Background: Good social connections have been identified as a factor that may be associated with healthy cognitive function in later life. In line with cognitive reserve theory, good social connections may provide mental stimulation through complex interaction with others and hence build cognitive reserve and maintain healthy cognitive function. However, there is considerable inconsistency in findings reported by studies that examine this association. Inconsistency in findings may be attributed to the heterogeneity of concepts potentially associated with social connections and to the variation in approaches to measuring and defining these concepts. Aims: To assess the association between aspects of social connections and cognitive function in later life. This thesis introduces a novel element by considering the moderating role of cognitive reserve in this association. Method: A scoping review was conducted to establish which concepts are used within the literature to describe social connections and how these are measured and defined. Next, a systematic review and meta-analysis was conducted to identify evidence regarding the association between social isolation and cognitive function in published studies. Empirical work was conducted using cross-sectional and longitudinal data from the Cognitive Function and Ageing Study-Wales (CFAS-Wales) to determine the associations between social isolation, cognitive reserve, and cognitive function in healthy older people. Extending this approach further, these associations were examined in two groups potentially at risk of social isolation: older people with depression or anxiety and older people living alone. Finally, empirical work was completed using the Platform for Research Online to investigate Genetics and Cognition in Ageing (PROTECT) to assess how satisfaction with social contact may be associated with cognitive function compared to a structural measure of isolation. Results: A lack of social connections was associated with poor cognitive function in later life. For people with depression or anxiety, these associations may be better explained by mood-related symptoms than social connections. People who live alone in later life were at no greater risk of poor cognitive function compared to those living with others. Satisfaction with social contact was associated with poor cognitive function but a structural measure of social isolation was not. Conclusions: Social connections play an important role in building cognitive reserve and protecting people against poor cognitive function in later life. People who are vulnerable to social isolation have different needs to those who are less vulnerable. Satisfaction with social contact is often neglected in measures that assess structural aspects of social connections but may be a better predictor of cognitive function.

這項研究利用多角度方法，去探索簡短靜觀訓練對因被排斥而導致的心理困擾有否影響;更會將靜觀訓練與自律鬆弛法和控制組作比較。本實驗的參加者包括了161名香港中文大學的本科生和研究生。並用了Cyberball遊戲去模擬社會排斥。所有參加者被隨機分成三組: 1) 靜觀、 2) 自律鬆弛、 3) 控制組。我根據他們的生理反應測量 (如皮膚電導、心率), 混合動機任務，隱含測試(i.e. lexical decision task)，和自我評估去度量情緒困擾、互動模式、對自己和他人的態度、和在靜觀能力及態度的改變。 本實驗採用了生理反應測量 (如皮膚電導, 心率), 混合動機任務、隱含測試、和自我報告去評估幾方面的反應:情緒困擾、 與別人互動樣式、對自己和他人的態度、和靜觀能力的改變。結果顯示，靜觀組 的自尊心和存在意義感相比其他兩組高; 然而，靜觀組和自律鬆弛組之間沒有顯著差異。在靜觀的改變上，我們使用兩套問卷: （一）Southampton Mindfulness Questionnaire (SMQ), （二）Self-Other Four Immeasurables (SOFI) 。結果顯示，靜觀組在 SOFI Positive-Self 方面有明顯的提升; 而在SOFI Positive-Other能維持不變，相反其他兩組就有顯著的下降。可是，SMQ 和 SOFI問卷的其餘部分 (例如：SOFI Negative-Self, SOFI-Negative-Other)，沒有發現顯著的組間差異。除了採用兩份靜觀問卷外，我也使用了Self-Compassion Scale (SCS) 來測量慈心品性對結果的影響。結果顯示，性格較為靜觀及慈心的人, 沒有那麽容易受排斥而引至有不良影響。有趣的是，不同層次的靜觀性格和自我慈悲能有不同的訓練受益。例如，慈心的人在靜觀後會更為慷慨。然而，對於那些SOFI Negative-Other 低分的人，在靜觀後會減少對別人指責的傾向。可是，其他結果 (例如: 混合動機任務、隱含測試 ) ，卻找不到跨組的差異。在生理反應測量方面，結果便與預期不太一致: 靜觀組和自律鬆弛組的皮膚電導，在post-Cyberball 期間甚至比控制組為高; 而其他時段則找不到跨組間差異。在這篇文章的總結中，我也提到這個實驗之不足之處以及可以改善的方法。總括而言，雖然實驗結果所發現的成效不是太明顯，但我們不能忽視靜觀簡化版本的實用價值。尤其是對那些被邊緣化的人來說，簡化版本能使他們更容易參與訓練並從中受益。在文章的末尾，我會提及更具體的意義和建議，希望能對今後的研究有所影響。 / This study utilized the experimental multimodal approach to explore the effectiveness of brief mindfulness training in reducing the psychological distress induced by ostracism, comparing with brief relaxation training and no intervention control. Participants included 161 undergraduate and graduate students from CUHK. Cyberball game paradigm was used to simulate social exclusion. All participants were randomized into 3 groups: 1) meditation, 2) relaxation, 3) no intervention control. Physiological measures (i.e., skin conductance, heart rate), mixed-motive task, and implicit test (i.e. lexical decision task), and self-reports were used to assess emotional distress, interactions styles, attitudes toward self and others, and change in mindfulness. Results indicated that meditation group expressed higher level of self-esteem and sense of meaningful existence despite of social rejection in comparison with no intervention control. However, there was no significant difference between meditation and relaxation group. In terms of the mindfulness qualities as measured by Southampton Mindfulness Questionnaire (SMQ) and Self-Other Four Immeasurables (SOFI), meditation group reported greater enhancement in SOFI Positive-Self, while other two groups remain statistically unchanged. For SOFI Positive-Other, only meditation group remained as positive as before while other two groups dropped. However, the result from other mindfulness measurement (i.e. SMQ) and dimensions (i.e. SOFI Negative-Self, SOFI Negative-Other) revealed no significant group difference. In addition to the two mindfulness scales, the Self-Compassion Scale (SCS) was also used to capture the baseline self-compassion disposition. Correlational result showed that, being more mindful and self-compassionate was in general associated with feeling less threatened by the exclusion task. Interestingly, people of various levels of mindfulness and self-compassion could benefit from mindfulness training differently. For example, participants who were more self-compassionate would display more generous behavior only if they were in meditation group. However, for those who scored low on baseline SOFI Negative-Other, meditation reduced their tendency to blame others. Contrary to expectation, no statistically significant difference was found across conditions in implicit self-other attitudes and interaction styles. For physiological arousal, no significant cross group difference was identified with the exception of during the post-Cyberball period, in which skin conductance was significantly higher for meditation and relaxation groups relative to no-intervention control. . Limitations in the Cyberball manipulation and intervention implementation were noted, which may impact the study findings. In sum, despite the small effect observed in the mindfulness training condition, the practical value of an abbreviated mindfulness format cannot be ignored, particularly for the socially ostracized population whom may not have the luxury to experience the full-scale mindfulness training. More specific implications and suggestions for future research were discussed. / Detailed summary in vernacular field only. / Chan, Tsz Ying Amy. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 73-79). / Abstracts also in Chinese; appendix A includes Chinese. / Abstract --- p.iv / Introduction --- p.1 / Stigmatization and Ostracism --- p.1 / Ways to Combat Stigma --- p.2 / Existing Efforts --- p.2 / Mindfulness Mechanism in Reducing the Negative Impacts of Being Socially Excluded --- p.3 / What is Mindfulness? --- p.3 / Suggested Mechanism on How Mindfulness Deals with Ostracism --- p.7 / Mindfulness Based Approach --- p.9 / Evidence of Mindfulness on Stigma Reduction --- p.10 / Constraint Of Current Mindfulness Intervention --- p.11 / Objectives of This Study --- p.13 / Main Hypothesis --- p.13 / Hypothesis 1 --- p.14 / Hypothesis 2 --- p.15 / Hypothesis 3 --- p.15 / Hypothesis 4 --- p.15 / Hypothesis 5 --- p.16 / Method --- p.17 / Pilot --- p.17 / Participants --- p.17 / Measures --- p.17 / Screening Measure --- p.17 / Baseline Measure --- p.18 / Procedure --- p.22 / Result --- p.28 / Hypothesis 1.1: Meditation Group Has the Lowest Physiological Arousal (i.e. HR, SC) Followed by Relaxation and Control Groups during and after Cyberball Game --- p.29 / Hypothesis 1.2: Meditation Group was Least Threatened by the Social Exclusion Effect of the Cyberball Game, Followed by Relaxation and Control. --- p.31 / Hypothesis 1.3: Meditation Group had the Most Positive Attitude and Least Negative Attitude toward Self and Other, Followed by Relaxation and Control. --- p.32 / Hypothesis 2: Meditation Group was the Most Mindful, Measured by SMQ and SOFI, Followed by Relaxation and Control Groups. --- p.33 / Hypothesis 3 Meditation Group has the Most Positive Communication Style (3.1) and Give the Largest Amount of Points to Opponents (3.2), Followed by Relaxation and Control Groups --- p.36 / Hypothesis 4: Mindfulness Trait’s Interaction with Group Assignment in Affecting Outcomes --- p.36 / Correlational Analysis --- p.36 / Group X Baseline Mindfulness Interaction Effect --- p.39 / Hypothesis 5: Trait Self-Compassion’s Interaction with Group Assignment in Affecting Outcomes --- p.41 / Discussion 43 / Was the Brief Mindfulness Training Successful in Reducing the Negative Effect of Ostracism? --- p.43 / Decrease in Physiological Arousal --- p.43 / Stronger Resilience toward Ostracism? --- p.44 / Does Mindfulness Increase Selfless Behavior? --- p.45 / Implicit Attitudes toward Self and Others --- p.46 / Was Brief Mindfulness Session Successful in Improving Mindfulness? --- p.47 / How Does the Mindfulness and Self-Compassionate Predisposition Affect One’s Receptivity toward Brief Mindfulness Training? --- p.48 / Limitations --- p.50 / Implications and Conclusion --- p.52 / APPENDIX A --- p.57 / Instruction for meditation group --- p.57 / Instruction for relaxation group --- p.60 / APPENDIX B --- p.64 / DASS 21 --- p.64 / APPENDIX C --- p.65 / Self-Compassion Scale (26 Items) --- p.65 / APPENDIX D --- p.66 / Southampton mindfulness questionnaire (SMQ) 16 item --- p.66 / APPENDIX E --- p.67 / Self-Other Four Immeasurable (SOFI) --- p.67 / APPENDIX F --- p.68 / Assessment of manipulations, need satisfaction, and mood following ostracism (31 items) --- p.68 / APPENDIX G --- p.70 / Communication Checklist-Key --- p.70 / REFERENCES --- p.72

Ng Kai-on. / "July 2001." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (p. 162-180). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

彭華民. / 呈交日期: 2005年11月. / 論文(哲學博士)--香港中文大學, 2006. / 參考文獻(p. 246-261). / Cheng jiao ri qi: 2005 nian 11 yue. / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in Chinese and English. / Lun wen (zhe xue bo shi)--Xianggang Zhong wen da xue, 2006. / Can kao wen xian (p. 246-261). / Peng Huamin.

O coronavírus canino (CCoV) causa gastroenterite em cães jovens, podendo ser letal, sobretudo quando há coinfecção com parvovírus canino (CPV). Os objetivos do presente projeto foram investigar a presença de CCoV e CPV em amostras fecais de cães jovens; estudar a diversidade molecular das amostras de CCoV com base em sequenciamento parcial dos genes M, S, 3b e N, incluindo amostras vacinais, e estudar a evolução in vitro de CCoV em células A72 de fibroma canino. Foram detectados 40,17% (47/117) animais positivos para CCoV e 13,68% (16/117) para CPV. Estudos filogenéticos demonstraram que oito amostras foram classificadas como CCoV-II, vinte e cinco como CCoV-I. Análises para o gene M destacaram alta identidade de CCoV-I com amostras de coronavírus da peritonite infecciosa felina (PIF) e uma possível amostra pantrópica foi demonstrada pela análise do gene S. O gene do nucleocapsídeo de CCoV é altamente conservado entre os tipos I e tipo II, com uma resolução mais baixa em relação a árvores para os genes M e S. Amostras dos tipos I e II apresentam um polimorfismo baixo para o gene 3b, sem marcadores estáveis para diferenciação dos tipos de CCoV. Uma amostra de CCoV-II putativamente pantrópica foi isolada em células A72, resultando em efeito citopático no 5o dia da 5a passagem. No estudo evolutivo, a amostra vacinal CCV 1-71 e nove passagens desta em células A72 foram submetidas a amplificação parcial e clonagem molecular do gene S seguida de sequenciamento de DNA. Os resultados mostraram mutações não silenciosas, silenciosas e três deleções de aminoácidos, mas nenhuma mutação compartilhada entre as diversas passagens. Amostras vacinais de CCoV-II adaptadas em células podem ser altamente geneticamente estáveis após passagens em série em uma mesma linhagem celular, acumulando substituições de nucleotídeos principalmente sinônimas no gene S devido a relação célula - hospedeiro estável. Estes resultados de epidemiologia molecular e processos evolutivos de CCoV podem servir para uma melhor compreensão da virologia básica e ser base de dados para estudos em outros coronavírus / Canine coronavirus (CCoV) causes gastroenteritis in young dogs and can be lethal, especially when there is co-infection with canine parvovirus (CPV). The objectives of this project were to investigate the presence of CCoV and CPV in stool samples from young dogs, the molecular diversity of CCoV strains based on partial sequencing of genes M, S, N and 3b, and the in vitro evolution of CCoV in A72 canine fibroma. Of the fecal samples studied, 40.17% animals (47/117) were positive for CCoV and 13.68% (16/117) for CPV. Phylogenetic studies have shown that eight strains were CCoV-II and twenty five CCoV-I. Phylogenetic analysis for the M gene highlighted the high identity of CCoV-I strains with a feline coronavirus strain (FCoV) that causes feline infectious peritonitis (FIP). Further analysis based on the spike gene showed a putative pantropic CCoV strain (CCoV-II/dog50). CCoV nucleocapsid gene is highly conserved among type I and type II, with a lower resolution relative to trees based on M and S genes. CCoV Types I and II had a low polymorphism for 3b gene, without any stable markers to differentiate thee types. Regarding the virus isolation trial, a putative pantropic CCoV-II strain was successfully isolated in A72 cells from, resulting in cytopathic effect on the 5th day of the 5th passage. In the evolutionary study, the vaccine strain CCV 1-71 and nine passages of this strain in A72 were submitted to partial S gene amplification and molecular cloning followed by DNA sequencing. Missense, silent, and three amino acids deletions were found amongst diverse clones of each passage, but no mutation was repeatedly found among passages. Cell culture-adapted CCoV-II vaccine strains can be highly genetically stable upon serial passage in a same cell line, accumulating primarily synonymous nucleotide substitutions in the S gene due to a stable cell-host relationship. In short, all data gathering herein on molecular epidemiology and evolutionary processes of CCoV can serve for a better understanding of basic virology and as a basis for studies on other coronaviruses

Made available in DSpace on 2015-04-22T22:18:37Z (GMT). No. of bitstreams: 1 DISSERTACAO-ADRIANA BRITO BARATA CABRAL.PDF: 5620980 bytes, checksum: a1dccce5a16ec2c748cbf144f42356a8 (MD5) Previous issue date: 2010-09-16 / Fundação de Amparo à Pesquisa do Estado do Amazonas / This search sought to analyze the leprosy fighting policies deployed in Manaus in the period 1921, time of installation of the Amazon Rural Prophylaxis Service, agency responsible for the State Public Health which fought for the giving of the Paricatuba lands to the construction of the first leper hospital in Manaus. Paricatuba, that time, is an island where the only access is by boat, these leper hospital internal were isolated for their whole lives. The leper hospital worked since 1930, it s inauguration year by mi-1980, when was vandalized, according to information obtained, ordered by the State Government itself to oblige the internal to live at Colônia Antônio Aleixo , the new leper hospital. Paricatuba which had the name Vila Belizário Pena was reformulated to meet the precepts of hygiene defined by the Departamento Nacional de Saúde Pública. The final point of this search is the year of 1942 when new leprosy fighting policies are inserted in the city and there s a new activity to the Colônia Antônio Aleixo which was located in the rural Manaus and its access routes were going to land. This date of 1942 also marks the end of compulsory isolation and leprosy, according to doctors, could be treated as a common disease. / Esta pesquisa procurou analisar as políticas de combate à lepra implantadas na cidade de Manaus no período de 1921 momento da instalação do Serviço de Profilaxia Rural do Amazonas órgão responsável pela Saúde Pública do Estado e que lutou para que as terras de Paricatuba fossem cedidas para a construção da primeira leprosaria da cidade de Manaus. Paricatuba nesse momento é uma ilha onde seu acesso se dá somente por via fluvial, os internos desta leprosaria ficariam isolados por toda a vida, a leprosaria funcionou de 1930 ano de sua inauguração até meados de 1980, onde foi depredada segundo informações colhidas por ordem do próprio Governo do Estado para obrigar os internos a irem morar na Colônia Antônio Aleixo, o novo leprosário. Paricatuba cujo nome era Vila Belisário Penna fora reformulada para atender aos preceitos de higiene ditados pelo Departamento Nacional de Saúde Pública. O marco final da pesquisa é o ano de 1942 onde novas políticas de combate a lepra são inseridas na cidade e há uma nova movimentação para a construção do novo leprosário a Colônia Antônio Aleixo que ficava situado na parte rural da cidade de Manaus e sua via de acesso era por via terrestre. Esta data de 1942 também marca o final do isolamento compulsório e a lepra, segundo os médicos, poderia ser tratada como uma doença comum.

Chan, Pui Ting. / Thesis submitted in: May 2007. / On t.p. "alpha" appears as the Greek letter. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 110-118). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.xi / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Peroxisome proliferater-activated receptors (PPARs) --- p.1 / Chapter 1.1.1 --- What are PPARs? --- p.1 / Chapter 1.1.2 --- PPAR isoforms --- p.1 / Chapter 1.1.3 --- PPARα ligands --- p.2 / Chapter 1.2 --- Biological role of PPARα --- p.3 / Chapter 1.2.1 --- Lipid metabolism --- p.3 / Chapter 1.2.2 --- Glucose metabolism --- p.5 / Chapter 1.2.3 --- Oxidative stress and carcinogenesis --- p.6 / Chapter 1.3 --- Discovery of PPARα-regulated and starvation-induced gene (PPSIG) --- p.7 / Chapter 1.4 --- Objectives of the present study --- p.9 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.10 / Chapter 2.1 --- Cloning of PPSIG cDNA into a pCMV-Tag epitope tagging mammalian expression vector --- p.10 / Chapter 2.1.1 --- Materials --- p.10 / Chapter 2.1.2 --- Methods --- p.10 / Chapter 2.2 --- Transient transfection of PPSIG cDNA into CHO-K1 and AML-12 cells --- p.16 / Chapter 2.2.1 --- Cell culture and transfection --- p.16 / Chapter 2.2.1.1 --- Materials --- p.16 / Chapter 2.2.1.2 --- Methods --- p.19 / Chapter 2.2.2 --- Western blot analysis --- p.20 / Chapter 2.2.2.1 --- Materials --- p.20 / Chapter 2.2.2.2 --- Methods --- p.20 / Chapter 2.3 --- Stable transfection of PPSIG cDNA into CHO-K1 and AML-12 cells --- p.22 / Chapter 2.3.1 --- Linearization of the pCMVT4B-PPSIG construct --- p.22 / Chapter 2.3.1.1 --- Materials --- p.22 / Chapter 2.3.1.2 --- Methods --- p.22 / Chapter 2.3.2 --- Cell culture and stable transfection --- p.23 / Chapter 2.3.2.1 --- Materials --- p.23 / Chapter 2.3.2.2 --- Methods --- p.23 / Chapter 2.3.3 --- Selection of the G418-resistant clones --- p.26 / Chapter 2.3.3.1 --- Materials --- p.26 / Chapter 2.3.3.2 --- Methods --- p.29 / Chapter 2.3.4 --- Picking and expanding the G418-resistant clones --- p.30 / Chapter 2.3.4.1 --- Materials --- p.30 / Chapter 2.3.4.2 --- Methods --- p.30 / Chapter 2.3.5 --- Screening and confirmation of the stable transfectants --- p.31 / Chapter 2.3.5.1 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.31 / Chapter 2.3.5.1.1 --- Materials --- p.31 / Chapter 2.3.5.1.2 --- Methods --- p.31 / Chapter 2.3.5.2 --- Northern blot analysis --- p.35 / Chapter 2.3.5.2.1 --- Materials --- p.35 / Chapter 2.3.5.2.2 --- Methods --- p.35 / Chapter 2.3.5.3 --- Western blot analysis --- p.37 / Chapter 2.3.5.3.1 --- Materials --- p.37 / Chapter 2.3.5.3.2 --- Methods --- p.37 / Chapter 2.3.5.4 --- Immunoprecipitation --- p.37 / Chapter 2.3.5.4.1 --- Materials --- p.37 / Chapter 2.3.5.4.2 --- Methods --- p.38 / Chapter 2.3.5.5 --- Matrix-assisted laser desorption / ionization-time of flight (MALDI-TOF) mass spectrometry analysis --- p.39 / Chapter 2.3.5.5.1 --- Materials --- p.39 / Chapter 2.3.5.5.2 --- Methods --- p.39 / Chapter 2.4 --- "Analysis of the all-trans-13,14-dihydroretinol saturase (RetSat) activity by high-performance liquid chromatography (HPLC) analysis" --- p.41 / Chapter 2.4.1 --- Materials --- p.41 / Chapter 2.4.2 --- Methods --- p.42 / Chapter 2.4.2.1 --- Preparation of all-trans-retinol --- p.42 / Chapter 2.4.2.2 --- Treatment of PPSIG-transfected cells with all-trans-retinol --- p.42 / Chapter 2.4.2.3 --- Retinoid analysis --- p.43 / Chapter 2.5 --- Analysis of fatty acid compositions by gas chromatography-mass spectrometry (GC-MS) --- p.43 / Chapter 2.5.1 --- Materials --- p.43 / Chapter 2.5.2 --- Methods --- p.44 / Chapter 2.5.2.1 --- Preparation of fatty acid-BSA complex --- p.44 / Chapter 2.5.2.2 --- Treatment of PPSIG-transfected cells with fatty acid-BSA complex --- p.44 / Chapter 2.5.2.3 --- Extraction of fatty acids --- p.45 / Chapter 2.5.2.4 --- Methylation of the fatty acids --- p.45 / Chapter 2.5.2.5 --- GC-MS analysis --- p.46 / Chapter 2.5.2.6 --- Statistical analysis --- p.47 / Chapter CHAPTER 3 --- RESULTS --- p.48 / Chapter 3.1 --- The PPSIG cDNA was subcloned into a pCMV-Tag epitope tagging mammalian expression vector --- p.48 / Chapter 3.2 --- The pCMVT4B-PPSIG expression construct was transiently transfected into CHO-K1 and AML-12 cells --- p.54 / Chapter 3.3 --- Stable transfection of the pCMVT4B-PPSIG expression construct into CHO-K1 and AML-12 cells --- p.54 / Chapter 3.3.1 --- PPSIG-transfected CHO-K1 and AML-12 cells were obtained after G418 selection --- p.54 / Chapter 3.3.2 --- PPSIG-transfected CHO-K1 and AML-12 cells had high PPSIG mRNA expression --- p.58 / Chapter 3.3.3 --- PPSIG-FLAG fusion protein was over-expressed in the PPSIG- transfected CHO-K1 and AML-12 cells --- p.61 / Chapter 3.3.4 --- The stable transfectants were immunoprecipitated and identified as PPSIG protein by the mass spectrometry analysis --- p.64 / Chapter 3.4 --- PPSIG protein posseses saturase activity towards all-trans-retinol --- p.66 / Chapter 3.5 --- PPSIG protein is not a fatty acid transporter --- p.78 / Chapter CHAPTER 4 --- DISCUSSION --- p.101 / FUTURE STUDIES --- p.107 / REFERENCES --- p.110 / Appendix A: Prediction of the molecular weight of pCMVT4B- PPSIG protein --- p.119 / Appendix B: Theoretical tryptic peptides of PPSIG --- p.120 / Appendix C: Protein-peptide mass reports --- p.122 / Chapter C1. --- Peptide mass summary of trypsin-digested PPSIG immunoprecipitated protein from clone L2H4B18 --- p.122 / Chapter C2. --- Peptide mass summary of trypsin-digested PPSIG immunoprecipitated protein from clone AL2L7 --- p.123 / Appendix D: HPLC spectrum of the RetSat activity towards all- trans retinol --- p.124 / Chapter D1. --- RetSat activity towards all-trans retinol according to the Moise's group study ((Moise et al. 2004) --- p.124

Ho, Sai Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 144-164). / Abstracts in English and Chinese. / TABLE OF CONTENTS --- p.ii / ABSTRACT --- p.xii / 撮要 --- p.xv / LIST OF ABBREIVIATIONS --- p.xvi / LIST OF TABLES --- p.xvii / LIST OF FIGURES --- p.xix / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Overview of lectins --- p.1 / Chapter 1.1.1 --- History of lectins --- p.1 / Chapter 1.1.2 --- Definitions of lectins --- p.2 / Chapter 1.1.3 --- Classification and nomenclature of lectins based on structure --- p.2 / Chapter 1.1.4 --- Classification and nomenclature of lectins based on carbohydrate-bindingspecificity --- p.4 / Chapter 1.1.5 --- Structure of plant lectins --- p.4 / Chapter 1.1.6 --- Biological function of plant lectins --- p.5 / Chapter 1.1.6.1 --- Anti-viral activity of plant lectiins --- p.5 / Chapter 1.1.6.2 --- Lectins as plant defense proteins --- p.6 / Chapter 1.1.6.3 --- Insecticidal activity of plant lectins --- p.7 / Chapter 1.1.6.4 --- Anti-fungal activity of plant lectins --- p.7 / Chapter 1.1.6.5 --- Mitogenic activity of plant lectins --- p.7 / Chapter 1.1.6.6 --- Anti-tumor and anti-proliferative activity of plant lectins --- p.9 / Chapter 1.1.7 --- Background of legume lectins --- p.11 / Chapter 1.1.7.1 --- Structure of legume lectins --- p.11 / Chapter 1.1.7.2 --- Functions and activities of legume lectins --- p.12 / Chapter 1.2 --- Overview of serine protease inhibitors in plants --- p.14 / Chapter 1.2.1 --- Classification of serine protease inhibitor --- p.15 / Chapter 1.2.2 --- The main functions of plant serine protease inhibitors --- p.17 / Chapter 1.2.3 --- Commercial application of serine protease inhibirtors --- p.19 / Chapter 1.2.3.1 --- Medical application --- p.19 / Chapter 1.2.3.2 --- Transgenic application in agriculture --- p.22 / Chapter 1.3 --- Overview of Pathogenesis-related proteins in plants --- p.25 / Chapter 1.3.1 --- Overview of PR-5 family Thaumatin-like proteins (TLPs) --- p.27 / Chapter 1.3.1.1 --- Structural similarities among TLPs --- p.28 / Chapter 1.3.1.2 --- Antifungal activity of TLP --- p.31 / Chapter 1.3.2 --- Overview of Chinase-like proteins (CLPs) --- p.33 / Chapter 1.3.2.1 --- Classification of chitinase --- p.34 / Chapter 1.3.2.1.1 --- On the basis of amino acid sequence of glycosyl hydrolase --- p.34 / Chapter 1.3.2.1.2 --- On the basis of amino acid sequence of plant chitinase --- p.35 / Chapter 1.3.2.2 --- Antifungal activity of CLP --- p.36 / Chapter 1.3.3 --- Anti-freeze property of PR proteins --- p.38 / Chapter 1.3.4 --- Application of PR proteins in agriculture --- p.40 / Chapter 1.4 --- Rationale of the present study --- p.42 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.43 / Chapter 2.2 --- Preparation of crude extract --- p.44 / Chapter 2.2.1 --- Hokkaido large black soybean --- p.44 / Chapter 2.2.2 --- Emperor banana --- p.45 / Chapter 2.3 --- Purification --- p.45 / Chapter 2.4 --- Chromatography --- p.46 / Chapter 2.4.1 --- DEAE-cellulose chromatography --- p.46 / Chapter 2.4.2 --- Affi-gel Blue gel --- p.47 / Chapter 2.4.3 --- SP-Sepharse --- p.48 / Chapter 2.4.4 --- Mono Q HR 5/5 and Mono S HR 5/5 --- p.49 / Chapter 2.4.5 --- Superdex 75 and superdex 200 --- p.50 / Chapter 2.5 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.50 / Chapter 2.6 --- Protein concentration determination --- p.54 / Chapter 2.7 --- Preparation of rabbit reticulocyte lysate --- p.54 / Chapter 2.8 --- Determination of N-terminal amino acid sequence --- p.56 / Chapter 2.9 --- Assay of inhibition of hemagglutinating activity by different carbohydrates --- p.56 / Chapter 2.10 --- Thermal stability determination assays --- p.57 / Chapter 2.10.1 --- Stability at various temperatures --- p.57 / Chapter 2.10.2 --- Stability at 100°C --- p.57 / Chapter 2.11 --- Assay of pH dependence of hemagglutinating activity --- p.58 / Chapter 2.12 --- Assay of ion dependence of hemagglutinating activity --- p.58 / Chapter 2.13 --- Assay of antifungal activity --- p.58 / Chapter 2.14 --- Assay of trypsin inhibitory activity --- p.60 / Chapter 2.15 --- Assay of antibacterial activity --- p.61 / Chapter 2.16 --- Assay for cytotoxic activity on cancer cell lines --- p.61 / Chapter 2.17 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.62 / Chapter 2.18 --- Assay of mitogenic activity --- p.63 / Chapter Chapter 3 --- Purification and Characterization of Defense-Related Proteins from their Respective Sources / Chapter 3.1 --- Purification and Characterization of a Lectin from the Seeds of Hokkaido large black soybean / Chapter 3.1.1 --- Introduction --- p.65 / Chapter 3.1.2 --- Results --- p.66 / Chapter 3.1.3 --- Purification --- p.68 / Chapter 3.1.3.1 --- Affinity chromatography on Affi-gel Blue gel --- p.69 / Chapter 3.1.3.2 --- Anion-exchange chromatography on DEAE-cellulose --- p.70 / Chapter 3.1.3.3 --- Anion-exchange chromatography on Mono Q column --- p.71 / Chapter 3.1.3.4 --- Gel filtration on Superdex 200 column --- p.72 / Chapter 3.1.3.5 --- Hemagglutinating activity at each purification step --- p.73 / Chapter 3.1.4 --- Characterization of Lectin --- p.74 / Chapter 3.1.4.1 --- Molecular mass determination --- p.74 / Chapter 3.1.4.2 --- N-terminal amino acid sequencing --- p.76 / Chapter 3.1.4.3 --- Assay of inhibition of hemagglutinating activity by different carbohydrates --- p.77 / Chapter 3.1.4.4 --- Thermal stability --- p.78 / Chapter 3.1.4.5 --- Assay of pH dependence of hemagglutinating activity --- p.80 / Chapter 3.1.4.6 --- Assay of ion dependence of hemagglutinating activity --- p.81 / Chapter 3.1.4.7 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.82 / Chapter 3.1.4.8 --- Assay of mitogenic activity --- p.83 / Chapter 3.1.4.9 --- Assay of antibacterial activity --- p.84 / Chapter 3.1.5 --- Discussion --- p.86 / Chapter 3.2 --- Purification and Characterization of a Trypsin inhibitor from the Seeds of Hokkaido large black soybean / Chapter 3.2.1 --- Introduction --- p.93 / Chapter 3.2.2 --- Results --- p.94 / Chapter 3.2.3 --- Purification --- p.95 / Chapter 3.2.3.1 --- Anion-exchange chromatography on Mono Q column --- p.96 / Chapter 3.2.3.2 --- Gel filtration on Superdex 75 column --- p.98 / Chapter 3.2.3.3 --- Trypsin inhibitory activity at each purification step --- p.99 / Chapter 3.2.4 --- Characterization of trypsin inhibitory --- p.100 / Chapter 3.2.4.1 --- Molecular mass determination --- p.100 / Chapter 3.2.4.2 --- N-terminal amino acid sequencing --- p.102 / Chapter 3.2.4.3 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.103 / Chapter 3.2.4.4 --- Antiproliferative effect on MCF-7 and Hep G2 cells --- p.104 / Chapter 3.2.4.5 --- pH and thermal stability --- p.105 / Chapter 3.2.5 --- Discussion --- p.106 / Chapter 3.3 --- Purification and Characterization of a Thaumatin-like protein and Chitinase-like protein from Emperor Banana / Chapter 3.3.1 --- Introduction --- p.108 / Chapter 3.3.2 --- Results --- p.109 / Chapter 3.3.3 --- Purification --- p.111 / Chapter 3.3.3.1 --- Affinity chromatography on Affi-gel Blue gel --- p.112 / Chapter 3.3.3.2 --- Cation exchange chromatography on Mono S column --- p.113 / Chapter 3.3.3.3 --- Gel filtration on Superdex 75 column --- p.114 / Chapter 3.3.3.3.1 --- Fraction MS 2 --- p.114 / Chapter 3.3.3.3.2 --- Fraction MS 4 --- p.115 / Chapter 3.3.3.3.3 --- Fraction MS 5 --- p.118 / Chapter 3.3.4 --- Characterization of the thaumatin-like protein --- p.121 / Chapter 3.3.4.1 --- N-terminal amino acid sequence determination --- p.121 / Chapter 3.3.4.2 --- Assay for antifungal activity --- p.122 / Chapter 3.3.4.3 --- Thermal stability --- p.124 / Chapter 3.3.4.4 --- pH stability --- p.125 / Chapter 3.3.4.5 --- Resistance to trypsin digestion --- p.125 / Chapter 3.3.4.6 --- Anti-HIV-1 reverse transcriptase activity --- p.126 / Chapter 3.3.4.7 --- Discussion --- p.127 / Chapter 3.3.5 --- Characterization of the two chitinase-like protein --- p.131 / Chapter 3.3.5.1 --- N-terminal amino acid sequence determination --- p.131 / Chapter 3.3.5.1.1 --- Emperor banana MS2 CLP --- p.131 / Chapter 3.3.5.1.2 --- Emperor banana MS4 CLP --- p.132 / Chapter 3.3.5.2 --- Assay for antifungal activity --- p.133 / Chapter 3.3.5.3 --- Discussion --- p.136 / Chapter Chapter 4 --- general discussion --- p.138 / References --- p.144

Os manguezais são considerados um ecossistema único, devido sua particular combinação de condições ambientais, influenciados pela sua localização na interface entre o continente e oceano. O estudo deste ecossistema se torna urgente uma vez que os manguezais estão desaparecendo em todo mundo, e sua diversidade de grupos microbianos é ainda pouco conhecida. Dentro dessa temática, o presente projeto visa descrever a possível funcionalidade de bactérias pertencentes ao filo Planctomycetes nos manguezais. Tais organismos são ainda pouco estudados, de difícil cultivo, sendo obtidos principalmente em ambientes marinhos, tratamento de água e locais de criação de peixes. Dentro desse filo são encontrados microrganismos pertencentes a gêneros descritos como capazes de realizar a oxidação anaeróbica do íon amônio (anammox), uma importante transformação do nitrogênio em ambientes com baixa disponibilidade de oxigênio. E ainda, visamos descrever a possível funcionalidade de bactérias pertencentes ao filo Planctomycetes nos manguezais. Para isso, foi inicialmente realizada uma comparação dos genomas de Planctomycetes com as sequências obtidas por análises de metagenômica e metatranscriptômica, onde foram encontradas sequências similares às afiliadas a funções desconhecidas (putative protein) e a sulfatases. Tais enzimas são descritas como hidrolíticas, que catalizam a clivagem de ésteres de sulfato, liberando o enxofre na forma assimilável. A análise de sequências de uma biblioteca metagenômica (obtida de um manguezal contaminado com petróleo) permitiu a visualização de fragmentos genômicos de Planctomycetes. Esta análise revelou também a ocorrência de genes relacionados a produção de sulfatases, além de indicar arranjos gênicos distintos dos descritos nos genomas de organismos deste grupo, possivelmente indicando a ocorrência de endemismo de Planctomycetes em manguezais. Esta observação foi reforçada pelo cultivo de isolados afiliados a este grupo, os quais tiveram sua afiliação confirmada pelo sequenciamento parcial do gene ribossomal 16S DNAr. Alguns destes isolados formaram clusters diferenciados dentro do filo Planctomycetes, indicando que podem ser estes novas espécies. Sendo assim, a caracterização desse grupo de microrganismos numa combinação de análises in sílico e in vivo, possibilitou a confirmação da ocorrência de tais organismos nos manguezais, e gerou as primeiras informações sobre sua funcionalidade neste sistema, onde parece ocorrer de forma diferenciada aos demais ambientes onde já foram descritos. / Mangroves are considered an unique ecosystem due to its particular combination of environmental conditions, influenced by its location at the interface between land and ocean. The study of this ecosystem becomes urgent since mangroves are disappearing worldwide, and its diversity of microbial groups is still poorly understood. Within this theme, this project aims to describe the possible functionality of bacteria belonging to the phylum Planctomycetes in mangroves. Such organisms are still poorly studied, difficult to cultivate and mainly isolated from marine environments, water treatment and fish farming sites. Within this phylum are found microorganisms belonging to genera described as capable of performing the anaerobic oxidation of ammonium (anammox), a major transformation of nitrogen in environments with low oxygen availability. Here we aimed to describe the possible functionality of bacteria belonging to the phylum Planctomycetes in the mangroves. In order to achieve the target, it was initially performed a comparison of the Planctomycetes genomes with sequences obtained by metagenomics and metatranscriptomics, revealing the presence of sequences with similarity to those affiliated to unknown function and sulfatases. These are hydrolytic enzymes, which catalyze the cleavage of sulfate esters, releasing sulfur on its available form. The analysis of sequences from a metagenomic library (obtained from an oil-contaminated mangrove) allowed the visualization of genomic fragments related to Planctomycetes. It also revealed the presence of genes related with the production of sulfatases, besides the indication of specific genome arrangements, distinct from those describe in organisms allocated in this group, possibly indicating the occurrence of endemicity for Planctomycetes in mangroves. It was reinforced by the cultivation of isolates affiliated to this groups, whose have their affiliation confirmed by the partial sequencing of the 16S rDNA gene. Some isolates clustered composed new clusters within the Planctomycetes phylum, indicating the occurrence of new species. In sum, the characterization of this group by combining in sílico and in vivo analyses, allowed the confirmation of the occurrence of these organisms in mangroves, and generated the first insights about its functioning on this system, where it seems to occur in a differentiated form from those observed in other environments where it has been described.

There are a vast number of factors that influence nucleosome formation, and consequently gene regulation. These factors include histone modifications, nucleotide composition, transcriptional region elements, and specific nucleotide motifs, among others. Although the amount we know now is limited, we are creating new techniques and discoveries to assist us in continued understanding of chromatin. To make a significant contribution to the field of chromatin, I conducted two hypothesis driven sets of experiments that address the topic of chromatin structure. First, I created a technique for tissue specific nucleosome isolation with the goal of observing the effect of single nucleotide polymorphisms (SNPs) on nucleosome formation. Second, I created and tested a method to recover lost in vitro nucleosome reconstitution data, which can improve this type of data, commonly used for observing nucleosome positioning. The first experiment needs a more specific antibody to complete the last step and function as designed. The second experiment shows that our nucleosome recovery method, when applied conservatively, can recover 90% of the lost nucleosome data.