Lam Kin Bong Hubert. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 139-161). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / (in chinese) --- p.iv / Acknowledgements --- p.vi / Publications --- p.ix / Table of Contents --- p.x / List of Tables --- p.xiii / List of Figures --- p.xiv / List of Abbreviations --- p.xvi / Chapter CHAPTER1： --- General Introduction --- p.1 / Chapter Section 1.1 --- Historical Background and Application of Arsenic Trioxide as an Anti-cancer Agent --- p.1 / Chapter Section 1.2 --- Arsenic Trioxide Induces Apoptosis in Cancer Cells --- p.3 / Chapter 1.2.1 --- The Intrinsic and Extrinsic Pathways of Apoptosis Initiation --- p.4 / Chapter 1.2.2 --- The Convergence of Pathways --- p.8 / Chapter 1.2.3 --- Induction of Apoptosis by Arsenic Trioxide --- p.9 / Chapter 1.2.3.1 --- Controversies in the Involvement of the Extrinsic Pathway --- p.9 / Chapter 1.2.3.2 --- "Arsenic Trioxide, Oxidative Stress and the Mitochondria" --- p.10 / Chapter 1.2.3.3 --- Caspase-3 Activation in Arsenic Trioxide-mediated Apoptosis --- p.12 / Chapter Section 1.3 --- Arsenic Trioxide Perturbs the Cell Division Cycle --- p.13 / Chapter 1.3.1 --- The Cell Cycle Oscillator --- p.14 / Chapter 1.3.2 --- DNADamage and Cell Cycle Checkpoints --- p.15 / Chapter 1.3.3 --- Induction of Cell Cycle Arrest by Arsenic Trioxide and its Association with Apoptosis --- p.17 / Chapter Section 1.4 --- Acute Megakaryocytic Leukemia and Arsenic Trioxide --- p.20 / Chapter CHAPTER 2： --- Objectives --- p.28 / Chapter CHAPTER 3： --- Methodology --- p.30 / Chapter Section 3.1 --- Materials --- p.30 / Chapter Section 3.2 --- Methods --- p.39 / Chapter 3.2.1 --- Culture of Megakaryocytic Cells and Their Treatment with Arsenic Trioxide --- p.39 / Chapter 3.2.1.1 --- Maintenance of Cell Lines --- p.39 / Chapter 3.2.1.2 --- Treatment with Arsenic Trioxide --- p.39 / Chapter 3.2.2 --- "Effects of Arsenic Trioxide on Cell Proliferation, Apoptosis, Mitochondrial Integrity and Cell Division Cycle Profiles of Human Megakaryocytic Leukemia Cell Lines" --- p.40 / Chapter 3.2.2.1 --- Trypan Blue Exclusion Assay --- p.40 / Chapter 3.2.2.2 --- Quantitation of Externalized Phosphatidylserine --- p.41 / Chapter 3.2.2.3 --- Quantitation of Active Caspase-3 Expression --- p.42 / Chapter 3.2.2.4 --- Assessment of Mitochondrial Intensity --- p.42 / Chapter 3.2.2.5 --- Analysis of Cell Division Cycle Profile --- p.43 / Chapter 3.2.2.6 --- Analysis of Cell Cycle Kinetics by BrdU Labeling --- p.43 / Chapter 3.2.2.7 --- Identification of Cell Cycle Specificity of Caspase-3 Expression --- p.45 / Chapter 3.2.3 --- Effects of Arsenic Trioxide on the Expression of Apoptotic Signals in Human Megakaryocytic Leukemia Cell Lines --- p.45 / Chapter 3.2.3.1 --- Effects of Arsenic Trioxide on mRNA Expression Levels of Apoptotic Regulators --- p.45 / Chapter 3.2.3.2 --- Effects of Arsenic Trioxide on Protein Expression Levels of Apoptotic Regulators --- p.50 / Chapter 3.2.3.2.1 --- Flow Cytometric Analysis --- p.50 / Chapter 3.2.3.2.2 --- Western Blot Analysis --- p.51 / Chapter 3.2.4 --- Effects of Arsenic Trioxide on Gene Expression Profiles of Human Megakaryocytic Leukemia Cell Lines By Microarray Analysis --- p.54 / Chapter 3.2.5 --- Statistical Analysis --- p.57 / Chapter CHAPTER 4： --- "Effects of Arsenic Trioxide on Cell Proliferation, Apoptosis, Mitochondrial Integrity and Cell Division Cycle Profiles of Human Megakaryocytic Leukemia Cell Lines" --- p.62 / Chapter Section 4.1 --- Introduction --- p.62 / Chapter Section 4.2 --- Results --- p.63 / Chapter 4.2.1 --- Effects of Arsenic Trioxide on Proliferation Kinetics --- p.63 / Chapter 4.2.2 --- Effects of Arsenic Trioxide on Cell Viability --- p.64 / Chapter 4.2.3 --- Apoptosis-inducing Capability of Arsenic Trioxide --- p.65 / Chapter 4.2.3.1 --- Quantitation of Externalized Phosphatidylserine --- p.65 / Chapter 4.2.3.2 --- Quantitation of Active Caspase-3 Expression --- p.66 / Chapter 4.2.4 --- Effects of Arsenic Trioxide on Mitochondrial Integrity --- p.67 / Chapter 4.2.5 --- Effects of Arsenic Trioxide on Cell Division Cycle Profiles --- p.69 / Chapter 4.2.6 --- Effects of Arsenic Trioxide on Cell Cycle Kinetics by Bromodeoxyuridine Labeling --- p.69 / Chapter 4.2.7 --- Identification of Cell Cycle Specificity of Arsenic Trioxide-Induced Caspase-3 Activation --- p.71 / Chapter Section 4.3 --- Discussion --- p.72 / Chapter CHAPTER 5： --- Effects of Arsenic Trioxide on Apoptotic Signal Expression in Human Megakaryocytic Leukemia Cell Lines --- p.91 / Chapter Section 5.1 --- Introduction --- p.91 / Chapter Section 5.2 --- Results --- p.92 / Chapter 5.2.1 --- Effects of Arsenic Trioxide on mRNA Expression Levels of Apoptotic Regulators --- p.92 / Chapter 5.2.2 --- Effects of Arsenic Trioxide on Protein Expression Levels of Apoptotic Regulators --- p.94 / Chapter 5.2.2.1 --- Flow Cytometric Analysis --- p.94 / Chapter 5.2.2.2 --- Western Blot Analysis --- p.96 / Chapter Section 5.3 --- Discussion --- p.96 / Chapter CHAPTER 6： --- Effects of Arsenic Trioxide on Gene Expression Profiles of Human Megakaryocytic Leukemia Cell Lines by Microarray Analysis --- p.119 / Chapter Section 6.1 --- Introduction --- p.119 / Chapter Section 6.2 --- Results --- p.119 / Chapter Section 6.3 --- Discussion --- p.122 / Chapter CHAPTER 7： --- General Discussion and Conclusions --- p.135 / BIblography --- p.139

Thesis (M. Phil.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

Thesis (M. Phil.)--University of Hong Kong, 2006. / Also available online.

Thesis (M. Phil.)--University of Hong Kong, 2009. / Includes bibliographical references (leaves 153-173). Also available online.

Thesis (Ph. D.)--University of Chicago. / "Reprinted from the Journal of the American Chemical Society, Vol. XLIII. No. 1. January 1921." Includes Abstract. Includes bibliographical references. Also available on the Internet.

Thesis (M.Sc.)--University of Wollongong, 2004. / Typescript. Includes bibliographical references: leaf 90-98.

Thesis (M. Phil.)--University of Hong Kong, 2009. / Includes bibliographical references (leaves 153-173). Also available in print.

Thesis (Doctoral)--University of Stockholm, 1983. / Added t.p. laid in. Includes papers on which the thesis is based. Includes bibliographical references.

Neste trabalho, foram caracterizados os estados eletrônicos fundamental e excitados de mais baixa energia dos sistemas 1;3[H,C,As]. Para isso, foram empregados vários métodos ab initio de estrutura eletrônica (MP2, CCSD(T), CCSD(T)-F12b e MRCISD) aliados a extensos conjuntos de funções de base consistentes na correlação (aug-cc-pVnZ, em que n = D, T, Q e 5). Buscando uma acurácia ainda maior, os resultados obtidos foram extrapolados para o limite do conjunto de base completa (CBS). O estado X1Σ+ da molécula HCAs e o estado eletrônico fundamental do sistema 1[H,C,As], com as seguintes distâncias internucleares: rHC=1,0748 Å e rCAs=1,6602 Å; para as frequências harmônicas, obtivemos ω1(&#963)=1068 cm-1, ω2(π)/ω3(&#960)= 626 cm-1 e ω4(σ)=3310 cm-1 no nível de teoria CCSD(T)-F12-CBS. A espécie 1Σ+ HAsC e um ponto de sela de segunda ordem sobre a superfície de energia potencial (localizado a 75,24 kcal.mol-1 do X1Σ+ HCAs), ou seja, chegamos a conclusão que esta espécie, neste estado eletrônico, em fase gasosa, não existe. Já para os estados tripletos, ha isomerização, sendo que o 13A\' HCAs e o 13A\' HAsC foram caracterizados como mínimos com uma energia relativa ao mínimo global de 59,27 kcal.mol-1 e 88,22 kcal.mol-1, respectivamente. Além disso, exploramos os canais de dissociação destas espécies no nível de teoria CCSD(T). Foram ainda calculadas as frequências fundamentais para os estados do HCAs e do HAsC, como também investigada a inclusão da correlação dos elétrons do caroço nos parâmetros estruturais, vibracionais e energéticos. Estimamos o calor de formação (ΔH0f) a 0 e 298,15 K para as espécies CH, AsH, CAs e HCAs, sendo que a maioria desses valores ainda não e conhecida na literatura. Para o X1Σ+ HCAs, obtivemos um valor de ΔH0f igual a 71,22 kcal.mol-1 a 0 K e 70,38 kcal.mol-1 a 298,15 K. Calculamos o potencial de ionização da molécula HCAs utilizando varias metodologias teóricas, obtendo valores muito próximos aos experimentais, por exemplo, o CCSD(T)-aVTZ forneceu 9,90 eV frente ao valor experimental de 9,8 eV. Os estados eletrônicos excitados singleto e tripleto das espécies HCAs e HAsC foram também caracterizados com a obtenção de dados estruturais, vibracionais e energéticos. A maioria dos dados das espécies HCAs e HAsC nesta dissertação são inéditos na literatura química. / In this work, the ground state and low-lying excited electronic states of system 1;3[H,C,As] were investigated theoretically. Several ab initio molecular electronic structure theory were employed (MP2, CCSD(T), CCSD(T)-F12b e MRCISD) along with extensive correlation-consistent basis sets (aug-cc-pVnZ, n= T, Q e 5). Seeking increasing accuracy, further extrapolation of the results to the complete-basis-set (CBS) limit were carried out. The ground electronic state of 1[H,C,As] is the 1Σ+ HCAs specie, with internuclear distances of rHC=1.0748 Å and rCAs=1.6602 Å, and with harmonic vibrational frequencies ω1(&#963)=1068 cm-1, ω2(π)/ω3(&#960)= 626 cm-1 ω4(σ)=3310 cm-1, at the CCSD(T)-F12-CBS level theory. The electronic state 1Σ+ HAsC is a second-order saddle point on the potential energy surface (located at 75.24 kcal.mol-1 above HCAs), thus providing evidence that this species does not exist in gas phase. However, there is isomerization for triplet electronic states 13A\' HCAs to 13A\' HAsC, with energy relative to global minimum of 59,27 kcal.mol-1 e 88,22 kcal.mol-1, respectively. Fundamental frequencies and the effects of correlation of core electrons in structural, vibrational, and energetic parameters were also evaluated for HCAs and HAsC. Furthermore, the dissociation channels of these species were also evaluated at the CCSD(T)-CBS level theory. The heats of formations (ΔH0f), at 0 and 298,15 K, for the species CH, AsH, CAs and HCAs, were estimated; for most of them these results are inexistent in the literature. For X1Σ+ HCAs, we obtained 71.22 kcal.mol-1 at 0 K and 70.38 kcal.mol-1 at 298.15 K for ΔH0f. The ionization potential was also calculated by several theoretical methodologies, and the results are close to the experimental data; using CCSD(T)-aVTZ, we predicted a value of 9,9 eV, in close agreement with experimental value of 9,8 eV. The singlet and triplet electronic excited states of HCAs and HAsC were investigated and their structural, vibrational and energetic properties evaluated. Most of the results of this work are new in the chemistry literature.

published_or_final_version / Medicine / Master / Master of Philosophy