Ganoderma species are one of the most widely researched fungi because of their reported potent bioactive properties. Although there is much information related to American, European and Asian isolates, little research has been conducted on Australian Ganoderma isolates. Ganoderma may only be imported into Australia under strict quarantine conditions, therefore, the isolation of a native strain that possesses bioactivity may be industrially and commercially significant. Three Australian species of this wood-decomposing fungus were isolated in northern Queensland. In this study, they have been identified as three separate species. Further, they have been studied to determine their optimal growth conditions in liquid culture and assessed for their antibacterial properties. Phylogeny inferred from the Internal Transcribed Spacer Regions (ITS) from the DNA sequences resolved the three Australian Ganoderma species into separate clades. Two isolates were identified to be isolates of Ganoderma cupreum (H1) and Ganoderma weberianum (H2). The third isolate could only be identified to the genus level, Ganoderma species, due to the lack of informative data that could be used for comparison. The effects of short term and long term storage on the viability of the fungi were investigated on agar plates, agar slants and balsa wood at varying temperatures ranging from 10 to 45�C. The most appropriate storage conditions were determined to be �80�C on balsa wood chips for periods of up to 2 years without subculture, and on agar slants at 4�C for up to a maximum of eight weeks. Light was observed to be detrimental to the survival of Ganoderma H1 and Ganoderma H2 during storage. Growth trials using potato dextrose agar plates determined the optimal temperature and pH for mycelial growth to be 30�C and a pH of 6, for all isolates. Subsequent growth trials in liquid media found that glucose, as the carbohydrate source, supported the greatest mycelial growth of Ganoderma H1 and Ganoderma H2 and that galactose and fructose supported the greatest growth of Ganoderma H3. Abstract ii Aqueous (hot water) and organic (hexane (HEX), dichloromethane (DCM), ethyl acetate (EtOAc), methanol (MeOH)) extracts from the liquid cultivated mycelium were assessed for their antibacterial activity using disc diffusion assays. Extracts from the mycelium of Ganoderma H1 exhibited activity against a greater number of Gram positive bacteria than those from Ganoderma H2 and H3. Subsequent studies on the DCM and EtOAc extracts from Ganoderma H1 determined the MIC and MBC against a number of Gram positive bacteria, including Bacillus cereus, B. subtilis, Enterococcus faecalis, Streptococcus pyogenes, Staphylococcus aureus, S. epidermidis and Listeria monocytogenes, as well as Clostridium species, including Clostridium perfringens, C. sporogenes and C. difficile, and some methicillin resistant Staphylococcus aureus (MRSA) strains. Time course growth assays confirmed that the DCM and EtOAc extracts predominantly exhibited bactericidal activity. Finally, the active compounds were determined to be terpenoid in structure with some phenolic groups attached.

Thesis (M. Med. Sc.)--University of Hong Kong, 2010. / Includes bibliographical references (p. 55-61).

Sun Baizhong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 75-79). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ABSTRACT (IN CHINESE) --- p.ii / TABLE OF CONTENTS --- p.iii / LIST OF FIGURES --- p.v / LIST OF TABLES --- p.vii / ABBREVIATIONS --- p.vi / ACKNOWLEDGEMENT --- p.ix / DECLARATION --- p.x / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1.1 --- Polysaccharides --- p.1 / Chapter 1.2 --- Pharmacological importance of polysaccharides --- p.2 / Chapter 1.3 --- Polysaccharides from Ganoderma Lucidum --- p.3 / Chapter 1.4 --- Characterization of polysaccharides --- p.3 / Chapter 1.5 --- Matrix-assisted laser desorption / ionization --- p.5 / Chapter 1.6 --- Matrix-assisted laser desorption / ionization of polysaccharides --- p.8 / Chapter 1.7 --- Outline of the project --- p.10 / Chapter CHAPTER TWO --- INSTRUMENTATION AND EXPERIMENTAL / Chapter 2.1 --- Time-of-flight mass spectrometry --- p.12 / Chapter 2.1.1 --- Instrumentation --- p.15 / Chapter 2.1.1.1 --- Laser system --- p.16 / Chapter 2.1.1.2 --- Ion source --- p.16 / Chapter 2.1.1.3 --- Reflector --- p.19 / Chapter 2.1.1.4 --- Detector --- p.19 / Chapter 2.1.1.5 --- Data acquisition and manipulation --- p.20 / Chapter 2.2 --- Ultraviolet-visible spectrometer --- p.21 / Chapter CHAPTER THREE --- OPTIMIZATION OF MALDI CONDITIONS FOR POLYSACCHARIDE ANALYSIS USING DMSO AS SOLVENT / Chapter 3.1 --- Introduction --- p.22 / Chapter 3.2 --- Experimental --- p.25 / Chapter 3.2.1 --- Selection of matrix materials --- p.26 / Chapter 3.2.2 --- Selection of co-matrix materials --- p.26 / Chapter 3.2.3 --- Ratios of matrix-to-analyte --- p.26 / Chapter 3.2.4 --- Effect of sample drying temperature --- p.27 / Chapter 3.2.5 --- Sample loading methods --- p.27 / Chapter 3.3 --- Results and discussion --- p.27 / Chapter 3.3.1 --- Selection of matrix materials --- p.28 / Chapter 3.3.2 --- Selection of co-matrix materials --- p.30 / Chapter 3.3.3 --- Effect of matrix-to-co-matrix ratio --- p.32 / Chapter 3.3.4 --- Effect of sample drying temperature --- p.37 / Chapter 3.3.5 --- Effect of matrix-to-analyte ratio --- p.39 / Chapter 3.3.6 --- Evaluation of sample preparation protocols --- p.43 / Chapter 3.4 --- Conclusion --- p.46 / Chapter CHAPTER FOUR --- MOLECULAR AND MOLECULAR WEIGHT DISTRIBUTION OF WATER-INSOLUBLE POLYSACCHARIDES FROM GANODERMA LUCIDUM / Chapter 4.1 --- Introduction --- p.47 / Chapter 4.2 --- Experimental --- p.48 / Chapter 4.2.1 --- Extraction of water-insoluble polysaccharides from Ganoderma Lucidum --- p.48 / Chapter 4.2.2 --- Gel permeation chromatography of water-insoluble polysaccharides --- p.49 / Chapter 4.2.3 --- Phenol sulfuric acid assay --- p.51 / Chapter 4.2.4 --- MALDI-TOF analysis --- p.51 / Chapter 4.2.5 --- Bradford assay --- p.52 / Chapter 4.3 --- Results and discussion --- p.52 / Chapter 4.3.1 --- Extraction of water-insoluble polysaccharides from Ganoderma Lucidum --- p.52 / Chapter 4.3.2 --- Fractionation of water-insoluble polysaccharides using gel permeation chromatography (GPC) --- p.54 / Chapter 4.3.3 --- MALDI-TOF analysis of fractionated polysaccharides --- p.57 / Chapter 4.3.4 --- Carbohydrate content analysis of fractionated polysaccharides --- p.62 / Chapter 4.3.5 --- MW and MWD of water-insoluble polysaccharides extracted from Ganoderma Lucidum --- p.69 / Chapter 4.4 --- Conclusion --- p.71 / Chapter CHAPTER FIVE --- CONCLUDING REMARKS / Chapter 5.1 --- Conclusion --- p.73 / REFERENCES --- p.76 / APPENDIX --- p.81 / Appendix A Reaction scheme of carbohydrate with phenol in phenol-sulfuric acid assay --- p.81

Thesis (Ph. D.)--University of Hong Kong, 2007. / Also available in print.

Thesis (Ph. D.)--University of Hong Kong, 2007. / Also available online.

Submitted by Ramon Santana (ramon.souza@ufpe.br) on 2015-03-04T14:42:08Z No. of bitstreams: 2 Tese Jose Antonio Pereira Jr.pdf: 3933563 bytes, checksum: 376666e17ed53d90fbfd8bacea4c41ce (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-04T14:42:08Z (GMT). No. of bitstreams: 2 Tese Jose Antonio Pereira Jr.pdf: 3933563 bytes, checksum: 376666e17ed53d90fbfd8bacea4c41ce (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2013 / CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) / O uso de cogumelos para fins medicinais é muito antigo, onde o basidiomiceto Ganoderma lucidum tem posição de destaque no Japão e na tradicional medicina chinesa, sendo relatadas atividades antitumorais, antivirais, antibacteriana, dentre outras. Várias espécies com superfície lacada são confundidas com G. lucidum, entre elas Ganoderma parvulum Murrill, uma espécie tropical que pode representar potencial econômico, respeitando-se a biodiversidade local. O presente trabalho objetivou avaliar a atividade antibacteriana, concentração inibitória mínima (CIM) e a atividade citotóxica dos extratos metanólico e em acetato de etila de G. parvulum, a atividade antibacteriana e a CIM do óleo essencial, bem como fornecer dados para caracterização da droga fúngica. Para os extratos estudados, tanto a atividade antibacteriana como citotóxica foram observadas apenas para o extrato em acetato de etila. A atividade antibacteriana foi considerada moderada frente a Staphylococcus aureus, Bacillus subtilis e Micrococcus luteus. A atividade citotóxica em dose única de 50 μg/ml de extrato foi de 71,94 ± 2,04% contra a linhagem de células HT-29 e 90,02 ± 2,17% contra a linhagem de células HEp-2. O óleo essencial teve atividade antimicrobiana contra as bactérias gram-positivas (Staphylococcus aureus, Bacillus subtilis e Micrococcus luteus) e gram-negativas (Escherichia coli, Klebsiella pneumoniae e Pseudomonas aeruginosa), contudo bactérias gram-positivas foram mais susceptíveis ao óleo essencial. Quanto à prospecção micoquímica, os compostos majoritários foram representados pelos esteróides e terpenóides, apresentando também flavonóides, derivados cinâmicos e fenilpropanoglicosídeos. A composição do óleo essencial foi analisada por cromatografia gasosa acoplada a espectroscopia de massas (CG-MS), onde os principais compostos foram o ácido benzóico, o sesquiterpeno aristolona mono-oxigenado, o 2,6-bis (1,1-dimetiletil)-4-metilfenol e o N-(2-hidroxi-4-oxo-4H-quinazolina-3-il)-benzamida. A morfoanatomia foi considerada fundamental para a identificação de G. parvulum, onde características como o tamanho do basidioma, tamanho do basidiósporo, o sistema hifálico, a coloração do contexto e linhas de deposição de material resinoso, foram características essenciais para distinção de G. parvulum. O tamanho das partículas do pó indicou se tratar de um pó moderadamente espesso por simples agregação das partículas, o teor de cinzas totais foi de 1,67 ± 0,14% e a perda por dessecação foi de 13% ± 0,07, dentro dos níveis encontrados para o gênero. Na análise microscópica do pó, a observação de hifas esqueléticas e de esporos de parede dupla e truncados, característicos do gênero, foram importantes para a identificação da droga. Os dados sobre teor de cinzas, perda por dessecação, análise granulométrica e microscópica do pó são inéditos para G. parvulum, bem como as atividades biológicas dos extratos estudados e do óleo essencial.

Ganoderma root rot disease has been reported world wide causing the death of affected hosts. The taxonomy of the genus Ganoderma is considered to be in disarray due to the use of basidiocarp morphological characters to differentiate the species which resulted in many synonyms, species complexes and possible misidentifications of species within the genus. The use of sexual compatibility tests and molecular techniques became powerful diagnostic tools to elucidate the taxonomy of Ganoderma species. Application of these techniques has resolved some of the taxonomic problems but the use of certain species names in the genus is still causing contention among taxonomists. The literature surrounding the taxonomy and techniques used in the taxonomy of the root rot fungus Ganoderma are considered in this thesis. It is clear that the taxonomy of Ganoderma is very difficult and it is still largely obscured by species complexes and incorrect species identifications. It is also evident that a single species concept will not aid in the identification of Ganoderma species. Rather, a combination of concepts based on morphology, mating tests and DNA sequence data should be used in elucidating the taxonomy of Ganoderma. Morphological characteristics as well as nucleotide sequence analysis of three gene regions; the internally transcribed spacer (ITS), the mitochondrial small subunit (mtSSU) and the intergenic spacer (IGS-1), were used to identify the causal agent of Ganoderma root rot of J. mimosifolia in the suburb of Brooklyn, Pretoria, South Africa. Morphological observations and DNA-based phylogenies revealed that all isolates collected from infected trees belong to a single species that reside in the G. lucidum sensu lato complex. Acacia mangium is a leguminous tree that is grown as an exotic plantation species in Indonesia. These economically important trees are threatened by Ganoderma root rot disease. This disease is considered to be the most important cause of losses in A. mangium plantations. Phylogenetic analysis of ITS sequence data showed that G. philippii is the primary agent of Ganoderma root rot in A. mangium in Sumatra, Indonesia. / Dissertation (MSc)--University of Pretoria, 2011. / Microbiology and Plant Pathology / Unrestricted

by Ma Suet-yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 119-131). / Abstracts in English and Chinese. / Acknowledgment --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Table of Contents --- p.vi / List of Tables --- p.x / List of Figures --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Ganoderma lucidum --- p.1 / Chapter 1.1.1 --- History of Ganoderma lucidum --- p.1 / Chapter 1.1.2 --- Classification --- p.1 / Chapter 1.1.3 --- Macroscopic and microscopic structure --- p.2 / Chapter 1.1.4 --- Ganoderma lucidum as a pathogen --- p.3 / Chapter 1.1.5 --- Availability of tree hosts in Hong Kong --- p.4 / Chapter 1.1.6 --- Medicinal effects --- p.5 / Chapter 1.2 --- Study of Populations in Fungi --- p.6 / Chapter 1.2.1 --- Definition of Population --- p.6 / Chapter 1.2.2 --- Study of Fungal Populations --- p.7 / Chapter 1.2.3 --- Techniques for Population Studies in Fungi --- p.7 / Chapter 1.2.3.1 --- Somatic Incompatibility Test / Chapter 1.2.3.2 --- Isozyme Analysis / Chapter 1.2.3.3 --- Restriction Fragment Length Polymorphisms (RFLPs) / Chapter 1.2.3.4 --- Polymerase Chain Reaction (PCR) Amplification / Chapter 1.3 --- Mitochondrial DNA (mt-DNA) in Fungi --- p.14 / Chapter 1.3.1 --- Inheritance in mt-DNA --- p.15 / Chapter 1.3.2 --- Mitochondrial DNA in Population Studies --- p.15 / Chapter 1.3.2.1 --- Mitochondrial small-subunit (mt-SSU) rDNA / Chapter 1.3.2.2 --- Cytochrome oxidase 3 (cox3) / Chapter 1.4 --- Biodiversity study on Ganoderma species --- p.19 / Chapter 1.5 --- Environment Pollutants in Hong Kong --- p.20 / Chapter 1.5.1 --- Air quality in Hong Kong --- p.20 / Chapter 1.5.2 --- Soil quality in Hong Kong --- p.20 / Chapter 1.5.3 --- Toxicity of pollutants --- p.23 / Chapter 1.5.4 --- Accumulation of heavy metals by G. lucidum --- p.26 / Chapter 1.6 --- Objectives of Study --- p.27 / Chapter 1.7 --- Project Strategies --- p.28 / Chapter 1.7.1 --- Survey on distribution and collection of Ganoderma lucidum in Hong Kong --- p.28 / Chapter 1.7.2 --- Genetic divergences of G. lucidum mitochondrial genes --- p.28 / Chapter 1.7.3 --- Contaminations on field collected G. lucidum --- p.29 / Chapter 1.8 --- Significance of Study --- p.29 / Chapter Chapter 2 --- Materials and Methods --- p.30 / Chapter 2.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.30 / Chapter 2.2 --- Tissue Isolation --- p.30 / Chapter 2.3 --- Somatic Incompatibility Test --- p.36 / Chapter 2.4 --- Molecular Identification --- p.40 / Chapter 2.4.1 --- Extraction of DNA --- p.40 / Chapter 2.4.2 --- Gel electrophoresis --- p.41 / Chapter 2.4.3 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.41 / Chapter 2.4.4 --- PCR of mt-SSU rDNA and --- p.43 / Chapter 2.4.5 --- Sequencing of mt-SSU rDNA and cox3 --- p.44 / Chapter 2.4.6 --- Comparison of G. lucidum complex with other Ganoderma and related species / Chapter 2. 4.7 --- Phylogenetic analyses --- p.46 / Chapter 2.5 --- Investigation of pollutants in Ganoderma lucidum collected in Hong Kong --- p.46 / Chapter 2.5.1 --- Metal analysis --- p.48 / Chapter 2.5.1.1 --- Acid digestion / Chapter 2.5.1.2 --- Statistical analysis / Chapter 2.5.2 --- Organic pollutant analysis --- p.49 / Chapter Chapter 3 --- Result --- p.52 / Chapter 3.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.52 / Chapter 3.1.1 --- Field observation --- p.52 / Chapter 3.1.2 --- Macroscopic characteristics --- p.52 / Chapter 3.1.3 --- Microscopic characteristics --- p.53 / Chapter 3.2 --- Somatic Incompatibility Test --- p.56 / Chapter 3.3 --- DNA fingerprints by Arbitrarily-Primed PCR --- p.57 / Chapter 3.4 --- Sequencing of mt-SSU rDNA region of G. lucidum and related species --- p.60 / Chapter 3.4.1 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.60 / Chapter 3.4.2 --- mt-SSU rDNA region of G. lucidum and other related species --- p.61 / Chapter 3.4.3 --- Phylogenetic analysis of mt-SSU rDNA region --- p.61 / Chapter 3.5 --- Sequencing of cox3 region --- p.71 / Chapter 3.5.1 --- Genetic variability in cox3 region of G. lucidum --- p.71 / Chapter 3.5.2 --- cox3 region of G. lucidum and other related species --- p.72 / Chapter 3.5.3 --- Phylogenetic analysis of cox3 region --- p.72 / Chapter 3.6 --- Metal content of field G. lucidum --- p.82 / Chapter 3.7 --- Organic pollutants in field collected G. lucidum --- p.90 / Chapter Chapter 4 --- Discussion --- p.93 / Chapter 4.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.93 / Chapter 4.1.1 --- Differentiation of G. lucidum and related species in the G lucidum species complex --- p.93 / Chapter 4.1.2 --- Field observation --- p.94 / Chapter 4.2 --- Biodiversity of populations of G. lucidum in Hong Kong --- p.95 / Chapter 4.2.1 --- Individualism of G. lucidum --- p.95 / Chapter 4.2.2 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.96 / Chapter 4.2.3 --- Genetic variability in cox3 region of G. lucidum --- p.98 / Chapter 4.2.4 --- Lineages of G. lucidum collected in Hong Kong --- p.100 / Chapter 4.2.5 --- Cryptic phylogenetic species --- p.101 / Chapter 4.3 --- Contamination of field collected Ganoderma lucidum in Hong Kong --- p.106 / Chapter 4.3.1 --- Metal contents in field collected G. lucidum in Hong Kong --- p.106 / Chapter 4.3.1.1 --- Metal contents of G. lucidum fruiting bodies collected at each site / Chapter 4.3.1.2 --- General discussion of metals / Chapter 4.3.1.3 --- Consumption of field collected G. lucidum fruiting bodies / Chapter 4.3.2 --- Comparison of metal contents between field collected Hong Kong G. lucidum with other mushrooms collected from other places --- p.112 / Chapter 4.3.3 --- Survey of organic pollutants in field collected G. lucidum in Hong Kong --- p.113 / Chapter Chapter 5 --- Conclusion --- p.116 / Chapter Chapter 6 --- Further investigation --- p.118 / Chapter Chapter 7 --- Reference --- p.119

published_or_final_version / abstract / Paediatrics and Adolescent Medicine / Doctoral / Doctor of Philosophy

by Fu Sai-chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 102-111). / Acknowledgements / List of Abbreviation / Abstract / Prologue --- p.1 / Chapter / Chapter 1. --- Introduction --- p.2 / Chapter 1.1 --- Historical background of Ling Zhi (靈芝） --- p.4 / Chapter 1.2 --- Biological description of Ling Zhi (靈芝） --- p.6 / Chapter 1.3 --- Chemical composition of Ganoderma lucidum --- p.6 / Chapter 1.3.1 --- The triterpenoids --- p.7 / Chapter 1.3.2 --- Matrix polysaccharides --- p.8 / Chapter 1.4 --- Medicinal properties and biological activities --- p.9 / Chapter 1.4.1 --- Medicinal properties --- p.9 / Chapter 1.4.2 --- Biological activities --- p.10 / Chapter 1.4.2.1 --- Anti-tumour activity --- p.10 / Chapter 1.4.2.2 --- Enhancement of protein and nucleic acid synthesis --- p.11 / Chapter 1.4.2.3 --- Effects on nervous system --- p.11 / Chapter 1.4.2.4 --- Effects on respiratory system --- p.12 / Chapter 1.4.2.5 --- Effects on cardiovascular system --- p.12 / Chapter 1.4.2.6 --- Effects on immune system --- p.14 / Chapter 1.4.2.7 --- Hepatoprotection and detoxicant actions --- p.14 / Chapter 1.4.3 --- Summary --- p.15 / Chapter 1.5 --- The relationship between antioxidant defense/immune system and disease --- p.15 / Chapter 2. --- Materials and Methods --- p.20 / Chapter 2.1 --- Cell culture reagents / Chapter 2.1.1 --- Preparation of RPMI-1640 medium --- p.20 / Chapter 2.1.2 --- Preparation of fetal calf serum (FCS) --- p.20 / Chapter 2.1.3 --- Preparation of phosphate-buffered saline (PBS) --- p.20 / Chapter 2.1.4 --- Mitogens and test samples --- p.21 / Chapter 2.1.5 --- Trypan blue exclusion test --- p.21 / Chapter 2.1.6 --- Liquid scintillation counting --- p.21 / Chapter 2.2 --- Animal care --- p.22 / Chapter 2.3 --- Fractionation of Ganoderma lucidum extracts --- p.22 / Chapter 2.4 --- Carbon tetrachloride (CC14)- induced hepatotoxicity in mice --- p.23 / Chapter 2.4.1 --- Pretreatment scheme and CCl4 treatment --- p.23 / Chapter 2.4.2 --- Preparation of liver homogenates --- p.23 / Chapter 2.4.3 --- Biochemical assays for assessing hepatotoxicity --- p.25 / Chapter 2.4.3.1 --- Determination of plasma alanine aminotransferase (ALT) --- p.25 / Chapter 2.4.3.2 --- Determination of tissue glutathione (GSH) content --- p.25 / Chapter 2.4.3.3 --- Depletion of tissue GSH in liver homogenates by tert- butylhydroperoxide (tBHP) --- p.26 / Chapter 2.4.3.4 --- Determination of tissue malondialdehyde (MDA) content --- p.26 / Chapter 2.5 --- Preparation of murine hepatocytes --- p.27 / Chapter 2.5.1 --- Liver perfusion --- p.27 / Chapter 2.5.2 --- Isolation of murine hepatocytes --- p.29 / Chapter 2.5.3 --- Incubation protocol --- p.29 / Chapter 2.5.4 --- Determination of cellular glutathione by HPLC method --- p.30 / Chapter 2.6 --- Preparation of splenocytes and lymphocytes --- p.31 / Chapter 2.6.1 --- Isolation of murine splenocytes --- p.31 / Chapter 2.6.2 --- Mitogenic assay by measuring 3H-thymidine uptake --- p.31 / Chapter 2.6.3 --- B cell- and T cell- enrichment in splenocyte preparation --- p.32 / Chapter 2.6.4 --- Isolation of human peripheral blood lymphocytes (HPBL) from cord blood --- p.34 / Chapter 2.7 --- Chromatographic methods --- p.34 / Chapter 2.7.1 --- Fast Protein Liquid Chromatography (FPLC) with Superose 12 column --- p.34 / Chapter 2.7.2 --- Removal of lipopolysaccharide (LPS) with Detoxi Gel --- p.35 / Chapter 2.7.3 --- Desalting by Sephadex G-25 --- p.35 / Chapter 2.7.4 --- Determination of FDNB-conjugated GSH and GSSG by HPLC reverse phase chromatography --- p.36 / Chapter 2.8 --- Measurement of in vivo antibody production --- p.39 / Chapter 2.8.1 --- Immunization and pretreatment scheme --- p.39 / Chapter 2.8.2 --- Direct plaque assay --- p.39 / Chapter 2.9 --- Biochemical analysis --- p.39 / Chapter 2.9.1 --- Determination of protein content --- p.40 / Chapter 2.9.2 --- Determination of hexose content --- p.40 / Chapter 2.9.3 --- Determination of uronic acid content --- p.40 / Chapter 2.9.4 --- Determination of sulphate content --- p.41 / Chapter 2.9.5 --- Determination of hexosamine content --- p.42 / Chapter 2.10 --- Statistical analysis --- p.42 / Chapter 3. --- Results --- p.44 / Chapter 3.1 --- Extraction and fractionation of Ganoderma lucidum --- p.44 / Chapter 3.2 --- Hepatoprotective effect of Ganoderma lucidum fractions aganist CCl4-induced hepatotoxicity --- p.47 / Chapter 3.2.1 --- Hepatoprotection of Ganoderma lucidum fractions --- p.47 / Chapter 3.2.2 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic malondialdehyde (MDA) level in CCl4-treated mice --- p.47 / Chapter 3.2.3 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic glutathione (GSH) level in CCl4-treated mice --- p.51 / Chapter 3.3 --- Effect of GL2 on isolated hepatocytes --- p.51 / Chapter 3.3.1 --- Protection against phorone-induced hepatocytotoxicity --- p.51 / Chapter 3.3.2 --- Protection against menadione-induced hepatocytotoxicity --- p.55 / Chapter 3.4 --- Immunomodulatory effect of Ganoderma lucidum fractions --- p.55 / Chapter 3.4.1 --- Discovery of a mitogenic principle from Ganoderma lucidum fractions --- p.55 / Chapter 3.4.2 --- B cell-specific mitogenic activity --- p.58 / Chapter 3.4.3 --- Mitogenic activity in Ganoderma lucidum distinguishable from possible LPS contamination --- p.65 / Chapter 3.4.3.1 --- The use of polymyxin B sulphate --- p.65 / Chapter 3.4.3.2 --- The use of Detoxi Gel --- p.69 / Chapter 3.4.3.3 --- The combined use of polymyxin B sulphate and Detoxi Gel --- p.69 / Chapter 3.5 --- Enhancement of humoral response to sheep red blood cells (SRBC) by the mitogenic GL2 fraction --- p.72 / Chapter 3.6 --- Physical-chemical characterization of GL2 fraction --- p.76 / Chapter 3.6.1 --- Estimation of molecular size by FPLC size exclusion chromatography with Superose 12 column --- p.76 / Chapter 3.6.2 --- Sugar composition of GL2 fraction --- p.76 / Chapter 3.6.3 --- Comparison of elution profiles in Superose12 column with respect to mitogenic activity and sugar composition --- p.80 / Chapter 4. --- Discussions --- p.83 / Chapter 4.1 --- Enhancement of antioxidant status by Ganoderma lucidum fractions --- p.83 / Chapter 4.2 --- The significance of enhancement of GSH status --- p.89 / Chapter 4.3 --- Immunomodulatory actions of Ganoderma lucidum --- p.95 / Conclusions --- p.99 / Appendix --- p.100 / Reference --- p.102