京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22832号 / 医博第4671号 / 新制||医||1047(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 林 康紀, 教授 安達 泰治, 教授 岩田 想 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Experimental design is a practice operating at the limit between craftmanship, art and technology. In collaboration with the studio breadeEscalope (Vienna, Austria) this project oscillates between research and production to study the link between industrial and experimental approach to design.  Hitherto initiated from an impulse or a problem requiring solution, the action of designing suggest a product or a service to fulfill needs. In industrial design, the creative process follows contextually anchored steps to frame, define, develop and deliver. For the sake of a pragmatic answer, the delivery is centred around the user and its needs, often leading to an opaque finished product.  Home’s basic, the coat rack was selected to support this study as a concrete product base for trend and consumer behavior analysis. From shared ornamental and functional object placed in the entrance of houses, the coat rack previously “hatstand”, is today an unnoticed domestic object. The final product was hand manufactured with recycled and raw component, allowing transparency on the production process and used material. Stacking up coats on the produced rack create a private space for the user to sit in, questioning the relationship between user and product (ownership, trust, value). Based on architectural concept and design theory, the product aims to bridge creative and technical disciplines to suggest a functional yet questionable product as dialogue starter.  How sustainable are the objects that surround us every day? What is the story of the product? With those question in head the experimental design thinking studio breadedEscalope based in Vienna (Austria), offers an alternative approach to design. Always starting projects with the production process as a story of formations, offering unique and playful outcome to the act of designing. / Experimentell design är en praxis som fungerar på gränsen mellan hantverk, konst och teknik. I samarbete med studio breadeEscalope (Wien, Österrike) pendlar detta projekt mellan forskning och produktion för att studera sambandet mellan industriell och experimentell strategi för design.  Hittills initierad från en impuls eller ett problem som kräver lösning, föreslår åtgärden att designa en produkt eller en tjänst för att uppfylla behoven. Inom industriell design följer den kreativa processen kontextuellt förankrade steg för att rama in, definiera, utveckla och leverera. För ett pragmatiskt svar är leveransen centrerad kring användaren och dess behov, vilket ofta leder till en ogenomskinlig färdig produkt.  Hemmets grund, klädhänget valdes för att stödja denna studie som en konkret produktbas för trend- och konsumentbeteendeanalys. Från delat prydnads- och funktionellt föremål placerat i ingången till hus är klädhänget tidigare "hatstand", idag ett obemärkt inhemskt föremål. Slutprodukten tillverkades för hand med återvunnen och rå komponent, vilket gav insyn i produktionsprocessen och använt material. Att stapla upp rockar på det producerade racket skapar ett privat utrymme för användaren att sitta i och ifrågasätter förhållandet mellan användare och produkt (ägande, förtroende, värde). Baserat på arkitektoniska koncept och designteori syftar produkten till att överbrygga kreativa och tekniska discipliner för att föreslå en funktionell men tvivelaktig produkt som dialogstartare.  Hur hållbara är de objekt som omger oss varje dag? Vad är historien om produkten? Med dessa frågor i huvudet erbjuder den experimentella designtänkande studion breadedEscalope baserad i Wien (Österrike), ett alternativt tillvägagångssätt för design. Alltid starta projekt med produktionsprocessen som en berättelse om formationer, erbjuder unikt och lekfullt resultat till handlingen att designa.

The Tobacco mosaic virus (TMV) is a simple and frequently used model virus which has been studied already more over than 130 years. Due to the intensive study of this virus the details of its infectious cycle, genomic information and also the structure of the created viral particle as well as the mechanism of its creation are known today. The process of encapsidation (viral particle formation) is sufficiently described in the in vitro conditions. In the in vitro conditions the origin of assembly (OAS) was also described. The OAS was identified in the coding sequence of the gene for the movement protein (MP). The importance of replication centers (replication factories) has also been supposed. The aim of the diploma thesis was to study the specificity of the interaction of RNA and coat protein in the process of the particle assembly taking place directly inside the plants. The experiments were performed to verify the necessity of presence of OAS sequence in process of initiation of viral encapsidation. The effect of the cell compartmentation on this process has also been studied. Based on several viral systems (the Tobacco mosaic virus, the Potato virus X, the Bean yellow dwarf virus and Cowpea mosaic virus) gene constructs were created. These constructs enables to study this idea at the molecular...

In mammalian cells, homeostasis and fate during development relies on the proper transport of membrane-bound cargoes to their designated cellular locations. The hetero-tetrameric adaptor protein complexes (APs) are required for sorting and concentration of cargo at donor membranes, a crucial step during targeted transport. AP2, which functions at the plasma membrane during clathrin-mediated endocytosis, is well characterized. In contrast, AP1 a clathrin adaptor mediating the delivery of lysosomal hydrolases via mannose 6-phosphate receptors (MPRs) and AP3 an adaptor ensuring the proper targeting of lysosomal membrane protein are difficult to study by classic cell biology tools. To gain new insights on these APs, our lab has previously designed an in vitro system. Reconstituted liposomes were modified with small peptides mimicking the cytosolic domains of bona fide cargoes for AP1 and AP3 respectively and thereby enabling the selective recruitment of these APs and the identification of the interacting protein network. In the study at hand we utilize above-described liposomes to generate supported lipid bilayers and Giant Unilamellar Vesicles (GUVs), large-scale membrane systems suited for analysis by fluorescence microscopy. By using cytosol containing fluorescently-tagged subunits, we visualized clathrin coats on artificial membranes under near physiological conditions for the first time. Moreover, we demonstrated clathrin-independent recruitment of AP3 coats on respective GUVs. Presence of active ARF1 was sufficient for the selective assembly of AP1-dependent clathrin coats and AP3 coats on GUVs. By using dye-conjugated ARF1, we show that ARF1 colocalized with AP3 coats on GUVs and that increased association of ARF1 with GUVs coincided with AP1-dependent clathrin coats. Our previous study identified members of the septin family together with AP3 coats on liposomes. Here we show on GUVs, that active ARF1 stimulated the assembly of septin7 filaments, which may constrain the size and mobility of AP3 coats on the surface. Subsequent cell biology studies in HeLa cells linked septins to actin fibers on which they may control mobility of AP3-coated endosomes and thus their maturation. An actin nucleation complex, based on CYFIP1 was identified together with AP1 on liposomes before. Here we show on GUVs, that CYFIP1 is recruited on the surface surrounding clathrin coats. Upon supply of ATP, sustained actin polymerization generated a thick shell of actin on the GUV surface. The force generated by actin assembly lead to formation of long tubular protrusions, which projected from the GUV surface and were decorated with clathrin coats. Thereby the GUV model illustrated a possible mechanism for tubular carriers formation. The importance of CYFIP1-reliant actin polymerization for the generation of MPR-positive tubules at the trans-Golgi network (TGN) of HeLa cells was subsequently demonstrated in our lab. The notion that tubulation of artificial membranes could be triggered by actin polymerization allowed us to perform a comparative mass spectrometry screen. By comparing the abundance of proteins on liposomes under conditions promoting or inhibiting actin polymerization, candidates possibly involved in stabilization, elongation or fission of membrane tubules could be identified. Among the proteins enriched under conditions promoting tubulation, we identified type I phosphatidylinositol-4-phosphate 5-kinases. Their presence suggested an involvement of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) in tubule formation. By cell biology studies in HeLa we show, that down regulation of these enzymes altered the dynamics of fluorescently-tagged MPRs, illustrating the importance of locally confined PI(4,5)P2 synthesis during formation of coated carriers at the TGN. Bin–Amphiphysin–Rvs (BAR) domains are known to sense membrane curvature and induce membrane tubulation. Among various BAR domain proteins, Arfaptin2 was enriched under conditions allowing tubulation of liposomes. By microscopy studies on HeLa cells we show, that Arfaptin2 as well as its close paralog Arfaptin1 were present on AP1-coated MPR tubules emerging from the TGN. We further show, that tubule fission occurred at regions were Arfaptin1 is concentrated and that simultaneous down regulation of both Arfaptins lead to increased number and length of MPR tubules. Since fission of coated transport intermediates at the TGN is poorly understood, our findings contribute a valuable component towards a model describing the entire biogenesis of coated post-Golgi carriers. In conclusion, combining artificial membrane systems and cell biology studies allowed us to propose new models for formation as wall as for fission of AP1-coated transport intermediates at the TGN. Further we gained new insights on AP3 coats and the possible involvement of septin filaments in AP3-dependent endosomal maturation.

Carbon fibre reinforced composites are known for their high specific strength-to-weight ratio and are of great interest to the aerospace industry. Incorporating these materials into the fuselage, like in Boeing's 787 "Dreamliner", offers considerable weight reduction which increases flying efficiency, and reduces the cost of flying. In flight, aircraft are often subject to lightning strikes which, in the case of composites, can result in localized melting given the high resistive nature of the material. Aerospace carbon fibre composites often incorporate a metallic mesh or foil within the composite layers to dissipate the electrical charge through the large aircraft. The damage to the aircraft is minimized but not always eliminated. This research aims to elaborate a practical technique to deposit thin layers of conductive material on the surface of aerospace grade composites. Using Cold Gas Dynamic Spray (CGDS), such coatings could be used to repair damaged components. An experimental research approach was used to develop metallic coated composites. Using the CGDS equipment of Centerline (SST-P), specific parameters (such as gas temperature and stagnation pressure) were determined for each type of metallic coating (tin-based & copper-based). The use of bond coats was explored in order to attain the desired coatings. Once optimized, these coatings were evaluated with respect to their corrosive, adhesive, and electrical properties following industry standards.

No description available.

Markéta Hrdličková, Ennoblement and it's process in Czech lands during the reign of emperor Leopold I., based on acts of Bohemian Court Chancellery Abstract This thesis is based on the research of files of Bohemian Court Chancellery during the reign of emperor Leopold I. (1657-1705). It resolves about the emperor Leopold and the events and affairs of his time, and those which preceded it and had a significant influence on the reasonings mentioned in the files and acts of ennoblement. It also contains chapters about nobility of the Lands of the Bohemian Crown, the process of ennoblements and its files. Addendum contains a list of ennoblement acts in alphabetical order regarding both persons and families.

Indiana University-Purdue University Indianapolis (IUPUI) / The single cell gel electrophoresis assay (comet assay) is a useful tool for monitoring individuals who may be at risk of DNA damage and the ensuing process of carcinogenesis or other disease states. Leukocytes in blood samples provide a means of obtaining cells for use in the comet assay. However instances may arise when samples must be stored for later analysis. The present study investigated the effects of storage conditions on DNA damage in the form of strand breaks and oxidized bases in rat and human leukocytes using the comet assay. Whole blood and buffy coat samples were stored at room temperature or 4ºC for 1, 2, 24, and 48 hours or cryopreserved at -80ºC for 1 day and 1, 2, 3, and 4 weeks. The results show that the time of storage is limited if the whole blood or buffy coat samples are stored at room temperature or 4ºC. However, if cryopreserved using glycerol or DMSO as the cryoprotectant, the samples may be stored for at least 4 weeks without DNA strand breaks or oxidative damage deviating significantly from the fresh samples.

Indiana University-Purdue University Indianapolis (IUPUI) / Automotive paints have used an ultraviolet (UV) absorbing clear coat system for nearly thirty years. These clear coats have become of forensic interest when comparing paint transfers and paint samples from suspect vehicles. Clear coat samples and their ultraviolet absorbers are not typically examined or characterized using Raman spectroscopy or microspectrophotometry (MSP), however some past research has been done using MSP. Chemometric methods are also not typically used for this characterization. In this study, Raman and MSP spectra were collected from the clear coats of 245 American and Australian automobiles. Chemometric analysis was subsequently performed on the measurements. Sample preparation was simple and involved peeling the clear coat layer and placing the peel on a foil-covered microscope slide for Raman or a quartz slide with no cover slip for MSP. Agglomerative hierarchical clustering suggested three classes of spectra, and principal component analysis confirmed this. Factor loadings for the Raman data illustrated that much of the variance between spectra came from specific regions (400 – 465 cm-1, 600 – 660 cm-1, 820 – 885 cm-1, 950 – 1050 cm-1, 1740 – 1780 cm-1, and 1865 – 1900 cm-1). For MSP, the regions of highest variance were between 230 – 270 nm and 290 – 370 nm. Discriminant analysis showed that the three classes were well-differentiated with a cross-validation accuracy of 92.92% for Raman and 91.98% for MSP. Analysis of variance attributed differentiability of the classes to the regions between 400 – 430 cm-1, 615 – 640 cm-1, 825 – 880 cm-1, 1760 – 1780 cm-1, and 1860 – 1900 cm-1 for Raman spectroscopy. For MSP, these regions were between 240 – 285 nm and 300 – 370 nm. External validation results were poor due to excessively noisy spectra, with a prediction accuracy of 51.72% for Raman and 50.00% for MSP. No correlation was found between the make, model, and year of the vehicles using either method of analysis.

This dissertation reports on research related to ultrafiltration (UF) membranes in drinking water applications. A pilot-scale investigation identified seasonal surface water quality impacts on UF performance and resulted in the development of a dynamic chemically enhanced backwash protocol for fouling management. Subsequent analysis of UF process data revealed limitations with the use of specific flux, transmembrane pressure (TMP), and other normalization techniques for assessing UF process fouling. A new TMP balance approach is presented that identifies the pressure contribution of membrane fouling and structural changes, enables direct process performance comparisons at different operating fluxes, and distinguishes between physically and chemically unresolved fouling. In addition to the TMP balance, a five component optimization approach is presented for the systematic improvement of UF processes on the basis of TMP variations. Terms are defined for assessing process event performance, a new process utilization term is presented to benchmark UF productivity, and new measures for evaluating maintenance procedures are discussed. Using these tools, a correlation between process utilization and operating pressures was established and a sustainable process utilization of 93.5% was achieved. UF process capabilities may be further enhanced by pre-coating media onto the membrane surface. Silicon dioxide (SiO2) and powdered activated carbon (PAC) are evaluated as precoating materials, and the applicability of the TMP balance for assessing pre-coated membrane performance is demonstrated. The first use of SiO2 as a support layer for PAC in a membrane pre-coating application is presented at the laboratory-scale. SiO2-PAC pre-coatings successfully reduced physically unresolved fouling and enhanced UF membrane organics removal capabilities.