the regulation of PE response, two pathways communicate with each other to mediate adaptation. Here we provided evidence to indicate that DifA does not undergo methylation during EPS regulation and PE chemotaxis. On the other hand, using mutants expressing the NarX-DifA chimera, it was found that signal transduction through DifA, DifC (CheW-like) and DifE (CheA-like) modulates FrzCD methylation. Surprisingly, the attractant 12:0 PE can modulate FrzCD methylation in two ways distinguishable by the dependency on DifA, DifC and DifE. The DifACE-independent mechanism, which may result from specific sensing of 12:0 PE by FrzCD, increases FrzCD methylation as expected. Unexpectedly, 12:0 PE decreases FrzCD methylation with the DifACE-dependent mechanism. This "opposite" FrzCD methylation by DifACE-dependent signaling was supported by results from NafA-expressing mutants because nitrate, which acts as a repellent, increases FrzCD methylation. Based on these findings, we proposed a model for chemotaxis toward 12:0 PE (and 16:1 PE). In this model, DifA and FrzCD both sense the same signal and activate the pathways of excitation (Dif) and adaptation (Frz) independently. The two pathways communicate with each other via methylation crosstalk between DifACE and FrzCD in such a way that processes of excitation and adaptation can be coordinated. / Ph. D.

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