Chan chi-wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 89-95). / Abstracts in English and Chinese. / Dedication --- p.i / Acknowledgments --- p.ii / Abstract --- p.iii / 摘要 --- p.v / Abbreviations --- p.vi / List of Figures --- p.ix / List of Tables --- p.xiii / Contents --- p.xiv / Chapter CHAPTER 1 --- Introduction --- p.1 / Chapter 1.1. --- Thesis synopsis --- p.1 / Chapter 1.2. --- hNP220 protein --- p.1 / Chapter 1.2.1. --- Domain organization --- p.1 / Chapter 1.2.2. --- Known splice variants --- p.5 / Chapter 1.2.3. --- Subcellular localization --- p.7 / Chapter 1.2.4. --- Proposed roles in transcriptional activation and RNA processing --- p.7 / Chapter 1.2.5. --- Interaction between C-terminal of hNP220 and FHL2 --- p.9 / Chapter 1.3. --- Hypothesis --- p.12 / Chapter 1.4. --- Principles of key methods --- p.14 / Chapter 1.4.1. --- RLM-RACE --- p.14 / Chapter 1.4.2. --- CytoTrap® two-hybrid system --- p.15 / Chapter CHAPTER 2 --- Materials and Methods --- p.18 / Chapter 2.1. --- Cloning protocol --- p.18 / Chapter 2.1.1. --- Amplification of DNA fragment --- p.18 / Chapter 2.1.2. --- Purification of PCR product --- p.19 / Chapter 2.1.3. --- Restriction endonuclease digestion --- p.20 / Chapter 2.1.4. --- Dephosphorylation of cloning vector 5'-termini --- p.20 / Chapter 2.1.5. --- Insert/vector ligation --- p.20 / Chapter 2.1.6. --- Preparation of chemically competent bacterial cells (E. coli strain DH5a) --- p.21 / Chapter 2.1.7. --- Transformation of ligation product into chemically competent bacterial cells --- p.22 / Chapter 2.1.8. --- Small-scale preparation of bacterial plasmid DNA --- p.22 / Chapter 2.1.9. --- Screening for recombinant clone --- p.24 / Chapter 2.1.10. --- Dideoxy DNA sequencing --- p.24 / Chapter 2.1.11. --- Midi-scale preparation of recombinant plasmid DNA --- p.25 / Chapter 2.2. --- Determination of the transcription start site (TSS) of hNP220 gene --- p.27 / Chapter 2.2.1. --- RNA ligase-mediated rapid amplification of cDNA 5'-end (5-RLM-RACE) --- p.27 / Chapter 2.3. --- Isolation and identification of the third splice variant of HNP220 (hNP220ε) --- p.29 / Chapter 2.3.1. --- PCR from human heart/testis cDNAs pool --- p.29 / Chapter 2.3.2. --- RT-PCR --- p.29 / Chapter 2.3.3. --- Northern hybridization --- p.30 / Chapter 2.4. --- Human tissue distribution of hNP220 --- p.31 / Chapter 2.4.1. --- RT-PCR --- p.31 / Chapter 2.4.2. --- Northern hybridization --- p.31 / Chapter 2.5. --- Visualization of the subcellular localization patterns of GFP-tagged hNP220ε in HepG2 cell line --- p.32 / Chapter 2.5.1. --- Cloning of hNP220a and hNP220s into vector pEGFP-Cl --- p.32 / Chapter 2.5.2. --- Transfection of GFP fusion constructs into HepG2 cell line --- p.32 / Chapter 2.5.3. --- Epi-fluorescence microscopy --- p.33 / Chapter 2.6. --- Identification of the protein-protein interaction between hNP220ε and MAPRE1 --- p.34 / Chapter 2.6.1. --- CytoTrap® XR HeLa Cell cDNA Library screening --- p.34 / Chapter 2.6.1.1. --- Cloning of hNP220ε into yeast two-hybrid bait vector pSos --- p.34 / Chapter 2.6.1.2. --- Preparation of cdc25Ha & cdc25Hα yeast competent cells --- p.34 / Chapter 2.6.1.3. --- Autonomous activation study of bait fusion construct pSos-hNP220ε --- p.36 / Chapter 2.6.1.4. --- Cotransformation of pSos-hNP220ε and CytoTrap® XR HeLa Cell cDNA Library --- p.36 / Chapter 2.6.1.5. --- Verification of interaction by yeast mating --- p.38 / Chapter 2.6.1.5.1. --- Generation of yeast plasmid segregant for mating --- p.38 / Chapter 2.6.1.5.2. --- Yeast mating in 96-well plate --- p.39 / Chapter 2.6.1.6. --- Identification of putative interaction partner --- p.39 / Chapter CHAPTER 3 --- Results --- p.42 / Chapter 3.1. --- Transcription start site of the HNP220 gene is located 312 nucleotides upstream the initiation codon --- p.42 / Chapter 3.2. --- Third splice variant of hNP220 gene hNP220s) is identified --- p.44 / Chapter 3.3. --- In silico analysis of hNP220ε --- p.54 / Chapter 3.4. --- hNP220a and hNP220s are ubiquitously expressed in human fetal and adult tissues --- p.65 / Chapter 3.5. --- hNP220ε shows a punctate subnuclear localization pattern in HepG2 cell line --- p.67 / Chapter 3.6. --- hNP220ε interacts with MAPRE1 --- p.69 / Chapter CHAPTER 4 --- Discussion --- p.71 / Chapter 4.1. --- "Identification of hNP220s, the third splice variant of hNP220 gene" --- p.71 / Chapter 4.2. --- Biological resemblance between hNP220α (hNP220) and hNP220ε --- p.73 / Chapter 4.3. --- Protein-protein interaction between hNP220ε and MAPRE1 --- p.74 / Chapter 4.3.1. --- MAPRE1 protein --- p.77 / Chapter 4.3.2. --- Wnt signaling pathway --- p.78 / Chapter 4.4. --- Potential roles of hNP220 in the regulation of chromosome stability and oncogenesis --- p.82 / Chapter 4.5. --- Summary --- p.85 / Chapter 4.6. --- Concluding questions --- p.86 / Chapter 4.7. --- Future work --- p.87 / References --- p.89 / Appendix --- p.96

Lau Man Tat. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 95-126). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Figures --- p.xi / List of Tables --- p.xiii / Symbols and Abbreviations --- p.xiv / Scientific Names --- p.xvii / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Gonadotropins / Chapter 1.1.1 --- Structure --- p.2 / Chapter 1.1.2 --- Function --- p.6 / Chapter 1.1.3 --- Regulation --- p.9 / Chapter 1.1.3.1 --- Hypothalamic regulators (GnRH) --- p.9 / Chapter 1.1.3.2 --- Endocrine regulators from gonads (steroids) --- p.12 / Chapter 1.1.3.3 --- Paracrine regulators (activin) --- p.14 / Chapter 1.2 --- Activin Family of Growth Factors / Chapter 1.2.1 --- Activin / Chapter 1.2.1.1 --- Structure --- p.14 / Chapter 1.2.1.2 --- Function --- p.14 / Chapter 1.2.1.3 --- Signaling --- p.18 / Chapter 1.2.2 --- Follistatin / Chapter 1.2.2.1 --- Structure --- p.21 / Chapter 1.2.2.2 --- Function --- p.21 / Chapter 1.3 --- Transcriptional regulation of pituitary gonadotropin subunit genes at the promoter level --- p.22 / Chapter 1.4 --- The project objectives and long-term significance --- p.26 / Chapter Chapter 2 --- "Cloning of Smad2, Smad3, Smad4 and Smad7 from the Goldfish Pituitary and Their Involvement in the FSHβ Transcription in LβT2 cells" / Chapter 2.1 --- Introduction --- p.28 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Chemicals --- p.31 / Chapter 2.2.2 --- Animals --- p.32 / Chapter 2.2.3 --- Isolation of total RNA --- p.32 / Chapter 2.2.4 --- "Cloning of cDNA fragments of Smad 2, 3, 4 and 7 from the goldfish pituitary" --- p.32 / Chapter 2.2.5 --- Rapid amplification of 5'-cDNA ends (5'-RACE) and full-length cDNA(3'-RACE) --- p.33 / Chapter 2.2.6 --- Primary pituitary cell culture --- p.34 / Chapter 2.2.7 --- "Validation of semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assays for goldfish Smad 2, 3, 4 and7" --- p.35 / Chapter 2.2.8 --- Construction of the reporter plasmid containing the goldfish FSHβ promoter --- p.36 / Chapter 2.2.9 --- Construction of expression plasmids --- p.37 / Chapter 2.2.10 --- Cell culture and transient transfection --- p.38 / Chapter 2.2.11 --- SEAP reporter gene assay --- p.39 / Chapter 2.2.12 --- β-galactosidase reporter gene assay --- p.40 / Chapter 2.2.13 --- Data analysis --- p.40 / Chapter 2.3 --- Results / Chapter 2.3.1 --- "Cloning and sequence characterization of goldfish Smad 2, 3,4 and7" --- p.41 / Chapter 2.3.2 --- "Tissue distribution of Smad 2，3, 4 and 7 expression" --- p.42 / Chapter 2.3.3 --- "Validation of semi-quantitative RT-PCR assays for Smad 2, 3,4 and7" --- p.43 / Chapter 2.3.4 --- Activin regulation of Smad 2，3，4 and 7 expression in cultured goldfish pituitary cells --- p.44 / Chapter 2.3.5 --- "Smad 2, 3 and 7 regulate basal and activin-induced FSHβ transcription in LβT2 cells" --- p.44 / Chapter 2.3.6 --- Autocrine regulation of the gfFSHβ transcription by activin in LβT2 cells --- p.45 / Chapter 2.4. --- Discussion --- p.47 / Chapter Chapter 3 --- Promoter Analysis for the Smad Responsive Element (SRE) in the Goldfish Follicle Stimulating Hormone β(FSHβGene / Chapter 3.1 --- Introduction --- p.71 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Chemicals --- p.74 / Chapter 3.2.2 --- Construction of expression plasmids --- p.74 / Chapter 3.2.3 --- Construction of SEAP reporter plasmids containing different lengths of gfFSHβ promoter --- p.74 / Chapter 3.2.4 --- Cell culture and transient transfection --- p.75 / Chapter 3.2.5 --- Reporter gene assays for SEAP and β-Gal --- p.75 / Chapter 3.2.6 --- Data Analyses --- p.76 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Localization of the proximal Smad-responsive Element (SRE) in the gfFHSβ promoter --- p.76 / Chapter 3.4 --- Discussion --- p.78 / Chapter Chapter 4 --- General Discussion / Chapter 4.1 --- Overview --- p.89 / Chapter 4.2 --- Contribution of the present research / Chapter 4.2.1 --- Cloning and characterization of Smad proteins from the goldfish pituitary --- p.90 / Chapter 4.2.2 --- Regulation of Smads in primary pituitary cell culture --- p.90 / Chapter 4.2.3 --- Identification of the Smad responsive element (SRE) on the gfFSHβ promoter --- p.91 / Chapter 4.3 --- Future research direction --- p.93 / References --- p.95

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Bibliography: leaves 97-114. "Self-incompatibility (SI) is an important genetic mechnism to prevent the inbreeding of flowering plants and also an excellent system for studying cell-cell recognition and signal transduction."

Bibliography: leaves 97-114. Electronic publication: Fulltext available in PDF format; abstract in HTML format. "Self-incompatibility (SI) is an important genetic mechnism to prevent the inbreeding of flowering plants and also an excellent system for studying cell-cell recognition and signal transduction." Electronic reproduction.[Australia] :Australian Digital Theses Program,2001.

Allorecognition, defined as the ability to discriminate between self and non-self, is ubiquitous to colonial metazoans and widespread in aclonal taxa. Invertebrate allorecognition phenomena are of broad interest and have long captured the attention of geneticists by virtue of the allotypic diversity they display, marine ecologists by virtue of their control of effector mechanisms determining the outcome of intraspecific competition, evolutionary biologists by virtue of their regulation of the level at which selection acts, and immunologists by virtue of their resemblance to the allogeneic interactions that characterize pregnancy and transplantation in vertebrates. Diverse histocompatibility modes have been described in the jawed vertebrates, protochordates, and cnidarians, which are to date the only three taxa for which a genetic model to study allorecognition has been developed. Outside of the MHC-based histocompatibility of vertebrates, allorecognition determinants have been recognized in only two invertebrates. In the tunicate Botryllus, two genes involved in the histocompatibility response were characterized, FuHc and fester. In Hydractinia, the loci controlling allorecognition, alr1 and alr2, were mapped to a single chromosomal region, the allorecognition complex, and alr2 was recently identified as a polymorphic immunoglobulin superfamily (IgSF) receptor. In this study, the identification of the second Hydractinia allodeterminant, alr1, was undertaken. Chapter I briefly reviews prominent allorecognition model organisms and details the phenomenon in the model organism, Hydractinia symbiolongicarpus studied here. Chapter II describes the isolation of a 300.8 kb alr1-containing chromosomal interval by positional cloning. The analysis of that interval for its gene content and the determination of a primary alr1 candidate, CDS4, are described in Chapter III. Chapter III also reveals the existence of a complex of IgSF-like genes, to which belongs CDS4. CDS4, a novel polymorphic IgSF receptor that encodes a type I transmembrane protein with two hypervariable immunoglobulin-like extracellular domains, was confirmed to be the alr1 allodeterminant in Chapter IV, based on the investigation of natural polymorphism. CDS4 allele sequences were found to largely predict the outcome of allorecognition responses within and between laboratory lines and wild-type colonies, confirming the identity of CDS4 as the classically defined locus alr1.

Abstract When it comes to the management of non-profit organizations, our country¡¦s academic sector has always placed an emphasis on the organization¡¦s decision making nuclei or those in command, and their effects on the fulfillment of organizational goals, resources placed into operation, efficiency, satisfaction, and other such self-management analysis. Seldom did they adopt an ¡§ethically-oriented¡¨ method of management. Instead they viewed the medical industry as a for-profit business, stressing business strategies and marketing, and therefore regarded patients as customers. This Study¡¦s Characteristics: 1. Focuses on medical and religious philosophies for investigation. Unlike single-issue studies, this study uses medicine as the primary basis for its perspective, and adds religious beliefs (Buddhism) to probe each health care morality issue. Health care morality issues include: abortion, cloning, euthanasia, organ transplants, and hospice care. In addition to ethical standpoints on medicine, using the sorted investigations on religious documents, we looked deeper into the viewpoints of Buddhism. 2. Feature: Besides investigating the morality of the body and flesh of ordinary medicine, we probed further into the root of the ethics of the mind. In retrospect to ¡§Medical Ethics¡¨, and the viewpoints of eastern and western medicine, we went beyond the perceptions of ordinary medicine, and explored a deeper plane of the mind, so as to increase the depth of health care ethics. Results: Through a factor analysis, we ended up with 10 factors: (hospice care, euthanasia, surrogate mothers, cloning and genetic issues, therapeutic art, religious care, stem cells, medical ethics, and abortion issues) and analyzed them according to 3 morality decision models to identify which is the best suited. Results: efficiency model and ethics model had positive effects on hospice care and therapeutic ethics, while as liability model had negative effects; liability model and ethics model had positive effects on religious care, medical ethics, and abortion issues; all 3 models, efficiency model, ethics model, and liability model, had positive effects on genetic issues and stem cells; efficiency model had positive effects on euthanasia, while as ethics model had negative effects; liability model had positive effects on surrogate mothers; liability model had positive effects on cloning issues, while as efficiency model and ethics model had opposite effects. Did people have different attitudes towards health care ethics? Results: the general public and medical personnel had significantly different attitudes towards hospice care, genetic issues, and therapeutic ethics; medical personnel, religious personnel, and the general public had significantly different attitudes towards religious care, stem cells, and abortion issues; medical personnel, patients, religious personnel, and the general public had significantly different attitudes towards medical ethics; medical personnel and religious personnel had significantly different attitudes towards euthanasia; the opinions of the groups were unanimous towards surrogate mothers and cloning issues. As to individual orientation, the higher the person¡¦s education level, the less he or she places emphasis on health care morality issues. Perhaps since medical ethics is only beginning to be drawn attention to in our country, most people with a higher degree of education have not yet concerned themselves to this level. In summary, this is worth the attention of our current education ministry. Key words¡GMedical Ethics¡AHuman Cloning¡AHospice

Thesis (M. Phil.)--University of Hong Kong, 2001. / Includes bibliographical references (leaves 92-101).

Thesis (Ph. D.)--Florida State University, 2003. / Advisor: Dr. Michael Ruse, Florida State University, College of Arts and Sciences, Dept. of Philosophy. Title and description from dissertation home page (viewed Apr. 9, 2004). Includes bibliographical references.

Thesis (Ph. D.)--University of Queensland, 2002. / Includes bibliographical references.