京都大学 / 0048 / 新制・課程博士 / 博士(生命科学) / 甲第21224号 / 生博第393号 / 新制||生||52(附属図書館) / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 西田 栄介, 教授 米原 伸, 教授 上村 匡 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21655号 / 医博第4461号 / 新制||医||1035(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 長船 健二, 教授 江藤 浩之, 教授 篠原 隆司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Indiana University-Purdue University Indianapolis (IUPUI) / There is a lack of existing research that investigates the feasibility of using consumers to evaluate the fidelity of evidence-based practices, including supported employment which is an intervention that helps people with severe mental illnesses to obtain competitive employment. Fidelity refers to the extent that the SE program adheres to the Individual Placement and Support (IPS) model of supported employment. The present study was a concordance study that investigated whether or not consumers’ self reports of IPS fidelity information agreed with administrative charts and employment specialists. Additionally, it was hypothesized that consumers’ program satisfaction ratings would be positively correlated with their self reported IPS fidelity scores. An additional purpose of this study was to examine what types of supported employment fidelity items consumers were able to report on. Participants included a volunteer sample of 30 consumers and 5 employment specialists from one IPS program in Indiana. Consumers in the IPS program were interviewed by telephone using a survey that included questions related to their program’s fidelity as well as their satisfaction with the program. Questions were based off of items from the IPS Fidelity Scale and were categorized into the following subscales: work incentives counseling, job search, engagement, organization, staffing, and job support. Similar questions were asked in an employment specialist survey and a chart review. All three sources (consumers, charts, and employment specialists) indicated high IPS fidelity responses on the researcher developed surveys. However, there was a low level of agreement between the sources at both the subscale level and item level. Although there was an overall low level of agreement between sources, there were several items that had a moderate or higher degree of agreement. Additionally, the present study did find a positive correlation between the consumer fidelity score percentage and consumer IPS program satisfaction ratings, supporting the researcher’s hypothesis. Among the items that consumers had difficulty answering were several tapping program level policies such as zero exclusion. Reasons for the discrepancy in agreement between sources as well as clinical implications of the findings are discussed.

京都大学 / 新制・課程博士 / 博士(理学) / 甲第23748号 / 理博第4838号 / 新制||理||1692(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 今井 啓雄, 准教授 大石 高生, 教授 古市 剛史 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

A reprogramação nuclear de uma célula somática a um estado embrionário tem diversas aplicações, como pesquisas básicas na biologia do desenvolvimento, terapia celular, melhoramento genético em animais de produção e conservação de espécies. As principais técnicas utilizadas para a reprogramação nuclear são a transferência nuclear de células somáticas (TNCS) e a geração de células tronco pluripotente induzidas (iPS). Muitos trabalhos têm mostrado uma baixa eficiência no processo de reprogramação nuclear nas duas técnicas, além disso, modificações epigenéticas tem sido apontada como a principal barreira para uma reprogramação nuclear eficiente. Por esse motivo, medidas como a utilização de células menos diferenciadas e/ou alteração do perfil epigenético das células somáticas podem aumentar a eficiência destas técnicas. Por isso, o objetivo deste trabalho foi investigar a influência de marcas epigenéticas em células bovinas utilizadas na reprogramação nuclear mediada por TNCS ou superexpressão de genes relacionados a pluripotêcia (iPS). Para isso, utilizamos 3 abordagens. Primeiro, analisamos marcações epigenéticas relacionadas ao desenvolvimento embrionário e pluripotência (H3K9me2, H3K9me3, H3K9ac, 5mC e 5hmC) em diferentes tipos celulares, analisamos a expressão gênica de genes responsáveis por essas marcações em células de diferentes tecidos (ex. células tronco mesenquimais (MSC) e fibroblastos) e as utilizamos como doadoras de núcleo na TNCS. Na segunda e a terceira abordagem, utilizamos células com menores níveis de H3K9me2 para a geração de iPS e na TNCS, respectivamente. Além disso, por se mostrar eficiente na TNCS, analisamos o efeito da sincronização do ciclo celular por privação de soro fetal bovino (SFB) na geração de células iPS. Com o intuito de diminuir os níveis de H3K9me2, as células foram tratadas com UNC0638, um inibidor especifico das metiltransferases de histona G9a/GLP. Nossos resultados do primeiro experimento mostraram que as MSC podem ser utilizadas como doadoras de núcleo na TNCS, no entanto, mesmo com algumas diferenças na expressão gênica em relação aos fibroblastos, a produção de blastocistos não foi diferente entre as duas células. No segundo experimento, as células privadas de SFB geraram mais colônias que as células controle, enquanto que as células tratadas não apresentaram diferença. Por último, as células tratadas com o UNC0638 apresentaram um menor nível de metilação no DNA em zigotos em relação às células controle. Os resultados encontrados neste trabalho podem contribuir para o melhor entendimento dos mecanismos epigenéticos envolvidos na reprogramação nuclear de bovinos / Nuclear reprogramming of somatic cells to embryonic state has several aplications, such as basic research on developmental biology, cell therapy, genetic improvement in livestock animals and preservation of endangered species. The principal techniques utilized to achieve nuclear reprogramming are Somatic Cell Nuclear Transfer (SCNT) and induced pluripotency. Several works has reported low efficiency rates of nuclear reprogramming when these techniques are used to reprogram somatic cells. Moreover, epigenetic modifications acquired during development act as epigenetic barrier to the complete reprogramming process. For this reason, strategies such as use of less differentiated cells and/or modification of epigenetic profile of somatic cells might increase the efficiency these techniques. The objective of this work was investigate the influence of epigenetic marks in bovine cells utilized on nuclear reprogramming experiments mediated by SCNT or induced pluripotency. To investigate it, we used three approaches. First, we analyzed the epigenetic marks related to the embryonic development and pluripotency (e.g H3K9me2, H3K9me3, H3K9ac, 5mC and 5hmC), gene expression of genes involved in these epigenetic marks in different tissues (i.e. mesenchymal stem cells (MSC) and fibroblasts) and their use as nuclear donor cells on SCNT procedure. Regarding the second and the third approach, we utilized cells with reduced levels of H3K9me2 to generate iPS cells and cloned embryos, respectively. Furthermore, since serum starvation has been demonstrated increase SCNT developmental rates, we assessed the effect of cell cycle synchronization mediated by serum starvation on nuclear reprogramming using iPS cells. Aiming decrease the levels of H3K9me2, cells were treated with UNC0638, a chemical probe that works as a specific inhibitor of the histone methyltransferases G9a and its counterpartner GLP. Our results showed that MSC are suitable to be used as nuclear donors on SCNT procedures, however, in spite of differences on gene expression comparing with fibroblasts, the embryonic developmental rates were not improved. On the second experiment, cells privated of fetal calf serum produced more iPS cells colonies than control cells, whereas cells treated with UNC did not show differences when compared with untreated cells. Lastly, UNC treated donor cells treated produced cloned zygotes with lower levels of DNA methylation compared to zygotes derivated from untreated cells. The results presented here will contribute to the better understanding of the epigenetic mechanisms involved on bovine nuclear reprogramming

The diploma thesis deals with the issues of the spruce monocultures in the relation to the incidence of the bark beetle (Ips typographus) in the Šumava National park. The aim of the thesis is not only to unify and arrange the information, but also find out the level of knowledge about this topic among the pupils of the ninth grade of the basic school. The work also shows the level of knowledge of the pupils about the related problems and what are their opinions and suggested solutions. The thesis is divided into the theoretical and practical part. The theoretical part consists of eight chapters, that are devoted to the history of the Šumava forests, pine tree monocultures with the closer focus on the Norway spruce (Picea abies), the differences among the "natural" and "economical" forest and to the uniqueness of the Boubín primeval forest. In one of the chapters we get information about the huriccane Kyrill and its effects on the Šumava forests and followed by the information about the bark beetle calamities and about the bark beetle on its own. The last two chapters are devoted the solution of the current situation in the Šumava forests and their future perspectives. The practical part of the thesis contains mainly of the questionnaire survey proceeded among the pupils of the ninth grade of the...

Terpenes and phenols are of importance in conifer defense against insects and pathogens. Knowledge about tree chemical defense is vital for developing practical methods to maintain healthy forests. With the aims of characterizing the defensive chemical induction in Norway spruce Picea abies and demonstrating its ecological function to spruce bark beetle Ips typographus, we measured the terpenoid and phenolic content in the bark of mature Norway spruce trees suffering windstorm, inoculated with Ceratocystis polonica, or treated with methyl jasmonate (MeJA), and investigated the colonization and pheromone emission of I. typographus. All three stressors altered the chemical profile in the bark of Norway spruce. Trees damaged by windstorm had lower proportions of (+)-3-carene and two unidentified stilbenes, and a higher taxifolin glycoside content than trees without apparent windstorm damage; C. polonica inoculation induced extremely strong quantitative terpene accumulation in the wound reaction zone, but only increased the levels of (+)-3-carene, sabinene and terpinolene in the bark near the reaction zone; MeJA treatment generally elicited quantitative terpene accumulation, but the induction differed extensively between individual trees. In addition, logs from MeJA-treated tree showed much stronger wounding response compared to control logs. The chemical profile of Norway spruce affected the colonization and pheromone emission of I. typographus. In response to fungal inoculation, terpene present in the reaction zone inhibited the colonization of I. typographus in a dose-dependent manner. Host defense elicited by MeJA treatment reduced emissions of 2-methyl-3-buten-2-ol and (S)-cis-verbenol, the two key aggregation pheromone components of I. typographus, and altered the ratio between the two components. / QC 20110503

Innomhuspositioneringssystem kan med fördel användas i många olika tillämpningar, allt från sjukhus till shoppingcenter. Denna rapport behandlar olika tekniker och lösningar för att designa ett positioneringssystem. Rapporten tar även upp i detalj hur ett system kan konstrueras av ZigBee kombinerat med dödräkning.

農林水産研究情報センターで作成したPDFファイルを使用している。

A reprogramação nuclear de uma célula somática a um estado embrionário tem diversas aplicações, como pesquisas básicas na biologia do desenvolvimento, terapia celular, melhoramento genético em animais de produção e conservação de espécies. As principais técnicas utilizadas para a reprogramação nuclear são a transferência nuclear de células somáticas (TNCS) e a geração de células tronco pluripotente induzidas (iPS). Muitos trabalhos têm mostrado uma baixa eficiência no processo de reprogramação nuclear nas duas técnicas, além disso, modificações epigenéticas tem sido apontada como a principal barreira para uma reprogramação nuclear eficiente. Por esse motivo, medidas como a utilização de células menos diferenciadas e/ou alteração do perfil epigenético das células somáticas podem aumentar a eficiência destas técnicas. Por isso, o objetivo deste trabalho foi investigar a influência de marcas epigenéticas em células bovinas utilizadas na reprogramação nuclear mediada por TNCS ou superexpressão de genes relacionados a pluripotêcia (iPS). Para isso, utilizamos 3 abordagens. Primeiro, analisamos marcações epigenéticas relacionadas ao desenvolvimento embrionário e pluripotência (H3K9me2, H3K9me3, H3K9ac, 5mC e 5hmC) em diferentes tipos celulares, analisamos a expressão gênica de genes responsáveis por essas marcações em células de diferentes tecidos (ex. células tronco mesenquimais (MSC) e fibroblastos) e as utilizamos como doadoras de núcleo na TNCS. Na segunda e a terceira abordagem, utilizamos células com menores níveis de H3K9me2 para a geração de iPS e na TNCS, respectivamente. Além disso, por se mostrar eficiente na TNCS, analisamos o efeito da sincronização do ciclo celular por privação de soro fetal bovino (SFB) na geração de células iPS. Com o intuito de diminuir os níveis de H3K9me2, as células foram tratadas com UNC0638, um inibidor especifico das metiltransferases de histona G9a/GLP. Nossos resultados do primeiro experimento mostraram que as MSC podem ser utilizadas como doadoras de núcleo na TNCS, no entanto, mesmo com algumas diferenças na expressão gênica em relação aos fibroblastos, a produção de blastocistos não foi diferente entre as duas células. No segundo experimento, as células privadas de SFB geraram mais colônias que as células controle, enquanto que as células tratadas não apresentaram diferença. Por último, as células tratadas com o UNC0638 apresentaram um menor nível de metilação no DNA em zigotos em relação às células controle. Os resultados encontrados neste trabalho podem contribuir para o melhor entendimento dos mecanismos epigenéticos envolvidos na reprogramação nuclear de bovinos / Nuclear reprogramming of somatic cells to embryonic state has several aplications, such as basic research on developmental biology, cell therapy, genetic improvement in livestock animals and preservation of endangered species. The principal techniques utilized to achieve nuclear reprogramming are Somatic Cell Nuclear Transfer (SCNT) and induced pluripotency. Several works has reported low efficiency rates of nuclear reprogramming when these techniques are used to reprogram somatic cells. Moreover, epigenetic modifications acquired during development act as epigenetic barrier to the complete reprogramming process. For this reason, strategies such as use of less differentiated cells and/or modification of epigenetic profile of somatic cells might increase the efficiency these techniques. The objective of this work was investigate the influence of epigenetic marks in bovine cells utilized on nuclear reprogramming experiments mediated by SCNT or induced pluripotency. To investigate it, we used three approaches. First, we analyzed the epigenetic marks related to the embryonic development and pluripotency (e.g H3K9me2, H3K9me3, H3K9ac, 5mC and 5hmC), gene expression of genes involved in these epigenetic marks in different tissues (i.e. mesenchymal stem cells (MSC) and fibroblasts) and their use as nuclear donor cells on SCNT procedure. Regarding the second and the third approach, we utilized cells with reduced levels of H3K9me2 to generate iPS cells and cloned embryos, respectively. Furthermore, since serum starvation has been demonstrated increase SCNT developmental rates, we assessed the effect of cell cycle synchronization mediated by serum starvation on nuclear reprogramming using iPS cells. Aiming decrease the levels of H3K9me2, cells were treated with UNC0638, a chemical probe that works as a specific inhibitor of the histone methyltransferases G9a and its counterpartner GLP. Our results showed that MSC are suitable to be used as nuclear donors on SCNT procedures, however, in spite of differences on gene expression comparing with fibroblasts, the embryonic developmental rates were not improved. On the second experiment, cells privated of fetal calf serum produced more iPS cells colonies than control cells, whereas cells treated with UNC did not show differences when compared with untreated cells. Lastly, UNC treated donor cells treated produced cloned zygotes with lower levels of DNA methylation compared to zygotes derivated from untreated cells. The results presented here will contribute to the better understanding of the epigenetic mechanisms involved on bovine nuclear reprogramming