Acquisition of full developmental competence of oocytes not only occurs during growth stage, and the final preparation during oocyte maturation is also critical. Previous studies have shown that nuclear maturation can occur spontaneously following culture in vitro; however, there may be some insufficiency in cytoplasmic maturation of the in vitro matured oocytes. But till now, the differences of the events of cytoplasmic maturation between in vitro and in vivo matured oocytes are still not clear. Ovarian stimulation by gonadotrophins is used to permits the growth and development of follicles, to time the initiation of pre-ovulatory oocyte maturation, and to increase the numbers of oocytes ovulated. It is one of the foundations of current treatments of human infertility. The success of clinical IVF has been depending on generation of matured oocytes at high frequency. However, ovarian stimulation with gonadotropins is associated with side effects and complications. / In order to illuminate mechanisms which affect the developmental competence of oocytes produced in vitro, in the present study, we have compared the difference of the quality of oocytes produced in vitro with that of the oocytes produced in vivo using mouse model. In order to understand the relationship between oocyte competence and ovarian responses to stimulation in the mouse, we also have compared difference of the quality of oocytes produced in vitro or in vivo from gonadotrophns stimulated ovaries with that of from natural cycling ovaries. / In-vitro matured oocytes were collected from (1) naturally ovulated mice and (2) superovulated (PMSG + hCG) mice. Immature oocytes were retrieved from (3) naturally cycling mice, and (4) from mice primed with PMSG. The results indicate that the percentages of cleavage and blastocyst formation are significantly different (P<0.05) between in-vivo and in-vitro matured oocytes. Blastocyst formation rate is significantly higher (P<0.05) in immature oocytes derived from PMSG primed mice compared to immature oocytes derived from naturally cycling mice. The percentages of oocytes with comet tails and the length of comet tails are significantly higher and longer respectively in in-vitro matured oocytes compared to in-vivo matured oocytes. Total cell numbers of blastocyst are also significantly different (P<0.05) between in-vivo and in-vitro matured oocytes. However, there are no differences in ratio of trophectoderm (TE)/inner cell mass (ICM) between in-vivo and in-vitro matured oocytes. In conclusion, in-vivo matured mouse oocytes are more competent than those of matured in-vitro, suggesting that it may be due to its less damage of DNA. Embryonic development capacity of in-vivo matured oocytes is not promoted by ovarian stimulation. Gonadotropin priming prior to immature mouse oocyte retrieval is beneficial to subsequent embryonic development. / Keywords. mouse oocyte, IVM, IVF, gonadotropin, development

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

Infants with bronchopulmonary dysplasia (BPD), showed impaired body growth when compared to control infants. In terms of changes in the biochemical composition of the lung, BPD infants had higher DNA, soluble protein, collagen and desmosine contents as well as increased concentrations of DNA, collagen and desmosine in their lungs when compared to the growth patterns obtained for the lungs of control infants. Pathologically BPD was classified into 4 grades. Grade I BPD, was a phase of acute lung injury, grades II and III were proliferative phases. In grade IV BPD, lung structure returned towards normal. Evidence of fibrosis was seen by a significant increase in collagen concentration in grades II and III while desmosine concentration was seen to increase in grades III and IV suggesting that the increase in collagen and desmosine contents in the lungs of BPD infants may be controlled by two different mechanisms. Collagen type I/III ratio was seen to decrease progressively from grade II to grade IV BPD in comparison to age matched controls, indicating a higher proportion of type III collagen in the lungs of infants with BPD. From the clinical analysis and the results obtained from discriminant analysis procedure, it was seen that there was a high degree of correlation between the continuation of the disease and collagen accumulation in the lungs suggesting that pulmonary fibrosis with excessive collagen accumulation is an integral part of BPD. This fibrotic process seemed to correlate significantly with assisted ventilation and high oxygen supplementation received by the infants, but it was difficult to assess the individual contribution of the two treatments in the pathogenesis of BPD. Other variables such as severity of the initial disease and the length of survival of the infants, made the assessment of individual contribution much more difficult. / Medicine, Faculty of / Pathology and Laboratory Medicine, Department of / Graduate

The morphological changes in arrangement of both cellular and extracellular material (ECM) during mouth formation in embryos of the asteroid Pisaster ochraceus, have been studied using LM, TEM, and SEM. In early gastrula, the ECM consists of small vesicles and amorphous material of an intermediate staining density, and "beaded strands" consisting of scattered intermediately stained fialments associated with densely stained granules. Initially, the ECM is found in the blastocoel immediately adjacent to the ectoderm and endoderm cells. In later stages, it extends further into the blastocoel, until it bridges the gap between the ectoderm and endoderm. As this occurs, less amorphous material is seen and increasing numbers of "beaded strands" are present throughout the region occupied by the ECM. These are particularly evident and appear better organized in the quadrant of the embryo in which the mouth will form. Between 59 and 90 hrs after fertilization, several events occur in rapid succession. The first involves the migration of cells from the endodermal epithelium at the tip of the archenteron into the blastocoel to form mesenchyme cells. Their departure leaves a hole in the endodermal epithelium of the archenteron tip which is covered by the endodermal basal lamina. Shortly after this, the presumptive stomodeal cells send filamentous and conical cellular processes into the blastocoel. The endodermal basal lamina covering the hole extends as a blister into the blastocoel. At the same time the archenteron has become bent toward the presumptive stomodeal region. In the blastocoel, mesenchyme cells are enmeshed in "beaded strands" located in the presumptive mouth region. At this stage, the strands are highly branched, and tend to be radially arranged, and have become almost completely encrusted with densely stained material.- Filopodia of the mesenchyme cells connect with the filamentous ectodermal processes described above. The cell bodies of the mesenchyme cells appear to be connected with either the basal lamina blister itself, or with scattered endoderm cells located within the blister, through small gaps in it. At a slightly later stage, the basal lamina blister is seen to be in contact with the conical ectodermal processes. Still later, a complete tube of basal lamina has formed between the ectoderm and endoderm. Initially, only scattered cells are associated with the surface of the tube. Later, the tube is occupied by endodermal cells and invaginated ectodermal cells forming the stomodeum . For approximately 24 hrs after it is formed, the mouth is plugged with an oral plate consisting of both ectodermal and endodermal cells. These cells eventually round up, loose their connection with their neighbors, and are lost to complete the formation of the mouth. The above observations suggest that the ECM components are secreted into the blastocoel by both the ectoderm and endoderm cells. The components then appear to undergo a self-assembly into a filamentous meshwork. This meshwork appears to form a scaffold in which the mesenchyme cells migrate. The change in orientation of the fibers in the meshwork and the increase in densely stained material associated with it after mesenchyme cells migrate through it, suggests that they may be responsible, at least in part, for these changes. Mesenchyme cells tend to be localized to the quadrant of the embryo in which the mouth will form, before and during mouth formation. This is also the region in which the ECM is highly organized. The increased organization of the ECM may guide and concentrate the mesenchyme cells to the presumptive mouth region where they form associations with both the filamentous processes of the stomodeal ectoderm and the blister of basal lamina, either directly or via connections through the basal lamina to cells within it. Once this contact has been made, contractile filaments, known to be located in the filopodia, could serve to pull the basal lamina blister across the blastocoel to make contact with the conical ectodermal spikes. This appears to be followed by fusion of the blister of the basal lamina with that of the stomodeum forming the endodermal tube. Once formed, the endodermal tube appears to provide a framework for the organization of both the ectodermal and endodermal cells which will form the mouth. / Medicine, Faculty of / Graduate

Retinal ganglion cell (RGC) axon arbors within the optic tectum are refined in development through a dynamic process of activity-dependent remodeling. The synaptic adhesion molecule N-cadherin is a candidate for mediating selective stabilization and elaboration of RGC axons due to its localization to perisynaptic sites and its modifiability by neural activity. RGCs of Xenopus tadpoles were co-transfected with plasmids encoding a dominant negative N-cadherin (N-cadDeltaE) and eGFP or eYFP. Using two-photon in vivo time-lapse imaging, we found that axons expressing N-cadDeltaE became less elaborate than controls over three days of daily live imaging. Shorter interval time-lapse imaging of axons expressing synaptophysin-GFP to visualize putative synaptic sites revealed that N-cadDeltaE expressing axons form fewer stable branches than controls and that stabilization of axonal branches at synaptic sites is altered. We conclude that N-cadherin participates in the stabilization of axonal branches in the Xenopus retinotectal system.

No description available.

Tsim Ting Yuk. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 119-135). / Abstracts in English and Chinese. / ABSTRACT --- p.i / 摘要 --- p.iii / 英漢譯名對照 --- p.v / ABBREVIATIONS --- p.vi / ACKNOWLEDGMENTS --- p.viii / Chapter 1. --- Introduction / Chapter 1.1. --- Olfactory system --- p.1 / Chapter 1.1.1. --- Olfactory bulb (OB) --- p.1 / Chapter 1.1.2. --- Accessory olfactory bulb (AOB) --- p.3 / Chapter 1.2. --- Stem cells --- p.5 / Chapter 1.3. --- Sexual differentiation --- p.8 / Chapter 1.3.1. --- Sexual dimorphic olfactory system --- p.8 / Chapter 1.3.2. --- Androgen receptor (AR) & estrogen receptor beta (ERβ) --- p.13 / Chapter 1.3.3. --- Aromatase --- p.15 / Chapter 1.3.4. --- Oligomycin sensitivity-conferringrotein (OSCP) --- p.18 / Chapter 1.4. --- rogrammed cell death (PCD) --- p.18 / Chapter 1.4.1. --- CD in the olfactory development --- p.18 / Chapter 1.4.2. --- Caspase 3 --- p.22 / Chapter 1.4.3. --- B cell leukemia/ Lymphoma 2 (Bcl-2) --- p.23 / Chapter 1.5. --- Axon guidance molecules --- p.25 / Chapter 1.5.1. --- Growth cone --- p.25 / Chapter 1.5.2. --- Mechanisms of growth cone advance --- p.26 / Chapter 1.5.3. --- Semaphorins --- p.28 / Chapter 1.5.4. --- Neuropilin --- p.31 / Chapter 1.5.5. --- lexin --- p.32 / Chapter 1.5.6. --- Collapsin response mediatorroteins (CRMPs) --- p.32 / Chapter 1.6. --- Olfactory markerroteins --- p.33 / Chapter 1.6.1. --- Markerroteins in ORNs --- p.33 / Chapter 1.6.2. --- Growth associatedrotein (GAP-43) --- p.34 / Chapter 1.6.3. --- Is the expression of GAP-43 in rat OB sexually dimorphic? --- p.36 / Chapter 1.6.4. --- Olfactory markerrotein (OMP) --- p.38 / Chapter 1.6.5. --- Golf --- p.39 / Chapter 1.7. --- Miscellaneous genes --- p.40 / Chapter 1.7.1. --- Substance (SP) --- p.40 / Chapter 1.7.2. --- Gonadotropin releasing hormone (GnRH) --- p.41 / Chapter 1.7.3. --- Metabotropic glutamate receptor 2 (mGluR2) --- p.42 / Chapter 1.7.4. --- Insulin-like growth factor binding protein-2 (IGFBP2) --- p.43 / Chapter 2. --- Materials and methods / Chapter 2.1. --- Animal study --- p.46 / Chapter 2.2. --- RNA extraction --- p.46 / Chapter 2.3. --- Quantitation of total RNA --- p.49 / Chapter 2.4. --- Reverse Transcription (RT) --- p.50 / Chapter 2.5. --- olymerase Chain Reaction (PCR) --- p.51 / Chapter 2.6. --- urification ofCRroducts --- p.55 / Chapter 2.7. --- Confirmation ofCRroducts --- p.56 / Chapter 2.8. --- Quantitation of cDNA --- p.57 / Chapter 2.9. --- Radioactive labeledCR --- p.58 / Chapter 2.10. --- Electrophoresis ofCRroducts --- p.59 / Chapter 2.11. --- Statistical analysis --- p.60 / Chapter 3. --- Results / Chapter 3.1. --- Standard curve construction --- p.61 / Chapter 3.2. --- β-actin --- p.62 / Chapter 3.3. --- Sexual differentiation related genes --- p.64 / Chapter 3.3.1. --- AR --- p.64 / Chapter 3.3.2. --- ERβ --- p.65 / Chapter 3.3.3. --- Aromatase --- p.65 / Chapter 3.3.4. --- OSCP --- p.66 / Chapter 3.4. --- CD related genes --- p.66 / Chapter 3.4.1. --- Bcl-2α --- p.66 / Chapter 3.4.2. --- Caspase 3 --- p.67 / Chapter 3.5. --- Axon guidance molecules and related genes --- p.67 / Chapter 3.5.1. --- SemaIII --- p.67 / Chapter 3.5.2. --- Neuropilin-1 --- p.68 / Chapter 3.5.3. --- lexin-1 --- p.68 / Chapter 3.5.4. --- CRMP-1 --- p.69 / Chapter 3.5.5. --- CRMP-2 --- p.70 / Chapter 3.5.6. --- CRMP-3 --- p.70 / Chapter 3.5.7. --- CRMP-4 --- p.71 / Chapter 3.6. --- Olfactory markerrotein genes --- p.71 / Chapter 3.6.1. --- GAP-43 --- p.71 / Chapter 3.6.2. --- OMP --- p.72 / Chapter 3.6.3. --- Golf --- p.72 / Chapter 3.7. --- Miscellaneous genes --- p.73 / Chapter 3.7.1. --- SubstanceP --- p.73 / Chapter 3.7.2. --- GnRH --- p.73 / Chapter 3.7.3. --- mGluR2 --- p.74 / Chapter 3.7.4. --- IGFBP-2 --- p.74 / Chapter 3.8. --- Graphs and tables --- p.75 / Chapter 4. --- Discussion / Chapter 4.1. --- Quantitation of cDNA and normalization of CR results --- p.97 / Chapter 4.2. --- Sexual differentiation related genes --- p.98 / Chapter 4.3. --- CD related genes --- p.100 / Chapter 4.4. --- Axon guidance molecule and related genes --- p.103 / Chapter 4.5. --- Olfactory markerrotein genes --- p.109 / Chapter 4.6. --- Miscellaneous genes --- p.112 / Chapter 5. --- References --- p.119

by Woo Yiu Ho, Anthony. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 162-171). / Abstract also in Chinese. / Abstract --- p.i / Acknowledgements --- p.iii / Table of Contents --- p.iv / List of Figures --- p.ix / List of Tables --- p.xi / List of Abbreviations --- p.xii / Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Bacterial Adhesion and Aggregation --- p.1 / Chapter 1.1.1 --- Significance of Bacterial Adhesion Studies --- p.1 / Chapter 1.1.2 --- Definitions --- p.4 / Chapter 1.1.3 --- Colonization Process --- p.7 / Chapter 1.1.4 --- Specific and Nonspecific Interactions --- p.8 / Chapter 1.1.5 --- Models of Bacterial Adhesion and Aggregation Processes --- p.14 / Chapter 1.1.6 --- Experimental Systems in Adhesion Research --- p.16 / Chapter 1.1.7 --- Experimental Systems in Aggregation Research --- p.19 / Chapter 1.2 --- Pseudomonas aeruginosa --- p.21 / Chapter 1.2.1 --- General Description and Clinical Significance --- p.21 / Chapter 1.2.2 --- Adhesins of Pseudomonas aeruginosa --- p.22 / Chapter 1.2.3 --- "Alginate, Mucoidity, Biofilm Formation and Cystic Fibrosis" --- p.23 / Chapter 1.2.4 --- Lipopolysaccharides --- p.26 / Chapter 1.2.5 --- Pili --- p.29 / Chapter 1.2.6 --- Flagella --- p.30 / Chapter 1.2.7 --- Lectins --- p.31 / Chapter 1.2.8 --- Other Adhesins --- p.31 / Chapter 1.2.9 --- Rhamnolipids --- p.32 / Chapter 1.3 --- Current Study --- p.33 / Chapter 2 --- MATERIALS AND EQUIPMENT --- p.35 / Chapter 2.1 --- Bacterial Strain --- p.35 / Chapter 2.2 --- Solid Surfaces --- p.35 / Chapter 2.3 --- Chemicals --- p.36 / Chapter 2.4 --- Recipes --- p.38 / Chapter 2.5 --- Equipment --- p.38 / Chapter 3 --- METHODS --- p.40 / Chapter 3.1 --- Maintenance and Culturation --- p.40 / Chapter 3.1.1 --- Maintenance of Bacterial Strains --- p.40 / Chapter 3.1.2 --- Seed Culture Preparation --- p.40 / Chapter 3.1.3 --- Culturation in Defined Growth Media --- p.40 / Chapter 3.2 --- Bacterial Adhesion and Aggregation Assay Methods --- p.41 / Chapter 3.2.1 --- Bacterial Adhesion on Glass Assay --- p.41 / Chapter 3.2.2 --- Bacterial Adhesion on Plastic Assay --- p.44 / Chapter 3.2.3 --- Bacterial Adhesion under Shear Assay --- p.44 / Chapter 3.2.4 --- Bacterial Aggregation Examination by Adhesion on Glass Assay --- p.45 / Chapter 3.2.5 --- Bacterial Aggregation Examination by Top-agar Assay --- p.45 / Chapter 3.2.6 --- Bacterial Aggregation Examination by Epi-fluorescence Microscopy --- p.46 / Chapter 3.2.7 --- Bacterial Aggregation Screening Test --- p.46 / Chapter 3.3 --- Determination of the Effects of Various Factors on Adhesion and Aggregation --- p.47 / Chapter 3.3.1 --- Culturation Period --- p.47 / Chapter 3.3.2 --- Osmotic Shock during the Washing Procedure --- p.47 / Chapter 3.3.3 --- Growth Media --- p.48 / Chapter 3.3.4 --- Assay Conditions --- p.48 / Chapter 3.3.5 --- Cell Pretreatments --- p.48 / Chapter 3.4 --- Isolation of Aggregation-deficient Mutants --- p.49 / Chapter 3.5 --- Outer Membrane Protein Profiles --- p.50 / Chapter 3.5.1 --- Isolation of Outer Membrane Fraction --- p.50 / Chapter 3.5.2 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.51 / Chapter 3.6 --- Determination of the Mobility of the Bacteria on Surfaces --- p.52 / Chapter 3.6.1 --- Subsurface Twitching Assay --- p.52 / Chapter 3.6.2 --- Soft-agar Swarm Assay --- p.53 / Chapter 3.7 --- Detection of Alginate Production --- p.53 / Chapter 3.7.1 --- Extraction of Alginate from Spent Growth Medium --- p.53 / Chapter 3.7.2 --- Releasing Cell Surface-associated Alginate --- p.54 / Chapter 3.8 --- Other Assay Methods --- p.55 / Chapter 3.8.1 --- Protein Assay --- p.55 / Chapter 3.8.2 --- Carbohydrate Determination --- p.55 / Chapter 3.8.3 --- Alginate Determination --- p.55 / Chapter 4 --- RESULTS --- p.57 / Chapter 4.1 --- Standardization of the Assays for Bacterial Adhesion and Aggregation --- p.57 / Chapter 4.1.1 --- Effects of Cell Density and Exposure Time on the Number of Adhered Bacteria Detected in Bacterial Adhesion on Glass Assay --- p.57 / Chapter 4.1.2 --- Characterization of Bacterial Aggregation by Different Examination Methods --- p.62 / Chapter 4.1.3 --- Effects of Culturation Period on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.67 / Chapter 4.1.4 --- Effects of Osmotic Shock during Washing on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.70 / Chapter 4.1.5 --- Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 as a Function of Time under the Standard Assay Condition --- p.71 / Chapter 4.1.6 --- Consistency of Bacterial Adhesion on Glass Assay --- p.74 / Chapter 4.2 --- Effects of Growth Media on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.77 / Chapter 4.3 --- Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 in Different Assay Media --- p.77 / Chapter 4.3.1 --- Effects of Various Buffers on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.77 / Chapter 4.3.2 --- Effects of pH on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.78 / Chapter 4.3.3 --- Effects of Various Electrolytes on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.81 / Chapter 4.3.4 --- Concentration Effects of Monovalent and Divalent Cations on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.88 / Chapter 4.3.5 --- Concentration Effects of Phosphate Buffers on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.93 / Chapter 4.3.6 --- Concentration Effects of Ammonium Sulfate and Cyclohexylammonium Sulfate on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.96 / Chapter 4.3.7 --- Effects of Cation Chelation on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.99 / Chapter 4.3.8 --- Effects of Sugars on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.100 / Chapter 4.3.9 --- Effects of Amino Acids on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.101 / Chapter 4.4 --- Adhesion and Aggregation after Pretreatments of the Cells --- p.103 / Chapter 4.4.1 --- Effects of Protease Treatments on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.103 / Chapter 4.4.2 --- Effects of Externally Added Proteins on Adhesion and Aggregation of Pronase-treated Cells --- p.107 / Chapter 4.4.3 --- Effects of Acid or Base Treatments on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.108 / Chapter 4.4.4 --- Effects of Heat Treatment on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.108 / Chapter 4.4.5 --- Effects of Extensive Washing on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.110 / Chapter 4.5 --- Isolation and Growth Characteristics of Aggregation-deficient Mutants --- p.111 / Chapter 4.6 --- Comparisons of the Adhesion and Aggregation Characters of Pseudomonas aeruginosa ATCC 10145 and Mutant 9 --- p.115 / Chapter 4.6.1 --- Under Standard Condition --- p.115 / Chapter 4.6.2 --- On Different Surfaces and in Different Electrolytes --- p.115 / Chapter 4.6.3 --- Under Shear --- p.118 / Chapter 4.6.4 --- Adhesion and Aggregation of Combined Suspensions of Pseudomonas aeruginosa ATCC 10145 and Mutant 9 --- p.122 / Chapter 4.7 --- Characterization of the Cell Surface Properties of Pseudomonas aeruginosa ATCC 10145 and Mutant 9 --- p.125 / Chapter 4.7.1 --- Outer Membrane Protein Profiles --- p.125 / Chapter 4.7.2 --- Pili-elicited Twitching Mobility --- p.125 / Chapter 4.7.3 --- Mobility Due to Flagella --- p.128 / Chapter 4.7.4 --- Production of Alginate --- p.128 / Chapter 5 --- DISCUSSIONS --- p.130 / Chapter 5.1 --- Choice of the Materials --- p.130 / Chapter 5.2 --- Development of the Assay Methods --- p.130 / Chapter 5.2.1 --- Development of the Procedures for Bacterial Adhesion Assays --- p.130 / Chapter 5.2.2 --- Development of the Assay Methods for Bacterial Aggregation --- p.132 / Chapter 5.2.3 --- Standardization of the Assays --- p.133 / Chapter 5.2.4 --- Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 as a Function of Time under the Standard Assay Condition --- p.134 / Chapter 5.2.5 --- Consistency of Bacterial Adhesion on Glass Assay --- p.135 / Chapter 5.2.6 --- Limits of Bacterial Adhesion on Glass Assay --- p.135 / Chapter 5.3 --- Effects of Growth Media on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.135 / Chapter 5.4 --- Effects of Various Chemicals in the Assay Media on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.136 / Chapter 5.4.1 --- Effects of Electrolytes on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.137 / Chapter 5.4.2 --- Effects of Aggregation on Adhesion --- p.140 / Chapter 5.4.3 --- Effects of Cyclohexylammonium Sulfate and Ammonium Sulfate on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.141 / Chapter 5.4.4 --- Effects of Sugars and Amino Acids on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.143 / Chapter 5.5 --- Effects of Various Cell-surface Modifications on Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 --- p.144 / Chapter 5.6 --- Isolation and Growth Characteristics of Aggregation-deficient Mutants --- p.146 / Chapter 5.7 --- Comparisons of the Adhesion and Aggregation Characters of Pseudonomas aeruginosa ATCC 10145 and Mutant 9 --- p.147 / Chapter 5.7.1 --- Adhesion and Aggregation of Pseudonomas aeruginosa ATCC 10145 and Mutant 9 on Different Surfaces In Different Electrolytes --- p.147 / Chapter 5.7.2 --- Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 and Mutant 9 Under Shear --- p.147 / Chapter 5.7.3 --- Adhesion and Aggregation of Combined Suspensions of Pseudomonas aeruginosa ATCC 10145 and Mutant 9 --- p.148 / Chapter 5.8 --- Characterization of the Cell Surface Properties of Pseudomonas aeruginosa ATCC 10145 and Mutant 9 --- p.148 / Chapter 5.9 --- General Discussions --- p.151 / Chapter 6 --- APPENDIX --- p.154 / Chapter 6.1 --- Visual Examination of Adhesion and Aggregation of Pseudomonas aeruginosa ATCC 10145 on Glass --- p.154 / Chapter 6.2 --- Fractal Analysis of Bacterial Aggregates --- p.154 / Chapter 7 --- REFERENCES --- p.162

Tjia Wai Mui. / Thesis submitted in: July 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 123-140). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Acknowledgement --- p.iv / Abbreviations --- p.v / Table of Contents --- p.vi / List of Figures --- p.x / List of Tables --- p.xii / Chapter Chapter One --- Literature Review / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Growth of L. edodes --- p.3 / Chapter 1.2.1 --- Life cycle of L. edodes --- p.3 / Chapter 1.2.2 --- Growth parameters of L. edodes --- p.6 / Chapter 1.2.2.1 --- Temperature --- p.6 / Chapter 1.2.2.2 --- Relative humidity --- p.7 / Chapter 1.2.2.3 --- Moisture content in substrate --- p.7 / Chapter 1.2.2.4 --- Light --- p.8 / Chapter 1.2.2.5 --- pH --- p.8 / Chapter 1.3 --- Cultivation of L. edodes --- p.9 / Chapter 1.3.1 --- History and development of artificial cultivation --- p.9 / Chapter 1.3.2 --- Use of forced fruiting --- p.11 / Chapter 1.4 --- Molecular studies of stress on fungi --- p.12 / Chapter 1.4.1 --- Studies of temperature stress in mushroom --- p.12 / Chapter 1.4.2 --- Studies of molecular chaperones in fungi --- p.13 / Chapter 1.4.2.1 --- Role of molecular chaperones --- p.13 / Chapter 1.4.2.2 --- Heat shock protein 70 (Hsp70) and their cochaperones --- p.13 / Chapter 1.4.2.3 --- Other chaperones --- p.15 / Chapter 1.4.2.4 --- Molecular chaperones and development --- p.16 / Chapter 1.5 --- Prospectus --- p.19 / Chapter Chapter Two --- The Effect of Stress on the Growth of L. edodes / Chapter 2.1 --- Introduction --- p.23 / Chapter 2.2 --- Materials and Methods --- p.24 / Chapter 2.2.1 --- Strain and culture conditions --- p.24 / Chapter 2.2.2 --- Stress treatments --- p.24 / Chapter 2.2.3 --- Data collection --- p.25 / Chapter 2.2.4 --- Data analysis --- p.25 / Chapter 2.3 --- Results --- p.27 / Chapter 2.3.1 --- Reliability analysis --- p.27 / Chapter 2.3.2 --- Descriptive statistics --- p.28 / Chapter 2.3.3 --- Independent t-test (ANOVA) --- p.33 / Chapter 2.4 --- Discussion --- p.37 / Chapter Chapter Three --- Sequence Analysis of selected Stress Genes / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.2 --- Materials and Methods --- p.40 / Chapter 3.2.1 --- Isolation of stress genes --- p.40 / Chapter 3.2.1.1 --- Construction of primordial cDNA library --- p.40 / Chapter 3.2.1.2 --- Screening of cDNA clones --- p.40 / Chapter 3.2.2 --- Sequence analyses of stress genes --- p.41 / Chapter 3.2.2.1 --- Amplification and purification of cDNA insert --- p.41 / Chapter 3.2.2.2 --- Full length DNA cycle sequencing --- p.42 / Chapter 3.2.2.3 --- Sequence analyses --- p.43 / Chapter 3.2.3 --- Screening of LeSSA (Inducible HSP70) --- p.45 / Chapter 3.2.3.1 --- PCR screening of LeSSA by degenerate primers and LeSSB specific primers --- p.45 / Chapter 3.2.3.2 --- Screening of LeSSA from cDNA library by hybridization --- p.49 / Chapter 3.3 --- Results --- p.51 / Chapter 3.3.1 --- Sequence analyses --- p.51 / Chapter 3.3.1.1 --- LeSSB --- p.51 / Chapter 3.3.1.2 --- LeMge1 --- p.57 / Chapter 3.3.1.3 --- LeSTI1 --- p.62 / Chapter 3.3.1.4 --- LeTCP1β --- p.69 / Chapter 3.3.1.5 --- LeTCP1γ --- p.74 / Chapter 3.3.2 --- Failure of isolating LeSSA (Inducible HSP70) --- p.80 / Chapter 3.4 --- Discussion --- p.82 / Chapter 3.4.1 --- Sequence analyses --- p.82 / Chapter 3.4.2 --- Screening of LeSSA --- p.84 / Chapter Chapter Four --- Characterization of stress genes upon different stresses / Chapter 4.1 --- Introduction --- p.86 / Chapter 4.2 --- Materials and Methods --- p.87 / Chapter 4.2.1 --- Strain and culture conditions --- p.87 / Chapter 4.2.2 --- Stress treatments --- p.87 / Chapter 4.2.3 --- Isolation of total RNAs --- p.87 / Chapter 4.2.4 --- Reverse transcriptase-polymerase chain reaction (RT-PCR) --- p.88 / Chapter 4.2.4.1 --- Reverse transcription --- p.88 / Chapter 4.2.4.2 --- PCR amplification by specific primers of stress genes --- p.89 / Chapter 4.2.5 --- Northern blot analyses --- p.91 / Chapter 4.2.5.1 --- RNA fractionation by formaldehyde gel electrophoresis --- p.91 / Chapter 4.2.5.2 --- Northern blotting --- p.91 / Chapter 4.2.5.3 --- Preparation of probes --- p.92 / Chapter 4.2.5.4 --- Hybridization and stringency washes --- p.93 / Chapter 4.2.6 --- Isolation of total protein --- p.94 / Chapter 4.2.7 --- Quantification of protein by Bradford method --- p.95 / Chapter 4.2.8 --- Western blot analyses --- p.95 / Chapter 4.2.8.1 --- Sodium dodecyl sulfate ´ؤ polyacrylamide gel electrophoresis (SDS-PAGE) --- p.95 / Chapter 4.2.8.2 --- Western blotting --- p.96 / Chapter 4.2.8.3 --- Immunodetection --- p.98 / Chapter 4.2.8.4 --- ECL detection --- p.98 / Chapter 4.3 --- Results --- p.99 / Chapter 4.3.1 --- Reverse transcriptase-polymerase chain reaction (RT-PCR) --- p.99 / Chapter 4.3.2 --- Northern blot hybridization --- p.106 / Chapter 4.3.2.1 --- Establishing an internal control --- p.106 / Chapter 4.3.2.2 --- Dig-labelling of stress genes --- p.106 / Chapter 4.3.2.3 --- Northern blot hybridizaton of stress genes --- p.106 / Chapter 4.3.3 --- Western blot hybridization --- p.111 / Chapter 4.4 --- Discussions --- p.113 / Chapter Chapter Five --- General Discussions --- p.118 / References --- p.123

Kong Fung Wong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 59-70). / Chapter CHATPER 1 --- GENERAL INTRODUCTION --- p.1-12 / Chapter CHATPER 2 --- EXAMINATION OF THE BEHAVIOR OF GROWTH CONE IN DIFFERENT REGIONS OF THE OPTIC CHIASM / Introduction --- p.13-14 / Materials and Methods --- p.15-18 / Results --- p.19-23 / Discussion --- p.24-27 / Chapter CHATPER 3 --- STUDY OF BINOCULAR INTERACTION AFTER UNILATERAL INTRA-UTERO ENUCLEATION / Introduction --- p.28-29 / Materials and Methods --- p.30-31 / Results --- p.32-35 / Discussion --- p.36-39 / Chapter CHATPER 4 --- ISOLATION OF DIFFERENTIALLY EXPRESSED mRNA IN DIFFERENT REGIONS OF THE RETINA / Introduction --- p.40-43 / Materials and Methods --- p.44-48 / Results --- p.48-50 / Discussion --- p.51-54 / Chapter CHATPER 5 --- GENERAL DISCUSSION --- p.56-58 / REFERENCE --- p.59-70 / FIGURES / TABLES