In this thesis, the effects of simple shear flow on macromolecular structure and interactions are investigated in detail via a combination of Nuclear Magnetic Resonance (NMR) spectroscopy and rheology, namely Rheo-NMR. A specially designed NMR couette shear cell and benchtop shear cell, developed in-house, demonstrated that the direct measurement of the above phenomena is possible. First, to determine whether the shear cells were creating simple shear flow, results were reproduced from literature studies of liquid crystal systems which report shear effects on: Cetyl Trimethyl Ammonium Bromide (CTAB) in deuterium oxide, and Poly(gamma-benzyl-L-glutamate) (PBLG) in m-cresol. Next, the possible conformational changes to protein structure brought about by shear were investigated by applying shear to Bovine -lactogobulin ( -Lg). As the protein was sheared, a small, irreversible conformational change was observed by means of one-dimensional and two-dimensional 1H NMR with reasonable reproducibility. However, no observable change was detected by means of light scattering. A large conformational change was observed after shearing a destabilized -Lg sample containing 10% Trifluoroethanol (TFE) (v/v). From an NMR point of view, the sheared state was similar to the structure of -Lg containing large amounts of -helices and, interestingly, similar to the structure of -Lg containing -sheet amyloid fibrils. Gel electrophoresis tests suggested that the changes were caused by hydrophobic interactions. Unfortunately, this proved to be difficult to reproduce. The effect of shear on an inter-macromolecular interaction was investigated by applying shear during an enzyme reaction of pectin methylesterase (PME) on pectin. Experimental method and analysis developments are described in detail. It was observed that under the conditions studied, shear does not interfere with the de-esterification of pectin with two types of PME, which have different action mechanisms at average shear rates up to 1570 s-1.

In this thesis, the effects of simple shear flow on macromolecular structure and interactions are investigated in detail via a combination of Nuclear Magnetic Resonance (NMR) spectroscopy and rheology, namely Rheo-NMR. A specially designed NMR couette shear cell and benchtop shear cell, developed in-house, demonstrated that the direct measurement of the above phenomena is possible. First, to determine whether the shear cells were creating simple shear flow, results were reproduced from literature studies of liquid crystal systems which report shear effects on: Cetyl Trimethyl Ammonium Bromide (CTAB) in deuterium oxide, and Poly(gamma-benzyl-L-glutamate) (PBLG) in m-cresol. Next, the possible conformational changes to protein structure brought about by shear were investigated by applying shear to Bovine -lactogobulin ( -Lg). As the protein was sheared, a small, irreversible conformational change was observed by means of one-dimensional and two-dimensional 1H NMR with reasonable reproducibility. However, no observable change was detected by means of light scattering. A large conformational change was observed after shearing a destabilized -Lg sample containing 10% Trifluoroethanol (TFE) (v/v). From an NMR point of view, the sheared state was similar to the structure of -Lg containing large amounts of -helices and, interestingly, similar to the structure of -Lg containing -sheet amyloid fibrils. Gel electrophoresis tests suggested that the changes were caused by hydrophobic interactions. Unfortunately, this proved to be difficult to reproduce. The effect of shear on an inter-macromolecular interaction was investigated by applying shear during an enzyme reaction of pectin methylesterase (PME) on pectin. Experimental method and analysis developments are described in detail. It was observed that under the conditions studied, shear does not interfere with the de-esterification of pectin with two types of PME, which have different action mechanisms at average shear rates up to 1570 s-1.

Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an attaching and effacing pathogen that upon attachment to host cells, induce characteristic attaching and effacing lesions and formation of F-actin rich pedestals beneath sites of bacterial attachment. EHEC harbors a Type III secretion system through which it delivers dozens of effectors into the host cell. The two secreted effectors critical for EHEC-mediated actin pedestal formation are the translocated intimin receptor (Tir) and EspFU. EspFU consists of an N-terminal secretion signal and a C-terminus containing six tandem 47-residue proline-rich repeats, each of which can bind and activate the actin nucleation promoting factor N-WASP. Structural and functional analyses described here have identified the mechanism of N-WASP activation by EspFU and the minimal domains and specific residues required for this activity. While EspFU and Tir are the only bacterial effectors required for F-actin pedestal formation, recruitment of EspFU to Tir is mediated by an unidentified putative host factor. To identify the host factor responsible for linking these two effectors, a combination of in vitro and functional assays were used to identify the host factor, IRTKS and the residues required for these interactions were defined. Further, the presence of at least two 47-residue repeats in all characterized clinical isolates of canonical EHEC strains led us to address the minimal requirements for EspFU functional domains to promote recruitment to Tir and N-WASP activation. Here we show that two proline-rich elements of EspFU are required for recruitment of EspFU by IRTKS to sites of bacterial attachment. Furthermore, once artificially clustered at the membrane, a single N-WASP binding element of EspFU can induce actin pedestal formation.

Hydroxyapatite (HAP) coatings are used in orthopaedic surgery for bone regeneration. Current methods of phase quantification of HAP coatings suffer from drawbacks. A novel methodology of quantitative phase analysis of HAP coatings has been devised and validated. This method, based on whole pattern fitting with a fundamental parameters approach, incorporates amorphous calcium phosphate (ACP) and apatite phases into structural refinements. A comparison of the structural and chemical properties of plasma sprayed (PS) and novel electrodeposited (ED) HAP coatings has been conducted. ED coatings contained less ACP and more preferred orientation than the PS coatings, although the stoichiometry was similar. In vitro investigations of PS and ED coatings in simulated body fluid and foetal calf serum revealed that both are bioactive. A carbonated apatite layer produced on the ED coatings was -0.7μm thick with a stoichiometry and chemical constituents similar to that of natural bone apatite. PS coatings produced a nanocrystalline carbonated apatite layer (-4μm). For the first time it has been possible to model crystalline HAP and nanocrystalline apatite as independent phases and obtain accurate lattice parameters for each. A positive linear correlation has been made between microstrain and the solubility of HAP and carbonated apatites. Dissolution studies have shown that the behaviour of HAP and carbonated apatite is dominated by crystallite size at low undersaturation and by crystallite size and microstrain at high undersaturation for crystallites between -30OA- 1000A. Metastable equilibrium occurred for crystallites <_400A at low undersaturation. Carbonate content did not affect the solubility or dissolution behaviour. A novel technology for coating polymeric tape with HAP for potential use in anterior cruciate ligament reconstruction has been devised. Mechanical tests have demonstrated that no adverse properties are induced by the coating technology. Cell culture studies have shown that the HAP layer is capable of enhanced attachment, proliferation and differentiation of osteoblast cells compared to uncoated tape.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The imminence of Panambi s dam construction, in the municipality of Alecrim, Rio Grande do Sul, affecting the municipality of Porto Mauá, which will overflow thousands of hectares of rainforest, Macluratinctoria s habitat and, the great exploitation of the specie in the region led to the initiation of this research work. The specie is commonly known as Tajuva, presents arboreal sized, and belongs to the botanical family Moraceae, native of Rio Grande do Sul. It is a semideciduous specie, with heights between 10 to 30 meters and, up to 100 cm of diameter; short trunk, with a smooth bark, grayish green with many yellowed lenticels, many times, with the presence of branched thorns. The study aimed to the realization of anatomical description, the chemical characterization and the basic and weighted densities of Maclura tinctoria(L) D. Don Ex Steud wood. The five studied exemplars were obtained through environmental license, in accordance with current legislation. Three wood samples, taken from BHD were used for the manufacture of histological slides and for the fibers quantification. To the determination of wood density and the chemical analysis of wood, disks were removed from the trunk and divided into opposing wedges. It were used opposing wedges removed from six pre-established heights (base, DAP, 25%, 50%, 75% and 100% of commercial height). It was used the ASTM standards to determinate the density, and from TAPPI to the chemical analysis of wood. The wood was anatomically described and illustrated with photomicrographs. The woody tissue consist mainly of fibers (60 ± 3,4%) and rays (20 ± 2,3%), totaling approximately 80%. The remaining, in other words, 20% are divided into vessels (12 ± 4%) and axial parenchyma (8 ± 3%). The mean values, to the basic density (pb) and weighted density (ppond), were 0,539 and 0,555 g/cm³, respectively. The macromolecular constituents of heartwood and sapwood were determinate separately. The average content found for extractives (18,92; 6,32%); Klason lignin (17,58; 17,55%); holocellulose (63,45; 78,59%); alpha-cellulose ( 35,44; 44,38%); hemicelluloses (28,01; 34,21%) and ashes (1,08; 1,35%), to the heartwood and sapwood, respectively, totaling 102,42 ± 3,1%. The Tajuva s wood brings together the anatomical characteristics prevalent in Moraceae, framing the specie in their respective botanical family. It is considered an evolved specie anatomically. The density presents a decrease in the values in relation to the axial position of the plant. The heartwood is constituted of phenolic compounds and acids in a higher proportion relative to the sapwood, which presents higher starch contents. Studies about the natural durability and the wood quality are fundamental to indicate the best application of wood. / A eminência da construção da barragem Panambi, no município de Alecrim, Rio Grande do Sul, afetando o município de Porto Mauá, a qual alagará milhares de hectares de mata atlântica, habitat da Maclura tinctoria e, a grande exploração da espécie na região motivaram o início desse trabalho de pesquisa. Popularmente, a espécie é conhecida como Tajuva, apresenta porte arbóreo, e pertence à família botânica Moraceae, nativa do Rio Grande do Sul. É uma espécie semicaducifólia, com altura entre 10 a 30 metros e, com até 100 cm de diâmetro; fuste curto, com casca lisa, verde acinzentada com numerosas lenticelas amareladas, muitas vezes, com presença de espinhos ramificados. O estudo teve como objetivo a descrição anatômica, a caracterização química e a determinação das densidades básica e ponderada da madeira de Maclura tinctoria (L) D. Don ex Steud. As cinco árvores estudadas foram obtidas através de licenciamento ambiental, de acordo com a legislação vigente. Três amostras do lenho, retiradas do DAP foram utilizadas para a confecção de lâminas histológicas e para a quantificação das fibras. Para a determinação da densidade da madeira e para as análises químicas da madeira, foram retirados discos do tronco e divididos em cunhas. Foram utilizadas cunhas opostas retiradas de seis alturas pré-estabelecidas (base, DAP, 25, 50, 75 e 100% da altura comercial). Utilizou-se as normas da ASTM para determinação da densidade, e da TAPPI para as análises químicas da madeira. O lenho foi anatomicamente descrito e ilustrado com fotomicrografias. O tecido lenhoso é constituído principalmente por fibras (60 ± 3,4%) e raios (20 ± 2,3%), totalizando aproximadamente 80%. O restante, ou seja, 20 % estão divididos entre vasos (12 ± 4%) e parênquima axial (8 ± 3%). Os valores médios, para a densidade básica média (ρb) e ponderada (ρpond), foram de 0,539 e 0,555 g/cm³, respectivamente. Os constituintes macromoleculares da madeira de cerne e alburno foram determinados separadamente. Os teores médios encontrados para extrativos totais (18,92; 6,32%); lignina Klason (17,58; 17,55%); holocelulose (63,45; 78,59%); alfa celulose (35,44; 44,38%); hemiceluloses (28,01; 34,21%) e cinzas (1,08; 1,35%), para cerne e alburno, respectivamente, totalizando 102,42 ± 3,1%. O lenho da Tajuva reúne as características anatômicas predominantes em Moraceae, enquadrando a espécie em sua respectiva família botânica. É considerada uma espécie evoluída anatomicamente. A densidade apresenta decréscimo de valores em relação à posição axial da planta. O cerne é constituído por compostos fenólicos e ácidos em maior proporção em relação ao alburno, que apresenta maiores teores de amido. Estudos sobre a durabilidade natural e a qualidade da madeira são fundamentais para indicar a melhor aplicação da madeira.

Pendant longtemps, la chromatine a été uniquement décrite comme un moyen de compacter près de deux mètres d’ADN dans un noyau de quelques micromètres de diamètre. On sait aujourd’hui que la chromatine représente en fait un élément majeur de régulation de toutes les fonctions nucléaires impliquant l’ADN. Dans le contexte de dommages de l’ADN induits par irradiations UV, la chromatine endommagée subit une décondensation rapide et transitoire qui l’amène à occuper un volume 1,5 fois plus grand que son volume initial. Cette relaxation chromatinienne est associée à une plus grande accessibilité de l’ADN. Néanmoins, le lien entre ces deux effets découlant de la présence de dommages, n’a pas été établi, ni caractérisé. En couplant l’imagerie de cellules vivantes à l’induction de dommages ciblés au sein de noyaux cellulaires par micro-irradiation laser, ces travaux ont permis de mettre en évidence le rôle majeur de PARP1 dans la réponse chromatinienne aux dommages de l’ADN. En effet, certaines conclusions contradictoires présentes dans la littérature scientifique concernant l’action de PARP1 sur la chromatine ont été réconciliées en démontrant que PARP1 seul peut se lier à la chromatine et entraîner une plus forte compaction de celle-ci, tandis que son activité catalytique de PARylation va, quant à elle, conduire à une décompaction de la structure chromatinienne. Cette étude s’est aussi intéressée à la dynamique particulière de l’histone H1 suite aux dommages de l’ADN. En effet, celui-ci est rapidement exclu des zones de dommages par un mécanisme encore inconnu, et les éléments apportés ici suggèrent que H1 pourrait jouer un rôle dans la décondensation de la chromatine suite aux dommages de l’ADN. Pour finir, des techniques de photo-perturbation et de spectroscopie de corrélation de fluorescence ont été employées pour comprendre et caractériser l’environnement moléculaire que constitue la chromatine endommagée et décondensée. Bien qu’une augmentation significative des interactions entre la chromatine et certains de ses partenaires d’interactions soit observée au sein des zones endommagées, aucun changement en termes d’encombrement moléculaire n’a pu être mis en évidence à ce niveau qui pourrait expliquer une plus grande accessibilité de l’ADN. / For a long time, chromatin was only described as a mean to fit the two-meters long DNA molecule into a nucleus of only a few microns. It is admitted today that chromatin actually represents a key element in the regulation of all nuclear functions dependent on DNA. In the context of UV-induced DNA damage, chromatin undergoes a rapid and transient relaxation which leads to an expansion of the damaged area to 1.5 times its original size. While this chromatin response to damage is associated with a higher DNA accessibility, the link between those two phenomena, as well as the mechanisms driving them, are still poorly understood. Using live-cell imaging and laser micro-irradiation to induce DNA damage on specific nuclear areas, this work allowed to gain hindsight on the predominant role played by PARP1 in the DNA damage-induced chromatin relaxation. Indeed, showing that PARP1 at DNA damage sites can both induce chromatin compaction through its recruitment at DNA breaks or chromatin decondensation through its PARylation activity helped reconcile its apparent opposite effects described in the literature. A focus was also made on the linker histone H1, as it displays a peculiar behavior upon DNA damage, being rapidly released from the site of DNA lesions. Even if the driving force behind H1 release from damaged chromatin areas has not been identified yet, its behavior suggests that H1 might play a part in chromatin relaxation or in increasing DNA accessibility upon DNA damage. Lastly, combining photo-activation techniques and fluorescence correlation spectroscopy, experiments were performed in order to understand the physical environment that damaged, relaxed chromatin constitutes. We report here that, while enhanced binding of random DNA binding factors is observed in the damaged chromatin area, no significant change is observed in the macromolecular crowding levels that could potentially explain this enhanced binding, as well as a higher DNA accessibility.

L’objectif de cette thèse consiste à développer des matériaux semi-conducteurs organiques efficaces pour le photovoltaïque organique. Le travail est focalisé sur l’optimisation de matériaux à caractère donneur d’électrons pour la préparation de dispositifs à hétérojonction volumique, en association avec un dérivé de fullerène comme matériau à caractère accepteur d’électrons. Plus particulièrement, il s’agit de réaliser une étude d’optimisation systématique de deux familles de référence (respectivement macromoléculaire et moléculaire) issus du laboratoire, qui ont déjà conduit à des performances photovoltaïques intéressantes. Pour cela, nous avons suivi une démarche rigoureuse et systématique en ciblant les paramètres chimiques les plus pertinents à faire varier. Afin de déterminer les propriétés des nouveaux matériaux ainsi synthétisés, des caractérisations spectroscopiques, électrochimiques, structurales, de transport de charge et photovoltaïque ont systématiquement été effectué. / The aim of this thesis is to develop efficient semi-conducting organic materials for organic photovoltaics. This work is focuses on the optimization of electron-donor organic semiconductors for the preparation of bulk heterojunction devices, in blend with a fullerene derivative used as electron-acceptor material. More specifically, it is to perform a systematic optimization study of two reference families (macromolecular and molecular respectively) from the laboratory, which have already led to interesting photovoltaic performances. For this, we followed a structured and systematic approach targeting the most relevant chemical parameters to be varied. To determine the properties of new materials synthesized, spectroscopic, electrochemical, structural, charge transport and photovoltaic characterizations were systematically made.

La démocratisation des technologies du collage dans le domaine aérospatial amène les industriels à exiger des adhésifs toujours plus performants. L’objectif de cette thèse est d’optimiser les propriétés thermomécaniques d’un adhésif polyépoxyde bi-composant qui devra présenter des propriétés structurales entre -150°C et +150°C, tout en respectant des exigences de mise en oeuvre.L’analyse de la littérature et des expériences préliminaires a permis de sélectionner le système polyépoxyde modèle binaire le plus pertinent en termes de réactivité à l’ambiante et de propriétés sur assemblage collé. Pour optimiser la résistance en température élevée de ce système, deux voies ont été explorées : l’incorporation d’époxydes multi-fonctionnalisés pour augmenter la densité de réticulation, et l’utilisation d’amines cycliques qui accroit la rigidité des segments macromoléculaires. Pour améliorer les propriétés de résistance à la propagation de fissure à basse température, les additifs de renforcement montrent un intérêt certain pour la formulation d’adhésifs structuraux polymérisés à température ambiante. Sur la base de ces résultats, plusieurs formulations complexes ont pu être élaborées. Des phénomènes de synergie et d’inhibition entre les différents éléments des formulations ont été constatés, sans pouvoir être totalement compris ce qui met en lumière la difficulté de prévoir le comportement d’adhésifs complexes, à l’image des formulations commerciales.Ces travaux de thèse ont permis de développer un adhésif structural répondant aux exigences strictes du secteur aérospatial, et de dégager une stratégie de formulation qui pourra être appliquée à d’autres secteurs d’activités. / The democratisation of bonding technologies in aerospace field leads industrials to demand more and more efficient adhesives. The aim of this thesis is to optimise thermomechanical properties of two-component epoxy adhesives which need to preserve structural properties on a large scale of temperature (from -150°C to +150°C) and observing application process requirements.The analysis of scientific literature and preliminary experiments enable to select the most relevant binary epoxy system in terms of reactivity and bonding mechanical properties. To optimise high temperature resistance of this epoxy system, two approaches have been explored: addition of multi-functionalised epoxy resin to extend the crosslink density, and use of cycloaliphatic amines which increase the rigidity of macromolecular segments. To improve crack propagation resistance in low temperature, tougheners reveal an interest to formulate room temperature structural adhesives. On the basis of these results, complex formulations have been developed. Synergy and inhibition phenomenon between the formulation blends have been observed without being able to fully understand them which highlight the difficulty to predict the behaviour of complex adhesives such as commercial formulations.This thesis works allowed to develop a two-component epoxy adhesive formulation which meets aerospace requirements, and to bring about a formulation strategy which could be adapted to others sectors activities.

Atherosclerosis is an arterial disease that occurs due to the build-up of lipids, cholesterol and other substances in the arterial wall, collectively called plaque, leading to narrowing of the vessel lumen and, in time, disruption to the blood supply. The study of flow through atherosclerotic vessels is especially important since plaques not only cause a reduction in the vessel lumen but can rupture from the arterial wall, causing a blood clot in the vessel that may ultimately lead to heart attack or a stroke. Elevated level of oxidated low density lipoprotein (LDL) is a known risk factor associated with the genesis of atherosclerosis in arterial walls. Previous studies reported in literature have explored the transport of LDL through the arterial wall using analytical and mathematical models. In this work, we have presented a computational framework for the study of LDL transport in the lumen and the porous arterial wall. We have employed a four-layer arterial wall model and used governing equations to model the transport of LDL. We have used physiological parameters for the wall layers from literature and have validated the model based on the calculated filtration velocities and LDL concentration profiles in the arterial wall. We have further used this established model to study the effect of change in permeability and pressure on the LDL concentration. We have also studied the effect of pulsatile flow on the transport of LDL through the porous walls to examine the validity of the initial assumption of steady state and have seen that pulsation increases the time averaged net flux of LDL species by about 20%. We have next modeled a drug-eluting stent (DES), which is one of the popular remedies to cure atherosclerosis. The validation of DES model is followed by a combined study to analyze the effect of stent placement on LDL transport. Although there is no chemical reaction between the drug and LDL, we have noticed recirculation zones near the stent strut which result in accumulation of LDL molecules in the arterial wall. Future studies are aimed at incorporating variable porosity and permeability and a stenosed region in the geometry. The deformation of arterial wall due to pulsatile blood flow may lead to alteration of wall properties, which can give a realistic view of macromolecular transport.

Le soudage par électrofusion est la technique majoritairement utilisée pour assembler les tubes et les accessoires en polyéthylène utilisés sur le réseau gazier. Ses très bons résultats initiaux ont été ternis ces dernières années par un certain nombre de dysfonctionnements relevés tant sur le terrain qu'en laboratoire. Ils se traduisent par des soudures de qualité médiocre du fait de la présence de zones de très faible cohésion. L'objectif de ce travail est de développer des outils experts tant expérimentaux que numériques permettant d'optimiser les conditions de soudage par électrofusion. L'originalité de notre étude a été de développer un modèle éléments finis tridimensionnel prenant en compte les différents mécanismes et phénomènes physiques sous-jacents, responsables de la formation de la soudure. Le modèle permet de calculer le taux de transformation de la matière, de prendre en compte les enthalpies de fusion et de cristallisation, de calculer le taux d'interdiffusion des macromolécules à l'interface entre les différents corps à souder pour finalement prédire la qualité de la soudure en fonction des conditions de chauffage imposées. Pour valider le modèle numérique, tout en facilitant l'accès aux grandeurs expérimentales, nous avons conçu et réalisé un dispositif expérimental se présentant sous la forme d'une géométrie relativement simple et plane mais respectant les caractéristiques d'un accessoire réel. La confrontation entre résultats numériques et expérimentaux a permis de démontrer les capacités du modèle numérique à reproduire fidèlement la réalité. Les différents outils ont alors été utilisés pour étudier l'influence des conditions de soudage sur la soudure et pour étudier le soudage de géométries plus complexes telles que celles rencontrées dans les pièces industrielles. / The Electrofusion welding process is widely used to join polyethylene components in gas distribution networks. Even trusty as a technique, the field feedback points out some divergences whose influence on the long term performance of the weld. One of the well-known consequences of these divergences is the “sticking” (aka “cold weld”) that is the result of an uncompleted or even inexistent interdiffusion of the macromolecules of the materials to join. Most numerical simulations are two-dimensional whereas the process is usually three-dimensional both in terms of heat transfer and mechanical aspects. The main objective of the work was to develop a 3D finite element model and to validate it by comparing to a real situation the temperature evolution and the thermally affected areas in a simple planar welding geometry with the same dimensional characteristics as a real fitting chosen to make easier the instrumentation. The numerical model takes into account the fitting parameters such as polyethylene thermal and mechanical properties (i.e. melting and crystallization kinetics, phase transition, thermal expansion) and the electrical and geometrical settings. It computes a criterion based on the macromolecular interdiffusion theory able to determine whether a good welding occurred or not at the end of the welding cycle. The computed results (temperature, melted and cold areas and fracture surfaces) were compared with experimental data and gave very good agreement in terms of temperature, liquid phase fraction distribution and fracture surfaces. Finally the numerical model and the experimental process were used to study the influence of welding conditions on the weld itself and to study the welding of complex geometries such as the industrial fittings.