京都大学 / 0048 / 新制・課程博士 / 博士(人間・環境学) / 甲第21862号 / 人博第891号 / 新制||人||213(附属図書館) / 2018||人博||891(吉田南総合図書館) / 京都大学大学院人間・環境学研究科共生人間学専攻 / (主査)教授 林 達也, 教授 石原 昭彦, 教授 久代 恵介 / 学位規則第4条第1項該当 / Doctor of Human and Environmental Studies / Kyoto University / DGAM

Submitted by ANA KARLA PEREIRA RODRIGUES (anakarla_@hotmail.com) on 2017-08-01T13:42:01Z No. of bitstreams: 1 arquivototal.pdf: 3885187 bytes, checksum: 43a97495626de95064ede3adad23bcd9 (MD5) / Made available in DSpace on 2017-08-01T13:42:01Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3885187 bytes, checksum: 43a97495626de95064ede3adad23bcd9 (MD5) Previous issue date: 2016-02-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In this work we performed ωB97XD, CAM-B3LYP, CASSCF, MR-CISD, MRCISD+ Q and NEVPT2 calculations. The basis sets aug-cc-pVDZ and cc-pVTZ were used to assess the stability of certain neutral and protonated N-nitrosamines in the ground and excited states. In addition, we analyzed the effect of methylation on proton transfer. The initial proposal was to characterize the stationary points and the potential energy curves in the ground and excited state, in order to suggest a strategy that results in lower computational demand to be applied in larger systems (cyclic and bicyclic N-nitrosamines). The results indicate that the methodology MR-CISD/cc-pVTZ//MR-CISD/aug-cc-pVDZ may be appropriate to study the N-nitrosamines. The used functionals provided good descriptions to vertical excitation energy, but were not capable to represent the states correctly to dissociation curves. MR-CID+Q/cc-pVTZ results for the system (CH3)2NNO protonated indicate that protonation is more favorable for NDMA_1A, increasing energy in 17.5 kcal mol-1, and that the methylation significantly alter the intramolecular proton transfer barriers. With the aim to find a methodology with lower computational cost, use a reference wave function CASSCF (12, 9) at the level NEVPT2/cc-pVTZ was used study the neutral NDMA and results were consistent with both the theoretical and experimental parameters in the literature. For studies involving larger systems, for example, cyclic and bicyclic N-nitrosamines, NEVPT2 may be used, since it is completely paralyzed, and involves the use of a CASSCF larger than CAS (14, 10), which is a fundamental characteristic to study larger systems. / Neste trabalho foram realizados cálculos ωB97XD, CAM-B3LYP, CASSCF, MRCISD, MR-CISD+Q e NEVPT2. Os conjuntos de base aug-cc-pVDZ e cc-pVTZ foram usados para avaliar a estabilidade de algumas N-nitrosaminas neutras e protonadas nos estados fundamental e excitado. Além disso, foi analisado o efeito da metilação nas transferências de próton. A proposta inicial foi caracterizar os pontos estacionários, as curvas de energia potencial no estado fundamental e excitado, com a finalidade de sugerir uma estratégia que resulte em menor demanda computacional para ser aplicada em sistemas maiores (N-nitrosaminas cíclicas e bicíclicas). Os resultados indicaram que a metodologia MR-CISD/cc-pVTZ//MR-CISD/aug-cc-pVDZ pode ser adequada para estudar as N-nitrosaminas. Os funcionais utilizados forneceram boas descrições para a energia de excitação vertical, mas para as curvas de dissociação não foram capazes de representar os estados corretamente. Os resultados com MR-CISD+Q/cc-pVTZ para o sistema (CH3)2NNO protonado indicam que a protonação é mais favorável para NDMA_1A, aumentando a energia em 17,5 kcal mol-1, e que a metilação altera significativamente as barreiras de transferência de próton intramoleculares. Com a proposta de encontrar uma metodologia com menor custo computacional, utilizou-se uma função de onda de referência CASSCF (12, 9) a nível NEVPT2/cc-pVTZ para estudar a NDMA neutra e os resultados foram compatíveis com os parâmetros experimentais e teóricos existentes na literatura. Para estudos envolvendo sistemas maiores, por exemplo, N-nitrosaminas cíclicas e bicíclicas, NEVPT2 poderá ser usado, uma vez que está totalmente paralelizado e comporta a utilização de um CASSCF maior que CAS (14, 10), característica fundamental para estudar sistemas maiores.

I used the C. elegans genetic model to examine the role of ceramide biosynthesis (sphingolipid pathway) and iron regulation and found that each process impacts germline development and function. Using a sphingolipid specific antibody mAb15B4, I found that sphingolipids are associated with germ granules (P granules) within C. elegans and zebrafish; thus, suggesting conservation of macromolecules associated with germ granules. Phenotype analysis of ceramide biosynthesis mutants in C. elegans revealed that this pathway is essential for normal germline function in the aging adult hermaphrodite; specifically, precocious germline senescence was observed. Furthermore, I found that disruption of ceramide biosynthesis, via the hyl-2 deletion mutation, negatively impacts mAb15B4 localization at the P granules. Through genetic suppression analysis, I determined that insulin signaling and lipid biosynthesis can modulate the mAb15B4 localization to P granules. Additional, phenotype analysis showed that ceramide biosynthesis dysfunction decreased fecundity, and led to germline structure defects and uterine tumors. Through suppression analysis, I determined that modulation of the insulin signaling pathway suppressed the precocious germline senescence due to ceramide biosynthesis dysfunction. Since the presence of uterine tumors is associated with reproductive senescence I concluded that ceramide biosynthesis has a role in germline maintenance in the aging of the germline (germline senescence). The other important fate of a germ cell is programmed cell death. Apoptosis, which occurs through a highly conserved molecular pathway, is a normal component of growth and homeostatic processes. I used C. elegans to gain a greater understanding of the cisd gene function. The C. elegans genome has three previously uncharacterized cisd genes which code for CISD-1 (homology to vertebrate mitoNEET/CISD1 and NAF-1/CISD2) and CISD-3.1 and CISD-3.2 (homology to vertebrate Miner2/CISD3). I determined that independent disruption of the cisd genes resulted in a significant increase in the number of cell corpses within the adult hermaphrodite germline. Genetic analysis was used to examine the dysfunction of cisd-1 relative to the cell death canonical pathway genes. The increased gamete cell death in the cisd-1 hermaphrodite is suppressed by the ced-9 (Bcl-2 homolog) gain-of-function and requires functional CED-3 (caspase) and CED-4 (APAF). Additionally, the increased germ cell programmed cell death is facilitated by the pro-apoptotic, CED-9-binding protein, CED-13. Further analysis of the cisd gene family members show that cisd-3.2 dysfunction leads to germline defects and reproductive dysfunction, suggesting defects in germline stem cell proliferation. Expression analysis using the cisd promoters to drive fluorescent protein reporters showed that the cisd gene family is expressed in various tissues including the germline; fusion protein analysis showed that CISD-3 is mitochondrial localized. I propose that cisd-3.2 germline defects are a result of abnormal mitochondrial function. Combined, this work is significant because it identifies sphingolipids as a new component of embryonic P granules, a role for ceramide biosynthesis in reproductive senescence, and places the cisd gene family members as regulators of physiological germline programmed cell death acting through CED-13 and the core apoptotic machinery. Furthermore, it is the first study to show that a CISD3 protein family member is required for normal germline function. These findings support the idea that ceramide biosynthesis and iron regulation are core components in germline development and function.

NEET proteins belong to a unique class of [2Fe-2S] cluster proteins that have been shown to participate in various biological processes such as regulating iron, reactive oxygen species and apoptosis within the cell and are localized to the mitochondria. Disruption of the mitochondrial NEET proteins are associated with different human diseases such as obesity, neurodegeneration, cancer and diabetes. In humans, a missense mutation in the CISD2 gene results in a heritable multisystem disorder termed Wolfram syndrome 2 (WFS2), a disease which displays an early onset of juvenile diabetes and various neuropsychiatric disorders. The C. elegans genome contains three previously uncharacterized cisd genes: cisd-1, which has homology to the human CISD1 and CISD2, and cisd-3.1 and cisd-3.2, both of which have homology to the human CISD3. Disruption of the cisd-3 gene(s) function results in mis-regulation of proteostasis in the mitochondria, whereas cisd-1 and cisd-3.1 disruption impacts proteostasis in the endoplasmic reticulum. Reduction of cisd-3.2 gene function also leads to a developmental delay in C. elegans. A knockout mutation of the cisd-3.2(pn68) gene function results in various germline defects including delayed development progression and morphological defects. Furthermore, I show the cisd gene(s) and protein expression profiles is present relative to sex, tissue type and developmental stages. This work is significant because it provides further insight of the essential role of CISD-3 relative to C. elegans. Furthermore, my studies can contribute to new genetic discoveries that will widen the scientific research relative to NEET protein family studies.

Magister Psychologiae - MPsych / South Africa is a country with one of the highest crime rates in the world, resulting in much violence and trauma. Trauma debriefing models are used with groups and individuals in an attempt to facilitate the trauma recovery process and prevent the development of post-traumatic stress disorder (PTSD). There has been much debate on the efficacy of trauma debriefing over the past two decades with few qualitative studies exploring the utility of debriefing. A phenomenological methodology was used in this study, with the aim of understanding how participants experienced the process of trauma debriefing and what aspects were perceived as helpful or unhelpful. Semi-structured in- depth interviews were conducted with six adults who received debriefing following a single traumatic event, by volunteer lay counsellors from the Community Intervention Centre (CIC) based at the Milnerton Medi-Clinic.

This study examined the validity of the South African Police Service (SAPS) multiple stressor intervention. The multiple stressor was developed for members of their specialised units to address Post Traumatic Stress Disorder (PTSD). Unfortunately, the SAPS multiple stressor intervention lacks scientific evidence to support its possible effectiveness in addressing PTSD. In the current study a deployment and intervention group was compared which employed a pre- test post-test design. The Davidson Trauma Scale and the Revised Impact of Event Scale measured PTSD globally, but also the PTSD dimensions of Intrusion, Avoidance/Numbing, and Hyperarousal. The Wilcoxon signed rank test results indicated that the intervention and deployment was both effective in addressing PTSD although the intervention group revealed the greatest improvement in their overall PTSD scores. The intervention group made significant progress in dealing with all three PTSD symptoms while the deployment group made less progress with their Intrusion and Avoidance/Numbing symptoms, but made significant progress with their Hyperarousal symptoms. The Mann-Whitney u test revealed no significant differences between the post intervention test scores of the two groups, either globally or on the three PTSD dimensions. It appears that deployment was just as effective as the multiple stressor intervention in addressing PTSD. / Psychology / M.A. (Psychology)

This study examined the validity of the South African Police Service (SAPS) multiple stressor intervention. The multiple stressor was developed for members of their specialised units to address Post Traumatic Stress Disorder (PTSD). Unfortunately, the SAPS multiple stressor intervention lacks scientific evidence to support its possible effectiveness in addressing PTSD. In the current study a deployment and intervention group was compared which employed a pre- test post-test design. The Davidson Trauma Scale and the Revised Impact of Event Scale measured PTSD globally, but also the PTSD dimensions of Intrusion, Avoidance/Numbing, and Hyperarousal. The Wilcoxon signed rank test results indicated that the intervention and deployment was both effective in addressing PTSD although the intervention group revealed the greatest improvement in their overall PTSD scores. The intervention group made significant progress in dealing with all three PTSD symptoms while the deployment group made less progress with their Intrusion and Avoidance/Numbing symptoms, but made significant progress with their Hyperarousal symptoms. The Mann-Whitney u test revealed no significant differences between the post intervention test scores of the two groups, either globally or on the three PTSD dimensions. It appears that deployment was just as effective as the multiple stressor intervention in addressing PTSD. / Psychology / M.A. (Psychology)