L'épissage alternatif des ARNs pré-messagers est le mécanisme majeur de diversification de l'information génétique chez les eucaryotes supérieurs. Près de 70% des gènes sont concernés par ce mécanisme. Au cours de la différenciation érythroïde, l'inclusion de l'exon 16 du pré-messager 4.1R représente un événement majeur pour la fonction érythrocytaire normale de la protéine 4.1R. Cet événement d'épissage est bloqué dans les cellules MEL surexprimant l'oncoprotéine Spi.1/PU.1. Mon travail de thèse avait pour but de comprendre ce mécanisme de blocage en étudiant le rôle de la voie de signalisation de la PI3K en relation avec la phosphorylation de Spi.1/PU.1 d'une part, et celle des facteurs d'épissage de la famille des protéines SR d'autre part. J'ai montré que l'inhibition de la PI3K active la production d'hémoglobine ainsi que l'épissage de l'exon 16 4.1R. Ensuite j'ai montré que la phosphorylation de Spi-1/PU.1 par la PI3K est nécessaire pour son activité d'inhibition de la différenciation et de l'épissage de l'exon 16 4.1R. Enfin, j'ai montré que le facteur d'épissage Srp40 de la famille des protéines SR est un activateur stade spécifique de l'inclusion de l'exon 16. La surexpression de Srp40 dans les cellules MEL active l'inclusion de l'exon 16 indépendamment de la différenciation.

The advanced fuels investigated in this thesis comprise fuels non− conventional in their design/form (TRISO), their composition (high content of plutonium and minor actinides) or their use in a reactor type, in which they have not been used before (e.g. nitride fuel in BWR). These fuels come with a promise of improved characteristics such as safe, high temperature operation, spent fuel transmutation or fuel cycle extension, for which reasons their potentialis worth assessment and investigation. Their possible use also brings about various challenges, out of which some were addressed in this thesis. TRISO particle fuels with their superior retention abilities enable safe, high−temperature operation. Their combination with molten salt in the Advanced High Temperature Reactor (AHTR) concept moreover promises high operating temperature at low pressure, but it requires a careful selection of the cooling salt and the TRISO dimensions to achieve adequate safety characteristic, incl. a negative feedback to voiding. We show that an AHTR cooled with FLiBe may safely operate with both Pu oxide and enriched U oxide fuels. Pu and Minor Actinides (MA) bearing fuels may be used in BWR for transmutation through multirecycling; however, the allowable amounts of Pu and MA are limited due to the degraded feedback to voiding or low reactivity.We showed that the main positive contribution to the void effect in the fuelswith Pu and MA content of around 11 to 15% consist of the decreased thermalcapture probability in Pu-240, Pu-239 and Am-241 and increased fast and resonance fission probability of U-238, Pu239 and Pu-240. The total void worthmoreover increases during multirecycling, limiting the allowable amount ofMA to 2.45% in uranium−based fuels. An alternative, thorium−based fuel allows for 3.45% MA without entering the positive voiding regime at any point of the multirecycling. The increased alpha−heating associated with the use of transmutation fuels, is at level 24−31 W/kgFUEL in the uranium based fuels and 32−37 W/kgFUEL in the thorium−based configurations. The maximum value of the neutron emission, reached in the last cycle, is 1.7·106 n/s/g and 2·106 n/s/g for uranium and for thorium−based fuels, respectively. Replacing the standard UO2 fuel with higher−uranium density UN orUNZrO2 fuels in BWR shows potential for an increase of the in-core fuelresidence time by about 1.4 year. This implies 1.4% higher availability of the plant. With the nitride fuels, the total void worth increases and the efficiency of the control rods and burnable poison deteriorates, but no major neutronics issue has been identified. The use of nitride fuels in the BWR environment is conditioned by their stability in hot steam. Possible methods for stabilizing nitride fuels in water and steam at 300◦ C were suggested in a recent patentapplication. / <p>QC 20121004</p>

<p>Pin1 is a ubiquitously expressed phosphorylation-specific prolyl isomerase that regulates substrate function by catalyzing the cis-trans isomerization of prolyl bonds. Through this modulation, Pin1 has been shown to influence the stability, localization, and/or activity of a diverse set of protein substrates that participate in a variety of cellular responses, such as cell cycle progression, modulation of cell stress, and apoptosis. In addition to extensive studies in non-hematopoietic cells, Pin1 has also been shown to regulate immune cell function. Indeed, Pin1 participates in germinal center B cell development and eosinophil granulocyte survival. It also facilitates cytokine production in T cells, eosinophil granulocytes, and plasmacytoid dendritic cells. Through specific activities such as these, Pin1 has been demonstrated to modulate responses to viral challenge, respiratory allergens, and organ transplantation. </p><p>Due to previously described functions of Pin1 in regulating cells of both the innate and adaptive immune system, we predicted that Pin1 would participate in systemic inflammatory responses. Upon inducing systemic inflammation in mice, we observed a profound reduction in circulating cytokine concentrations in Pin1-null mice compared to WT mice. This result prompted further investigations, which are described in chapter 3 and chapter 4 of this dissertation. In chapter 3, we evaluate the potential contribution of macrophages to the defects we observe in LPS-challenged Pin1-null mice. Using primary macrophages, bone marrow-derived macrophages, and MEF, we ultimately exclude a role for Pin1 in modulating LPS-induced production of pro-inflammatory cytokines in these cells. In chapter 4, we uncover a defect in the accumulation of conventional dendritic cells (cDC) in LPS-challenged Pin1-null mice. Upon more careful examination of spleen cDC subsets in Pin1-null mice, we discovered a defect in the CD8+ subset. Experiments described in this chapter collectively indicate a role for Pin1 in preferentially modulating late stages of development of the CD8+ subset of cDC. Consistent with such a defect, the expansion of adoptively transferred WT CD8+ T cells was less robust in Pin1-null mice than WT mice upon infection with the bacterium Listeria monocytogenes . At the end of chapter 4, we provide evidence that Pin1 facilitates the degradation of the hematopoietic transcription factor PU.1, and propose that deregulation of PU.1 expression may be one mechanism by which Pin1 modulates CD8+ cDC development. The work described in this dissertation began by evaluating a potential role for Pin1 in modulating pro-inflammatory cytokine production in macrophages; ultimately, however, we uncovered a novel role for Pin1 in preferentially modulating the development of the CD8+ subset of cDC. The results presented herein expand the current understanding of DC development and further implicate Pin1 as an important modulator of both innate and adaptive immune responses.</p> / Dissertation

The goal for this thesis was to apply electrospun biomimetic coatings on implantable glucose biosensors and test their efficacy as mass-transport limiting and tissue engineering membranes, with special focus on achieving reliable and long sensing life-time for biosensors when implanted in the body. The 3D structure of electrospun membranes provides the unique combination of extensively interconnected pores, large pore volumes and mechanical strength, which are anticipated to improving sensor sensitivity. Their structure also mimics the 3D architecture of natural extracellular matrix (ECM), which is exploited to engineer tissue responses to implants. A versatile vertical electrospinning setup was built in our workshop and used to electrospin single polymer - Selectophore™ polyurethane (PU) and two polymer (coaxial) – PU and gelatin (Ge) fibre membranes. Extensive studies involving optimization of electrospinning parameters (namely solvents, polymer solution concentration, applied electric potential, polymer solution feed flow rate, distance between spinneret and collector) were carried out to obtain electrospun membranes having tailorable fibre diameters, pore sizes and thickness. The morphology (scanning electron microscopy (SEM) and optical microscopy), fibre diameter (SEM), porosity (bubble point and gravimetry methods), hydrophilicity (contact angle), solute diffusion (biodialyzer) and uniaxial mechanical properties (tensile tester) were used to characterize certain shortlisted electrospun membranes. Static and dynamic collector configurations for electrospinning fibres directly on sensor surface were optimized of which the dynamic collections system helped achieve snugly fit membranes of uniform thickness on the entire surface of the sensor. The biocompatibility and the in vivo functional efficacy of electrospun membranes off and on glucose biosensors were evaluated in rat subcutaneous implantation model. Linear increase in thickness of electrospun membranes with increasing electrospinning time was observed. Further, the smaller the fibre diameter, smaller was the pore size and higher was the fibre density (predicted), the hydrophilicity and the mechanical strength. Very thin membranes showed zero-order (Fickian diffusion exponent ‘n’ ~ 1) permeability for glucose transport. Increasing membrane thickness lowered ‘n’ value through non-Fickian towards Fickian (‘n’ = 0.5) diffusion. Thin electrospun PU membranes (~10 μm thick) did not affect, while thicknesses between 20 and 140 μm all decreased sensitivity of glucose biosensor by about 20%. PU core - Ge shell coaxial fibre membranes caused decrease in ex vivo sensitivity by up to 40%. The membranes with sub-micron to micron sized pore sizes functioned as mass-transport limiting membranes; but were not permeable to host cells when implanted in the body. However, PU-Ge coaxial fibre membranes, having <2 μm pore sizes, were infiltrated with fibroblasts and deposition of collagen in their pores. Such tissue response prevented the formation of dense fibrous capsule around the implants, which helped improve the in vivo sensor sensitivity. To conclude, this study demonstrated that electrospun membrane having tailorable fibre diameters, porosity and thickness, while having mechanical strength similar to the natural soft tissues can be spun directly on sensor surfaces. The membranes can function as mass-transport limiting membranes, while causing minimal or no effect on sensor sensitivity. With the added bioactive Ge surfaces, evidence from this study indicates that reliable long-term in vivo sensor function can be achieved.

The ability to quickly and accurately quantify the plutonium (Pu) content in pressurized water reactor (PWR) spent nuclear fuel (SNF) is critical for nuclear forensics purposes. One non-destructive assay (NDA) technique being investigated to detect bulk Pu in SNF is measuring the self-induced x-ray fluorescence (XRF). Previous XRF measurements of Three Mile Island (TMI) PWR SNF taken in July 2008 and January 2009 at Oak Ridge National Laboratory (ORNL) successfully illustrated the ability to detect the 103.7 keV x ray from Pu using a planar high-purity germanium (HPGe) detector. This allows for a direct measurement of Pu in SNF. Additional gamma ray and XRF measurements were performed on TMI SNF at ORNL in October 2011 to measure the signal-to-noise ratio for the 103.7 keV peak. Previous work had shown that the Pu/U peak ratio was directly proportional to the Pu/U content and increased linearly with burnup. However, the underlying Compton background significantly reduced the signal-to-noise ratio for the x-ray peaks of interest thereby requiring a prolonged count time. Comprehensive SNF simulations by Stafford et al showed the contributions to the Compton continuum were due to high-energy gamma rays scattering in the fuel, shipping tube, cladding, collimator and detector1. The background radiation was primarily due to the incoherent scattering of the 137Cs 661.7 keV gamma. In this work methods to reduce the Compton background and thereby increase the signal-to-noise ratio were investigated. To reduce the debilitating effects of the Compton background, a crystal x-ray spectrometer system was designed. This wavelength-dispersive spectroscopy technique isolated the Pu and U x rays according to Bragg's law by x-ray diffraction through a crystal structure. The higher energy background radiation was blocked from reaching the detector using a customized collimator and shielding system. A flat quartz-crystal x-ray spectrometer system was designed specifically to fit the constraints and requirements of detecting XRF from SNF. Simulations were performed to design and optimize the collimator design and to quantify the improved signal-to-noise ratio of the Pu and U x-ray peaks. The proposed crystal spectrometer system successfully diffracted the photon energies of interest while blocking the high-energy radiation from reaching the detector and contributing to background counts. The spectrometer system provided a higher signal-to-noise ratio and lower percent error for the XRF peaks of interest from Pu and U. Using the flat quartz-crystal x-ray spectrometer and customized collimation system, the Monte Carlo N-Particle (MCNP) simulations showed the 103.7 keV Pu x-ray peak signal-to-noise ratio improved by a factor of 13 and decreased the percent error by a factor of 3.3.

Espumas de assentos personalizados para cadeiras de rodas se adaptam a anatomia do usuário, no entanto, ao passo que melhoram o posicionamento, podem influenciar negativamente no conforto térmico por aumentar a superfície de contato. O conforto térmico pode ser melhorado com materiais de mudança de fase (PCM), que devem ser aplicados protegidos por um invólucro para impedir seu escape na fase líquida. O invólucro mais comum são as microcápsulas. Assim, o objetivo dessa pesquisa é obter, aplicar e caracterizar microcápsulas com núcleo de material de mudança de fase, mais especificamente o eicosano, em espumas de PU flexível após sua expansão e tecido que recobre o assento, ampliando o efeito termorregulador desse material, reduzindo o tempo de aplicação e desperdício das microcápsulas. Essa é uma pesquisa de natureza aplicada, com técnica de pesquisa de documentação direta através de pesquisa em laboratório, objetivo descritivo e explicativo, com procedimento técnico experimental e abordagem quantitativa. Foram obtidas microcápsulas de eicosano com a variação de diferentes parâmetros e caracterizadas através de MEV, FTIR, granulometria, TGA e DSC. As microcápsulas foram inseridas em amostras de espuma com duas técnicas diferentes (filtração e aerografia) e essas amostras foram analisadas com um termógrafo para verificação do efeito termorregulador gerado. Pode-se concluir que a aerografia proporciona melhor aproveitamento de tempo e de material, ampliando os efeitos térmicos gerados na espuma flexível de PU. Também, o efeito térmico depende não apenas do tipo de microcápsula, mas da quantidade aplicada e sua distribuição pelo material. / Wheelchair custom seats foams can be adapt to the user’s anatomy nonetheless, although it can improve positioning, it influences negatively in thermal comfort as it increases the contact surface. Thermal comfort can be improved with phase change materials (PCM) that must be applied in a casing to prevent its escape in the liquid phase. The most common casing is the microcapsules. The main objective of this research is to obtain, apply and characterize PCM microcapsules, more specifically eicosane, into flexible PU foams after expansion and fabric that recovers the seat, enlarging the thermoregulatory effect of this material, reducing application time and microcapsule waste. It is a study of applied nature, research technique of direct documentation through laboratory investigation, descriptive and explanatory objective, and experimental technical procedure generating quantifiable data. Eicosane microcapsules were obtained varying different parameters and characterizes through SEM, FTIR, granulometry, TGA and DSC. The microcapsules were inserted into foam samples with two different techniques (filtration and aerography). These samples were analyzed through a thermographer to verify its thermoregulatory effect. It can be concluded that aerography has better use of time and material, improving the overall thermal effect in PU foam. Additionally, the thermal effect depends not only in the type of microcapsule, but also in quantity applied and distribution through out the material.

Espumas de assentos personalizados para cadeiras de rodas se adaptam a anatomia do usuário, no entanto, ao passo que melhoram o posicionamento, podem influenciar negativamente no conforto térmico por aumentar a superfície de contato. O conforto térmico pode ser melhorado com materiais de mudança de fase (PCM), que devem ser aplicados protegidos por um invólucro para impedir seu escape na fase líquida. O invólucro mais comum são as microcápsulas. Assim, o objetivo dessa pesquisa é obter, aplicar e caracterizar microcápsulas com núcleo de material de mudança de fase, mais especificamente o eicosano, em espumas de PU flexível após sua expansão e tecido que recobre o assento, ampliando o efeito termorregulador desse material, reduzindo o tempo de aplicação e desperdício das microcápsulas. Essa é uma pesquisa de natureza aplicada, com técnica de pesquisa de documentação direta através de pesquisa em laboratório, objetivo descritivo e explicativo, com procedimento técnico experimental e abordagem quantitativa. Foram obtidas microcápsulas de eicosano com a variação de diferentes parâmetros e caracterizadas através de MEV, FTIR, granulometria, TGA e DSC. As microcápsulas foram inseridas em amostras de espuma com duas técnicas diferentes (filtração e aerografia) e essas amostras foram analisadas com um termógrafo para verificação do efeito termorregulador gerado. Pode-se concluir que a aerografia proporciona melhor aproveitamento de tempo e de material, ampliando os efeitos térmicos gerados na espuma flexível de PU. Também, o efeito térmico depende não apenas do tipo de microcápsula, mas da quantidade aplicada e sua distribuição pelo material. / Wheelchair custom seats foams can be adapt to the user’s anatomy nonetheless, although it can improve positioning, it influences negatively in thermal comfort as it increases the contact surface. Thermal comfort can be improved with phase change materials (PCM) that must be applied in a casing to prevent its escape in the liquid phase. The most common casing is the microcapsules. The main objective of this research is to obtain, apply and characterize PCM microcapsules, more specifically eicosane, into flexible PU foams after expansion and fabric that recovers the seat, enlarging the thermoregulatory effect of this material, reducing application time and microcapsule waste. It is a study of applied nature, research technique of direct documentation through laboratory investigation, descriptive and explanatory objective, and experimental technical procedure generating quantifiable data. Eicosane microcapsules were obtained varying different parameters and characterizes through SEM, FTIR, granulometry, TGA and DSC. The microcapsules were inserted into foam samples with two different techniques (filtration and aerography). These samples were analyzed through a thermographer to verify its thermoregulatory effect. It can be concluded that aerography has better use of time and material, improving the overall thermal effect in PU foam. Additionally, the thermal effect depends not only in the type of microcapsule, but also in quantity applied and distribution through out the material.

Espumas de assentos personalizados para cadeiras de rodas se adaptam a anatomia do usuário, no entanto, ao passo que melhoram o posicionamento, podem influenciar negativamente no conforto térmico por aumentar a superfície de contato. O conforto térmico pode ser melhorado com materiais de mudança de fase (PCM), que devem ser aplicados protegidos por um invólucro para impedir seu escape na fase líquida. O invólucro mais comum são as microcápsulas. Assim, o objetivo dessa pesquisa é obter, aplicar e caracterizar microcápsulas com núcleo de material de mudança de fase, mais especificamente o eicosano, em espumas de PU flexível após sua expansão e tecido que recobre o assento, ampliando o efeito termorregulador desse material, reduzindo o tempo de aplicação e desperdício das microcápsulas. Essa é uma pesquisa de natureza aplicada, com técnica de pesquisa de documentação direta através de pesquisa em laboratório, objetivo descritivo e explicativo, com procedimento técnico experimental e abordagem quantitativa. Foram obtidas microcápsulas de eicosano com a variação de diferentes parâmetros e caracterizadas através de MEV, FTIR, granulometria, TGA e DSC. As microcápsulas foram inseridas em amostras de espuma com duas técnicas diferentes (filtração e aerografia) e essas amostras foram analisadas com um termógrafo para verificação do efeito termorregulador gerado. Pode-se concluir que a aerografia proporciona melhor aproveitamento de tempo e de material, ampliando os efeitos térmicos gerados na espuma flexível de PU. Também, o efeito térmico depende não apenas do tipo de microcápsula, mas da quantidade aplicada e sua distribuição pelo material. / Wheelchair custom seats foams can be adapt to the user’s anatomy nonetheless, although it can improve positioning, it influences negatively in thermal comfort as it increases the contact surface. Thermal comfort can be improved with phase change materials (PCM) that must be applied in a casing to prevent its escape in the liquid phase. The most common casing is the microcapsules. The main objective of this research is to obtain, apply and characterize PCM microcapsules, more specifically eicosane, into flexible PU foams after expansion and fabric that recovers the seat, enlarging the thermoregulatory effect of this material, reducing application time and microcapsule waste. It is a study of applied nature, research technique of direct documentation through laboratory investigation, descriptive and explanatory objective, and experimental technical procedure generating quantifiable data. Eicosane microcapsules were obtained varying different parameters and characterizes through SEM, FTIR, granulometry, TGA and DSC. The microcapsules were inserted into foam samples with two different techniques (filtration and aerography). These samples were analyzed through a thermographer to verify its thermoregulatory effect. It can be concluded that aerography has better use of time and material, improving the overall thermal effect in PU foam. Additionally, the thermal effect depends not only in the type of microcapsule, but also in quantity applied and distribution through out the material.

The master‘s thesis is focused on the study of biodegradation of polyhydroxyalkanoates, namely polymer polyhydroxybutyrate. The first part of the thesis is focused on the study of biodegradation of polyhydroxybutyrate in the form of crystalline granules of PHB and PHB films using selected species of microorganisms from bacteria, yeasts and fungi. As a representative of bacteria was chosen microorganism Delftia acidorovans, as yeast was selected Aureobasidium pullulans and Aspergillus fumigatus as fungi. PHB depolymerase activity was measured employing turbidemtiric method with suspension of PHB granules as substrate. The results showed that D. acidorovans can partially degrade PHB. On the contrary A. pullulans cannot effectively degrade PHB. The most significant degradation ability revealed A. fumigatus, which was able to degrade PHB completely. Extracellular enzymes excreted by these microorganisms when cultivated on PHB materials as sole carbon sources were analyzed by SDS-PAGE. The second part of the thesis deals with the biodegradation of PHB in the form of PHB film, PHB hardened foil and PHB Nanoul fabric using standard composting test. Semi-solid cultivation showed positive results. In the interval from 14 days to two months were all forms of the PHB completely biodegraded. With semi-solid cultivation was also studied biodegradation rate of the polyurethane elastomeric films which were modified by partial replacement of polyester polyol by PHB. The test samples were prepared using PHB from Sigma and the PHB samples prepared at the Faculty of chemistry VUT. Samples with different concentrations of the dispersed PHB (1 %, 5 % and 10 %) in the polyurethane were also object of the study. At the end of the cultivation (after 2 months) were measured mechanical properties in tension of the material, then efficiency of biodegradation by gravimetric analysis and modification of the material surface by microscopic analysis.

This work is focused on verifying the permeability and olfactometric assessment of emissions from selected polymeric foam materials and foam rubber. The olfactometric assessment is based on the analysis of identified and identified volatile organic compounds. The research was conducted by taking samples of the emissions from a small-space sampling chamber. Emissions were analyzed by gas chromatography with mass spectrometry and their assessment by the assessor was performed by dynamic olfactometry. The work was supplemented by designing its own experimental procedure of mechanical stress of foam materials and changes in climatic conditions in the air-conditioning chamber in connection with release of emissions. Standard testing procedures according to European standards (EN ISO 16000-9, ČSN 13725 and EN ISO 7231) have been used to test these properties. The aim of the work is to verify selected properties of polymeric foam materials ? permeability and olfactometric assessment related to the analysis of volatile organic compounds. It also involves permeability is a change in breathability, depending on mechanical stress and release of emissions influencing given properties in terms of time, mechanical stress, and changing climatic conditions.