Thesis (Ph. D.)--University of Lund, 1995. / Published dissertation.

Thesis (Ph. D.)--University of Lund, 1995. / Published dissertation.

by Leung Yuet Kin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 86-100). / Abstract also in Chinese. / List of Figures --- p.ix / List of Tables --- p.xii / List of Abbreviations --- p.xiii / Chapter Chapter One: --- Introduction / Chapter 1.1 --- Carcinogenicity of the tobacco products --- p.1 / Chapter 1.2 --- Biochemical Pathway involved in NNK metabolism --- p.5 / Chapter 1.2.1 --- Metabolic activation of NNK --- p.6 / Chapter 1.2.2 --- Detoxification of NNK --- p.8 / Chapter 1.3 --- Modulation of NNK carcinogenesis --- p.9 / Chapter 1.4 --- Mediation of NNK oxidation : chemoprevention by isothiocyanates --- p.11 / Chapter 1.5 --- Aim of study --- p.13 / Chapter 1.5.1 --- Experimental approaches --- p.15 / Chapter Chapter Two : --- Modulation of α-carbon hydroxylations in NNK metabolism / Chapter 2.1 --- Background --- p.17 / Chapter 2.2 --- Materials and Methods --- p.17 / Chapter 2.2.1 --- Chemicals --- p.17 / Chapter 2.2.2 --- Methods --- p.18 / Chapter 2.2.2.1 --- Preparation of rat microsomes --- p.18 / Chapter 2.2.2.2 --- Assay of NNK metabolism --- p.18 / Chapter 2.2.2.3 --- Determination of solvent extraction --- p.19 / Chapter 2.2.2.4 --- Determination of detergent effects --- p.19 / Chapter 2.2.2.5 --- HPLC analysis of NNK metabolites --- p.20 / Chapter 2.2.2.6 --- Study of strain differences between SD rats and F344 rats --- p.20 / Chapter 2.3 --- Results --- p.21 / Chapter 2.3.1 --- HPLC analysis of NNK metabolites --- p.21 / Chapter 2.3.2 --- Determination of solvent extraction --- p.22 / Chapter 2.3.3 --- Effects of detergents --- p.25 / Chapter 2.3.4 --- Study of strain differences using F344 rats and SD rats --- p.26 / Chapter 2.4 --- Discussion --- p.27 / Chapter Chapter Three : --- Modulation by potentiation of detoxification process in NNK metabolism / Chapter 3.1 --- Background --- p.30 / Chapter 3.2 --- Materials and Methods --- p.31 / Chapter 3.2.1 --- Chemicals --- p.31 / Chapter 3.2.2 --- Methods --- p.32 / Chapter 3.2.2.1 --- Preparation of rat microsomes --- p.32 / Chapter 3.2.2.2 --- Analysis of NNK metabolism --- p.32 / Chapter 3.2.2.3 --- UDP-glucuronosyltransferase Assay --- p.33 / Chapter 3.2.2.4 --- Cytochrome P450 2E1 Assay --- p.34 / Chapter 3.3 --- Results --- p.34 / Chapter 3.3.1 --- Screening tests of the effects of vitamins and drugs --- p.34 / Chapter 3.3.1.1 --- Male liver microsomes --- p.35 / Chapter 3.3.1.2 --- Female liver microsomes --- p.37 / Chapter 3.3.1.3 --- Male lung microsomes --- p.39 / Chapter 3.3.1.4 --- Female lung microsmes --- p.41 / Chapter 3.3.2 --- Kinetic analysis of vitamin C-palmitate on NNK reduction --- p.43 / Chapter 3.3.3 --- Effects of vitamin C-palmitate on UDP- glucuronosyltransferase --- p.47 / Chapter 3.3.4 --- Effects of vitamin C-palmitate on cytochrome P4502E1 (CYP2E1) --- p.48 / Chapter 3.4 --- Discussion --- p.50 / Chapter Chapter Four: --- Purification of carbonyl reductase from rat liver microsomes / Chapter 4.1 --- Background --- p.58 / Chapter 4.2 --- Materials and Methods --- p.58 / Chapter 4.2.1 --- Chemicals --- p.58 / Chapter 4.2.2 --- Methods --- p.59 / Chapter 4.2.2.1 --- Preparation of male rat liver microsomes --- p.59 / Chapter 4.2.2.2 --- Purification of carbonyl reductase from rat liver microsomes --- p.59 / Chapter A. --- Solubilization of microsomes --- p.59 / Chapter B. --- Chromatographic separation by octyl-Sepharose CL-4B --- p.60 / Chapter C. --- Chromatographic separation by DEAE-cellulose --- p.60 / Chapter 4.2.2.3 --- SDS polyacrylamide gel electrophoresis --- p.61 / Chapter 4.3 --- Results --- p.61 / Chapter 4.4 --- Discussion --- p.63 / Chapter Chapter Five : --- Summary and Discussion / Chapter 5.1 --- Summary --- p.66 / Chapter 5.2 --- Discussion --- p.67 / Chapter 5.2.1 --- Anticarcinogenesis of vitamin C in NNK-induced cancer through smoking --- p.67 / Chapter 5.2.2 --- Future studies on microsomal carbonyl reductase --- p.68 / Chapter 5.2.3 --- Future studies on UDP-glucuronidation of NNAL --- p.72 / Appendix A Effects of atropine on tolbutamide metabolism --- p.74 / Appendix B Recovery of keto acid and keto alcohol from the assay mixture ( without solvent extraction ) --- p.75 / Appendix C Calibration Curve of NNAL --- p.76 / Appendix D Effects of methanol on NNK carbonyl reduction --- p.77 / Appendix E Time course study of NNAL production at various concentrations of vitamin C-palmitate --- p.78 / Appendix F Time course assay of UDP-glucuronosyltransferase at various concentrations of vitamin C-palmitate --- p.80 / Appendix G Calculation of specific activity of UDP-Glucuronosyl transferase --- p.82 / Appendix H Calculation of specific activity of cytochrome P450 2E1 --- p.84 / References --- p.86

Chow, Cheung-ming. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 149-155). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgement --- p.vi / General abbreviations --- p.viii / Abbreviations of chemicals --- p.ix / List of figures --- p.x / List of tables --- p.xiv / Table of contents --- p.xv / Chapter 1. --- Literature review --- p.1-26 / Chapter 1.1 --- General introduction of nitrogen metabolism in plants --- p.1 / Chapter 1.2 --- Interaction between nitrogen metabolism and other metabolic and developmental pathways in plants --- p.2 / Chapter 1.2.1 --- Carbon metabolism --- p.2 / Chapter 1.2.2 --- Development --- p.2 / Chapter 1.2.3 --- Flowering --- p.3 / Chapter 1.3 --- Metabolic signalling in plants --- p.5 / Chapter 1.3.1 --- Nitrogen signalling in plants --- p.5 / Chapter 1.3.1.1 --- Inorganic N signalling --- p.5 / Chapter 1.3.1.2 --- Organic N signalling --- p.6 / Chapter 1.3.2 --- Carbon signalling --- p.7 / Chapter 1.3.2.1 --- Signalling pathways --- p.7 / Chapter 1.3.2.2 --- Gene expression regulated by sugar --- p.8 / Chapter 1.3.2.3 --- Role of sugar signalling in growth and development --- p.9 / Chapter 1.4 --- Ways to elucidate a new signal transduction pathway --- p.10 / Chapter 1.4.1 --- Carbon signalling as a paradigm to provide hints for exploring nitrogen signalling in plants --- p.10 / Chapter 1.4.2 --- Designing long-term approach to tackle nitrogen signalling in plants --- p.14 / Chapter 1.5 --- "Molecular tools available to change the ""signal"" and the proposed ""sensor""" --- p.15 / Chapter 1.5.1 --- ASN1 overexpressing lines (35S-ASN1) --- p.15 / Chapter 1.5.2 --- PII overexpressing lines (PII ox) & PII truncated lines (PII trunc.) --- p.16 / Chapter 1.5.2.1 --- Plant PII and its possible role --- p.16 / Chapter 1.5.2.2 --- Nitrogen/carbon sensing as the proposed in vivo function of PII-like protein in Arabidopsis thaliana by the in planta bioassay of PII overexpressing (PII ox) lines --- p.17 / Chapter 1.5.2.3 --- Changes in physiological and transcriptional expression of nitrogen assimilatory genes in PII transgenic lines --- p.17 / Chapter 1.6 --- Review on nitrogen controls and sensing mechanism of microbial organism and higher plants --- p.21 / Chapter 1.6.1 --- Nitrogen sensing in enteric bacteria --- p.21 / Chapter 1.6.2 --- Nitrogen sensing in cyanobacteria --- p.21 / Chapter 1.6.3 --- Nitrogen sensing in fungi --- p.22 / Chapter 1.6.4 --- Implication of the nitrogen sensing mechanisms in microorganisms to nitrogen sensing in plants --- p.23 / Chapter 1.7 --- "Hypothesis, objectives and outlines of this thesis work" --- p.25 / Chapter 2 --- Materials and Methods --- p.27-50 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- "Plants,bacterial strains and vectors" --- p.27 / Chapter 2.1.2 --- Chemicals and Regents --- p.28 / Chapter 2.1.3 --- "Buffer, solution and gel" --- p.28 / Chapter 2.1.4 --- Commercial kits --- p.28 / Chapter 2.1.5 --- Equipments and facilities used --- p.29 / Chapter 2.1.6 --- Growth medium --- p.29 / Chapter 2.1.7 --- Primers --- p.29 / Chapter 2.2 --- Methods --- p.29 / Chapter 2.2.1 --- Growth condition for plant materials --- p.29 / Chapter 2.2.1.1 --- General conditions --- p.29 / Chapter 2.2.1.2 --- Mature Arabidopsis for gene expression profile --- p.30 / Chapter 2.2.1.3 --- Arabidopsis seedlings for physiological experiment --- p.30 / Chapter 2.2.2 --- Molecular Techniques --- p.31 / Chapter 2.2.2.1 --- Bacterial cultures for recombinant DNA --- p.31 / Chapter 2.2.2.2 --- Preparation of pBluescript II KS(+) T-vector for cloning --- p.31 / Chapter 2.2.2.3 --- Cloning techniques --- p.32 / Chapter 2.2.2.4 --- Transformation of DH5a competent cell --- p.32 / Chapter 2.2.2.5 --- Gel electrophoresis --- p.33 / Chapter 2.2.2.6 --- DNA and RNA extractions from plant tissues --- p.34 / Chapter 2.2.2.7 --- First strand cDNA synthesis --- p.35 / Chapter 2.2.2.8 --- PCR techniques --- p.35 / Chapter 2.2.2.9 --- Sequencing --- p.37 / Chapter 2.2.3 --- Analysis of sequences and homology search --- p.37 / Chapter 2.2.4 --- Biochemical analysis --- p.41 / Chapter 2.2.4.1 --- Sugar content analysis --- p.41 / Chapter 2.2.4.2 --- Anthocyanin content analysis --- p.42 / Chapter 2.2.4.3 --- Fresh weight measurement --- p.43 / Chapter 2.2.4.4 --- Statistic analysis --- p.43 / Chapter 2.2.5 --- Generation of crossing progenies --- p.44 / Chapter 2.2.5.1 --- Artificial crossing of A. thaliana --- p.44 / Chapter 2.2.5.2 --- PCR screening for successful crossing --- p.44 / Chapter 2.2.6 --- Construction of subtractive libraries --- p.45 / Chapter 2.2.7 --- Reverse-dot blot screening --- p.45 / Chapter 2.2.7.1 --- in vitro transcription for making ampicillin cRNA --- p.46 / Chapter 2.2.7.2 --- PCR amplification --- p.47 / Chapter 2.2.7.3 --- Dot-blotting of PCR products on nylon membrane --- p.48 / Chapter 2.2.7.4 --- P probe preparation --- p.49 / Chapter 2.2.7.5 --- Hybridization --- p.49 / Chapter 2.2.7.6 --- Signal detection --- p.50 / Chapter 3 --- Results --- p.51-124 / Chapter 3.1 --- Differential growth behaviour and sugar content in 35S-ASNI lines --- p.51 / Chapter 3.1.1 --- Growth of the seedlings of 35S-ASN1 lines under different N and C supplementations --- p.51 / Chapter 3.1.2 --- Lowered reducing sugar content in 35S-ASN1 lines --- p.52 / Chapter 3.2 --- Development of markers for nitrogen signalling events related to altered N status in 35S-ASN1 lines --- p.60 / Chapter 3.2.1 --- Sugar-induced anthocyanin levels as common morphological marker shared by 35S-ASN1 lines and PII transgenic lines --- p.60 / Chapter 3.2.2 --- Expression markers related to altered N status in 35-ASN1 lines --- p.64 / Chapter 3.3 --- Generation of transgenic plants constitutively expressing ASN1 and GLB1 (or ASN1 and truncated GLB1) through crossing --- p.74 / Chapter 3.4 --- Search for homologs of well-known microbial nitrogen signalling components in Arabidopsis thaliana --- p.78 / Chapter 3.4.1 --- Homologs of yeast general amino acid control components --- p.80 / Chapter 3.4.1.1 --- Arabidopsis thaliana GCN2-like protein --- p.80 / Chapter 3.4.1.2 --- Arabidopsis thaliana GCN1 -like protein --- p.84 / Chapter 3.4.1.3 --- Arabidopsis thaliana GCN20-like protein --- p.84 / Chapter 3.4.1.4 --- Plant eIF2α --- p.86 / Chapter 3.4.1.5 --- Arabidopsis thaliana GCN4-CRE like sequences --- p.87 / Chapter 3.4.2 --- Homologs of fungi nitrogen sensing components: Globally acting factor in nitrogen control in fungi --- p.89 / Chapter 3.4.3 --- Homologs of cyanobacteria nitrogen control components: IF7 & IF 17 (Negative regulators of GS activity) --- p.89 / Chapter 4 --- Discussion --- p.125-147 / Chapter 4.1 --- Differential physiological and morphological behaviours found in the comparative studies between control lines and 35S-ASN1 lines --- p.125 / Chapter 4.1.1 --- in planta promotive effect of ASN1 overexpression on the seedlings growth under low nitrogen and in the absence of exogenous applied metabolizable sugar --- p.125 / Chapter 4.1.2 --- Modulation of sugar level in 35S-ASN1 lines --- p.126 / Chapter 4.2 --- Development of morphological marker and gene expression markers --- p.128 / Chapter 4.2.1 --- Anthocyanin accumulation as a morphological marker for epistatic analysis --- p.128 / Chapter 4.2.2 --- Differential expressed genes as candidates for gene expression markers of nitrogen signalling event --- p.131 / Chapter 4.3 --- Arabidopsis homolog search for well-known microbial signalling components --- p.132 / Chapter 4.3.1 --- "Possible amino acid sensing system in Arabidopsis constructed by homologs of yeast GCN2, GCN1, GCN20 and eIF2a" --- p.132 / Chapter 4.3.1.1 --- Arabidopsis GCN2-like (A. thaliana GCN2-like) protein --- p.132 / Chapter 4.3.1.2 --- Arabidopsis GCNl-like (A. thaliana GCNl-like) & GCN20-like (A. thaliana GCNl-like) proteins --- p.136 / Chapter 4.3.1.3 --- Plant eIF2a phosphorylation pathway --- p.139 / Chapter 4.3.1.4 --- GCN4 related transcriptional factors and GCN4-like motif (GLM) cis-element in plants --- p.140 / Chapter 4.3.1.5 --- Implication of the presence of plant homologs of fungi regulatory proteins involved in the general control of amino acid biosynthesis --- p.142 / Chapter 4.3.2 --- Failure in identifying homologs of nitrogen regulators responsible for switching of nitrogen source in Arabidopsis --- p.144 / Chapter 4.4 --- Overview of research platform construction --- p.146 / Chapter 5 --- Conclusion and Perspectives --- p.148 / Chapter 6 --- References --- p.149-155 / Chapter 7 --- Appendix --- p.156-167

"July 2004" / Bibliography: leaves 108-144. / v, 187 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, School of Earth and Environmental Sciences, Discipline of Environmental Biology, 2004

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Bibliography: leaves 179-231. / xv, 234 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / The aim of this thesis is to characterise the oophorectomised rat model for post menopausal bone loss by determining the effect of oophorectomy on calcium balance and the components of calcium balance in young and adult rats. The study utilises the metabolic calcium balance technique for this purpose. Many of the characteristics of the animal model are described particularly in terms of the effects of oophorectomy on bone histomorphometry and metabolic markers of bone turnover. The study characterises the changes in calcium balance and its components through the growth period. / Thesis (Ph.D.)--University of Adelaide, Dept. of Physiology, 1996