by Kwok Fai (Simon) Leung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1990. / Bibliography: leaves 72-99. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / Chapter SECTION 1: --- Literature Review / Chapter 1.1 --- Historical perspectives of ANP --- p.1 / Chapter 1.2 --- Nature of ANP --- p.5 / Chapter 1.3 --- Release of ANP --- p.13 / Chapter 1.4 --- Biological effects of ANP --- p.17 / Chapter 1.5 --- Clinical implications --- p.24 / Chapter SECTION 2: --- Effect of ANP on Left Atrium / Chapter 2.1 --- Introduction --- p.29 / Chapter 2.2 --- Methods --- p.32 / Chapter 2.3 --- Results --- p.37 / Chapter 2.4 --- Discussion --- p.44 / Chapter SECTION 3: --- Effect of ANP on Mesenteric Artery / Chapter 3.1 --- Introduction --- p.47 / Chapter 3.2 --- Methods --- p.51 / Chapter 3.3 --- Results --- p.62 / Chapter 3.4 --- Discussion --- p.66 / Chapter SECTION 4: --- General Discussion --- p.67 / Chapter SECTION 5: --- References --- p.72

Transition metals are crucial components of several metabolic pathways and are critical for DNA, RNA and protein synthesis. However, when found in excess, these metal ions are toxic. To maintain the physiological concentration of metal ions at non-toxic concentration, bacteria rely on members of the Ferric uptake regulator (Fur) family of metalloregulators. Intriguingly, despite being coined as “metalloregulator”, specific members of the Fur family activate and repress gene expression in presence or absence of regulatory metals. Based on these observations, we hypothesized that the ability of these transcription factors to adopt different structural conformations underlies their ability to display different functions in presence and absence of metal ions. To address this important question, we solved the crystal structure of apo-Fur protein from Campylobacter jejuni. Structural analysis revealed that the protein adopts a V-shaped conformation harboring an evolutionary conserved cluster of positively charged residues on the surface. Using an extensive library of mutants and electrophoretic mobility shift analysis, we found that substituting residues forming the positively charged surface is detrimental for Fur interaction with DNA. Furthermore, our in vivo studies suggest that these positively charged residues are important for the regulation of CjFur target genes and that different mechanisms modulate the activity of Fur family of metalloregulators depending on the number of occupied metal binding sites. We showed that the disruption of metal binding sites of CjFur significantly reduces DNA binding in vitro and is deleterious for the repression of Fur target genes and gut colonization by C. jejuni. Finally, based on initial findings that adding a tag at the N-terminus of CjFur significantly reduces its ability to incorporate regulatory metal ions and bind DNA, we developed a new protocol for the purification of a highly active untagged CjFur protein. Overall, our studies shed new lights on the mechanistic basis controlling Fur gene regulatory activity in C. jejuni.

在哺乳動物中，表皮生長因子(EGF)家族及其同源受體(EGFR)家族是卵泡生成的重要調節者。表皮生長因子受體信號轉導不但在促黃體激素誘導的卵母細胞成熟和排卵中是必不可少的，而且也通過與促性腺激素互相作用而影響卵泡的類固醇生產和生存。表皮生長因子對卵母細胞成熟的促進作用，也保存在金魚和斑馬魚等硬骨魚類。然而，在硬骨魚類中，有關表皮生長因子家族功能的知識仍然是非常有限的。在本研究中，我們的目的如下: (1)試圖找出那些表皮生長因子受體信號轉導路徑傳達表皮生長因子對卵母細胞成熟; (2)表皮生長因子受體和促黃體激素的潛在關係; (3)表皮生長因子和其他身體激素之間的相互作用。所有發現都可以幫助我們更系統地更了解表皮生長因子在斑馬魚卵泡發育中的作用。 / 我們實驗室先前的研究表明，表皮生長因子促進斑馬魚卵母細胞的成熟是通過激活素系統，但不知道其通過什麼細胞內信號轉導路徑調節激活素系統的表達。在這裡，我們使用斑馬魚卵泡培養和藥理抑製劑來研究幾個熟知的表皮生長因子受體轉導信號在表皮生長因子刺激激活素亞基表達的作用。我們發現，有絲分裂原活化蛋白質激酶3/1(MAPK3/1)，p38有絲分裂原活化蛋白質激酶 (p38MAPK)，蛋白激酶C(PKC)和蛋白激酶A(PKA)在抑制素βAA(inhbaa)和抑制素βB(inhbb)亞基表達中起到刺激作用，但對抑制素βAB(inhbab)的表達起抑制作用。另一方面，只有磷脂酰肌醇3激酶(PI3K)調節表皮生長因子對抑制素βB的表達，而它沒有對抑制素βAA和抑制素βB亞基的表達起任何淨影響。 / 除了表皮生長因子在斑馬魚卵巢自己產生的作用， 探討表皮生長因子和其他因素之間的相互作用也是很重要的，因為卵泡是由多種激素和彼此互動的因素控制。 / 在雌性哺乳動物的生殖中，表皮生長因子受體轉導促黃體激素的作用是十分重要的，這使我們很感興趣研究這種情況是否在斑馬魚一樣發生。 和哺乳動物不同，人绒毛膜促性腺激素(hCG)沒有激活斑馬魚表皮生長因子受體。另一方面，和哺乳動物相似，人绒毛膜促性腺激素誘導有絲分裂原活化蛋白質激酶3 / 1在斑馬魚卵泡細胞中的磷酸化，但它是通過蛋白激酶A不是表皮生長因子受體的。此外，在雌性哺乳動物中，人绒毛膜促性腺激素對有絲分裂原活化蛋白質激酶3 / 1的作用是使卵母細胞成熟, 但這作用可能不在斑馬魚中發生，因為它負調控抑制素βAB的表達。 / 表皮生長因子在斑馬魚卵巢的另一個有趣地方是胰島素的存在改變它對調節芳香化酶表達中的作用。表皮生長因子對芳香化酶表達起輕微的抑制效果，但它提高了胰島素對芳香化酶表達的刺激作用。此外，胰島素，促性腺激素和表皮生長因子一起的時候，更加增加了芳香化酶的表達。然而，表皮生長因子抑制佛司可林(FK)對芳香化酶和抑制素αmRNA表達的刺激作用。 / 兩者合計，我們提供了一些基本但重要的信息有關表皮生長因子及其受體在斑馬魚卵巢的功能，包括每個表皮生長因子受體信號轉導信號路徑的分子機制及它們參與調節激活素亞基的表達；表皮生長因子，促黃體激素和胰島素之間的相互作用。這些信息可以讓我們進一步探討表皮生長因子在斑馬魚卵泡以及其他脊椎動物的重要角色。 / In mammals, epidermal growth factor (EGF) family and its cognate receptor (EGFR) family are the key regulators of folliculogenesis. EGFR signaling not only is indispensable in luteinizing hormone-induced oocyte maturation and ovulation, but also influences steroidogenesis and survival of follicles via interacting with gonadotropins. The positive role of EGF on oocyte maturation is shown to conserve in teleosts such as goldfish and zebrafish. However, the knowledge about the functions of EGF family in teleosts is still very limited. In the present study, we sought to find out the EGFR signaling pathways that mediate the action of EGF on oocyte maturation, the potential relationship of EGFR and LH and the interaction between EGF and systemic hormones. All these findings help us to understand more about the role of EGF in zebrafish folliculogenesis in a more systematic manner. / Our lab previously demonstrate that EGF promotes oocyte maturation in zebrafish via activin system but the contribution of the EGFR signaling pathways in regulating the activin system is not known. Here, we investigated the role of several well-known EGFR signaling cascades in EGF-stimulated activin subunit expression using cultured zebrafish follicle and pharmacological inhibitors. We found that MAPK3/1, p38 MAPK, PKC and PKA played stimulatory role in the expression of inhbaa and inhbb but suppressive role in inhbab. On the other hand, only PI3K was found to mediate the action of EGF on the expression of inhbab while it did not have any net effect on the expression of inhbaa and inhbb. / Apart from its own action in zebrafish ovary, it is essential to investigate the interaction or cross-talk between EGF and other factors because folliculogenesis is controlled by many hormones and factors interacting with each other. / The vital mediatory action of EGFR on LH in female reproduction of mammals was led us to ask whether the situation is the same as in zebrafish. However, unlike in mammals, hCG did not transactivate zebrafish EGFR. On the other hand, similar to mammals, hCG did induce MAPK3/1 phosphorylation in zebrafish follicle cells but it was via PKA dependent but not EGFR dependent manner. Moreover, hCG-induced MAPK3/1 activation is involved in oocyte maturation in mammals while in zebrafish, the function of this activation might not be the same as in mammals because it negatively regulated the expression of inhbab. / Another interesting issue about EGF in zebrafish ovary is that its role in regulating the expression of aromatase in response to insulin. EGF alone has slight suppressive effect on aromatase expression but it boosted the stimulatory effect of insulin on aromatase expression. Furthermore, insulin, hCG and EGF together even increased the expression of aromatase. However, EGF always suppressed the stimulatory effect of forskolin on aromatase and inhibin alpha mRNA expression. / Taken together, we provided some fundamental but important information about the function of EGF and its receptor in zebrafish ovary by elucidating the molecular mechanism of each EGFR signaling pathways involved in regulating the expression of activins subunits, the interaction between EGF, LH and insulin. All these studies not only allow us to further explore the elegant roles of EGF in zebrafish folliculogenesis, but also in other vertebrates in the future. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Chung, Chi Kin. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 116-132). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract in English --- p.I / Abstract in Chinese --- p.III / Acknowledge --- p.V / Table of contents --- p.VI / List of figures and tables --- p.XI / Symbols and abbreviations --- p.XVI / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Folliculogenesis --- p.1 / Chapter 1.1.1 --- In mammals --- p.1 / Chapter 1.1.2 --- In teleosts --- p.2 / Chapter 1.1.2.1 --- Vitellogenesis --- p.3 / Chapter 1.1.2.2 --- Oocyte maturation --- p.3 / Chapter 1.2 --- Epidermal growth factor receptor family --- p.4 / Chapter 1.2.1 --- Structure --- p.4 / Chapter 1.2.2 --- Signal transduction pathways --- p.5 / Chapter 1.2.2.1 --- MEK1/2-MAPK3/1 pathway --- p.6 / Chapter 1.2.2.2 --- p38 MAPK pathway --- p.6 / Chapter 1.2.2.3 --- PI3K pathway --- p.7 / Chapter 1.2.2.4 --- PKC pathway --- p.7 / Chapter 1.2.3 --- Reproductive function of EGFR signaling --- p.8 / Chapter 1.2.3.1 --- In mammals - mediator of luteinizing hormone --- p.9 / Chapter 1.2.3.2 --- In teleosts --- p.10 / Chapter 1.3 --- Interaction of EGF family with other hormones --- p.11 / Chapter 1.4 --- Objectives of the present study --- p.12 / Chapter Chapter 2 --- The Involvement of Diverse EGFR Signaling Pathways in Regulating the Expression of Activin Subunits in Zebrafish / Chapter 2.1 --- Introduction --- p.15 / Chapter 2.2 --- Materials and methods --- p.16 / Chapter 2.2.1 --- Animals and chemicals --- p.16 / Chapter 2.2.2 --- Primary zebrafish follicle cell culture --- p.17 / Chapter 2.2.3 --- Follicle incubation --- p.18 / Chapter 2.2.4 --- Immunoblotting --- p.18 / Chapter 2.2.5 --- Immunohistochemistry --- p.18 / Chapter 2.2.6 --- RNA extraction and reverse transcription --- p.19 / Chapter 2.2.7 --- Real-time quantitative polymerase chain reaction (qPCR) --- p.20 / Chapter 2.2.8 --- Data analysis --- p.20 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Recombinant human EGF up-regulated the expression of all known activin subunits in zebrafish ovary via specific activation of zebrafish EGFR --- p.21 / Chapter 2.3.2 --- MAPK3/1, p38 MAPK, PKC and PKA played similar roles in regulating EGF-stimulated expression of activin subunits --- p.22 / Chapter 2.3.3 --- PI3K pathway was involved in EGF-stuimulated expression of inhbab but not inhbaa and inhbb in zebrafish follicle cells --- p.24 / Chapter 2.3.4 --- MAPK3/1, p38 MAPK and PKC all together were sufficient to mediate the action of EGF on the expression of all activin subunits in zebrafish follicle cells --- p.24 / Chapter 2.3.5 --- Signaling cross-talk between PKA, MAPK3/1 and PI3K in zebrafish follicle cells --- p.25 / Chapter 2.3.6 --- EGFR signaling in the intact zebrafish follicles --- p.25 / Chapter 2.3.7 --- EGF and hCG togher up-regulatory the expression of inhbab synergistically in zebrafish follicle cells --- p.25 / Chapter 2.4 --- Discussion --- p.26 / Chapter Chapter 3 --- hCG-induced MAPK3/1 Phosphorylation in the Zebrafish Follicle Cells is Independent of EGFR Activation / Chapter 3.1 --- Introduction --- p.52 / Chapter 3.2 --- Materials and methods --- p.54 / Chapter 3.2.1 --- Animals and chemicals --- p.54 / Chapter 3.2.2 --- Primary zebrafish follicle cell culture --- p.54 / Chapter 3.2.3 --- Immunoblotting --- p.54 / Chapter 3.2.4 --- RNA extraction and reverse transcription --- p.54 / Chapter 3.2.5 --- Real-time quantitative polymerase chain reaction (qPCR) --- p.54 / Chapter 3.2.6 --- Data analysis --- p.54 / Chapter 3.3 --- Results --- p.54 / Chapter 3.3.1 --- hCG stimulated MAPK3/1 phosphorylation in zebrafish follicle cells --- p.54 / Chapter 3.3.2 --- hCG-induced MAPK3/1 phosphorylation was PKA-dependent but EGFR-independent --- p.55 / Chapter 3.3.3 --- Role of hCG-induced MAPK3/1 activation in regulating the expression of activin subunits --- p.55 / Chapter 3.4 --- Discussion --- p.56 / Chapter Chapter 4 --- Interaction between EGF, hCG and insulin on the Expression of inha and cyp19a1a in Zebrafish Follicles - a Potential Link between Nutrition and Reproduction / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Materials and chemicals --- p.69 / Chapter 4.2.1 --- Animals and chemicals --- p.69 / Chapter 4.2.2 --- Primary zebrafish follicle cell culture --- p.69 / Chapter 4.2.3 --- Follicle incubation --- p.69 / Chapter 4.2.4 --- Immunhistochemistry --- p.69 / Chapter 4.2.5 --- RNA extraction and reverse transcription --- p.70 / Chapter 4.2.6 --- Real-time quantitative polymerase chain reaction (qPCR) --- p.70 / Chapter 4.2.7 --- Data analysis --- p.70 / Chapter 4.3 --- Results --- p.70 / Chapter 4.3.1 --- Recombinant human insulin up-regulated the expression of inha and cyp19a1a in zebrafish follicles --- p.70 / Chapter 4.3.2 --- Interaction between insulin and hCG on the expression of cyp19a1a and inha in intact zebrafish follicles and follicle cells --- p.71 / Chapter 4.3.3 --- Interaction of cAMP pathway and insulin in the regulation of cyp19a1a and inha in zebrafish follicle and zebrafish follicle cells --- p.72 / Chapter 4.3.4 --- Differential effect of EGF on insulin, hCG and FK-regulated expression of cyp19a1a and inha in zebrafish follicles and follicle cells --- p.72 / Chapter 4.3.5 --- Insulin activated Akt pathway in both follicle cells and oocyte in zebrafish follicles --- p.73 / Chapter 4.4 --- Discussion --- p.73 / Chapter Chapter 5 --- General Discussion --- p.96 / Chapter Appendix --- Chapter Phosphorylation of MAPK3/1 in the Follicle in Vivo Evidence for Roles of MAPK3/1 in the Two Compartments of Follicles during Final Maturation and Ovulation / Introduction --- p.104 / Materials and methods --- p.105 / Animals and chemicals --- p.105 / Immunohistochemistry --- p.105 / Results --- p.105 / Discussion --- p.106 / References --- p.116

"The MCNP6 radiation transport code was used to quantify changes in the absorbed dose tissue conversion factors for LiF, Al2O3, and silicon-based electronic dosimeters. While normally calibrated in-air and applied to all general geometric measurements, tissue conversion factors for each dosimeter were obtained at various depths in a simulated water phantom and compared against the standard in-air calibration method. In these experiments, a mono-energetic photon source was modeled at energies between 30 keV and 300 keV for a point-source placed at the center of a water phantom, a point-source placed at the surface of the phantom, and for a 10-cm radial field geometry. Again, mono-energetic photon source was modeled up to 1300 keV for a disk-source placed at the surface of the phantom and dosimetric calculations were obtained for water, LiF, Al2O3, and silicon at depths of 1 mm to 35 cm from the source. The dosimeter’s absorbed dose conversion factor was calculated as a ratio of the absorbed dose to water to that of the dosimeter measured at a specified phantom depth. The dosimeter’s calibration value also was obtained for both mono and polyenergetic source and the calibration value from poly-energetic source was compared with the intensity weighted average calibration value from mono-energetic photon. The calculated changes in the tissue conversion factors are significant because the American Association of Physicists in Medicine (AAPM) recommend that measurements of a brachytherapy or diagnostic source be made with an overall uncertainity of 5% or better. Yet, based on results, the absorbed dose tissue conversion factor for a LiF dosimeter was found to deviate from its calibration value by up to 9%, an Al2O3 dosimeter by 43%, and a silicon dosimeter by 61%. These uncertainties are in addition to the normal measurement uncertainties. By applying these tissue correction factors, these data may be used to meet the AAPM measurement requirements for mono-energetic and poly-energetic sources at measurement depths up to 35 cm under the irradiation geometries investigated herein. "

In analysis of the contingency tables, the Fisher's exact test is a very important statistical significant test that is commonly used to test independence between the two variables. However, the Fisher' s exact test is based upon the assumption of the fixed margins. That is, the Fisher's exact test uses information beyond the table so that it is conservative. To solve this problem, we allow the margins to be random. This means that instead of fitting the count data to the hypergeometric distribution as in the Fisher's exact test, we model the margins and one cell using multinomial distribution, and then we use the likelihood ratio to test the hypothesis of independence. Furthermore, using Bayesian inference, we consider the Bayes factor as another test statistic. In order to judge the test performance, we compare the power of the likelihood ratio test, the Bayes factor test and the Fisher's exact test. In addition, we use our methodology to analyse data gathered from the Worcester Heart Attack Study to assess gender difference in the therapeutic management of patients with acute myocardial infarction (AMI) by selected demographic and clinical characteristics.

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Factor H (FH) is a 155-kDa plasma protein that regulates the alternative pathway of the complement system. Its 20 CCP modules, of 51-62 amino acid residues each, are linked by short stretches (“linkers’) of three to eight residues. We set out to test the hypothesis that long linkers towards the middle of FH play a role in ensuring that its architecture allows binding sites near its N- and C-termini to engage cooperatively with the main target, C3b, which is the key complement pathway-triggering product of C3 cleavage. In initial work, site-directed mutagenesis was used to test whether two mutations, R53H and R78G, located within CCP 1 and linked to the kidney disease atypical hemolytic uremic syndrome, are functionally deficient. Mutant versions and a native-sequence version of CCPs 1-4 of FH (i.e. FH 1-4) were tested for their ability to act as a cofactor for the FI-mediated cleavage of C3b, and accelerate the decay of the C3 convertase. It was shown that FH 1-4 R53H binds normally to C3b but has no regulatory activity while FH 1-4 R78G binds very poorly and is also deficient in cofactor and decay-accelerating activities. In subsequent work, mutagenesis was used to make the eight-residue CCPs 12-13 linker shorter (SL), or more flexible through introduction of glycine residues (3xGLY), within recombinant (r) module pair FH 12-13, and in rFH 10-15 and rFH 8-15 as well as full-length rFH. NMR showed CCPs 12 and 13 remain intact following mutation of the linker but (in FH 12-13) are more flexibly mutually disposed, as expected. SAXS indicated that both FH 10-15 SL and FH 10-15 3xGLY nonetheless have similar compact structures to native sequence (WT) FH 10-15. On the other hand, FH linker mutants interact with C3b (according to surface plasmon resonance) somewhat less well than WT FH and in the case of FH SL, affinity is similar to that of FH 19-20, i.e. there is no evidence that both C3b-binding sites in this mutant bind to the target simultaneously. Nonetheless, the bacterial protein PspCN boosts binding of linker mutants to C3b by a similar factor (three-to-fivefold) to that observed for FH WT. Thus, while interactions between non-sequential CCPs are important for FH architecture, a bend at the 12-13 linker is needed for full-length FH to adopt a fully biological activity confirmation. The use of EPR for structural studies of rFH and its mutants was explored. Free cysteines were engineered in so they could have spin labels site-specifically attached. Alternatively, a recognition site for transglutaminase was introduced so a spin label could be incorporated. These strategies were applied to rFH 12-13 and rFH 10-15 as a prelude to studies of full-length FH. Several suitably engineered proteins were prepared but only one paramagnetically labeled sample (of FH 12-13) made it for EPR; this yielded results commensurate with the NMR-derived structure. Taken together, these promising data lay the groundwork for a future, potentially very insightful, combined mutagenesis and EPR study of FH architecture and its role in complement activation.

Tse, Chung Kwan Anna. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 86-105). / Abstracts in English and Chinese. / Abstract in English --- p.i / Abstract in Chinese --- p.iii / Acknowledgement --- p.v / Table of content --- p.vii / List of figures and tables --- p.xi / Symbols and abbreviation --- p.xiii / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Folliculogenesis --- p.1 / Chapter 1.1.1 --- In Mammals --- p.1 / Chapter 1.1.2 --- In Teleosts --- p.2 / Chapter 1.1.2.1 --- Vitellogenesis --- p.3 / Chapter 1.1.2.2 --- Oocyte Maturation --- p.4 / Chapter 1.1.3 --- Oocyte Control of Folliculogenesis --- p.5 / Chapter 1.2 --- Epidermal Growth Factor Family --- p.7 / Chapter 1.2.1 --- Epidermal Growth Factor Ligand Family --- p.7 / Chapter 1.2.1.1 --- Discovery --- p.7 / Chapter 1.2.1.2 --- Structure --- p.8 / Chapter 1.2.1.3 --- Shedding --- p.8 / Chapter 1.2.1.4 --- Functions --- p.9 / Chapter 1.2.2 --- Epidermal Growth Factor Receptor Family --- p.10 / Chapter 1.2.2.1 --- Structure --- p.10 / Chapter 1.2.2.2 --- Ligand Binding and Dimerization --- p.11 / Chapter 1.2.2.3 --- Signaling and Internalization --- p.12 / Chapter 1.2.3 --- EGF Family in Reproduction --- p.13 / Chapter 1.2.3.1 --- Reproductive Function of EGF Family --- p.13 / Chapter 1.2.3.2 --- EGF Network as Downstream Mediator of Gonadotropins --- p.15 / Chapter 1.3 --- Objectives of the Present Study --- p.18 / Chapter Chapter 2 --- Spatiotemporal Expression of the EGF Family in Zebrafish Ovary / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Materials and Methods --- p.22 / Chapter 2.2.1 --- Chemicals --- p.22 / Chapter 2.2.2 --- Animals --- p.22 / Chapter 2.2.3 --- Isolation of Different Tissues and Ovarian Follicles --- p.22 / Chapter 2.2.4 --- Separation of Oocyte and Follicle layers --- p.23 / Chapter 2.2.5 --- Embryo Collection --- p.23 / Chapter 2.2.6 --- Tyrphostin AG 1478 Treatment of Embryos --- p.23 / Chapter 2.2.7 --- Total RNA Isolation and Reverse Transcription --- p.24 / Chapter 2.2.8 --- Semi-quantitative and Real-time Polymerase Chain Reaction (PCR) --- p.24 / Chapter 2.2.9 --- Data Analysis --- p.25 / Chapter 2.3 --- Results --- p.27 / Chapter 2.3.1 --- Validation of Semi-quantitative PCR Quantification --- p.27 / Chapter 2.3.2 --- Tissue Distribution of the EGF Family --- p.27 / Chapter 2.3.3 --- Spatial Expression of the EGF and the Activin Families within the Follicles --- p.27 / Chapter 2.3.4 --- Temporal Expression of the EGF Family in Folliculogenesis --- p.29 / Chapter 2.3.5 --- Temporal Expression of the EGF Family in Embryogenesis --- p.29 / Chapter 2.3.6 --- Effect of Blocking EGFR on Embryonic Development --- p.30 / Chapter 2.4 --- Discussion --- p.31 / Chapter Chapter 3 --- Regulation of the EGF Family in Cultured Follicle Cells and Their Effects on the Expression of Activin Subunits / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Materials and Methods --- p.58 / Chapter 3.2.1 --- Chemicals --- p.58 / Chapter 3.2.2 --- Primary Follicle Culture --- p.58 / Chapter 3.2.3 --- Ovarian Fragment Incubation --- p.58 / Chapter 3.2.4 --- Total RNA Isolation and Reverse Transcription --- p.58 / Chapter 3.2.5 --- Semi-quantitative and Real-time Polymerase Chain Reaction (PCR) --- p.59 / Chapter 3.2.6 --- Microinjection --- p.59 / Chapter 3.2.7 --- Data analysis --- p.59 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Expression of the EGF Family in Cultured Follicle Cells --- p.61 / Chapter 3.3.2 --- Effects of EGF on the EGF Family Expression in Cultured Follicle Cells --- p.61 / Chapter 3.3.3 --- Effects of BTC and HB-EGF on the Expression of EGF Ligands in Cultured Follicle Cells --- p.61 / Chapter 3.3.4 --- Effect of EGF Ligands on the Expression of Activin Subunits in Cultured Follicle Cells --- p.62 / Chapter 3.3.5 --- Effects of Actinomycin D on Ovarian Fragments --- p.62 / Chapter 3.3.6 --- Effects of Microinjecting Anti-EGF Morpholino on Full Grown Follicles --- p.62 / Chapter 3.4 --- Discussion --- p.64 / Chapter Chapter 4 --- General Discussion --- p.81 / References --- p.86

Duan, Xin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 36-43). / Abstracts in English and Chinese. / Abstract --- p.i-ii / Acknowledgement --- p.iii / Table of Contents --- p.iv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Related Literature --- p.6 / Chapter 2.1 --- Exchange Market Pressure Index --- p.6 / Chapter 2.2 --- The Sentiment Index --- p.11 / Chapter Chapter 3 --- Stock market sentiment --- p.16 / Chapter 3.1 --- Data --- p.16 / Chapter 3.2 --- Methodology --- p.23 / Chapter 3.3 --- Estimated Results --- p.25 / Chapter Chapter 4 --- Application to the Hong Kong stock market --- p.28 / Chapter 4.1 --- Threshold Model Estimation --- p.28 / Chapter 4.2 --- Trading Strategy --- p.30 / Chapter Chapter 5 --- Conclusion --- p.32 / Appendix: Principle Component --- p.34 / References --- p.36

The activation and effector function of CD4+ T cells are critical points of regulation during an antigen specific T cell response. Dysregulation of these processes can lead to the development of human diseases, encompassing both immunodeficiency and autoimmunity. Members of the TNF superfamily have recently emerged as important regulators of T cell responses, with their overexpression causing autoimmune inflammation in animal models. As overproduction of the novel TNF superfamily ligand BAFF is associated with several autoimmune conditions, we sought to examine the potential role of BAFF as a regulator of T cell activation and effector function. We initially demonstrated BAFF costimulation of T cell activation in vitro. Generation of specific monoclonal antibodies identified BAFF-R as the only BAFF receptor present on T cells, and showed that it was expressed in an activation-dependent and subset-specific manner. Impaired BAFF costimulation in BAFF-R deficient mice indicated that BAFF-R was crucial for mediating BAFF effects in T cells. Analysis of T cell responses in vivo revealed that BAFF transgenic mice have increased T cell priming and recall responses to protein antigens, and showed a corresponding increase in the DTH model of Th1 cell-dependent inflammation. In addition, Th2-dependent allergic airway responses are suppressed in BAFF transgenic mice. Crossing to a B cell deficient background revealed that the proinflammatory effects of BAFF on T cell priming and DTH rely on the presence of B cells, while the suppressive effects during allergic airway inflammation are B cell independent. These data demonstrated that BAFF regulated the outcome of T cell responses in vivo and identified BAFF dependent crosstalk between T and B cells. Stimulation of B cells with BAFF induced the upregulation of MHC class II and ICOS-L both in vitro and in vivo. Induction of these cell surface molecules was associated with an increased capacity to induce T cell proliferation, however this effect was independent of ICOS-L expression. Thus it was demonstrated that BAFF regulated T cell activation and effector function both directly, via stimulation of BAFF-R, and indirectly, by altering the function of B cells. These data suggest that BAFF dependent alterations in T cell function may be an additional causative factor in the association between elevated BAFF levels and the generation of autoimmunity.