Li Pik Ha Ivy. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 83-87). / Abstracts in English and Chinese. / TABLE OF CONTENTS --- p.i / LIST OF FIGURES --- p.v / LIST OF TABLES --- p.vii / ABSTRACT --- p.viii / ACKNOWLEDGEMENT --- p.x / DECLARATION --- p.xi / ABBREVIATIONS --- p.xii / Chapter Chapter one --- Introduction / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Polysaccharides isolated from Ganoderma Lucidum --- p.4 / Chapter 1.3 --- Conventional methods for molecular weight (MW) determination of polysaccharides --- p.6 / Chapter 1.3.1 --- Osmometry --- p.7 / Chapter 1.3.2 --- Light Scattering --- p.8 / Chapter 1.3.3 --- Intrinsic Viscosity --- p.8 / Chapter 1.3.4 --- Size Exclusion Chromatography (SEC) --- p.9 / Chapter 1.3.5 --- Mass Spectrometry --- p.10 / Chapter 1.4 --- Matrix-assisted Laser Desorption / Ionization Mass Spectrometry --- p.12 / Chapter 1.5 --- MALDI-TOF Mass Spectrometry of polysaccharides --- p.13 / Chapter 1.6 --- Outline of project --- p.15 / Chapter Chapter two --- Instrumental and experimental / Chapter 2.1 --- Instrumentation --- p.18 / Chapter 2.1.1 --- Laser-based ion source --- p.18 / Chapter 2.1.2 --- Time-of-flight (TOF) analyzer --- p.19 / Chapter 2.1.3 --- Ion deflector --- p.23 / Chapter 2.1.4 --- Detector and data acquisition system --- p.23 / Chapter 2.2 --- Experimental --- p.26 / Chapter 2.2.1 --- Isolation of water-soluble polysaccharides from Ganoderma Lucidum by water extraction --- p.26 / Chapter 2.2.2 --- Isolation of water-soluble polysaccharides from Ganoderma Lucidum by dimethyl ssulfoxide (DMSO) extraction --- p.26 / Chapter 2.2.3 --- Fractionation of water-soluble polysaccharides by Gel Permeation Chromatography (GPC) --- p.27 / Chapter 2.2.4 --- Bradford protein assay --- p.28 / Chapter 2.2.5 --- Phenol / sulfuric acid assay --- p.28 / Chapter 2.2.6 --- Sample preparation in MS --- p.28 / Chapter 2.2.7 --- Calibration of MALDI-TOF-MS --- p.29 / Chapter 2.2.8 --- Data analysis --- p.29 / Chapter Chapter three --- Extraction and purification of water-soluble polysaccharides from Ganoderma Lucidum / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Experimental --- p.32 / Chapter 3.2.1 --- Extraction efficiency --- p.32 / Chapter 3.2.2 --- Dialysis --- p.32 / Chapter 3.2.3 --- Signal suppression effect --- p.32 / Chapter 3.2.4 --- Sevag method --- p.33 / Chapter 3.2.5 --- Trichloroacetic acid (TCA) precipitation --- p.33 / Chapter 3.2.6 --- Bradford Protein Assay --- p.34 / Chapter 3.2.7 --- Phenol sulfuric acid assay --- p.34 / Chapter 3.3 --- Results and discussion --- p.35 / Chapter 3.3.1 --- Extraction efficiency --- p.35 / Chapter 3.3.2 --- Purification of crude polysaccharides --- p.37 / Chapter 3.3.3 --- Desalting --- p.38 / Chapter 3.3.4 --- Deproteination --- p.40 / Chapter 3.3.4.1 --- Monitoring of protein contents --- p.40 / Chapter 3.3.4.2 --- Monitoring of carbohydrate contents --- p.45 / Chapter 3.3.4.3 --- Deproteination using Sevag and TCA procipitation method --- p.47 / Chapter 3.4 --- Conclusions --- p.54 / Chapter Chapter four --- Evaluation of MW and MWD of water-soluble Polysaccharides extracted from Ganoderma Lucidum / Chapter 4.1 --- Introduction --- p.55 / Chapter 4.2 --- Experimental --- p.58 / Chapter 4.2.1 --- Aqueous DHB matrix --- p.58 / Chapter 4.2.2 --- Aqueous DHB/NH4F matrix --- p.58 / Chapter 4.2.3 --- Aqueous DHB matrix in TA solution --- p.58 / Chapter 4.2.4 --- Aqueous DHB/NH4F matrix in TA solution --- p.59 / Chapter 4.2.5 --- Aqueous DHB/NH4F matrix with sodium salt in TA solution --- p.59 / Chapter 4.2.6 --- Aqueous DHB/NH4F matrix with potassium salt in TA solution --- p.59 / Chapter 4.2.7 --- Fractionation of water-soluble polysaccharide extracted by DMSO by Gel Permeation Chromatography (GPC) --- p.59 / Chapter 4.2.8 --- Ultra-violet absorption spectrometry (UV-VIS) --- p.60 / Chapter 4.3 --- Results and discussion --- p.60 / Chapter 4.3.1 --- Development of matrix and solvent system for water-soluble polysaccharides --- p.60 / Chapter 4.3.2 --- MW and MWD of water-soluble polysaccharides extracted from Ganoderma Lucidum --- p.64 / Chapter 4.3.2.1 --- Water extraction --- p.65 / Chapter 4.3.2.2 --- DMSO extraction followed by water-back extraction --- p.70 / Chapter 4.4 --- Conclusions --- p.79 / Chapter Chapter five --- Concluding remarks --- p.82 / References --- p.84

Thesis (M.Med.Sc.)--University of Hong Kong, 2003. / Includes bibliographical references (leaves 83-93). Also available in print.

No description available.

Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Físicas e Matemáticas. Programa de Pós-Graduação em Química / Made available in DSpace on 2012-10-21T03:36:34Z (GMT). No. of bitstreams: 1 200130.pdf: 466439 bytes, checksum: e3f134f496b0341130a270664fd0832a (MD5) / Ganoderma Karsten é um gênero de fungo pertencente a família

Ganoderma lucidum is a popular herbal medicine used in China to promote health. Modern studies have disclosed that the active ingredients of Ganoderma can exhibit several effects, including antitumor effects and immunomodulation. The present study evaluated the antitumor effects of self-prepared Ganoderma extracts and spores oil, and investigated the possible underlying mechanisms by observing the effects of the extracts and oil on topoisomerases and the cell cycle. The results showed that Ganoderma extracts and spores oil presented dose-dependent inhibitory effects on tumor cells. The half maximal inhibitory concentration (IC50) values of Ganoderma extracts on HL60, K562 and SGC-7901 cells for 24 h were 0.44, 0.39 and 0.90 mg/ml, respectively; for Ganoderma spores oil, the IC50 values were 1.13, 2.27 and 6.29 mg/ml, respectively. In the in vivo study, the inhibitory rates of Ganoderma extracts (4 g/kg/d, intragastrically) on S180 and H22 cells were 39.1 and 44.6%, respectively, and for Ganoderma spores oil (1.2 g/kg/d, intragastrically) the inhibitory rates were 30.9 and 44.9%, respectively. Ganoderma extracts and spores oil inhibited the activities of topoisomerase I and II. Ganoderma spores oil was shown block the cell cycle at the transition between the G1 and S phases and induce a marked decrease in cyclin D1 levels in K562 cells, with no significant change in cyclin E level. These results suggest that the Ganoderma extracts and spores oil possessed antitumor effects in the in vitro and in vivo studies. The antitumor mechanisms of the extracts and spores oil were associated with inhibitory effects on topoisomerase I and II activities, and for Ganoderma spores oil, the antitumor effects may also be associated with decreased cyclin D1 levels, thus inducing G1 arrest in the cell cycle.

by Cheung Ka Wan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 121-128). / Abstracts in English and Chinese. / Chapter Chapter 1 - --- Introduction --- p.1 / Chapter 1.1 --- Plant Pathogens --- p.1 / Chapter 1.1.1 --- Virus --- p.2 / Chapter 1.1.2 --- Viroids --- p.2 / Chapter 1.1.3 --- Bacteria --- p.3 / Chapter 1.1.4 --- Fungi --- p.3 / Chapter 1.1.5 --- Mycoplasma like organisms --- p.3 / Chapter 1.1.6 --- Nematodes --- p.4 / Chapter 1.1.7 --- Insects --- p.4 / Chapter 1.1.8 --- Mammals --- p.4 / Chapter 1.2 --- Pathogenicity --- p.5 / Chapter 1.3 --- Disease Development --- p.6 / Chapter 1.3.1 --- Primary infection --- p.6 / Chapter 1.3.2 --- Penetration to host --- p.6 / Chapter 1.3.2.1 --- Entry through wounds ^ / Chapter 1.3.2.2 --- Entry through natural openings --- p.8 / Chapter 1.3.2.3 --- Direct penetration / Chapter 1.3.3 --- Colonization of pathogen --- p.9 / Chapter 1.3.4 --- Mechanisms of attack --- p.9 / Chapter 1.3.5 --- Symptom expression --- p.11 / Chapter 1.3.6 --- Spread of disease --- p.11 / Chapter 1.4 --- Detection of Pathogen --- p.12 / Chapter 1.4.1 --- Traditional diagnostic methods --- p.12 / Chapter 1.4.2 --- Molecular diagnostic methods --- p.13 / Chapter 1.4.3 --- Advantages of molecular diagnostic tools over traditional detection methods --- p.14 / Chapter 1.4.4 --- Sensitivity of molecular diagnostic tools --- p.14 / Chapter 1.4.5 --- Polymerase chain reaction (PCR) --- p.15 / Chapter 1.4.5.1 --- Mechanism of PCR --- p.16 / Chapter 1.4.5.2 --- Application of PCR --- p.17 / Chapter 1.4.6 --- Designation of specific primers in pathogen detection --- p.17 / Chapter 1.4.6.1 --- Nuclear ribosomal DNA genes --- p.18 / Chapter 1.4.6.2 --- Sequencing of ITS regions of rDNA --- p.19 / Chapter 1.5 --- Ganoderma lucidum Complex --- p.19 / Chapter 1.5.1 --- History of Ganoderma lucidum complex --- p.19 / Chapter 1.5.2 --- Classification --- p.20 / Chapter 1.5.3 --- Macroscopic and microscopic structure --- p.21 / Chapter 1.5.4 --- Species identification in G. lucidum complex --- p.22 / Chapter 1.5.5 --- Ganoderma species in Hong Kong --- p.23 / Chapter 1.5.6 --- Act as pathogen --- p.25 / Chapter 1.5.7 --- Availability of tree hosts in Hong Kong --- p.25 / Chapter 1.5.7.1 --- Acacia confusa --- p.26 / Chapter 1.5.7.2 --- Listea cubeba --- p.26 / Chapter 1.5.7.3 --- Leucaena leucocephala --- p.27 / Chapter 1.5.8 --- Disease control for Ganoderma lucidum --- p.27 / Chapter 1.6 --- Aims of Study --- p.29 / Chapter 1.7 --- Significance of the Study --- p.29 / Chapter 1.8 --- Project Strategies --- p.30 / Chapter 1.8.1 --- Survey on Ganoderma lucidum complex in Hong Kong --- p.30 / Chapter 1.8.2 --- Artificial infection --- p.30 / Chapter 1.8.3 --- Detection of pathogen --- p.30 / Chapter Chapter 2 - --- Materials and Methods --- p.31 / Chapter 2.1 --- Collection of Ganoderma lucidum Species Complex in Hong Kong --- p.31 / Chapter 2.2 --- Tissue Isolation --- p.31 / Chapter 2.3 --- Molecular Identification --- p.45 / Chapter 2.3.1 --- Extraction of DNA --- p.45 / Chapter 2.3.2 --- Gel Electrophoresis --- p.45 / Chapter 2.3.3 --- Sequencing of ITS 1 and ITS2 --- p.46 / Chapter 2.3.4 --- Comparison of G. lucidum complex with other Ganoderma and related species --- p.48 / Chapter 2.3.5 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.49 / Chapter 2.4. --- Mating Compatibility for Species Delimitation --- p.49 / Chapter 2.4.1 --- Protoplast isolation --- p.49 / Chapter 2.4.2 --- Mon-Mon mating --- p.50 / Chapter 2.4.3 --- Di-Mon mating --- p.50 / Chapter 2.5 --- Preparation of Samples for Scanning Electron Microscope (SEM) --- p.51 / Chapter 2.6 --- Cytological Studies of Basidiocarps of G. lucidum --- p.52 / Chapter 2.7 --- Pathogenicity Study --- p.53 / Chapter 2.7.1 --- Growth and spread of G. lucidum in soil --- p.53 / Chapter 2.7.2 --- Colonization of G. lucidum on different organs of plants --- p.53 / Chapter 2.7.2.1 --- Determination of dry weight loss --- p.53 / Chapter 2.7.2.2 --- Chitin assay --- p.54 / Chapter 2.7.3 --- Artificial infection to tree seedlings --- p.54 / Chapter 2.7.3.1 --- Artificial infection of vegetative mycelia --- p.54 / Chapter 2.7.3.2 --- Artificial infection with basidiospores --- p.56 / Chapter Chapter 3 - --- Results --- p.57 / Chapter 3.1 --- Collection of Ganoderma lucidum Complex in Hong Kong --- p.57 / Chapter 3.1.1 --- Macroscopic characteristics --- p.57 / Chapter 3.1.2 --- Microscopic characteristics --- p.57 / Chapter 3.1.3 --- G. lucidum under scanning electron microscopy --- p.59 / Chapter 3.2 --- Field Observation --- p.62 / Chapter 3.3 --- Sequencing of ITS Region of G. lucidum Complex and Related Species --- p.64 / Chapter 3.3.1 --- ITS 1 Region of G. lucidum --- p.66 / Chapter 3.3.2 --- ITS 2 Region of G. lucidum --- p.68 / Chapter 3.3.3 --- Relationship between Ganoderma and related species --- p.71 / Chapter 3.4 --- Species Delimitation of G. lucidum --- p.74 / Chapter 3.4.1 --- Arbitrarily-Primed PCR --- p.74 / Chapter 3.4.2 --- Di-Mon mating --- p.77 / Chapter 3.5 --- Pathogenicity of G. lucidum --- p.81 / Chapter 3.5.1 --- Growth and spread in soil --- p.81 / Chapter 3.5.2. --- Preference in colonization on different organs of plants --- p.81 / Chapter 3.5.3 --- Artificial infection --- p.87 / Chapter Chapter 4 - --- Discussion --- p.98 / Chapter 4.1 --- Ganoderma lucidum Complex in Hong Kong --- p.98 / Chapter 4.1.1 --- Macroscopic and microscopic characteristics of G. lucidum complex and related species --- p.98 / Chapter 4.1.2 --- Cytological studies --- p.99 / Chapter 4.1.3 --- Field observation --- p.100 / Chapter 4.1.4 --- Sequences of ITS regions of G. lucidum complex and related species --- p.101 / Chapter 4.1.5 --- Species identification within G. lucidum --- p.104 / Chapter 4.2 --- Pathogenicity Test for G. lucidum --- p.108 / Chapter 4.2.1 --- Growth and spread of G. lucidum --- p.108 / Chapter 4.2.2 --- Colonization of G. lucidum on plants --- p.110 / Chapter 4.2.3 --- Artificial infection by G. lucidum --- p.111 / Chapter 4.3 --- Further Investigation --- p.116 / Chapter Chapter 5 - --- Summary --- p.118 / Chapter Chapter 6- --- Conclusion --- p.120 / References --- p.121

La compréhension de la structure génétique et de la dynamique des populations, ainsi que les mécanismes moléculaires régissant les interactions entre l’hôte et le pathogène sont des éléments clefs pour la gestion des maladies. Au cours de cette étude nous avons cherché à développer dans un premier temps des marqueurs microsatellites à partir de données issues du séquençage d’un isolat de Ganoderma boninense. Ces marqueurs microsatellites nous ont permis d’étudier la structuration génétique et l’histoire démographique de G. boninense. Le génotypage d’un sous-échantillonnage issus de missions de récolte en Malaisie et à Sumatra nous a permis de mettre en évidence deux groupes principaux au sein de l’Asie du Sud-Est. Nous avons ensuite concentré notre attention sur la région regroupant la Malaisie péninsulaire et l’île de Sumatra, zone historique de développement de la culture industrielle du palmier à huile et d’apparition de la pourriture basale du stipe due à G. boninense, qui semble former une seule population. Nous avons examiné à une échelle géographique plus restreinte cette zone géographique afin de mettre en évidence une potentielle sous structure au sein de cette population. En testant l’effet du fond génétique du palmier d’origine de chaque individu, du nombre de génération de palmier précédent le moment de la récolte de l’individu, ou de la distance géographique sur une possible sous-structure des individus au sein de la zone historique, nous avons mis en évidence qu’aucune sous-structure génétique n’émergeait. En revanche, La comparaison de plusieurs scénarios d’évolution démographique a permis de mettre en évidence un phénomène d’expansion très ancien bien antérieur au début du développement de la culture industrielle du palmier à huile. Pour finir, la comparaison des données transcriptomiques entre des isolats agressifs et non agressifs a permis de souligner la présence d’une centaine de gènes différentiellement exprimés possédant une annotation fonctionnelle. Les résultats de ces deux approches pourront permettre une meilleure gestion de la maladie ainsi que l’amélioration des programmes développement et de gestion des résistances. / The understanding of genetic structuration and population dynamic, as well as the molecular mechanisms ruling host/pathogen interaction, are key elements for disease management. During this study, as first step, we were looking to develop microsatellites markers from genomic data obtained from sequencing of a Ganoderma boninense pure strain. Those markers allowed studying genetic structuration and demographic history of G. boninense. Genotyping of a subset of samples from sampling mission in Malaysia and Sumatra have highlighted two main groups in South-East Asia. Then, we focused on a region gathering peninsular Malaysia and Sumatra together, as it is both historical region of industrial oil palm culture development and the first region of basal stem rot observation caused by G. boninense, and to appear as a single population. We examine this region at a lower scale in order to highlight a potential genetic substructure in this population. We tested for effect of genetic tree background of each sample, number of planting generation before sampling and geographical distance between sample in order to observe a potential correlation between genetic substructure and one of those factors. As no correlation appeared, we concluded that this population does not have a genetic substructure. On the other hand, the comparison between several demographic evolution scenarios have shown a strong support for a past expansion event further back in time from the beginning of industrial oil palm culture development. To conclude, the exploration of transcriptomic data between strains owning aggressive or non-aggressive profile showed the differential expression of a hundred genes owning a functional annotation. Results from both approaches will allow the development of better disease management and a better resistance selection and management program.

published_or_final_version / Paediatrics and Adolescent Medicine / Master / Master of Medical Sciences

Fung Pik Ha. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 116-121). / Acknowledgments --- p.i / Table of Contents --- p.ii / List of Figures --- p.v / List of Tables --- p.x / Abstract --- p.xi / Chapter Chapter I --- Introduction --- p.1 / Chapter Chapter II --- Literature Reviews / Chapter II.l --- Glycosidase --- p.3 / Chapter II.2 --- Biosynthesis of N-linked Glycoprotein --- p.4 / Chapter II.3 --- Mechanism of Enzyme Catalysed Reaction --- p.8 / Chapter II.4 --- Types of Glycosidase Inhibitors --- p.12 / Chapter II.5 --- Cyclophellitol and Aminocyclitols / Chapter II.5.1 --- General background on cyclophellitol --- p.17 / Chapter II.5.2 --- Mode of inhibition of cyclophellitol --- p.20 / Chapter II.5.3 --- General background on aminocyclitols --- p.24 / Chapter Chapter III --- Characterization of Synthetic Glycosidase Inhibitors / Chapter III.1 --- Covalent-based Inactivator (Cyclophellitol and its Analogues) / Chapter III.1.1 --- Introduction --- p.28 / Chapter III.1.2 --- Materials --- p.32 / Chapter III.1.3 --- Methods / Chapter III.1.3.1 --- Inhibitory assay of commercially available glycosidases --- p.33 / Chapter III.1.3.2 --- Partial purification of β-D-mannosidase from A. oryzae --- p.34 / Chapter III.1.3.3 --- Protein assay in purification of β-D-mannosidase --- p.38 / Chapter III.1.3.4 --- Inhibitory assay for partially purified β-D- mannosidase (A . oryzae) --- p.38 / Chapter III.1.3.5 --- Influence of dialysis on glycosidase inhibition --- p.39 / Chapter III.1.3.6 --- Inactivation experiment on glycosidases --- p.39 / Chapter III.1.4 --- Results / Chapter III.1.4.1 --- Inhibitory activities of cyclophellitol and its analogues against glycosidases --- p.41 / Chapter III.1.4.2 --- Effect of dialysis on glycosidase inhibition --- p.44 / Chapter III.1.4.3 --- The kinetic studies of glycosidase inactivation --- p.47 / Chapter III. 1.5 --- Discussion --- p.50 / Chapter III.1.6 --- Further studies --- p.55 / Chapter III.2 --- Reversible Competitive Inhibitors (Aminocyclitols) / Chapter III.2.1 --- Introduction --- p.56 / Chapter III.2.2 --- Materials --- p.58 / Chapter III.2.3 --- Methods / Chapter III.2.3.1 --- Assay of glucoside hydrolase inhibition activity --- p.60 / Chapter III.2.3.2 --- Glucose oxidase method for determination of released D-glucose --- p.60 / Chapter III.2.3.3 --- Inhibitory assay of aminocyclitols on other glycosidases --- p.61 / Chapter III.2.3.4 --- Influence of dialysis on the glycosidase inhibition --- p.62 / Chapter III.2.3.5 --- Lineweaver-Burk plot --- p.63 / Chapter III.2.4 --- Results / Chapter III.2.4.1 --- Inhibitory activities of valiolamine and related aminocyclitols against six glycosidases --- p.64 / Chapter III.2.4.2 --- Characterization the aminocyclitols as reversible competitive inhibitors --- p.69 / Chapter III.2.5 --- Discussion --- p.80 / Chapter Chapter IV --- Isolation of the Naturally Occurring Glycosidase Inhibitor from Mushrooms / Chapter IV.1 --- Introduction --- p.83 / Chapter IV.2 --- Materials --- p.84 / Chapter IV.3 --- Methods / Chapter IV.3.1 --- Preparation of Ganoderma lucidum --- p.86 / Chapter IV.3.2 --- Preparation of V. volvacea --- p.86 / Chapter IV.3.3 --- Inhibitory assay of aqueous extract of mushrooms on glycosidases --- p.87 / Chapter IV.3.4 --- Anthrone method for determination of reducing sugars --- p.87 / Chapter IV.3.5 --- Flash liquid chromatography for purification of putative inhibitors in G. lucidum --- p.88 / Chapter IV.4 --- Results / Chapter IV.4.1 --- Prescreening of Inhibitory effects of Various Fungal Extracts --- p.90 / Chapter IV.4.2 --- Inhibitory Effects of Partially Purified G. lucidum Extract on Glycosidase --- p.92 / Chapter IV.4.3 --- Effect of Endogenous Substrates on Glycosidase Activities --- p.93 / Chapter IV.4.4 --- Results of Liquid Column Chromatography --- p.93 / Chapter IV.4.5 --- Structure Determination and Characterization of purified compounds --- p.95 / Chapter IV.4.6 --- Inhibitory Activities of Compounds A and B against Brewers yeast a- glucosidase --- p.96 / Chapter IV.5 --- Discussion --- p.98 / Chapter Chapter V --- Conclusions --- p.113 / References --- p.116

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences