Typescript (photocopy). / Digitized by Kansas Correctional Industries

Lo Kit Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 193-208). / Abstracts in English and Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT (Chinese version) --- p.v / CONTENT --- p.vii / LIST OF TABLES --- p.xiii / LIST OF FIGURES --- p.xviii / LIST OF ABBREVIATIONS --- p.xx / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter 1.1 --- An introduction of natural antioxidants --- p.1 / Chapter 1.1.1 --- Definition of antioxidants --- p.1 / Chapter 1.1.2 --- Application of natural antioxidants in foods --- p.3 / Chapter 1.1.2.1 --- Oxidation of foods --- p.3 / Chapter 1.1.2.1.1 --- Autoxidation of food --- p.3 / Chapter 1.1.2.1.2 --- Photo-oxidation of food --- p.4 / Chapter 1.1.3 --- Free radicals and antioxidants --- p.6 / Chapter 1.1.3.1 --- Free radicals and reactive oxygen species --- p.6 / Chapter 1.1.3.1.1 --- Superoxide anion radical --- p.8 / Chapter 1.1.3.1.2 --- Hydrogen peroxide --- p.9 / Chapter 1.1.3.1.3 --- Hydroxyl radical --- p.10 / Chapter 1.1.3.1.4 --- Peroxyl radical --- p.12 / Chapter 1.1.3.1.5 --- Lipid peroxidation of cell membranes --- p.13 / Chapter 1.1.3.1.6 --- Oxidation of LDL and atherosclerosis --- p.16 / Chapter 1.1.4 --- Natural antioxidants and their mechanisms --- p.18 / Chapter 1.1.4.1 --- Carotenoids --- p.18 / Chapter 1.1.4.2 --- Phenolic compounds --- p.20 / Chapter 1.1.4.2.1 --- Flavonoids --- p.21 / Chapter 1.1.4.2.2 --- Phenolic acids --- p.22 / Chapter 1.1.4.3 --- Sterols --- p.24 / Chapter 1.1.4.4 --- Vitamins --- p.25 / Chapter 1.2 --- Antioxidant in mushrooms --- p.27 / Chapter 1.2.1 --- Antioxidant properties of mushrooms --- p.27 / Chapter 1.2.2 --- Characterization of mushroom phenolic antioxidants --- p.30 / Chapter 1.2.3 --- Biosynthesis of phenolic compounds in mushrooms or fungi --- p.33 / Chapter 1.3 --- Assays for evaluation of antioxidants --- p.35 / Chapter 1.3.1 --- Beta-carotene bleaching method --- p.35 / Chapter 1.3.2 --- Scavenging activity of DPPH radical --- p.36 / Chapter 1.3.3 --- Erythrocyte hemolysis --- p.36 / Chapter 1.3.4 --- Scavenging activity of ABTS ´Ø + radical cation --- p.37 / Chapter 1.3.5 --- Scavenging activity of hydroxyl radical --- p.37 / Chapter 1.3.6 --- Assay for lipid peroxidation of rat brain homogenate --- p.38 / Chapter 1.3.7 --- Inhibition of low-density lipoproteins (LDLs) oxidation --- p.39 / Chapter 1.4 --- Analysis of phenolic antioxidants --- p.41 / Chapter 1.4.1 --- Extraction of phenolic compounds --- p.41 / Chapter 1.4.2 --- Determination of total phenolic content --- p.43 / Chapter 1.4.3 --- Chromatographic fractionation of phenolic compounds --- p.44 / Chapter 1.4.4 --- Characterization of phenolic compounds --- p.45 / Chapter 1.4.4.1 --- Thin-layer chromatography (TLC) --- p.45 / Chapter 1.4.4.2 --- High performance liquid chromatography (HPLC) --- p.46 / Chapter 1.4.4.3 --- Liquid chromatography-Mass spectrometry (LC-MS) --- p.47 / Chapter 1.5 --- Objectives --- p.49 / Chapter CHAPTER 2: --- MATERIALS AND METHODS --- p.50 / Chapter 2.1 --- Sample preparation --- p.50 / Chapter 2.2 --- Sample extraction. --- p.51 / Chapter 2.2.1 --- Small-scale methanol and water extraction --- p.51 / Chapter 2.2.2 --- Large-scale methanol and water extraction and fractionation --- p.54 / Chapter 2.2.2.1 --- Large-scale methanol and water extraction --- p.54 / Chapter 2.2.2.2 --- Fractionation of crude extracts --- p.55 / Chapter 2.2.2.2.1 --- Fractionation of methanol crude extract --- p.55 / Chapter 2.2.2.2.2 --- Fractionation of water crude extract --- p.55 / Chapter 2.3 --- Fractionation by column chromatography --- p.58 / Chapter 2.4 --- Assays for measuring antioxidant activity --- p.60 / Chapter 2.4.1 --- Beta-carotene bleaching method --- p.60 / Chapter 2.4.2 --- Scavenging activity of DPPH radical --- p.62 / Chapter 2.4.3 --- Erythrocyte hemolysis --- p.63 / Chapter 2.4.4 --- Scavenging activity of ABTS ´Ø + radical cation --- p.64 / Chapter 2.4.5 --- Scavenging activity of hydroxyl radical --- p.65 / Chapter 2.4.6 --- Assay for lipid peroxidation of rat brain homogenate --- p.66 / Chapter 2.4.7 --- Inhibition of human low-density lipoproteins (LDLs) oxidation --- p.67 / Chapter 2.4.7.1 --- Isolation of human LDLs --- p.67 / Chapter 2.4.7.2 --- Calculation of density --- p.68 / Chapter 2.4.7.3 --- Lowry's method for determination of protein content --- p.69 / Chapter 2.4.7.4 --- Preparation of reagents --- p.69 / Chapter 2.4.7.5 --- Determination of thiobarbituric acid reactive substance (TBARS) --- p.70 / Chapter 2.5 --- Total phenolic content --- p.70 / Chapter 2.6 --- Total carbohydrate content --- p.71 / Chapter 2.7 --- Determination of protein content- the Biuret method --- p.71 / Chapter 2.8 --- Thin-layer chromatography (TLC) --- p.72 / Chapter 2.9 --- High performance liquid chromatography (HPLC) --- p.73 / Chapter 2.9.1 --- Analysis of subfractions of methanol crude extract --- p.73 / Chapter 2.9.2 --- Analysis of fractionated subfractions and subfractions of Pevf and Aa mushrooms --- p.74 / Chapter 2.10 --- Liquid chromatography- Mass spectrometry (LC-MS) --- p.74 / Chapter 2.10.1 --- Liquid chromatography --- p.74 / Chapter 2.10.2 --- Mass spectrometry --- p.75 / Chapter 2.11 --- Data analysis --- p.75 / Chapter CHAPTER 3: --- RESULTS AND DISCUSSION --- p.77 / Chapter 3.1 --- Small-scale extraction scheme --- p.77 / Chapter 3.1.1 --- Extraction yield --- p.77 / Chapter 3.1.2 --- Assays for measuring antioxidant activity --- p.80 / Chapter 3.1.2.1 --- Beta-carotene bleaching method --- p.80 / Chapter 3.1.2.2 --- Scavenging activity of DPPH radical --- p.91 / Chapter 3.1.2.3 --- Erythrocyte hemolysis --- p.98 / Chapter 3.1.2.4 --- Summary for small-scale extraction --- p.105 / Chapter 3.2 --- Large-scale extraction and fractionation scheme --- p.107 / Chapter 3.2.1 --- Extraction yield for crude extracts and subfractions --- p.107 / Chapter 3.2.2 --- Antioxidant activity of subfractions and crude extracts of Aa and Pevf mushrooms --- p.111 / Chapter 3.2.2.1 --- Scavenging activity of ABTS ´Ø + radical cation --- p.111 / Chapter 3.2.2.2 --- Scavenging activity of hydroxyl radical --- p.114 / Chapter 3.2.2.3 --- Assay for lipid peroxidation of rat brain homogenate --- p.125 / Chapter 3.2.2.4 --- Summary for large-scale extraction --- p.135 / Chapter 3.2.3 --- Chemical characterization of the crude extracts and their sub fractions of Aa and Pevf mushrooms --- p.13 8 / Chapter 3.2.3.1 --- Total phenolic content --- p.139 / Chapter 3.2.3.1.1 --- Total phenolic content of crude extract and their sub fractions of Aa and Pevf mushrooms --- p.139 / Chapter 3.2.3.1.2 --- Correlation between total phenolic content and antioxidant activity --- p.140 / Chapter 3.2.3.2 --- Total carbohydrate content --- p.144 / Chapter 3.2.3.2.1 --- Total carbohydrate content of water crude extract and their sub fractions of Aa and Pevf mushrooms --- p.144 / Chapter 3.2.3.2.2 --- Correlation between total carbohydrate content and antioxidant activity --- p.144 / Chapter 3.2.3.3 --- Determination of protein content- the Biuret method --- p.146 / Chapter 3.2.3.3.1 --- Protein content of water crude extract and their sub fractions of Aa and Pevf mushrooms --- p.146 / Chapter 3.2.3.3.2 --- Correlation between protein content and antioxidant activity --- p.147 / Chapter 3.2.3.4 --- Summary of correlation between chemical components and antioxidant activity --- p.148 / Chapter 3.3 --- Column fractionation of ethyl acetate and butanol subfractions of Aa mushroom --- p.151 / Chapter 3.3.1 --- Rf value in TLC and yield of fractionated subfractions --- p.151 / Chapter 3.3.2 --- Total phenolic content of fractionated subfractions --- p.153 / Chapter 3.3.3 --- Antioxidant activity of fractionated subfractions --- p.155 / Chapter 3.3.3.1 --- Scavenging activity of ABTS ´Ø + radical cation --- p.155 / Chapter 3.3.3.2 --- Scavenging activity of DPPH radical --- p.158 / Chapter 3.3.3.3 --- Inhibition of human low-density lipoprotein (LDL) oxidation --- p.163 / Chapter 3.4 --- Chromatographic characterization of the subfractions of methanol crude extract of Aa and Pevf mushrooms --- p.167 / Chapter 3.4.1 --- Thin-layer chromatography (TLC) --- p.167 / Chapter 3.4.2 --- High performance liquid chromatography (HPLC) --- p.172 / Chapter 3.5 --- Chromatographic and spectrometric characterization of the fractionated subfractions of the ethyl acetate and butanol subfractions of Aa --- p.177 / Chapter 3.5.1 --- High performance liquid chromatography (HPLC) --- p.177 / Chapter 3.5.2 --- Liquid chromatography- Mass spectrometry (LC-MS) --- p.186 / Chapter CHAPTER 4: --- CONCLUSION --- p.189 / REFERENCES --- p.193 / RELATED PUBLICATION --- p.209

Cheung Lai Ming. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 153-161). / Abstracts in English and Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT (Chinese version) --- p.v / CONTENTS --- p.vi / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xiii / LIST OF ABBREVIATIONS --- p.xv / Chapter CHAPTER ONE: --- INTRODUCTION --- p.1 / Chapter 1.1 --- Free radical --- p.2 / Chapter 1.1.1 --- Definition --- p.2 / Chapter 1.1.2 --- Reaction mechanism --- p.3 / Chapter 1.1.3 --- Sources of oxygen reactive species --- p.4 / Chapter 1.1.3.1 --- Enzymes --- p.4 / Chapter 1.1.3.2 --- The auto-oxidation of small molecules --- p.4 / Chapter 1.1.3.3 --- Haem proteins --- p.5 / Chapter 1.1.3.4 --- Endoplasmic reticulum sources --- p.5 / Chapter 1.1.3.5 --- Mitochondrial sources --- p.5 / Chapter 1.1.3.6 --- Nucleus --- p.6 / Chapter 1.1.4 --- Lipid peroxidation --- p.6 / Chapter 1.1.4.1 --- Initiation of lipid peroxidation --- p.7 / Chapter 1.1.4.2 --- Propagation of lipid peroxidation --- p.8 / Chapter 1.1.4.3 --- Products of lipid peroxidation --- p.9 / Chapter 1.1.5 --- Human diseases associated with free radicals --- p.10 / Chapter 1.2 --- Antioxidants --- p.12 / Chapter 1.2.1 --- Definition --- p.12 / Chapter 1.2.2 --- Defence against free radical damage --- p.13 / Chapter 1.2.2.1 --- Catalytic free radical removal --- p.13 / Chapter 1.2.2.2 --- Free radical scavenging --- p.14 / Chapter 1.2.2.3 --- Removal of catalytic iron and copper ions --- p.14 / Chapter 1.2.3 --- Synthetic vs. natural antioxidant --- p.15 / Chapter 1.2.3.1 --- Synthetic antioxidants --- p.15 / Chapter 1.2.3.2 --- Natural antioxidants --- p.16 / Chapter 1.3 --- Measurement of antioxidant activity --- p.17 / Chapter 1.3.1 --- Loss of substrate --- p.17 / Chapter 1.3.1.1 --- Beta-carotene bleaching method --- p.17 / Chapter 1.3.2 --- Measurement of free radical scavenging --- p.17 / Chapter 1.3.2.1 --- "Scavenging of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH´Ø)" --- p.17 / Chapter 1.3.2.2 --- Superoxide scavenging --- p.18 / Chapter 1.3.2.3 --- Hydrogen peroxide scavenging --- p.18 / Chapter 1.3.2.4 --- Hydroxyl radical scavenging --- p.19 / Chapter 1.3.2.5 --- Peroxyl radical --- p.19 / Chapter 1.3.3 --- Measurement of end product --- p.21 / Chapter 1.3.3.1 --- Diene conjugation --- p.21 / Chapter 1.3.3.2 --- Light emission --- p.21 / Chapter 1.3.3.3 --- The thiobarbituric acid (TBA) test --- p.22 / Chapter 1.3.4 --- Low-density lipoprotein oxidation --- p.22 / Chapter 1.4 --- Phenolic antioxidant --- p.24 / Chapter 1.4.1 --- Chemistry --- p.24 / Chapter 1.4.2 --- Mechanism of action of phenolic antioxidants --- p.25 / Chapter 1.4.3 --- Isolation and characterization --- p.25 / Chapter 1.4.3.1 --- Extraction --- p.25 / Chapter 1.4.3.2 --- Analysis of phenolic compounds --- p.27 / Chapter 1.4.3.2.1 --- Colorimetric method --- p.27 / Chapter 1.4.3.2.2 --- Enzymatic method --- p.28 / Chapter 1.4.3.2.3 --- Paper chromatography --- p.28 / Chapter 1.4.3.2.4 --- Thin-layer chromatography --- p.29 / Chapter 1.4.3.2.5 --- UV-Vis absorption spectroscopy --- p.29 / Chapter 1.4.3.2.6 --- High-performance liquid chromatography --- p.30 / Chapter 1.4.4 --- Natural sources of phenolic antioxidants --- p.31 / Chapter 1.4.4.1 --- Olive oil --- p.31 / Chapter 1.4.4.2 --- Berry --- p.32 / Chapter 1.4.4.3 --- Cherry --- p.32 / Chapter 1.4.4.4 --- Red wine --- p.32 / Chapter 1.4.4.5 --- Herb --- p.33 / Chapter 1.4.4.6 --- Vegetables --- p.33 / Chapter 1.5 --- Mushroom Sample --- p.34 / Chapter 1.5.1 --- Pleurotus tuber-regium --- p.34 / Chapter 1.5.2 --- Lentinus edodes --- p.34 / Chapter 1.5.3 --- Volvariella volvacea --- p.35 / Chapter 1.5.4 --- Antioxidants in fungi or mushroom --- p.37 / Chapter 1.5.5 --- Phenolic compounds in mushrooms --- p.39 / Chapter 1.6 --- Objectives --- p.42 / Chapter CHAPTER TWO: --- MATERIALS AND METHODS --- p.43 / Chapter 2.1 --- Sample Collection --- p.43 / Chapter 2.2 --- Sample Preparation --- p.43 / Chapter 2.3 --- Moisture Content --- p.43 / Chapter 2.4 --- Solvent Extraction --- p.44 / Chapter 2.4.1 --- Scheme I (Aqueous extraction only) --- p.44 / Chapter 2.4.2 --- Scheme II (Methanol and water extraction) --- p.45 / Chapter 2.4.3 --- Scheme III (Differential solvent extraction) --- p.46 / Chapter 2.4.4 --- Scheme IV (Scaled-up extraction) --- p.47 / Chapter 2.5 --- Antioxidant activity assays --- p.50 / Chapter 2.5.1 --- Beta-carotene bleaching method --- p.50 / Chapter 2.5.2 --- "Scavenging activity on 1,1 -diphenyl-2-picrylhydrazyl radicals" --- p.51 / Chapter 2.5.3 --- Assay for erythrocyte hemolysis --- p.51 / Chapter 2.5.4 --- Assay of lipid peroxidation using rat brain --- p.52 / Chapter 2.5.5 --- LDL oxidation (TBARS) --- p.53 / Chapter 2.5.5.1 --- LDL Isolation --- p.53 / Chapter 2.5.5.2 --- Calculation of density --- p.54 / Chapter 2.5.5.3 --- Lowry Method for Protein Determination --- p.55 / Chapter 2.5.5.4 --- Reagents for TBARS assay --- p.55 / Chapter 2.5.5.5 --- TBARS formation --- p.56 / Chapter 2.6 --- Determination of total polyphenolic compounds --- p.56 / Chapter 2.7 --- Fractionation --- p.57 / Chapter 2.7.1 --- Fractionation of the methanol crude extracts obtained under reflux by solvent --- p.57 / Chapter 2.7.2 --- Fractionation of boiling water crude extracts by ultrafiltration --- p.57 / Chapter 2.8 --- Crude Protein Content (Kjeldahl method) --- p.58 / Chapter 2.9 --- Total carbohydrate content --- p.59 / Chapter 2.10 --- Thin-layer chromatography --- p.59 / Chapter 2.11 --- High performance liquid chromatography --- p.60 / Chapter 2.11.1 --- Analysis of methanol fractions --- p.60 / Chapter 2.11.2 --- Analysis of water fractions --- p.61 / Chapter 2.12 --- Liquid chromatography-Mass spectrometry --- p.61 / Chapter 2.12.1 --- Liquid chromatography --- p.61 / Chapter 2.12.2 --- Mass spectrometric analysis --- p.62 / Chapter 2.13 --- Data analysis --- p.62 / Chapter CHAPTER THREE: --- RESULTS AND DISCUSSION --- p.63 / Chapter 3.1 --- Mushroom sample --- p.63 / Chapter 3.2 --- Extraction scheme I --- p.65 / Chapter 3.2.1 --- Antioxidant activity --- p.65 / Chapter 3.2.1.1 --- Effect of extraction temperature --- p.65 / Chapter 3.2.1.2 --- Effect of concentration of extracts --- p.66 / Chapter 3.3 --- Extraction scheme II --- p.69 / Chapter 3.3.1 --- Antioxidant activity --- p.69 / Chapter 3.3.1.1 --- Effect of extraction temperature --- p.69 / Chapter 3.3.1.2 --- Effect of concentration of extracts --- p.72 / Chapter 3.3.1.3 --- Effect of solvent --- p.72 / Chapter 3.4 --- Extraction scheme III --- p.75 / Chapter 3.4.1 --- Extraction yield --- p.75 / Chapter 3.4.2 --- Total phenolic content --- p.76 / Chapter 3.4.3 --- Antioxidant activity --- p.80 / Chapter 3.4.3.1 --- Beta-carotene bleaching method --- p.80 / Chapter 3.4.3.1.1 --- Effect of extract concentration --- p.80 / Chapter 3.4.3.1.2 --- Relation between total phenolic content and antioxidant activity --- p.82 / Chapter 3.4.3.2 --- "Scavenging activity of 1,1 -diphenyl-2-picrylhydrazyl (DPPH) radical" --- p.85 / Chapter 3.4.3.3 --- Assay for erythrocyte hemolysis --- p.88 / Chapter 3.5 --- Extraction scheme IV --- p.91 / Chapter 3.5.1 --- Yield and Fractionation --- p.91 / Chapter 3.5.2 --- Chemical characterization of fractions --- p.93 / Chapter 3.5.2.1 --- Protein content --- p.93 / Chapter 3.5.2.2 --- Total carbohydrate content --- p.93 / Chapter 3.5.2.3 --- Total phenolic content --- p.94 / Chapter 3.5.3 --- Antioxidant activity --- p.99 / Chapter 3.5.3.1 --- Assay for lipid peroxidation of rat brain --- p.99 / Chapter 3.5.3.2 --- LDL oxidation --- p.118 / Chapter 3.5.4 --- Identification of antioxidant by chromatographic methods --- p.126 / Chapter 3.5.4.1 --- Thin-layer chromatography --- p.126 / Chapter 3.5.4.2 --- High-performance liquid chromatography --- p.132 / Chapter 3.5.4.3 --- Liquid chromatography-Mass spectrometry --- p.142 / Chapter CHAPTER FOUR: --- CONCLUSION --- p.148 / REFERENCES --- p.153 / RELATED PUBLICATION --- p.161

No description available.

Whey protein isolates (WPI) have been indicated to have potent cholesterol lowering and antioxidative properties. Such effects, however, are not consistently observed, which could be the result of major differences in the processing, isolation and composition of WPI. Moreover, the mechanisms of action or the bioactive component(s) in WPI are poorly understood although the relatively high cysteine content in WPI has been suggested to play an important role. Although high dietary cysteine has been shown to lower plasma homocysteine concentrations, the impact of WPI in this regard has not been investigated. The overall objective of this thesis was to examine the antioxidative and plasma cholesterol and homocysteine lowering properties of two WPI that were produced via different industrial processing and isolation techniques with the milk protein, casein, used as the control protein. We also examined for the mechanism(s) of action of WPI in terms of possible antioxidative, and plasma cholesterol and homocysteine lowering effects. In this regard, the intake of bovine serum albumin (BSA), a major cysteine-rich whey protein was also studied since this protein has been implicated as a key bioactive component for the antioxidant effects of WPI. Four studies were performed. The first involved the characterization of a variety of commercially prepared WPI by high performance capillary electrophoresis for identification of two WPI products that showed major differences in protein composition for subsequent feeding trials. Most of the WPI had similar characteristic electrophoretic profiles, however, significant differences in protein and macronutrient (Ca, Mg, P) composition were noted in two commercial WPI that were chosen as the test proteins in subsequent feeding trials. In the first two feeding studies, hamsters were fed different commercial WPI or milk protein (BSA or casein) containing diets that were either matched or unmatched in terms of macro

Reduction of milk production following acute bovine mastitis causes important economic losses. In this study, two experiments were conducted to asses the ability of different antioxidants to prevent neutrophil (PMN)-induced mammary damage in acute bovine mastitis. First, a co-culture model composed of bovine mammary epithelial cell line (MAC-T cells) and bovine PMN activated by phorbol myristate acetate was used. Activated PMN release reactive oxygen species that are cytotoxic for bovine epithelial cells. Addition of dimethylthiourea or bathocuproinic acid did not induce any protective effect. On the other hand, addition of catechin, deferoxamine or glutathione ethyl ester (GEE) significantly reduced PMN-induced cytotoxicity in a dose-dependent manner as demonstrated by lower levels of released lactate dehydrogenase (LDH). The second experiment was undertaken with the last three antioxidants to evaluate their protective effects in vivo. A model of LPS-induced mastitis on dairy cows was used. The extent of cell damages was evaluated by measuring quarter milk levels of LDH and 4-methylumbelliferyl N-acetyl beta-D-glucosaminidase ( NAGase) at varying intervals before and after intramammary infusions of LPS, with or without antioxidants. Milk levels of haptoglobin and bovine serum albumin were also analysed. Catechin and GEE did not induce any protective effect whereas infusions of deferoxamine, a chelator of iron, decreased milk levels of LDH, NAGase and haptoglobin hence suggesting a protective effect against PMN-induced damage. Deferoxamine did not interfere with PMN migration into the mammary gland. Additionally, deferoxamine inhibited bacterial growth in vitro but did not affect PMN's ability to phagocytize live Escherichia coli. Overall, our results suggest that local infusion of deferoxamine may be an effective tool to protect mammary tissue against PMN-induced oxidative stress during bovine mastitis.

No description available.

Lee Chung-hung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 113-133). / Abstracts in English and Chinese. / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- History of soybean --- p.1 / Chapter 1.2 --- Health benefits of soybean --- p.2 / Chapter 1.3 --- Introduction to flavonoids --- p.3 / Chapter 1.4 --- Bioavailability of flavonoids --- p.5 / Chapter 1.5 --- Chemistry of isoflavones --- p.6 / Chapter 1.6 --- Estrogenic property of isoflavones --- p.8 / Chapter 1.7 --- Nutritional significance of isoflavones and their glycosides --- p.8 / Chapter 1.7.1 --- Anticarcinogenic activity --- p.9 / Chapter 1.7.2 --- Antioxidative activity --- p.10 / Chapter 1.7.3 --- Cardioprotective activity --- p.13 / Chapter 1.7.4 --- Osteoprotective activity --- p.14 / Chapter 1.7.5 --- Neuroprotective activity --- p.15 / Chapter 1.7.6 --- Antiangiogenic activity --- p.16 / Chapter Chapter 2 --- Composition of Soybean Isoflavones / Chapter 2.1 --- Introduction --- p.17 / Chapter 2.2 --- Objective --- p.19 / Chapter 2.3 --- Materials and Methods --- p.20 / Chapter 2.3.1 --- Extraction and isolation --- p.20 / Chapter 2.3.1.1 --- Preparation of soybean butanol extract --- p.20 / Chapter 2.3.1.2 --- Preparation of isoflavones and their glycosides from soybean butanol extract --- p.20 / Chapter 2.3.2 --- HPLC analysis --- p.21 / Chapter 2.3.2.1 --- Sample preparation for the HPLC analysis --- p.21 / Chapter 2.3.2.2 --- HPLC analysis --- p.22 / Chapter 2.3.2.3 --- Quantification of the flavonoids and their glycosides --- p.24 / Chapter 2.4 --- Results --- p.25 / Chapter 2.4.1 --- Structural identification --- p.25 / Chapter 2.4.1.1 --- Compound 1 --- p.25 / Chapter 2.4.1.2 --- Compound 2 --- p.26 / Chapter 2.4.1.3 --- Compound 3 --- p.26 / Chapter 2.4.1.4 --- Compound 4 --- p.27 / Chapter 2.4.1.5 --- Compound 5 --- p.27 / Chapter 2.4.1.6 --- Compound 6 --- p.28 / Chapter 2.4.1.7 --- Compound 7 --- p.28 / Chapter 2.4.1.8 --- Compound 8 --- p.29 / Chapter 2.4.2 --- Quantification of isoflavones in traditional Chinese foods --- p.29 / Chapter 2.5 --- Discussion --- p.32 / Chapter Chapter 3 --- Hypocholesterolemic Effects of Soymilkin Hamsters / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.1.1 --- Lipoproteins and their functions --- p.35 / Chapter 3.1.2 --- Risk factors of cardiovascular disease --- p.36 / Chapter 3.1.3 --- Hamster as an animal model of cholesterol metabolism --- p.38 / Chapter 3.2 --- Objective --- p.39 / Chapter 3.3 --- Materials and Methods --- p.40 / Chapter 3.3.1 --- Preparation of soymilk --- p.40 / Chapter 3.3.2 --- Animals --- p.40 / Chapter 3.3.2.1 --- Experiment one - Hypocholesterolemic effect of soymilk in hamsters --- p.40 / Chapter 3.3.2.1 --- Experiment two 一 The effect of fluid cross-over between soymilk and cow´ة s milk on serum cholesterol in hamsters --- p.41 / Chapter 3.3.3 --- Serum lipid and lipoprotein determinations --- p.42 / Chapter 3.3.4 --- Determination of cholesterol in organs --- p.42 / Chapter 3.3.5 --- Statistics --- p.43 / Chapter 3.4 --- Results --- p.46 / Chapter 3.4.1 --- Experiment one-Hypocholesterolemic effect of soymilk in hamsters --- p.46 / Chapter 3.4.1.1 --- Growth and food intake --- p.46 / Chapter 3.4.1.2 --- "Effect of SM and CM on TG, TC and HDL-C" --- p.46 / Chapter 3.4.1.3 --- Effect of SM and CM on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.46 / Chapter 3.4.1.4 --- Effect of SM and CM on concentration of hepatic cholesterol --- p.47 / Chapter 3.4.1.5 --- "Effect of SM and CM on brain, heart and kidney cholesterol" --- p.47 / Chapter 3.4.2 --- Experiment two - The effect of fluid cross-over between soymilk and cow´ةs milk on serum cholesterol in hamsters --- p.52 / Chapter 3.4.2.1 --- Growth and food intake --- p.52 / Chapter 3.4.2.2 --- Effect of fluid cross-over on serum TC --- p.52 / Chapter 3.5 --- Discussion --- p.55 / Chapter Chapter 4 --- Antioxidant Activities of Soybean Isoflavones and Their Glycosides / Chapter 4.1 --- Introduction --- p.58 / Chapter 4.1.1 --- Role of low density lipoprotein oxidation in the development of atherosclerosis --- p.59 / Chapter 4.1.2 --- LDL oxidation --- p.61 / Chapter 4.1.3 --- Thiobarbituric acid reactive substances (TBARS) as an index of LDL oxidation --- p.62 / Chapter 4.1.4 --- "The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power""" --- p.65 / Chapter 4.1.5 --- "1,1-diphenyl-2-picrylhydrazyl (DPPH) as a measure of free radical scavenging capacity" --- p.65 / Chapter 4.1.6 --- Antioxidant and LDL oxidation --- p.65 / Chapter 4.2 --- Objective --- p.67 / Chapter 4.3 --- Materials and Methods --- p.68 / Chapter 4.3.1 --- Preparation of samples --- p.68 / Chapter 4.3.2 --- Isolation of LDL from human serum --- p.68 / Chapter 4.3.3 --- LDL oxidation --- p.69 / Chapter 4.3.4 --- TBARS assay --- p.69 / Chapter 4.3.5 --- FRAP assay --- p.70 / Chapter 4.3.6 --- DPPH assay --- p.71 / Chapter 4.3.7 --- Statistics --- p.72 / Chapter 4.4 --- Results --- p.73 / Chapter 4.4.1 --- Effects of seven individual soybean isoflavones and their glycosides on LDL oxidation --- p.73 / Chapter 4.4.2 --- The antioxidant power of individual soybean isoflavones and their glycosides in the FRAP assay --- p.73 / Chapter 4.4.3 --- Activity of individual soybean isoflavones and their glycosides as radical scavenging antioxidants --- p.74 / Chapter 4.5 --- Discussion --- p.78 / Chapter Chapter 5 --- Hypocholesterolemic Effects of Soybean Isoflavones in Ovariectomized Golden Syrian Hamsters / Chapter 5.1 --- Introduction --- p.83 / Chapter 5.1.1 --- Coronary heart disease in women --- p.83 / Chapter 5.1.2 --- Menopause as a risk factor in CHD --- p.84 / Chapter 5.1.3 --- Dietary soy in treating postmenopausal hypercholesterolemia --- p.85 / Chapter 5.2 --- Objective --- p.87 / Chapter 5.3 --- Materials and Methods --- p.88 / Chapter 5.3.1 --- Preparation of soymilk --- p.88 / Chapter 5.3.2 --- Preparation of soybean extract --- p.88 / Chapter 5.3.3 --- Animals --- p.89 / Chapter 5.3.4 --- Serum lipid determinations --- p.90 / Chapter 5.3.5 --- Determination of tissue cholesterol content --- p.90 / Chapter 5.3.6 --- Extraction of neutral and acidic sterols from fecal samples --- p.90 / Chapter 5.3.6.1 --- Determination of neutral sterols --- p.91 / Chapter 5.3.6.2 --- Determination of acidic sterols --- p.92 / Chapter 5.3.6.3 --- GLC analysis of neutral and acidic sterols --- p.92 / Chapter 5.3.7 --- Statistics --- p.93 / Chapter 5.4 --- Results --- p.96 / Chapter 5.4.1 --- Growth and food intake --- p.96 / Chapter 5.4.2 --- Effect of ovariectomy on serum TC --- p.96 / Chapter 5.4.3 --- "Effect of soymilk and soybean extract on serum TC,TG and HDL-C" --- p.96 / Chapter 5.4.4 --- Effect of soymilk and soybean extract on non-HDL-C and ratio of non- HDL-C to HDL-C --- p.97 / Chapter 5.4.5 --- Effect of soymilk and soybean extract on concentration of hepatic cholesterol --- p.97 / Chapter 5.4.6 --- Effect of soymilk and soybean extract on heart and kidney cholesterol --- p.97 / Chapter 5.4.7 --- Effect of soymilk and soybean extract on fecal neutral and acidic sterols --- p.103 / Chapter 5.5 --- Discussion --- p.106 / Chapter Chapter 6 --- Conclusion --- p.110 / References --- p.113

Ng Yuk Fan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 145-162). / Abstracts in English and Chinese. / Thesis Committee: --- p.i / Acknowledgements --- p.ii / Abstract --- p.iii / 摘要 --- p.v / Content --- p.vii / List of Tables --- p.xiii / List of Figures --- p.xvi / Abbreviations --- p.xviii / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Antioxidants --- p.1 / Chapter 1.1.1 --- Definition and mode of actions of antioxidants --- p.1 / Chapter 1.1.2 --- Synthetic antioxidants --- p.2 / Chapter 1.1.3 --- Natural antioxidants --- p.3 / Chapter 1.2 --- Changes of antioxidant activity in food processing --- p.4 / Chapter 1.2.1 --- Blanching --- p.4 / Chapter 1.2.2 --- Drying --- p.5 / Chapter 1.2.3 --- Microwave and Infrared energy --- p.7 / Chapter 1.2.4 --- Freezing --- p.8 / Chapter 1.3 --- Lipid oxidation and antioxidant --- p.8 / Chapter 1.3.1 --- Free radicals --- p.8 / Chapter 1.3.1.1 --- Superoxide --- p.10 / Chapter 1.3.1.2 --- Hydrogen peroxide --- p.11 / Chapter 1.3.1.3 --- Hydroxyl radical --- p.13 / Chapter 1.3.2 --- Mechanism of lipid oxidation --- p.14 / Chapter 1.3.3 --- Oxidation of low-density-liporoproteins (LDLs) and coronary heart disease --- p.15 / Chapter 1.3.4 --- Role of antioxidants in inhibiting lipid oxidation --- p.16 / Chapter 1.4 --- Hypocholesterolemic and antioxidant activity of phenolics --- p.19 / Chapter 1.5 --- Medicinal properties of mushrooms --- p.21 / Chapter 1.5.1 --- Background information of mushrooms --- p.21 / Chapter 1.5.2 --- Phenolics in mushrooms --- p.22 / Chapter 1.5.3 --- Hypocholesterolemic effect in mushroom --- p.23 / Chapter 1.5.4 --- Previous studies in Agrocybe aegerita --- p.25 / Chapter 1.6 --- Animal model for hypocholesteroliemic study --- p.27 / Chapter 1.6.1 --- General requirements --- p.27 / Chapter 1.6.2 --- Hamster model --- p.27 / Chapter 1.7 --- Principles of assays that involved in antioxidant activity --- p.30 / Chapter 1.7.1 --- ABTS + radical cation scavenging activity --- p.30 / Chapter 1.7.2 --- Beta carotene bleaching method --- p.31 / Chapter 1.7.3 --- Ferric reducing antioxidant power (FRAP) --- p.31 / Chapter 1.7.4 --- Scavenging activity of hydroxyl radical --- p.32 / Chapter 1.7.5 --- Inhibition of low-density lipoproteins (LDLs) oxidation --- p.33 / Chapter 1.7.6 --- Total phenolic content determination --- p.33 / Chapter 1.8 --- Principles of assays in hypocholesterolemic study --- p.34 / Chapter 1.8.1 --- HDL-Cholesterol determination --- p.34 / Chapter 1.8.2 --- Total cholesterol determination --- p.34 / Chapter 1.8.3 --- Determination of plasma total triglyceride --- p.35 / Chapter 1.9 --- Objectives --- p.36 / Chapter Chapter 2: --- Materials and Methods --- p.37 / Chapter 2.1 --- Sample preparation --- p.37 / Chapter 2.2 --- Proximate Analysis of FAa and DAa --- p.38 / Chapter 2.2.1 --- Determination of crude protein --- p.38 / Chapter 2.2.2 --- Determination of ash --- p.39 / Chapter 2.2.3 --- Total dietary fiber --- p.39 / Chapter 2.2.4 --- Determination of fat --- p.41 / Chapter 2.2.5 --- Moisture content --- p.42 / Chapter 2.3 --- Sample extraction --- p.42 / Chapter 2.3.1 --- Small-scale extraction --- p.42 / Chapter 2.3.2 --- Large-scale extraction --- p.43 / Chapter 2.4 --- Total phenolic content of DAa and FAa extract --- p.44 / Chapter 2.5 --- Chemical assays for in vitro antioxidative properties determination --- p.45 / Chapter 2.5.1 --- Hydroxyl free radical scavenging activity --- p.45 / Chapter 2.5.2 --- Beta-carotene bleaching method --- p.46 / Chapter 2.5.3 --- Inhibition of human low-density-lipoproteins (LDLs) oxidation --- p.47 / Chapter 2.5.4 --- Scavenging activity of ABTS+radical cation --- p.50 / Chapter 2.6 --- In vivo tests for antioxidative and hypocholesterolemic effect of DAa --- p.51 / Chapter 2.6.1 --- Feeding experiments --- p.51 / Chapter 2.6.2 --- Collection of plasma --- p.52 / Chapter 2.6.3 --- Liver sample preparation --- p.52 / Chapter 2.6.4 --- Determination of in vivo antioxidative effect --- p.54 / Chapter 2.6.4.1 --- FRPA assay --- p.54 / Chapter 2.6.4.2 --- ABTS + radical cation scavenging activity --- p.55 / Chapter 2.6.5 --- Determination of plasma lipid profiles --- p.55 / Chapter 2.6.5.1 --- Plasma total cholesterol (TC) --- p.55 / Chapter 2.6.5.2 --- Plasma total triglyceride (TG) --- p.56 / Chapter 2.6.5.3 --- Plasma high density lipoprotein cholesterol (HDL-C) determination --- p.57 / Chapter 2.6.5.4 --- Hepatic cholesterol determination by gas chromatography analysis --- p.57 / Chapter 2.7 --- Statistical analysis --- p.59 / Chapter Chapter 3: --- Results and discussion --- p.61 / Chapter 3.1 --- Proximate analysis --- p.61 / Chapter 3.2 --- Small-scale extraction scheme --- p.63 / Chapter 3.2.1 --- Extraction yield --- p.63 / Chapter 3.2.2 --- Antioxidant assays --- p.65 / Chapter 3.2.2.1 --- Hydroxyl free radical scavenging activity --- p.65 / Chapter 3.2.2.2 --- Beta-carotene bleaching method --- p.68 / Chapter 3.2.2.3 --- The formation of TBARS in human LDL oxidation --- p.75 / Chapter 3.2.2.4 --- Total phenolic content (TPC) in DAa and FAa ethanolic and water extracts --- p.81 / Chapter 3.2.2.5 --- Correlation between total phenolic content and antioxidant activity of mushroom extracts --- p.84 / Chapter 3.2.2.6 --- Comparison of antioxidant activity and TPC in DAa and FAa ethanolic and water extracts in the small-scale extraction scheme --- p.88 / Chapter 3.3 --- Large-scale extraction scheme --- p.91 / Chapter 3.3.1 --- Extraction yield --- p.91 / Chapter 3.3.2 --- Antioxidant assays --- p.91 / Chapter 3.3.2.1 --- Hydroxyl free radical scavenging activity --- p.91 / Chapter 3.3.2.2 --- Beta-carotene bleaching method --- p.94 / Chapter 3.3.2.3 --- ABTS + radical cation scavenging activity --- p.96 / Chapter 3.3.2.4 --- Formation of TBARS in human LDL oxidation in the DAa_E_l and Daa_W_1 --- p.97 / Chapter 3.3.2.5 --- Total phenolic content (TPC) of DAa_E_l and DAa_W_l --- p.97 / Chapter 3.3.2.6 --- Correlation between total phenolic content and antioxidant activity --- p.101 / Chapter 3.3.2.7 --- Summary of large-scale extraction scheme --- p.103 / Chapter 3.4 --- In vivo antioxidant activity and hypocholesterolemic effect of DAa studied by animal model --- p.104 / Chapter 3.4.1 --- Effect of DAa´ؤE_1 and DAa_W_l on body weight and food intake --- p.105 / Chapter 3.4.2 --- Effect of DAa一E´ؤ1 and DAa_W_l on plasma total cholesterol (TC) in hamsters --- p.108 / Chapter 3.4.3 --- Effect of DAa´ؤE_1 and DAa W l on plasma total triglycerides (TG) in hamsters --- p.114 / Chapter 3.4.4 --- Effect of DAa_E_l and DAa_W_l on plasma high-density-lipoprotein cholesterol (HDL-C) in hamsters --- p.119 / Chapter 3.4.5 --- Effect of DAa_E_l and DAa一W_1 on hepatic cholesterol (HC) profile in hamsters --- p.124 / Chapter 3.4.6 --- Effect of DAa_E_l and DAa W l on ferric reducing antioxidant power (FRAP) in hamsters (FRAP) --- p.128 / Chapter 3.4.7 --- Effect of DAa_E_l and DAa_W_l on ABTS + cation radical scavenging activity --- p.131 / Chapter 3.4.8 --- The antioxidant activity and hypocholesterolemic effect of DAa extracts --- p.134 / Chapter 3.4.9 --- Summary of in vivo antioxidant activity and hypocholesterolemic effect of DAa studied by animal model --- p.140 / Chapter Chapter 4: --- Conclusions --- p.142 / References --- p.145

Ho, Ka Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 111-124). / Abstracts in English and Chinese. / Thesis committee --- p.i / Acknowledgements --- p.ii / Abstract --- p.iii / 摘要 --- p.v / List of Tables --- p.vii / List of Figures --- p.viii / Abbreviations --- p.x / Content --- p.xiii / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Reactive oxygen species (ROS) --- p.1 / Chapter 1.1.1 --- Definition and examples --- p.1 / Chapter 1.1.2 --- Generation of ROS in biological systems --- p.2 / Chapter 1.1.3 --- Features of specif ic ROS --- p.3 / Chapter 1.1.3.1 --- Superoxide anion --- p.3 / Chapter 1.1.3.2 --- Peroxyl radical --- p.4 / Chapter 1.1.3.3 --- Hydrogen peroxide --- p.4 / Chapter 1.1.3.4 --- Hydroxyl radical --- p.5 / Chapter 1.1.4 --- Damaging effects of ROS on biomolecules --- p.5 / Chapter 1.1.4.1 --- Lipid peroxidation --- p.6 / Chapter 1.1.4.2 --- DNA damage --- p.8 / Chapter 1.1.4.3 --- Protein oxidation --- p.9 / Chapter 1.2 --- Antioxidants --- p.11 / Chapter 1.2.1 --- Introduction --- p.11 / Chapter 1.2.2 --- Mode of action --- p.11 / Chapter 1.2.3 --- Endogenous Antioxidants --- p.12 / Chapter 1.2.3.1 --- Antioxidant enzymes --- p.12 / Chapter 1.2.3.2 --- Antioxidant compounds --- p.15 / Chapter 1.2.4 --- Exogenous antioxidants --- p.16 / Chapter 1.3 --- Oxidative stress --- p.17 / Chapter 1.3.1 --- Balance between ROS and antioxidants --- p.17 / Chapter 1.3.2 --- Diseases associated with oxidative stress --- p.18 / Chapter 1.4 --- Previous studies on edible mushroom antioxidants --- p.19 / Chapter 1.4.1 --- Previous studies on Agrocybe aegerita --- p.20 / Chapter 1.5 --- Cell culture models for antioxidant research --- p.21 / Chapter 1.6 --- Objectives --- p.23 / Chapter Chapter 2 --- Materials and Methods --- p.24 / Chapter 2.1 --- Materials --- p.24 / Chapter 2.1.1 --- Mushroom fruiting bodies --- p.24 / Chapter 2.1.2 --- Cell lines and their subcultures --- p.24 / Chapter 2.2 --- Principle of Methods and Procedures --- p.26 / Chapter 2.2.1 --- Sample preparation and extraction --- p.26 / Chapter 2.2.2 --- Chemical assays for in vitro antioxidative properties of mushroom extracts --- p.28 / Chapter 2.2.2.1 --- ABTS + scavenging activity --- p.28 / Chapter 2.2.2.2 --- Hydroxyl radical scavenging activity --- p.30 / Chapter 2.2.2.3 --- Hydrogen peroxide scavenging activity --- p.32 / Chapter 2.2.3 --- Total phenolic content --- p.34 / Chapter 2.2.4 --- Cytotoxicity of hydrogen peroxide --- p.36 / Chapter 2.2.5 --- Cytoprotectivity of mushroom extracts --- p.36 / Chapter 2.2.6 --- "Colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay" --- p.37 / Chapter 2.2.7 --- Lactate dehydrogenase (LDH) assay --- p.39 / Chapter 2.2.8 --- Total cellular protein loss --- p.40 / Chapter 2.2.9 --- Comet assay (Single cell gel electrophresis assay) --- p.41 / Chapter 2.2.10 --- Thiobarbituric Acid Reactive Substances (TBARS) assay ..… --- p.44 / Chapter 2.2.11 --- Preparation of cell lysate for evaluating cellular antioxidant defense system --- p.45 / Chapter 2.2.12 --- Total Glutathione level --- p.46 / Chapter 2.2.13 --- Enzyme activity --- p.49 / Chapter 2.2.13.1 --- Catalase (CAT) --- p.49 / Chapter 2.2.13.2 --- Glutathione peroxidases (GPx) --- p.51 / Chapter 2.2.13.3 --- Glutathione Reductase (GR) --- p.53 / Chapter 2.2.13.4 --- Superoxide dismutase (SOD) --- p.54 / Chapter 2.2.14 --- Determination of protein --- p.56 / Chapter 2.2.15 --- Statistical analysis --- p.56 / Chapter Chapter 3 --- Results and discussions --- p.57 / Chapter 3.1 --- Extraction yield --- p.57 / Chapter 3.2 --- Chemical assays for in vitro antioxidative properties of mushroom extracts --- p.60 / Chapter 3.2.1 --- ABTS + scavenging activity --- p.60 / Chapter 3.2.2 --- Hydroxyl radicals scavenging activity --- p.61 / Chapter 3.2.3 --- Hydrogen peroxide scavenging activity --- p.64 / Chapter 3.3 --- Total phenolic content --- p.67 / Chapter 3.4 --- Cytotoxicity of hydrogen peroxide --- p.69 / Chapter 3.4.1 --- "Colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay" --- p.71 / Chapter 3.4.2 --- Lactate dehydrogenase (LDH) assay --- p.72 / Chapter 3.4.3 --- Total cellular protein loss --- p.73 / Chapter 3.4.4 --- Residual hydrogen peroxide level --- p.76 / Chapter 3.4.5 --- Lipid peroxidation --- p.77 / Chapter 3.4.6 --- DNA damage --- p.79 / Chapter 3.5 --- Cytotoxicity of extracts --- p.85 / Chapter 3.6 --- Protection of H2()2-induced oxidative damage in HDFa cells --- p.88 / Chapter 3.6.1 --- Protective effect of mushroom water extracts --- p.88 / Chapter 3.6.2 --- Protective effect of CfAa on H2()2-incluced damage to HDFa --- p.93 / Chapter 3.6.3 --- Protective effect of CfAa on DNA damage in HDFa cells --- p.96 / Chapter 3.7 --- Modulation of cellular antioxidant defense system by CfAa --- p.99 / Chapter 3.7.1 --- Intracellular total glutathione --- p.100 / Chapter 3.7.2 --- Enzyme activities --- p.102 / Chapter 3.8 --- Speculation on the possible components in CfAa --- p.108 / Chapter Chapter 4 --- Conclusion and further works --- p.109 / References --- p.111