Please read the abstract in the section 00front of this document / Dissertation (MSc (Reproductive Biology))--University of Pretoria, 2005. / Urology / unrestricted

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21638号 / 医博第4444号 / 新制||医||1034(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 木村 剛, 教授 浅野 雅秀, 教授 山下 潤 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

yes / Type 2 diabetes (T2DM) promotes premature atherosclerosis and inferior prognosis after arterial reconstruction. Vascular smooth muscle cells (SMC) respond to patho/physiological stimuli, switching between quiescent contractile and activated synthetic phenotypes under the control of microRNAs (miRs) that regulate multiple genes critical to SMC plasticity. The importance of miRs to SMC function specifically in T2DM is unknown. This study was performed to evaluate phenotype and function in SMC cultured from non-diabetic and T2DM patients, to explore any aberrancies and investigate underlying mechanisms. Saphenous vein SMC cultured from T2DM patients (T2DM-SMC) exhibited increased spread cell area, disorganised cytoskeleton and impaired proliferation relative to cells from non-diabetic patients (ND-SMC), accompanied by a persistent, selective up-regulation of miR-143 and miR-145. Transfection of premiR-143/145 into ND-SMC induced morphological and functional characteristics similar to native T2DM-SMC; modulating miR-143/145 targets Kruppel-like factor 4, alpha smooth muscle actin and myosin VI. Conversely, transfection of antimiR-143/145 into T2DM-SMC conferred characteristics of the ND phenotype. Exposure of ND-SMC to transforming growth factor beta (TGFβ) induced a diabetes-like phenotype; elevated miR-143/145, increased cell area and reduced proliferation. Furthermore, these effects were dependent on miR-143/145. In conclusion, aberrant expression of miR-143/145 induces a distinct saphenous vein SMC phenotype that may contribute to vascular complications in patients with T2DM, and is potentially amenable to therapeutic manipulation.

yes / Type 2 diabetes mellitus prevalence is growing globally, and the leading cause of mortality in these patients is cardiovascular disease. Epigenetic mechanisms such as microRNAs (miRs) and DNA methylation may contribute to complications of type 2 diabetes mellitus. We discovered an aberrant type 2 diabetes mellitus–smooth muscle cell phenotype driven by persistent up-regulation of miR-145. This study aimed to determine whether elevated expression was due to changes in methylation at the miR-145 promoter. Smooth muscle cells were cultured from saphenous veins of 22 non-diabetic and 22 type 2 diabetes mellitus donors. DNA was extracted, bisulphite treated and pyrosequencing used to interrogate methylation at 11 CpG sites within the miR-145 promoter. Inter-patient variation was high irrespective of type 2 diabetes mellitus. Differential methylation trends were apparent between non-diabetic and type 2 diabetes mellitus–smooth muscle cells at most sites but were not statistically significant. Methylation at CpGs −112 and −106 was consistently lower than all other sites explored in non-diabetic and type 2 diabetes mellitus–smooth muscle cells. Finally, miR-145 expression per se was not correlated with methylation levels observed at any site. The persistent up-regulation of miR- 145 observed in type 2 diabetes mellitus–smooth muscle cells is not related to methylation at the miR-145 promoter. Crucially, miR-145 methylation is highly variable between patients, serving as a cautionary note for future studies of this region in primary human cell types.

Yes / Increased cardiovascular morbidity and mortality in individuals with type 2 diabetes (T2DM) is a significant clinical problem. Despite advancements in achieving good glycaemic control, this patient population remains susceptible to macrovascular complications. We previously discovered that vascular smooth muscle cells (SMC) cultured from T2DM patients exhibit persistent phenotypic aberrancies distinct from those of individuals without a diagnosis of T2DM. Notably, persistently elevated expression levels of microRNA-145 co-exist with characteristics consistent with aging, DNA damage and senescence. We hypothesised that increased expression of microRNA-145 plays a functional role in DNA damage signalling and subsequent cellular senescence specifically in SMC cultured from the vasculature of T2DM patients. In this study, markers of DNA damage and senescence were unambiguously and permanently elevated in native T2DM versus non-diabetic (ND)-SMC. Exposure of ND cells to the DNA-damaging agent etoposide inflicted a senescent phenotype, increased expression of apical kinases of the DNA damage pathway and elevated expression levels of microRNA-145. Overexpression of microRNA-145 in ND-SMC revealed evidence of functional links between them; notably increased secretion of senescence-associated cytokines and chronic activation of stress-activated intracellular signalling pathways, particularly the mitogen-activated protein kinase, p38a. Exposure to conditioned media from microRNA-145 overexpressing cells resulted in chronic p38a signalling in naïve cells, evidencing a paracrine induction and reinforcement of cell senescence. We conclude that targeting of microRNA-145 may provide a route to novel interventions to eliminate DNA-damaged and senescent cells in the vasculature and to this end further detailed studies are warranted.

University of Macau / Faculty of Arts and Humanities. / Department of Chinese

Made available in DSpace on 2016-04-26T20:25:19Z (GMT). No. of bitstreams: 1 Dissertacao Roberta Maria Rangel.pdf: 1342468 bytes, checksum: 1d2c3d9591757daabd96e130d1de500c (MD5) Previous issue date: 2005-05-30 / ABSTRACT The present dissertation investigated the limits of the text of the article 145, II, of the Federal Constitution, in order to narrow the criteria contained there, which are supplied to the political people for the construction of its rule matrices of tributary incidence of police and services taxes. After a brief chronology of the evolution of the tax legal theories in the positive law, Brazilian jurisprudence and doctrine, one concludes that the theory that prevails between us and that is deduced of the article 145, II, of the Federal Constitution is the one of the binding of the incidence hypothesis. This is the theory that supplies the three criteria for the construction of the rule matrices of the taxes by contributors: the criterion of the binding to a state activity; the criterion of the reference to the contributor and the criterion of the direct reference to the contributor. Here it is the positive limitation of the article 145, II, of the Federal Constitution. However, one realized that the same article has a negative limitation, when it does not indicate the content and the form of organization of the State activities contained there: the exercise of police power and the supply or provision of public services. Therefore, tracing a parallel with the norms of tributary legislation competence, from the analysis of the logical structure of the juridical norm, one extracts of the Federal Constitution the norms of administrative and legislative competence, which confer permission to the political people to organize the supply of their administrative activities specially the public services of varied forms, which will withdraw or attract the rule of the remuneration by tax, inserted in the article 145, II, of the Federal Constitution. / A presente dissertação investigou os limites constitucionais da redação do artigo 145, II, da Constituição Federal, buscando precisar os critérios ali contidos e que são fornecidos às pessoas políticas para construção de suas regras-matrizes de incidência tributária das taxas de polícia e das taxas de serviço. Traçou um breve histórico da evolução, no direito positivo, jurisprudência e doutrina brasileiras, das teorias jurídicas da taxa, onde concluiu que a teoria que hoje prevalece entre nós e que é deduzida do artigo 145, II, da Constituição Federal a teoria da vinculabilidade da hipótese de incidência fornece três critérios para construção das regras-matrizes das taxas pelos entes tributantes: o critério da vinculabilidade a uma atividade estatal; o critério da referibilidade ao contribuinte e o critério da referibilidade direta ao contribuinte. Aqui está a limitação positiva do artigo 145, II, da Constituição Federal. Entretanto, percebeu-se que o mesmo artigo tem uma limitação negativa, quando não indica o conteúdo e a forma de organização das atividades estatais ali constantes: exercício do poder de polícia e a prestação ou disponibilização de serviços públicos. Desta forma, traçando-se um paralelo com as normas de competência legislativa tributária, a partir da análise da estrutura lógica da norma jurídica, extraiu-se da Constituição Federal as normas de competência legislativa administrativa, as quais conferem permissão às pessoas políticas para que organizem a prestação de suas atividades administrativas em especial os serviços públicos de variadas formas, o que afastará ou atrairá a regra da remuneração por taxa inserida no artigo 145, II, da Constituição Federal.

梁德華. / "2007年8月" / "附錄1: 《漢紀》、《漢書》、《史記》三書比勘表"--P. 1-873 (2nd group). / 論文(哲學碩士)--香港中文大學, 2007. / 參考文獻(leaves 197-209). / "2007 nian 8 yue" / "Fu lu 1: 'Han ji', 'Han shu', 'Shi ji' san shu bi kan biao"--P. 1-873 (2nd group). / Abstract also in English. / Liang Dehua. / Can kao wen xian (leaves 197-209). / Lun wen (zhe xue shuo shi)--Xianggang Zhong wen da xue, 2007. / Chapter 第一章： --- 引言 --- p.1 / Chapter 第二章： --- 荀悦生平及其學術背景 --- p.2 / Chapter 第一節： --- 荀悦的生平 --- p.2 / Chapter 第二節： --- 《漢紀》、《申鑒》引書硏究 --- p.5 / Chapter 第三節： --- 「荀悦曰」與儒家典籍之關係 --- p.35 / Chapter 第三章： --- 《漢紀》用《史記》考 --- p.48 / Chapter 第一節： --- 前人硏究《漢紀》史源槪況 --- p.48 / Chapter 第二節： --- 《漢紀》用《史記》集證 --- p.53 / Chapter i. --- 《漢紀》用《史記》官名、數字及人物例 --- p.54 / Chapter ii. --- 《漢紀》據《史記》補充史料及文句例 --- p.61 / Chapter iii. --- 《漢紀》直用《史記》文辭例 --- p.69 / Chapter iv. --- 《漢紀》據《史記》改寫文辭例 --- p.77 / Chapter V. --- 《漢紀》與徐廣所見古本《史記》相合例 --- p.88 / Chapter 第三節： --- 結語 --- p.91 / Chapter 第四章： --- 《漢紀》用《漢書》考 --- p.100 / Chapter 第一節： --- 化紀傳體爲編年體 --- p.100 / Chapter 第二節： --- 通比其事 --- p.104 / Chapter 第三節： --- 《漢紀》、《資治通鑑》比事硏究 --- p.106 / Chapter i. --- 取材問題. --- p.109 / Chapter a. --- 《漢紀》弓懺《漢´書Ø志》、〈表〉之文句不見於《通鑑》例 --- p.110 / Chapter b. --- 《漢紀》引錄《漢´書Ø傳》之內容不見於《通鑑》例 --- p.118 / Chapter ii. --- 紀年問題 --- p.125 / Chapter a. --- 論《漢紀》記事以人物爲中心而《通鑑》以時間爲中心 --- p.127 / Chapter b. --- 論《漢紀》記事連類相及而《通鑑》繫年於確實年份 --- p.131 / Chapter 第四節： --- 《漢紀》整理《漢書》之方法 --- p.133 / Chapter i. --- 《漢紀》迻錄《漢書》原文例 --- p.133 / Chapter ii. --- 《漢紀》以通假、同義、近義詞改易《漢書》文句例 --- p.135 / Chapter iii. --- 《漢紀》删削《漢書》人物對白例 --- p.139 / Chapter iv. --- 《漢紀》删削《漢書》情節例 --- p.145 / Chapter v. --- 《漢紀》因删削而失《漢書》原文文意例 --- p.152 / Chapter vi. --- 《漢紀》槪括《漢書》文句例 --- p.156 / Chapter vii. --- 《漢紀》修飾《漢書》文辭例 --- p.160 / Chapter viii. --- 《漢紀》改寫《漢書》文句例 --- p.165 / Chapter ix. --- 《漢紀》補充《漢書》例 --- p.172 / Chapter X. --- 《漢紀》倂合《漢書》不同篇章例 --- p.178 / Chapter Xi. --- 《漢紀》據其他古籍改易《漢書》文句例 --- p.186 / Chapter 第四節： --- 結語 --- p.192 / Chapter 第五章： --- 結論 --- p.195 / 參考書目 --- p.197 / Chapter 附錄一： --- 《漢紀》、《漢書》、《史記》三書比勘表 --- p.1-837

The chemokine receptor CCR5 has been implicated in the pathogenesis of prostate cancer (PCa). A novel series of piperazine derivatives have been designed and synthesized as CCR5 antagonists and their activity as inhibitors of PCa cell lines proliferation was explored. A lead compound has been identified which induced 100% inhibition of PCa cell proliferation at micromolar concentrations. Anibamine, the only natural product CCR5 antagonist, was also examined for its anti-proliferative activity and was found to inhibit proliferation of PCa cells at micromolar concentrations as well. The expression of RANTES mRNA was observed in DU-145, M12 and P69 cells via RT-PCR, while the expression of CCR5 mRNA was observed only in M12 cells. A CHO-CCR5 stable cell line was prepared for the CCR5 ligand competition binding assays. Both anibamine and the newly identified lead compound will serve as leads in the development of novel CCR5 antagonists as anti-prostate cancer agents.

Les récoltes de céréales sont souvent contaminées par des moisissures qui se développent pendant la récolte et l’entreposage et produisent des métabolites secondaires appelés mycotoxines. Le porc est reconnu pour être sensible au déoxynivalénol (DON). L’infection virale la plus importante chez le porc est causée par le virus du syndrome reproducteur et respiratoire porcin (VSRRP). Celui-ci provoque un syndrome grippal et des troubles de reproduction. L’objectif du présent projet était de déterminer l'effet in vitro de DON sur la réplication du VSRRP dans de lignées cellulaires permissives, MARC-145 et PAM, et déterminer in vivo l'impact de DON dans des aliments naturellement contaminés sur l’infection au VSRRP chez le porcelet. Tout d’abord, les cellules ont été incubées avec des doses croissantes de DON et ont été infectées avec du VSRRP pour évaluer la viabilité et la mortalité cellulaire, la réplication virale et l’expression de cytokines. Les résultats ont montré que les concentrations de DON de 560ng/ml et plus affectaient significativement la survie des cellules MARC-145 et PAM infectées par le VSRRP. En revanche, il y avait une augmentation significative de la viabilité et une réduction de la mortalité cellulaire à des concentrations de DON de 140 à 280 ng/ml pour les cellules PAM et de 70 à 280 ng/ml pour les cellules MARC-145 avec une réduction de l'effet cytopathique provoqué parle VSRRP. Au niveau in vivo, 30 porcelets divisés en 3 groupes de 10 porcelets et nourris pendant 2 semaines avec 3 différentes diètes naturellement ont été contaminées avec DON (0; 2,5 et 3,5 mg/kg). Les porcelets ont été subdivisés en 6 groupes, 3 groupes de 6 porcelets et ont été exposés au DON pendant 2 semaines et infectés par voie intratrachéale et intramusculaire avec le virus. Les 3 autres groupes de 4 porcelets servaient de contrôle non infectés. Les signes cliniques ont été enregistrés pendant 21 jours. La virémie a été évaluée par PCR. À la fin de l’expérimentation, les porcelets ont été euthanasiés et les lésions pulmonaires ont été évaluées. Les résultats ont montré que l’ingestion de DON à 3,5 mg/kg a augmenté l’effet du VSRRP sur la sévérité des signes cliniques, les lésions pulmonaires et la mortalité. L’ingestion de DON à 2,5 mg/kg a entrainé une augmentation de la virémie au jour 3 après l’infection mais sans impact sur les signes cliniques et les lésions pulmonaires. Mot clés: DON, VSRRP, MARC-145, PAM, effet cytopathique, cytokines, PCR / Cereal crops are often contaminated with moulds that grow during harvest and storage and produce secondary metabolites called mycotoxins. Pig is known to be sensitive to deoxynivalenol (DON). On the other hand, infection by porcine reproductive and respiratory syndrome virus (PRRSV) causes a flu-like syndrome and reproductive disorders. The objectives of this project were to determine the in vitro effect of DON on the replication of PRRSV in permissive cell lines, MARC-145 and PAM and the in vivo impact of DON-naturally contaminated feed on PRRSV infection in piglets. Firstly, cells were incubated with gradually increasing doses of DON and were infected with PRRSV to evaluate cytopathic effect and to assess cell viability, virus replication and cytokine mRNA expression on infected and uninfected cells. Results showed that DON concentrations of 560 ng/ml and higher were significantly detrimental to the survival of MARC-145 cells infected with PRRSV. In contrast, there was a significant increase of cell viability and decreased of cell mortality at DON concentrations within 140 to 280 ng/ml for PAM cells and 70 to 280 ng/ml ranges for MARC-145 showing a reduced cytopathic effect (CPE) caused by PRRSV. In vivo study was carried out on 30 piglets divided into 3 groups of 10 piglets fed naturally contaminated diets with different levels of DON; 0, 2.5 and 3.5 mg/kg. After 2 weeks, pigs were further divided into 6 subgroups, 3 subgroups of 6 piglets were infected intra tracheally and intramuscularly with PRRSV. The other 3 subgroups of 4 piglets were used as uninfected controls. Clinical signs were recorded for 21 days post-infection (p.i.). Sera were evaluated for viremia by PCR. At the end of the experiment, piglets were euthanized and pulmonary lesions were evaluated. Results showed that ingestion of diet highly contaminated with DON at 3.5 mg/kg increased the effect of PRRSV infection on the severity of clinical signs, weight loss, lung lesions and mortality. Diet with DON at 2.5 mg/kg showed an increase of viremia at day 3 but had not significant impact on clinical signs and lung lesions. Keywords: DON, PRRSV, MARC-145, PAM, cytopathic effect, cytokines, PCR