京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20977号 / 医博第4323号 / 新制||医||1026(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 小柳 義夫, 教授 小川 誠司, 教授 朝長 啓造 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

The study of signaling pathways that modulate multidrug resistance The theme of cancer cell resistance to anti-cancer drugs including the common mechanisms of resistance development and the theory of cancer stem cells was introduced in the Introduction to the doctoral thesis. The theoretical part was focused more deeply on the two topics - the role of ATP-binding cassette (ABC) transport proteins and chromosomal abnormalities in the development of cancer chemoresistance. The possible therapeutic potential for the treatment of cancer was stressed for both topics. The Results were composed of the commentaries on the five published works, which the author of the thesis conducted as the main author. The first work brought the evidence supporting the hypothesis of the existence of ABC gene expression profiles (signatures), which are common to multiple types of tumors and are associated with significant clinical consequences. These general ABC gene expression profiles could possibly form a new hallmark of cancer. The second work studied more closely a group of acute myeloid leukemia patients, who did not achieve complete cytogenetic remission after two attempts to maintain remission of the malignant disease. The new entity, consisting of patients with the translocation t(2;11)(p21;q23) without the...

Syftet med mitt examensarbete är att göra en ABC-bok, ett läromedel för skriftspråkslärande och bokstavsbearbetning. Bokstavsbearbetningen är en del av skriftspråkslärandet och ska vara en del i elevernas vardag. Genom ABC-boken vill jag åskådliggöra hur man kan integrera olika uttrycksformer i undervisningen. Jag har studerat olika teorier om skriftspråkslärande, bokstavsbearbetning och estetiska läroprocesser. Tillsammans med teorier, samtal med lärare och med bakgrund i Lpo 94 har jag gjort en ABC-bok där jag har författat övningar, fotograferat föremål och gjort layouten. Min förhoppning med boken är att göra skriftspråkslärandet och bokstavsbearbetningen i skolår 1, lärorik på ett roligt och kreativt sätt. Resultatet finner jag tilltalande.

Heart disease is leading cause of death in the United States. High-density lipoprotein (HDL) levels are inversely correlated with the prevalence of coronary heart disease. The anti-atherogenic properties of HDL are associated with its role in the pathway of the reverse cholesterol transport, which removes cholesterol from peripheral cells for transport back to hepatocytes. The formation of HDL is facilitated by ATP cassette transporter ABCA1 and apolipoprotein A-I, the major protein of HDL particles. However, the underlying molecular mechanism behind the biogenesis of HDL is not well understood. To provide further understanding of this mechanism, we developed two ABCA1 expression systems in both Sf9 insect cells and FreestyleTM 293-F human cells for functional and structural studies. We designed all constructs of ABCA1 to contain a C-terminal rho1d4 tag that bound to an affinity matrix of rho1d4 antibodies for successful purification. To reconstitute ABCA1 in a detergent-free environment that models the native membrane, we developed three reconstitution systems for ABCA1: saposin A nanodiscs, peptidisc, and amphipol A8-35. Biochemical and structural studies were carried out to understand the mechanism behind ABCA1’s function. We demonstrated a potential direct interaction of ABCA1 and apolipoprotein A-I with a pull-down experiment. Two cryo-electron microscopy data collections were obtained of ABCA1 in a detergent environment in the presence of ATP with the goal of determining the structure of ABCA1’s active state. We produced a 12 Å reconstruction of ABCA1 from this first data collection. This low-resolution structure confirmed the general structure that currently exists for ABCA1. Processing the data helped us streamline and troubleshoot the electron microscopy workflow pipeline for future data collections. Unfortunately, the second data collection had astigmatism issues that prevented particle alignment during data analysis. However, these data collections provided considerable insight into the ideal sample freezing and grid preparation conditions that affect data collection and data processing. More transmembrane protein structures are being solved each year but there remain many obstacles and challenges in ABCA1 purification and grid preparation that affect the ability to perform functional studies and high-resolution structure determination. Our developmental work has helped move forward our biochemical understanding of ABCA1 to achieve these aims. The more that is learned about this important membrane protein the more likely it is that future consistent production of ABCA1 will be accomplished to answer the questions of how ABCA1 mediates the formation of HDL particles.

No description available.

Syftet i den här studien är att undersöka vad ABC-klubben läseböcker på A-nivå (från årkurs 1 till 3) har för progression. För att uppfylla syftet har studien använt sig av både kvantitativa och kvalitativa metoder. Det görs genom att fokusera på ord och meningar i de valda kapitlen i varje bok med hjälp av läsbarhetsindex (LIX-värde). Den kvantitativa metoden kompletterats med en textanalys på meningsbyggnad för att se hur meningarna i varje bok utvecklas. Förhållande mellan text och bild har också analyserats med fokus på vilken funktion de två har och hur de samspelar med varandra. Dessutom har huvudkaraktärerna i läsebokserien analyserats för att finna tecken på utvecklingen över tid. Analysresultatet visar att det finns progression både i och mellan de tre böckerna. Enligt LIX-värdet har bok 1A tydligast progression medan bok 2A har mindre och 3A har minst. Samspelet mellan text och bild har en omvänd funktion. I början är det bilden som bär ansvar för att driva berättelsen framåt sedan är det texten som tar över ansvaret och kan stå på sina egna ben. Progressionen sker även när det gäller huvudkaraktärerna. Genom att undersöka berättelsen så kunde huvudkaraktärernas progression delas in i två teman, emotionell utveckling och utveckling av hur karaktärerna löser komplexa problem. Avslutningsvis diskuteras analysresultatet utifrån didaktiska perspektiv.

This thesis deals with control of stock in an inventory, focusing on inventory placement. The purpose of this thesis is to reduce the transport distance within the main stock house while gathering inventory. This will be achieved by reconstructing the inventory placement in consideration with how frequently the inventories get picked and mass of the inventory. In particular, the literature and the data that is collected from the company´s business system have laid the foundation for the thesis. In general, interviews and observations also contributed to the data collection. To fulfill the aim and to produce arbitrary results, two issues have been developed regarding which attributes that should determine the position of the inventory in the stock house and how to obtain a more effective inventory structure? The authors have jointly produced a result of suggestions for future inventory placement in terms of picking frequency and weight. Initially a situation analysis was conducted to identify known problems with the inventory´s placement and storage systems. The problems that were identified were that the inventory placement has no consideration regarding picking frequency. To determine the most frequent picked inventory an ABC analysis was conducted. All of the inventories were spread out throughout the whole stock house. To take in account, the additional criterion, which was weight, a multi-criteria analysis was performed in combination with the ABC analysis. The results of the combined analysis provided that the basis for drawing up concepts for future inventory placement. The proposal includes optimized inventory placements in different zones of the most frequently picked inventory with weight as an additional criterion. / Arbetet behandlar området lagerstyrning med inriktning på optimering av artikelplacering. Syftet med arbetet är att minska transportsträckor inom centrallagret, detta genom en omstrukturerad artikelplacering med hänsyn till plockfrekvens och tyngd. I synnerhet har litteratur samt insamlad data från företagets affärssystem lagt grunden för arbetet. I allmänhet har intervjuer och observationer också bidragit till datainsamling. För att uppfylla arbetets syfte och ta fram ett godtyckligt resultat har två frågeställningar tagits fram angående vilka attribut som bör avgöra en artikels position i lagret samt hur en effektivare lagerstruktur kan erhållas. Författarna har gemensamt arbetat fram ett resultat i form av ett förslag för framtida artikelplacering sett till plockfrekvens och tyngd. Inledningsvis genomfördes en nulägesanalys för att identifiera kända problem med artikelplacering och lagersystem. De problem som identifierades var att artikelplaceringen sker utan underlag för plockfrekvens. För att utröna de mest frekventa artiklarna genomfördes en ABC-analys. Samtliga artiklar lokaliserades utspridda över hela lagret. För att ta hänsyn till ytterligare ett kriterium, vilket var tyngd, genomfördes multikriterieanalys i kombination med ABC-analys. Resultatet av den kombinerade analysen gav underlag för att ta fram förslag för framtida artikelplacering. Förslaget innehåller optimerade artikelplaceringar inom olika zoner för de mest plockfrekventa artiklarna med tyngd som ytterligare kriterium.

Breast cancer is a leading cause of death among women in many countries. In the treatment of the breast cancer cytotoxic drugs (chemotherapy) are often used. Interindividual differences of drug response are an important cause of treatment failures. Bioavailability also depends on a major extent from the expression and activity of drug transport across biomembranes. In particular efflux transporters of the ATP-binding cassette family such as ABCB1, ABCC1 and ABCC2 have been identified as major determinants of chemoresistance in tumor cells. It was hypothesized that variance in the gene expression of membrane transporters and their genetic variance could explain at least in part interindividual differences of pharmacokinetics and clinical outcome of a variety of drugs. This thesis focuses on the functional significance of gene expression of ABCB1, ABCC1 and ABCC2 and single nucleotide polymorphisms in ABCC1 gene.

Estrogen signalling pathway plays crucial role in carcinogenesis of breast cancer. Estrogen receptor (ER) is a prototypical hormone receptor that upon binding its ligand, estradiol, translocates into the nucleus and turns on target genes related to cellular proliferation and survival. Although estrogen signalling physiologically supports normal breast tissue development, deregulations of this pathway contribute to development of breast tumours that are estrogen receptor dependent. One of the main obstacles in breast cancer treatment is acquired resistance to common anticancer drugs also known as multidrug resistance (MDR). The switch between chemotherapy responsive to chemotherapy resistant cell phenotype is usually accompanied by increased expression of ABC transporters, special membrane proteins responsible for export of various kinds of commonly used anticancer drugs from the intracellular to extracellular space and is also linked to the existence of cancer stem cells (CSCs). ABC transporters can not only export chemotherapeutic drugs but may modulate tumour microenvironment through the transport of endogenous intracellular substrates such as leukotrienes (LTs), sphingolipids and prostaglandins PGs). This function may also play important role in carcinogenesis. The aim of the thesis was to...

Polar auxin transport provides essential directional and positional information for many developmental processes in plants. At the cellular level, it is realized by both passive diffusion and the active transport through the membrane proteins - AUX1/LAXes, PINs and ABCBs. The aim of this thesis was to characterize the role of ABCB1, ABCB4 and ABCB19 proteins in polar auxin transport using transformed tobacco BY-2 cell lines. It was shown that the plasma membrane (PM) localization of the ABCB1, 4 and 19 is not polar. The ABCB4 was also more stable on PM after the treatment with auxin influx inhibitors; making use of ABCB4-cell line helped to uncover new characteristics of markers of endocytosis - the FM- dyes. The induction of ABCB19 has led to a decrease in 3 H-NAA accumulation with characteristic auxin starvation phenotype, similar to PIN7 overexpressing cell line, that could be rescued in case of PIN7 cell line by application of the auxin efflux inhibitor NPA; however, the accumulation of auxin in ABCB19-overexpressing cell line was less sensitive to NPA and the rescue of the auxin starvation phenotype was ineffective. Importantly, unique property of the ABCB4 was demonstrated: It displayed dual, auxin-concentration-dependent auxin transport activity in Arabidopsis roots, tobacco BY-2 and yeast cells. The...