Thesis (Ph. D.)--Cornell University, May, 2007. / Vita. Includes bibliographical references (leaves 120-133).

by Hiu-ming Li. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 81-88). / SUMMARY --- p.1 / Chapter CHAPTER 1. --- INTRODUCTION --- p.3 / Chapter CHAPTER 2. --- LITERATURE REVIEW --- p.7 / Chapter 2.1. --- Retinal pigment epithelium --- p.7 / Chapter 2.1.1. --- Embryology / Chapter 2.1.2. --- Anatomy and histology / Chapter 2.1.3. --- Growth and aging / Chapter 2.1.4. --- Macular region / Chapter 2.2. --- The photoreceptor outer segment --- p.12 / Chapter 2.3. --- Lipofuscin --- p.13 / Chapter 2.4. --- Lipofuscin in retinal pigment epithelium and retinal photoreceptor disc shedding --- p.14 / Chapter 2.5. --- Possible mechanism for lipofuscin formation in the RPE --- p.22 / Chapter 2.6. --- Age-related lipofuscin accumulation in the RPE --- p.22 / Chapter 2.7. --- Racial difference of RPE lipofuscin concentration --- p.25 / Chapter 2.8. --- RPE lipofuscin and age-related macular degeneration --- p.26 / Chapter CHAPTER 3. --- MATERIALS AND METHODS --- p.28 / Chapter 3.1. --- Histologic Specimens --- p.28 / Chapter 3.2. --- Measuring equipment --- p.28 / Chapter 3.3. --- Software of measurements --- p.31 / Chapter 3.4. --- Light source and filters --- p.31 / Chapter 3.5. --- Control --- p.31 / Chapter 3.6. --- Measurement of autofluorescent Intensity --- p.32 / Chapter 3.7. --- Bleaching (oxidation) of melanin --- p.39 / Chapter CHAPTER 4. --- RESULTS --- p.42 / Chapter 4.1. --- Bleaching test --- p.42 / Chapter 4.2. --- RPE autofluorescence observation in different age --- p.43 / Chapter 4.3. --- RPE autofluorescence observation within individual eyes --- p.45 / Chapter 4.4. --- Topographic distribution of lipofuscin --- p.49 / Chapter 4.5. --- Lipofuscin content at the Foveola --- p.50 / Chapter 4.6. --- The relationship of age and lipofuscin content in total RPE --- p.51 / Chapter 4.7. --- The relationship of age and lipofuscin content in the macular RPE --- p.57 / Chapter 4.8. --- Relationship of age and lipofuscin content in the posterior pole of RPE --- p.59 / Chapter 4.9. --- Relationship of age and lipofuscin content in the temporal RPE --- p.61 / Chapter 4.10. --- Relationship of age and lipofuscin content in the nasal RPE --- p.63 / Chapter 4.11. --- Age related topographic changes --- p.65 / Chapter 4.12. --- The relationship of age and lipofuscin content in the RPE of male --- p.66 / Chapter 4.13. --- The relationship of age and lipofuscin content in the RPE of female --- p.68 / Chapter 4.14. --- Relationship of lipofuscin content in different sex --- p.70 / Chapter CHAPTER 5. --- DISCUSSION --- p.71 / Chapter 5.1. --- Evaluation of method --- p.71 / Chapter 5.2. --- RPE lipofuscin content in different age --- p.71 / Chapter 5.3. --- Topographic distribution of lipofuscin --- p.74 / Chapter 5.4. --- Lipofuscin and age-related macular degeneration in Chinese --- p.78 / REFERENCES --- p.81

An in vitro phagocytosis assay system was developed and characterized for studying the specific receptor-mediated phagocytosis of bovine ROS by bovine RPE cells. The phagocytosis of ROS was detected qualitatively by electron microscopy and quantitatively by treating RPE cells with radioiodinated ROS or by probing ROS-treated RPE cells with a radiolabeled antirhodopsin monoclonal antibody. The binding sites for various antirhodopsin monoclonal antibodies were localized as an essential step in their application as immunochemical probes for analysis of the structure and function of rhodopsin. Five monoclonal antibodies raised against rhodopsin have been shown to be directed against the N-terminal regions on the basis of their reactivity to an immunoaffinity purified 2-39 glycopeptide, a 2-16 tryptic glycopeptide and a 1-16 synthetic peptide as measured by radioimmune competition assays. Limited proteolysis, immunogold-dextran labeling and competitive inhibition studies identified two antirhodopsin monoclonal antibodies which bound to internal cytoplasmic loop regions of rhodopsin. Finally, the binding sites for these and other C-terminal specific antirhodopsin monoclonal antibodies were used to elucidate the proposed transmembrane helical model of rhodopsin. An antirhodopsin monoclonal antibody (rho 4D2), which bound to rhodopsin in glutaraldehyde-fixed ROS plasma membranes, was employed as an immunocytochemical probe in studying the possible role of rhodopsin in the binding and phagocytosis of rod outer segments. An immunoaffinity purified 2-39 N-terminal rhodopsin glycopeptide, a synthetic 1-16 peptide analogue of rhodopsin and phospholipid vesicles reconstituted with rhodopsin were all found to be ineffective in inhibiting the phagocytosis of ¹²⁵I-labeled ROS by RPE cells. In essence, these results provided compelling evidence that rhodopsin in the ROS plasma membrane does not function as the ligand for recognition by RPE cells. The molecular properties of the ROS cell surface ligand(s), which are involved in recognition by bovine RPE cells, were studied by limited-proteolytic digestion in conjunction with quantitative phagocytosis assays. Mildly trypsin-treated ROS were found to be less effectively phagocytized than untreated ROS by bovine RPE cells. Moreover, the glycopolypeptides (34kD and 24kD) released from the ROS cell surface by trypsin were capable of inhibiting ROS phagocytosis. The ROS plasma membrane specific, ricin-binding, 230kD glycoprotein was observed by SDS-gel electrophoresis and western blotting to be highly trypsin sensitive under these conditions. Hence, ricin affinity chromatography and immunoaffinity chromatography were employed in an attempt to purify this 230kD glycoprotein from ROS membranes. Enriched preparations of the 230kD glycoprotein were reconstituted into phospholipid vesicles and effectively used to inhibit the phagocytosis of ROS by RPE cells. In summary, a ROS plasma membrane specific, 230kD glycoprotein has been identified and isolated; this protein may act as a ligand in specific ligand-receptor interactions between ROS and RPE cells. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

Thesis (Ph. D.)--University of Virginia, 1999. / Spine title: Structure - function analysis of IRBP. Includes bibliographical references (p. 167-172). Also available online through Digital Dissertations.

by Keung Wing Ying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 115-128). / Abstract also in Chinese. / Abstract --- p.I / Acknowledgments --- p.IV / Abbreviations --- p.V / List of Tables --- p.VII / List of Figures --- p.VIII / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Familial Adenomatous Polyposis (FAP) --- p.1 / Chapter 1.1.1 --- Occurrence and prevalence --- p.1 / Chapter 1.1.2 --- Clinical features --- p.2 / Chapter 1.1.3 --- Laboratory studies --- p.5 / Chapter 1.1.4 --- Diagnosis --- p.6 / Chapter 1.1.5 --- Management --- p.8 / Chapter 1.2 --- Congenital Hypertrophy of the Retinal Pigment Epithelium (CHRPE) --- p.8 / Chapter 1.2.1 --- Clinical features --- p.9 / Chapter 1.2.2 --- Pathogenesis --- p.11 / Chapter 1.2.3 --- Histology --- p.12 / Chapter 1.2.4 --- Differential diagnosis --- p.13 / Chapter 1.2.5 --- CHRPE as an early clinical marker for FAP --- p.14 / Chapter 1.3 --- The Adenomatous Polyposis Coli (APC) Gene --- p.16 / Chapter 1.3.1 --- Discovery --- p.16 / Chapter 1.3.2 --- Structure and function --- p.17 / Chapter 1.3.3 --- Sequence alterations in the APC gene --- p.18 / Chapter 1.3.4 --- APC mutations associated with specific clinical features --- p.21 / Chapter 1.3.5 --- APC gene mutations in Chinese --- p.22 / Chapter 1.3.6 --- Methods for detecting mutation in the APC gene and linkage analysis --- p.23 / Chapter Chapter 2 --- Study Objectives --- p.44 / Chapter Chapter 3 --- Methodology --- p.45 / Chapter 3.1 --- Subjects --- p.45 / Chapter 3.2 --- CHRPE analysis --- p.45 / Chapter 3.2.1 --- Ophthalmoscopic examination --- p.45 / Chapter 3.2.2 --- Diagnostic criteria of CHRPE --- p.45 / Chapter 3.3 --- Materials and Equipment --- p.46 / Chapter 3.3.1 --- Enzymes --- p.46 / Chapter 3.3.2 --- DNA markers --- p.46 / Chapter 3.3.3 --- Reagent kits --- p.46 / Chapter 3.3.4 --- Primers for PCR --- p.46 / Chapter 3.3.5 --- Chemicals and reagents --- p.47 / Chapter 3.3.6 --- Radioisotopes --- p.47 / Chapter 3.3.7 --- Solutions and buffers --- p.47 / Chapter 3.3.8 --- Equipment --- p.48 / Chapter 3.4 --- Methods --- p.49 / Chapter 3.4.1 --- Blood collection --- p.49 / Chapter 3.4.2 --- DNA extraction --- p.49 / Chapter 3.4.3 --- DNA quantitation --- p.50 / Chapter 3.4.4 --- Polymerase Chain Reaction (PCR) --- p.50 / Chapter 3.4.5 --- Agarose gel electrophoresis --- p.51 / Chapter 3.4.6 --- Single Strand Conformation Polymorphism (SSCP) --- p.52 / Chapter 3.4.7 --- Direct DNA sequencing --- p.52 / Chapter 3.4.8 --- Analysis of microsatellite markers --- p.54 / Chapter Chapter 4 --- Results --- p.59 / Chapter 4.1 --- Study subjects --- p.59 / Chapter 4.1.1 --- FAP index patients --- p.59 / Chapter 4.1.2 --- FAP families --- p.59 / Chapter 4.1.3 --- Control subjects with CHRPE only --- p.60 / Chapter 4.1.4 --- Normal control subjects --- p.60 / Chapter 4.2 --- CHRPE analysis --- p.60 / Chapter 4.2.1 --- CHRPE in FAP index patients --- p.60 / Chapter 4.2.2 --- CHRPE in family members --- p.61 / Chapter 4.2.3 --- CHRPE in controls subjects --- p.61 / Chapter 4.2.4 --- Statistical analysis --- p.61 / Chapter 4.3 --- PCR optimization --- p.62 / Chapter 4.4 --- SSCP analysis of the APC gene --- p.62 / Chapter 4.5 --- Direct DNA sequencing analysis --- p.63 / Chapter 4.5.1 --- Nonsense mutations --- p.63 / Chapter 4.5.2 --- Novel silent mutations --- p.64 / Chapter 4.5.3 --- Polymorphisms --- p.65 / Chapter 4.6 --- Haplotype analysis --- p.67 / Chapter 4.7 --- Family studies --- p.67 / Chapter 4.7.1 --- Family A --- p.67 / Chapter 4.7.2 --- Family B --- p.68 / Chapter 4.7.3 --- Family C --- p.68 / Chapter 4.7.4 --- Family D --- p.69 / Chapter 4.7.5 --- Family E --- p.70 / Chapter 4.7.6 --- Family F --- p.70 / Chapter Chapter 5 --- Discussion --- p.104 / Chapter 5.1 --- The predictive value of CHRPE in FAP patients and family members --- p.104 / Chapter 5.2 --- The laboratory techniques in this study --- p.105 / Chapter 5.2.1 --- PCR optimization --- p.105 / Chapter 5.2.2 --- Single Strand Conformation Polymorphism (SSCP) --- p.106 / Chapter 5.2.3 --- Direct DNA sequencing --- p.107 / Chapter 5.3 --- Novel mutation in the APC gene --- p.108 / Chapter 5.4 --- Reported mutations in the APC gene --- p.108 / Chapter 5.4.1 --- 3183del5 --- p.108 / Chapter 5.4.2 --- R216X and R283X --- p.109 / Chapter 5.5 --- Novel silent mutations and polymorphisms in the APC gene --- p.109 / Chapter 5.5.1 --- Novel silent mutations --- p.109 / Chapter 5.5.2 --- Polymorphisms --- p.110 / Chapter 5.6 --- The relationship between APC gene mutations and CHRPE --- p.111 / Chapter 5.7 --- Haplotype analysis --- p.112 / Chapter Chapter 6 --- Conclusion --- p.114 / Chapter Chapter 7 --- References --- p.115

Zheng Tingting. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 98-124). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Table of Contents --- p.vi / List of Tables --- p.viii / List of Figures --- p.ix / Abbreviations --- p.xi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Triamcinolone acetonide(TA) --- p.1 / Chapter 1.1.1 --- Application in ophthalmology --- p.1 / Chapter 1.1.2 --- Mechanism of anti-inflammatory effect --- p.2 / Chapter 1.1.3 --- Side effects of TA --- p.4 / Chapter 1.1.4 --- Toxicity of TA --- p.5 / Chapter 1.2 --- Retinal pigment epithelial (RPE) cell --- p.6 / Chapter 1.3 --- Mechanism of cell death --- p.8 / Chapter 1.3.1 --- Apoptosis --- p.9 / Chapter 1.3.2 --- caspase --- p.11 / Chapter 1.3.3 --- Mitogen-activated protein kinase (MAPK) --- p.13 / Chapter 1.3.4 --- Activator Protein-1 (AP-1) --- p.15 / Chapter 1.4 --- Potassium channel (K+）） --- p.15 / Chapter 1.4.1 --- Molecular structure of KAtp channel --- p.16 / Chapter 1.4.2 --- Regulation of Katp channel --- p.17 / Chapter 1.4.3 --- Pinacidil --- p.18 / Chapter 1.5 --- Calcium channel --- p.20 / Chapter 1.5.1 --- VDCCs and subtypes --- p.21 / Chapter 1.5.2 --- Calcium channel blocker --- p.22 / Chapter 1.5.3 --- Verapamil --- p.23 / Chapter 1.6 --- Study objectives --- p.25 / Chapter Chapter 2 --- Methodology --- p.37 / Chapter 2.1 --- Cell biology --- p.37 / Chapter 2.1.1 --- Materials --- p.37 / Chapter 2.1.1.1 --- Culture related material --- p.37 / Chapter 2.1.1.2 --- Drugs --- p.37 / Chapter 2.1.1.3 --- Cell line and instrument --- p.37 / Chapter 2.1.2 --- Preparations --- p.38 / Chapter 2.1.2.1 --- Working medium --- p.38 / Chapter 2.1.2.2 --- Drugs --- p.38 / Chapter 2.1.2.3 --- MTT solution --- p.39 / Chapter 2.1.3 --- Cell culture and treatment process --- p.40 / Chapter 2.1.3.1 --- Seed cell --- p.40 / Chapter 2.1.3.2 --- Treatment --- p.40 / Chapter 2.1.4 --- MTT-Cell Proliferation Assay --- p.41 / Chapter 2.2 --- Molecular biology --- p.42 / Chapter 2.2.1 --- Materials --- p.42 / Chapter 2.2.1.1 --- "Chemicals, reagents, and kits" --- p.42 / Chapter 2.2.1.2 --- Solutions and Buffers --- p.42 / Chapter 2.2.1.3 --- Primers and Enzymes --- p.43 / Chapter 2.2.1.4 --- Equipment --- p.43 / Chapter 2.2.1.5 --- Software --- p.43 / Chapter 2.2.2 --- Reverse transcription 226}0ؤ Polymerase Chain Reaction (RT-PCR) --- p.44 / Chapter 2.2.2.1 --- Cell collection and RNA Isolation --- p.44 / Chapter 2.2.2.2 --- Reverse Transcription (RT) --- p.45 / Chapter 2.2.2.3 --- PCR Reaction --- p.46 / Chapter 2.3 --- Immunocytochemistry --- p.48 / Chapter 2.3.1 --- Materials and instrumentation --- p.49 / Chapter 2.3.1.1 --- Antibodies and Equipment --- p.49 / Chapter 2.3.1.2 --- Chemicals and other useful items --- p.49 / Chapter 2.3.2 --- Preparations --- p.50 / Chapter 2.3.2.1 --- Preparation of coverslips --- p.50 / Chapter 2.3.2.2 --- Prepations of solutions --- p.50 / Chapter 2.3.3 --- Procedures --- p.51 / Chapter 2.4 --- Expression of results and statistics --- p.52 / Chapter Chapter 3 --- Results --- p.57 / Chapter 3.1 --- Effects of TA on RPE cell culture --- p.57 / Chapter 3.1.1 --- Cell morphology --- p.57 / Chapter 3.1.2 --- MTT assay --- p.57 / Chapter 3.1.3 --- Gene expressions --- p.57 / Chapter 3.2 --- Effects of PIN/VP on TA treated RPE cells --- p.58 / Chapter 3.2.1 --- MTT assay --- p.58 / Chapter 3.2.2 --- Gene expression --- p.59 / Chapter 3.2.2.1 --- Expression of housekeeping gene --- p.59 / Chapter 3.2.2.2 --- Expression of apoptosis-related gene --- p.59 / Chapter 3.2.2.3 --- Expression of early-response genes --- p.60 / Chapter 3.2.3 --- Immunofluorescence --- p.61 / Chapter Chapter 4 --- Discussion --- p.86 / Chapter Chapter 5 --- References --- p.98

Thesis (Ph. D.)--University of Virginia, 1999. / Spine title: IRBP expression & turnover. Includes bibliographical references (p. 176-177). Also available online through Digital Dissertations.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 6 uppsatser.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Oct. 13, 2008). Includes bibliographical references (p. 56-64).