No presente trabalho foram estudados os fenômenos de encruamento e, principalmente, a formação e reversão da martensita alfa-linha (a\', cúbica de corpo centrado, CCC, ferromagnética) induzida por deformação em um aço inoxidável dúplex UNS S31803 e um super dúplex UNS S32520. Inicialmente, as microestruturas dos dois materiais na condição solubilizada foram caracterizadas com auxílio de várias técnicas complementares de análise microestrutural. Foram determinadas fração volumétrica, estrutura cristalina, composição química, tamanho e morfologia das duas fases (ferrita e austenita). Posteriormente, os dois aços foram deformados por dois métodos: a laminação a frio, dividida em vários estágios, com menores graus de deformação e a limagem, sendo que o cavaco limado resultante apresenta altos graus de deformação. Algumas amostras deformadas foram recozidas. Os fenômenos de encruamento, formação e reversão de martensita induzida por deformação na austenita, recuperação, recristalização da austenita e da ferrita no cavaco limado foram estudados predominantemente por difratometria de raios X e usando o método de Rietveld. A difratometria de raios X também foi utilizada para determinação das microdeformações residuais e tamanhos de cristalito (subgrão), calculadas a partir do alargamento dos picos de difração causado pelas deformações. Desta forma, puderam-se comparar os níveis de deformação da laminação e limagem. Qualitativamente, a formação e reversão da martensita induzida por deformação também foi estudada por meio de medidas magnéticas utilizando-se dados de saturação magnética das curvas de histerese obtidas com o auxílio de um magnetômetro de amostra vibrante. Observou-se que para o aço inoxidável dúplex, tanto a laminação quanto a limagem causaram a formação de martensita induzida por deformação e para o aço inoxidável super dúplex, apenas a limagem promoveu essa transformação. Em comparação com o aço dúplex, o aço super dúplex apresentou maior resistência à formação de martensita induzida por deformação, pois apresenta uma austenita mais rica em nitrogênio e uma maior propensão à formação de fase sigma durante o recozimento, pois apresenta uma ferrita mais rica em cromo e nitrogênio. / In the present work the phenomena of strain hardening, formation and reversion of the strain induced alpha-prime martensite (a\', body centered cubic, BCC, Ferromagnetic) in an UNS S31803 duplex and UNS S32520 super duplex stainless steels have been studied. Firstly, the microstructures of both materials in the solution annealed condition were characterized with the aid of several microstructural analysis complementary techniques. The volume fraction, crystalline structure, chemical composition, size and morphology of the two phases (ferrite and austenite) have been determined. Further, both steels were deformed by two methods: cold rolling, divided into several stages, with lower strain levels than filing, which the chips resulting had higher strain levels. The phenomena of strain hardening, formation and reversion of strain induced martensite in the austenite phase, recovery and recrystallization of austenite and ferrite phases have been studied, mainly using X-ray diffraction and the Rietveld method. X-ray diffraction was also used to determine the residual microstrain and crystallite size (sub grain), calculated from the diffraction peak broadening caused by straining. Thus, the levels of cold rolling and filing strains could be compared. Qualitatively, the formation and reversion of strain induced martensite was also studied by magnetic measurements using data from magnetic saturation of hysteresis curves obtained with the aid of a vibrating sample magnetometer. It has been observed that for the duplex stainless steel, both filing as well as cold rolling promoted strain induced martensite. On the other hand, for the super duplex stainless steel, just filing promoted this transformation. In the comparing with duplex, the super duplex stainless steel austenite is more stable that is why is richer in nitrogen, so, the strain induced martensite formation is more difficult. The easier sigma phase precipitation during annealing as well in the super duplex stainless steel is due higher levels of chrome and molybdenum than the duplex stainless steel.

Les propriétés finales des alliages métalliques sont directement liées à la microstructure de fin de mise en forme. Les mécanismes de recristallisation dynamique (DRX) et post-dynamique (PDRX) jouent un rôle important sur les évolutions microstructurales intervenant pendant et après les étapes de déformation à chaud. Dans ce contexte, un défi majeur pour les industriels et les chercheurs est de prédire la microstructure obtenue en fonction des conditions de mise en forme. Cela implique de bien connaître les mécanismes de DRX et PDRX et leur cinétique. Les modèles en champ complet permettent de modéliser explicitement la microstructure des alliages métalliques et ses possibles évolutions à l’échelle du polycristal. Ces modèles sont précis comparativement aux modèles œuvrant aux plus grandes échelles, mais ils sont généralement très couteux en termes de temps de calcul. Les modèles à champ moyen sont quant à eux basés sur une description implicite de la microstructure, conduisant à des temps de calcul considérablement réduits, mais ils reposent sur un grand nombre d’hypothèses, notamment topologiques. Cette thèse propose un nouveau modèle champ complet de DRX/PDRX et croissance de grains, capable de fonctionner en 2D comme en 3D, et une nouvelle approche en champ moyen, s'appuyant sur ces simulations en champ complet. La nouvelle approche champ moyen prend notamment mieux en compte les effets topologiques pour une meilleure prédiction des distributions de tailles de grains. Ce travail inclut une procédure de calibration et une validation des deux modèles s'appuyant sur une campagne d’essais expérimentaux sur un acier austénitique 304L. / Final properties of metal alloys are directly related to their microstructure, inherited from the processing route. Dynamic (DRX) and post-dynamic recrystallization (PDRX) mechanisms play a primordial role in microstructure evolutions occurring during and after hot-deformation. Within this context, predicting microstructures depending on the applied thermomechanical conditions is a major challenge for both industrials and researchers. This requires a good knowledge of recrystallization mechanisms and kinetics. Full field models are based on an explicit description of the microstructure of a metallic alloy, and its possible evolutions at a polycrystalline scale. These models are accurate compared to models operating at larger scales, but they generally lead to prohibitive numerical costs. On the other hand, mean field models are based on an implicit description of the microstructure, leading to considerably reduced numerical costs, but they are based on many assumptions, notably with regards to topology. The outcome of this PhD work is a new full field model of DRX/PDRX and grain growth, working in 3D as well as in 2D, and a new DRX/PDRX mean field approach which better accounts for topological effects, and provides better predictions for grain size distributions. This work also includes a calibration procedure and a validation of these two new models, using experimental data obtained from compression tests performed on the 304L austenitic steel.

Placing recrystallization annealing in a manufacturing proces is often necessary in order to restore plasticity of worked blanks right before working products to its final shape. For better understanding of processes which are taking place in a material during annealing the use of in-situ techniques is favorable. Such observations enable us to watch changes in the materiál in real time at the original spot. Observations at grain-level are ideally made with scanning electron microscope – SEM by electron back-scattered diffraction – EBSD. The technique makes watching nucleation, growth and potential coarsening of recrystallized grains possible. Special construction of a tensile stage allows us to watch changes of a sample during tensile tests which can be used in observing influence of MnS inclusions on plasticity of free-cutting steels. Materials studied in this work are commonly used free-cutting steel 11SMnPb30 and its lead-free alternative 11SMnBi30.

Supramolecular structure of eight commercial types of iPP, its blends with poly(L-lactide) (PP-PLLA) and copolymers with ethylene-propylene rubber (PP-EPR) together with four types of polyethylene (LDPE, HDPE) was uncovered and observed in this work. The spherulitic structure of iPP and PE homopolymers and PP copolymers was uncovered by etching of six different solutions of mineral acids with KMnO4 and by dissolution in four selected solvents. The latter was found to be ineffective. The structure of PP-PLLA blends was uncovered only after recrystallization (the change of structure by annealing and cooling rate). The uncovered crystalline structure was observed directly by confocal laser scanning microscope (CLSM) and SEM. Crystallinity together with a size distribution of crystallites was determined by DSC. It was proved that spherulitic supramolecular structure was easy to uncover by chemical etching only for molded samples and for fracture surfaces of injected samples. The efficiency of etchants varied but, generally, the samples with high degree of crystallinity were etched earlier. The etchants containing nitric acid and high content of sulphuric acid uncovered fine details of shperulites with respect to mixtures containing orthophosporic acid. It was also observed that not only the etchant (especially with nitric acid) itself but also its vapors were effective. The structure was revealed later but the appearance was plastic (3D) and the structure was very fine. The supramolecular structure of PP-PLLA blends differed with amount of each component. The ethylene-rubber phase came forth with increasing time of etching while PP spherulites disappeared in PP-EPR samples. CLSM was shown to be very good tool for observing supramolecular structure of studied samples with respect to SEM, which was proved to be entirely unsuitable for polyolefines. DSC is recommended to be performe before uncovering supramolecular structure for basic information about proportion of amorphous/crystalline phase and size of crystallites.

Les substrats métalliques ont été élaborés par des traitements thermomécaniques (laminages et recuits)sur des alliages Fe48%Ni et Ni5%W dans le but d'obtenir une hypertexture Cube indispensable à l'épitaxie de l'YBaCuO et du silicium dans la fabrication des câbles supraconducteurs et des cellules photovoltaïques à couches minces. Le rôle des éléments d'alliages tels que le soufre et le niobium sur la recristallisation et la formation de l'hypertexture Cube a été étudié dans le Fe48%Ni. Il a été montré que l'ajout du soufre favorise le développement de la texture Cube alors que l'ajout du niobium empêche la formation de la texture Cube. Le soufre se combine avec le Mn pour former les précipités MnS qui contribuent à l'augmentation de la différence d'énergie stockée entre l'orientation Cube et les orientations de laminage à froid (ECube/autres) quand le soufre augmente. Ce gap d'énergie explique explique l'acuité de la texture Cube avec l'ajout du soufre. Contrairement au soufre, l'ajout du niobium empêche la formation de la texture Cube, ce résultat s'explique par le fait la différence d'énergie stockée entre l'orientation Cube et les orientations de laminage diminue avec l'ajout du niobium. Pour expliques tous ces résultats, les analyses de microstructures et textures ont été faites par la technique EBSD et l'énergie stockée a été estimée à partir de la diffraction des neutrons sur les états déformés. / Substrate tapes were prepared by cold rolling and annealing of a Fe48%Ni and Ni5%W alloys in order to obtain Sharp Cube {100}<001> oriented substrate for photovoltaic thin films and superconductor cables in particular.The effect of microalloying elements sulfur and niobium on recrystallization and sharp Cube formation was studied in Fe48%Ni. It was shown that the addition of sulfur promotes the formation of Cube grains while the addition of niobium prevents the Cube grains formation. Regarding sulfur, it combines with manganese to form the MnS precipitates wich increases the stored energy difference between Cube component and others cold rolled components ECube/other when sulfur is added. This stored energy difference explains the sharpness of the Cube texture when sulfur is added. On the contrary the niobium microalloying element addition prevents the formation of Cube grains. This could be explained by the fact that stored energy of cold-rolled components decreases with the addition of niobium and thus decreases Cube grains fraction when niobium is added. In order to explain these results, the development of Cube texture during recrystallization has been investigated in detail by EBSD, furthermore, the effect of stored energy has been studied by carrying out neutron diffraction measurements on the deformed states.

Les raccords et connexions de caténaires ont un rôle important dans l’intégrité électrique et mécanique des caténaires. Ces pièces sont pour la plupart constituées d’un alliage de cuivre Cu-Ni-Si mis en forme par forgeage à chaud. Dans un contexte de fiabilisation du réseau ferroviaire, l’amélioration de leurs performances et de leurs procédés de fabrication est recherchée. Ces travaux se concentrent sur l’étape de mise en forme de ces pièces, et ont pour objectif de mieux comprendre les couplages matériaux-procédés des alliages Cu-Ni-Si.Une campagne expérimentale de compression est menée. Elle permet de développer un modèle rhéologique à base physique décrivant le comportement d’un alliage Cu-Ni-Si dans les gammes de température 800 - 950 °C et de vitesse de déformation 0.1 - 10 s−1. Il tient compte des phénomènes d’adoucissement par restauration et recristallisation dynamiques. Le modèle ainsi créé est ensuite implémenté dans un logiciel éléments finis.Un nouvel essai de formabilité est proposé. Il permet de reproduire les sollicitations mécaniques rencontrées lors d’un procédé de forgeage et vise à déterminer les conditions limites de formabilité d’un matériau. La mise en forme d’une pièce de référence est étudiée par le biais de cet essai. Sa caractérisation précise en matière de champ de déformations et de contraintes permet d’identifier les paramètres expérimentaux. Les résultats identifient une température limite de forgeage en-deçà de laquelle des fissures sont prédites dans la pièce. / Contact wire splices play a leading role in the electric and mechanical integrity of the catenary systems. Most of these parts are made of Cu-Ni-Si alloys and manufactured by a hot forging process. With the aim of increasing the reliability of the railway network, the improvement of their performances and their manufacturing processes is sought. This work is focused on the forming stage of these parts and aims to give a better understanding of the materials-processes couplings for the Cu-Ni-Si alloys.An experimental campaign including compression tests is conducted. It allows the development of a physically-based model to describe the rheological behavior of a Cu-Ni-Si alloy deformed in the temperature range between 800 °C and 950 °C, at strain rates in the range of 0.1 - 10 s−1. It takes into account the work-softening implied by dynamic recovery and dynamic recrystallization. The model thus created is then implemented in a finite element software.A new workability test is proposed. Its purpose is to reproduce the mechanical conditions encountered in a forming process and to determine the critical conditions of workability related to a material. The forging process of a reference part is studied by means of this test. An accurate characterization of the process in terms of strain magnitude and stress field allows the identification of the experimental parameters. The results lead to the identification of a limit temperature of workability below which fractures are predicted in the part.

In numerous Gram-negative and Gram-positive bacteria as well as in Archaea SL proteins form the outermost layer of the cell envelope. SL (glyco)monomers self-assemble with oblique (p2), tetragonal (p4), or hexagonal (p3, p6) symmetries [12]. SL subunits interact with each other and with the underlying cell surface by relatively weak non-covalent forces such as hydrogen-bonds, ionic bonds, salt-bridges or hydrophobic interactions. This makes them easy to isolate by applying chaotropic agents like urea and guanidine hydrochloride (GuHCl), chelating chemicals, or by changing the pH of the environment [10]. Upon dialysis in an ambient buffer monomers recrystallize into regular arrays that possess the forms of flat sheets, open ended cylinders, or spheres on solid substrates, at air-water intefaces and on lipid films, making them appealing for nanobiotechnological applications [3, 18]. The aim of this study was to investigate the structure, thermal stability, in vivo self-assembly process, recrystallization and metallization of three different recombinant SL proteins (SslA-eGFP, mSbsC-eGFP and S13240-eGFP) expressed in yeast S. cerevisiae BY4741 which could be further used in nanobiotechnological applications. In order to fulfill this aim, I investigated the in vivo expression of SL proteins (SslA, SbsC, S13240) tagged with eGFP (SL-eGFP) in the yeast S. cerevisiae BY4141. First, I characterized the heterologous expression of SL fusion constructs with growth and fluorescence measurements combined with Western blot analyses. Fluorescence microscopy investigations of overnight grown cultures showed that SslA-eGFP fusion protein was expressed as fluorescent patches, mSbsC-eGFP as tubular networks, and S13240-eGFP as hollow-like fibrillar network structures, while eGFP did not show any distinct structure Thermal stability of in vivo expressed SL-eGFP fusion proteins were investigated by fluorescence microscopy and immunodetection. In vivo self-assembly kinetics during mitosis and meiosis was the second main issue. In parallel, association of in vivo mSbsC-eGFP structures with the cellular components was of interest. A network of tubular structures in the cytosol of the transformed yeast cells that did not colocalize with microtubules or the actin cytoskeleton was observed. Time-resolved analysis of the formation of these structures during vegetative growth and sporulation was investigated by live fluorescence microscopy. While in meiosis ascospores seemed to receive assembled structures from the diploid cells, during mitosis surface layer structures were formed de novo in the buds. Surface layer assembly always started with the appearance of a dot-like structure in the cytoplasm, suggesting a single nucleation point. In order to get these in vivo SL assemblies stably outside the cells (in situ), cell distruption experiments were conducted. The tubular structures formed by the protein in vivo were retained upon bursting the cells by osmotic shock; however their average length was decreased. During dialysis, monomers obtained by treatment with chaotropic agents recrystallized again to form tube-like structures. This process was strictly dependent on calcium ions, with an optimal concentration of 10 mM. Further increase of the Ca2+ concentration resulted in multiple non-productive nucleation points. It was further shown that the lengths of the S-layer assemblies increased with time and could be controlled by pH. After 48 hours the average length at pH 9.0 was 4.13 µm compared to 2.69 µm at pH 5.5. Successful chemical deposition of platinum indicates the potential of recrystallized mSbsC-eGFP structures for nanobiotechnological applications. For example, such metalized protein nanotubes could be used in conductive nanocircuit technologies as nanowires.

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第20703号 / 工博第4400号 / 新制||工||1684(附属図書館) / 京都大学大学院工学研究科材料工学専攻 / (主査)教授 辻 伸泰, 教授 乾 晴行, 教授 安田 秀幸 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

Given the rising demand for diverse cell types in regenerative and transfusion medicines, such as human induced pluripotent stem cell-derived neurons (iPSC-Ns), human T/chimeric antigen receptor (CAR) T cells, and human natural killer (NK) cells, the ability to cryopreserve cells has become increasingly important. In regenerative medicine, iPSC-Ns are powerful tools for treating and modelling neurodegenerative diseases. Moreover, transplants/transfusions of T/CAR T cells or NK cells offer promising treatment for numerous types of tumors, such as leukemia and multiple myeloma. Cryopreservation of cells at sub-zero temperatures (-80 to -196 °C) allows for the development of master cell banks that can be used for clinical applications. Conventional cryoprotective agents (CPAs), such as dimethylsulfoxide (DMSO) and glycerol, are utilized to protect cells from cryoinjuries associated with the freezing process. However, the use of high concentrations of DMSO (i.e., 10 to 20%) has been shown to be accompanied with toxic effects on patients receiving cell therapies if it is not removed or diluted prior to transfusion. Moreover, DMSO does not prevent the occurrence of the cryoinjury associated with ice recrystallization, which is one of the major causes of cell death/damage during cryopreservation. As a result, there is a surge of attention toward developing new non-toxic cryo-additives that inhibit ice recrystallization during cryopreservation to permit future advancement in regenerative and transfusion medicines. Moreover, the use of ice recrystallization inhibitors (IRIs) as novel CPAs has become a promising strategy to improve cell viability and function post-thaw. The Ben laboratory heavily invested in synthesizing several classes of carbohydrate-based small molecule IRIs (i.e., O-linked alkyl and aryl glycosides, and N-aryl-D-gluconamides), and studying the correlation between their IRI activity and molecular properties, such as polar surface area to molecular surface area (PSA/MSA) ratio. Moreover, compounds that belong to the O-linked aryl glycosides and N-aryl-D-gluconamides classes of IRIs have been shown to enhance the viability and functionality of red blood cells (RBCs), hematopoietic stem cells (HSCs), and induced pluripotent stem cells (iPSCs) after thawing. Part of the research presented throughout this thesis focuses on structure-activity relationship (SAR) studies of alkyl pyranoses with modified alkyl chain lengths to explore any correlations between the IRI activity and the net polarity (i.e., PSA/MSA ratio) of the IRI candidates. O- and C-linked alkyl pyranose derivatives with different alkyl chain lengths were synthesized and their IRI activity was assessed using the modified splat cooling assay. While the IRI activity of the O- and C-linked alkyl glucosides did differ as the length of the alkyl chain increased, no correlation between the PSA/MSA ratios and their IRI activity was observed. In addition, this work allowed for investigation into the effect of the type of the glycosidic bond (i.e., C-O and C-C bonds) at the anomeric position, on the IRI activity of the different compounds. The O-linked alkyl glucosides appeared to be more IRI active than the C-linked compounds, suggesting the nature of the glycosidic bond is important for IRI activity. The second part of the research presented in this thesis focuses on examining the potential for IRIs to cryopreserve iPSC-Ns, T/CAR T cells, and NK cells. 2-fluorophenyl-D-gluconamides (2FA), which is one of the most active IRIs from the N-aryl-Dgluconamides, has shown promising results in maintaining a high number of viable and functional HSCs and iPSCs post-thaw, and therefore it was employed in the cryopreservation protocol of iPSC-Ns, human-derived T/CAR T cells, and human-derived NK cells. The efficacy of the cryopreservation protocol being constructed was evaluated by assessing the post-thaw viability and recovery rate, as well as the functionality of iPSCNs, T/CAR T cells, and NK cells post-thaw. These studies showed that protecting against ice recrystallization during cryopreservation with IRIs increases the number of viable and functional iPSC-Ns, and T/CAR T cells. It was also observed that employing IRI technology in the cryopreservation protocol of NK cells does not compromise their functionality compared to fresh, non-frozen NK cells. Overall, inhibition of ice recrystallization using IRIs appeared to enhance the cryopreservation outcomes of the different cell types, which will allow for the development of off-the-shelf cell therapy products and improvement of the delivery of efficacious cell products to clinics and hospitals.

Цель работы – изучить изменения текстуры и структуры в медной капиллярной трубке в процессе ее получения. Теоретическое исследование проводилось методом анализа литературных источников. Прикладное исследование – методами растровой электронной микроскопии, а также дюрометрическим анализом. По результатам исследования выявлено изменение структуры и текстуры в изделии. / The aim of the work is to study the changes in the texture and structure in the copper capillary tube during its production. The theoretical study was carried out by analyzing literary sources. Applied research – by scanning electron microscopy, as well as by durometric analysis. The results of the study revealed a change in the structure and texture of the product.