The Best PhD Thesis in the Faculties of Dentistry, Engineering, Medicine and Science (University of Hong Kong), Li Ka Shing Prize,2008-2009 / published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

Thesis (MTech (Biomedical Technology))--Cape Peninsula University of Technology, 2006 / Cells living under aerobic conditions constantly face the oxygen paradox i.e. oxygen is indispensable for supporting life; however, its metabolites such as reactive oxygen species (ROS) can modify cell function. Oxidative stress (OS) arises as a consequence of excessive ROS production and/or impaired antioxidant defence mechanisms. Environmental and physiological factors have been implicated in poor sperm function. Excessive ROS generation results in oxidative damage and consequently decreased sperm function. The objectives of this study are threefold: (i) To measure the production of O 2 -. by sperm by means of flow cytometry. (ii) To determine effects of O2 -. on sperm motility and viability in the presence or absence of commercially available scavengers and RPO. (iii) To determine effects of O2 -. on selective apoptotic markers in ejaculated sperm in the presence of absence of commercially available scavengers and RPO. In the first part of the study, suitable solvents were investigated in order to introduce RPO (because of its hydrophobic nature) as a possible scavenger of ROS in human spermatozoa. Secondly, the O2 -. donor; 2, 3-dimetoxy-1-naphthoquinone (DMNQ) (2.5~M-100~M, 60 min.) was added to normozoospermic post swim-up samples in the absence or presence of Mn(lIl)TMPyP (50~M) or SOD (501U) or RPO at 0.1% and 0.5%. CASA was used to analyse motility parameters, while FACS was used to determine viability (PI, 1mM, 15 min.) and O2 -. levels (DHE, 30~M, 15min.).

Poon, Chui Wa Christina. / "August 2011." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 123-131). / Abstracts in English and Chinese. / ABSTRACT --- p.i / 論文摘要 --- p.vi / ACKNOWLEDGEMENTS --- p.ix / PUBLICATIONS --- p.x / Abstracts --- p.x / ABBREVIATIONS --- p.xii / Chapter 1. --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1. --- Diabetes --- p.1 / Chapter 1.1.1. --- Overview --- p.1 / Chapter 1.1.2. --- Diagnostic Criteria of Type-2 Diabetes --- p.2 / Chapter 1.1.3. --- Type-2 Diabetes (T2DM) --- p.3 / Chapter 1.1.3.1. --- Impaired Insulin Synthesis and Insulin Secretory Defects in Type-2 Diabetes --- p.3 / Chapter 1.1.3.2. --- β-Cell Dysfunction --- p.5 / Chapter 1.1.3.3. --- Insulin Resistance --- p.5 / Chapter 1.1.4. --- Glucose Toxicity --- p.6 / Chapter 1.1.4.1. --- Fasting Hyperglycemia --- p.8 / Chapter 1.1.4.2. --- Postprandial Hyperglycemia --- p.8 / Chapter 1.2. --- Oxidative Stress --- p.8 / Chapter 1.2.1. --- ROS and Mitochondria --- p.8 / Chapter 1.2.2. --- ROS Production by Mitochondria --- p.9 / Chapter 1.2.3. --- The Relationship of Glucose Recognition by β-cells and Oxidative Stress --- p.11 / Chapter 1.2.4. --- Important Roles of Glutathione in Pancreatic β-cells and Glutathione Synthesis --- p.14 / Chapter 1.2.5. --- N-acetyl-L-cysteine - A Potential Drug Treatment for Type-2 Diabetes? --- p.17 / Chapter 1.3. --- Role of F-actin Cytoskeleton on Glucose-induced Insulin Secretion --- p.18 / Chapter 1.4. --- Current Clinical Treatments for Type-2 Diabetes Mellitus --- p.21 / Chapter 1.4.1. --- Metformin --- p.22 / Chapter 1.4.2. --- Sulfonylureas --- p.22 / Chapter 1.4.3. --- Thiazolidinediones --- p.23 / Chapter 1.4.4. --- Glinides (Meglitinide Analogues) --- p.23 / Chapter 1.4.5. --- α-Glucosidase (AG) Inhibitors --- p.24 / Chapter 1.4.6. --- Dipeptidyl Peptidase-4 (DPP-4) Inhibitors --- p.24 / Chapter 1.4.7. --- (Clinical) Antioxidant Treatment --- p.24 / Chapter 1.5. --- Animal Models Used in Type-2 Diabetes Research --- p.25 / Chapter 1.6. --- Aims of Study --- p.27 / Chapter 2. --- RESEARCH DESIGN & METHODS --- p.28 / Chapter 2.1. --- Materials --- p.28 / Table 1. Sources and concentrations of drugs tested in this study: --- p.28 / Culture Medium - --- p.29 / General Reagents --- p.29 / Chapter 2.2. --- Isolation of Islets of Langerhans and Single Pancreatic β-Cells --- p.31 / Chapter 2.3. --- Measurement of Mitochondrial ROS Levels --- p.32 / Chapter 2.4. --- Measurement of Islets Insulin Release and Insulin Content --- p.34 / Chapter 2.4.1. --- Preparation of Samples --- p.34 / Chapter 2.4.2. --- Enzyme-Link Immunosorbent Assay (ELISA) --- p.35 / Chapter 2.5. --- Immunocytochemistry --- p.35 / Chapter 2.6. --- Data and Statistical Analysis --- p.37 / Chapter 3. --- RESULTS --- p.38 / Chapter 3.1. --- "Effects of L-NAC, Various Oxidative Stress Inducers/Reducers and Actin Polymerisation/Depolymerisation Inducers on Releasable Insulin Levels and Insulin Contents in Response to Low Glucose (5 mM) and High Glucose (15 mM) of Isolated Pancreatic Islets of (db+/m+) and (db+/db+) Mice" --- p.38 / Chapter 3.1.1. --- Effect of L-NAC on Insulin Secretion and Insulin Contents --- p.38 / Chapter 3.1.2. --- Effect of Cytochalasin B on Insulin Secretion and Insulin Contents --- p.39 / Chapter 3.1.3. --- Effect of 4-Phenyl Butyric Acid on Insulin Secretion and Insulin Contents --- p.43 / Chapter 3.1.4. --- Effect of Ursodeoxycholic Acid on Insulin Secretion and Insulin Contents --- p.46 / Chapter 3.1.5. --- Effect of Hydrogen Peroxide on Insulin Secretion and Insulin Contents --- p.49 / Chapter 3.1.6. --- Effect of Jasplakinolide on Insulin Secretion and Insulin Contents --- p.53 / Chapter 3.1.7. --- Effect of Thapsigargin on Insulin Secretion and Insulin Contents --- p.57 / Chapter 3.1.8. --- Effect of BSO on Insulin Secretion and Insulin Contents --- p.61 / Chapter 3.2. --- "Effects of L-NAC, Various Oxidative Stress Inducers/Reducers and Actin Polymerisation/Depolymerisation Inducers on Mitochondrial ROS Levels in Response to High Glucose (15 mM) Challenge in Isolated Single Pancreatic β-Cells of (db +/m+) and (db +/db +) Mice" --- p.65 / Chapter 3.2.1. --- "Effects of L-NAC (20 mM), 4-Phenyl Butyric Acid (4-PBA) (1 mM), Ursodeoxycholic Acid (UA) (500 μg/ml), H202 (200 μM), Thapsigargin (0.5 μM) and DL-Buthionine-[S,R]-Sulfoximine (BSO) (0.1 μM) Pre-treatments on Mitochondrial ROS Level in Response to High Glucose (15 mM) Challenge" --- p.65 / Chapter 3.2.2. --- "Effects of L-NAC (20 mM), Cytochalasin B (10 μM) and Jasplakinolide (5 μM) Pre-treatments on Mitochondrial ROS Level in Response to High Glucose (15 mM) Challenge_" --- p.76 / Chapter 3.3. --- "Effects of L-NAC, Various Oxidative Stress Inducers/Reducers and Actin Polymerisation/Depolymerisation Inducers on F-actin Cytoskeleton Levels Incubated in Low Glucose (5 mM) and High Glucose (15 mM) Medium in Single Pancreatic β-Cells of Non-Diabetic (db +/m+) and Diabetic (db +/db +) Mice" --- p.81 / Chapter 4. --- DISCUSSION --- p.100 / Chapter 4.1. --- General Discussion --- p.100 / Chapter 5. --- SUMMARY --- p.120 / Chapter 6. --- FUTURE PERSPECTIVES --- p.121 / Chapter 7. --- REFERENCES --- p.123

Chan Chi Chung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 100-109). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / List of Abbreviations --- p.v / List of Tables --- p.vi / List of Figures --- p.vii / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- "Free radicals, oxidative stress and antioxidants" --- p.2 / Chapter 1.1.1 --- Free radical and ROS --- p.2 / Chapter 1.1.2 --- Oxidative stress --- p.6 / Chapter 1.1.3 --- Antioxidants --- p.8 / Chapter 1.2 --- Plant as a source of antioxidants --- p.13 / Chapter 1.2.1 --- Common food sources of antioxidants --- p.13 / Chapter 1.2.2 --- Legume seeds as antioxidant sources --- p.15 / Chapter 1.3 --- Methods used to evaluate the antioxidant activity --- p.16 / Chapter 1.3.1 --- β-carotene bleaching method --- p.17 / Chapter 1.3.2 --- DPPH. scavenging method --- p.17 / Chapter 1.3.3 --- High-performance liquid chromatograph (HPLC) --- p.18 / Chapter 1.3.4 --- Single cell gel electrophoresis (SCGE) --- p.20 / Chapter 1.4 --- Objectives of the study --- p.22 / Chapter 2 --- Materials and Methods --- p.23 / Chapter 2.1 --- Plant materials and chemicals --- p.23 / Chapter 2.2 --- Sample preparation --- p.23 / Chapter 2.3 --- Determination of antioxidant activity using β-carotene bleaching method --- p.24 / Chapter 2.4 --- Evaluation of free radical scavenging ability --- p.26 / Chapter 2.5 --- HPLC separation of seed extract --- p.27 / Chapter 2.6 --- Evaluation of protective effects of legumes on the DNA damage using the comet assay --- p.28 / Chapter 2.6.1 --- Preparation of reagents --- p.28 / Chapter 2.6.2 --- Blood sample --- p.29 / Chapter 2.6.3 --- Hydrogen peroxide (H2O2) treatment --- p.29 / Chapter 2.6.4 --- Ethidium bromide-acridine orange (Et-Ac) viability determination --- p.31 / Chapter 2.6.5 --- Slide preparation --- p.31 / Chapter 2.6.6 --- Alkaline comet assay --- p.31 / Chapter 2.6.7 --- Quantification of DNA damage --- p.33 / Chapter 2.6.8 --- Statistical analysis --- p.33 / Chapter 3 --- Results --- p.40 / Chapter 3.1 --- General description of 24 selected legume seeds --- p.40 / Chapter 3.1 --- Determination of antioxidant activity using β-carotene bleaching method --- p.40 / Chapter 3.2 --- Evaluation of free radical scavenging ability --- p.43 / Chapter 3.3 --- Evaluation of protective effects of legumes on the DNA damage using the comet assay --- p.45 / Chapter 3.3.1 --- Optimal assay conditions --- p.46 / Chapter 3.3.2 --- Protective effects of seed extract and vitamin C --- p.47 / Chapter 3.3.3 --- Effects of heat treatment on the protective effect --- p.48 / Chapter 4 --- Discussion --- p.84 / Chapter 4.1 --- Methanolic extraction --- p.84 / Chapter 4.2 --- Antioxidant activities determined with β-carotene bleaching method and DPPH' scavenging method --- p.84 / Chapter 4.3 --- Evaluation of protective effects of legumes on the DNA damage using the comet assay --- p.93 / Chapter 4.3.1 --- H202-mediated DNA damage --- p.93 / Chapter 4.3.2 --- Protective effects of seed extracts and vitamin C --- p.94 / Chapter 5 --- Conclusion --- p.98 / References --- p.100

In Africa, non-communicable diseases such as diabetes mellitus have been generally neglected. This problem has worsened over the years owing to continuous threats from infectious diseases such as HIV/AIDS, tuberculosis and malaria. Despite this, statistics have shown that by 2030, the African region will have the highest proportional increase in diabetes prevalence. Over 80% of all diabetic deaths occur in developing countries probably not only due to poor equity of access to medication but also due to limited efficacy and side effects associated with the commonly available anti-diabetic agents. Therefore, this creates the desperate need for the development of new anti-diabetic agents that are more efficacious and can be sourced from within the continent. With oxidative stress as a suggested mechanism underlying the cause of diabetes mellitus and diabetic complications, the discovery of natural anti-oxidants that prevent free radical mediated damage is important for developing new treatment strategies. Marine algae have been identified as good sources for natural anti-oxidants. Unfortunately, very few studies have embarked on the discovery of marine-derived anti-oxidant compounds with potential anti-diabetic activity. In this project, we investigated the potential anti-oxidant activity of the South African endemic algae Stypopodium multipartitum, Dictyopterus ligulata, Cystophora fibriosa, Bifurcariopsis capensis, Sargassum sp. and Sargassum heterophyllum. From these studies, Sargassum heterophyllum yielded prenylated compounds, the main compound being sargahydroquinoic acid (3.6) and the carotenoid metabolite fucoxanthin (3.8), which are in part responsible for the radical scavenging activity of the crude extract. Sargahydroquinoic acid (3.6) and fucoxanthin (3.8) also exhibited significant anti-inflammatory activity. Sargaquinoic acid (3.1), sargachromenoic acid (3.9) and sarganaphthoquinoic acid (3.10) were then semi-synthesized from sargahydroquinoic acid (3.6) and their in-vitro cytotoxicity profiles evaluated using Chang Liver, HT-29, Caco-2 and 3T3-L1 cell lines prior to antidiabetic testing. From the semi-synthetic derivatives, sargachromenoic acid (3.9) exhibited the most potent anti-oxidant activity (IC₅₀ = 6.99 μg/mL). After the evaluation of antidiabetic activity using 3T3-L1 preadipocyte differentiation, sarganaphthoquinoic acid (3.10) showed the most potent insulinomimetic activity at 1.19 μM by inducing a PPARγ response similar to that of rosiglitazone at 1 μM.

Thesis (M.Sc. (Microbiology)) -- University of Limpopo, 2015 / Antioxidants play an important role in living organisms to control level of free radicals and other reactive molecules in the body to reduce oxidative damage. Synthetic antioxidant compounds are used in food industries as food additives to boost our immune systems. These compounds are associated with a number of critical side effects including liver damage and carcinogenesis. Scientists are also concerned about microorganisms that have developed resistant genes against current antibiotics used in hospitals. The aim of the study was to isolate and characterize bioactive compounds from Ricinus communis leaves with activity against Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853). Consequently, medicinal plants are studied and considered for their efficacy and safety, because they possess bioactive compounds with various biological activities. Leaves of R. communis were collected at the University of Limpopo, Turfloop campus in Limpopo province, South Africa. The leaves were dried and milled to a fine powder. A number of trial extraction methods were employed using various solvents of different polarities on a fine powder leaves to identify the best extraction method. Plant extracts were analyzed by thin layer chromatography (TLC) developed in four mobile phases. To detect separated phytochemical compounds, TLC plates were sprayed with vanillin- sulphuric acid in methanol and heated at 110oC for optimal colour development. Qualitative antioxidant activity was determined by using 2, 2–diphenyl-1-picrylhydrazyl (DPPH) assay on TLC plates. Quantitative antioxidant activity was determined by measuring percentages scavenging activity of DPPH and 2, 2’-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) free radical molecules by plant extracts. Antibacterial activity of all extracts was quantified by a serial microbroth dilution method while bioautography was used in qualitative analysis of the active compounds. Cytotoxicity effect of R. communis extracts was evaluated using tetrazolium-based calorimetric assay on human Caucasian skin fibroblast (Bud-8) cell line. Anti-inflammatory activity was assessed using phagoburst kit on Raw 264.7 macrophages cell line. Pure compounds were subjected to nuclear magnetic resonance spectroscopy for 1H, 13C and DEPT experiments to elucidate structures of compounds. 2 During extraction process, methanol was the best extractant, extracting greater amount of extracts than any of the other solvents. Serial exhaustive extraction method was selected as the best extraction method for extracting compounds from ground plant materials. In quantitative antioxidant assays, chloroform and methanol extracts had highest percentage scavenging activity against DPPH free radicals compared to other extracts and vitamin C. Methanol extract had the highest percentage scavenging activity of ABTS free radicals and minimum percentage scavenging activity was in hexane extract. Acetone, ethyl acetate and ethanol extracts showed strong antioxidant activity against DPPH free radicals in qualitative antioxidant assay on TLC plates. In quantitative antibacterial assay, crude extracts showed lowest minimum inhibitory concentration value of 0.13 mg/ml against all tested organisms and the highest was 1.05 mg/ml. Hexane extracts revealed potent antibacterial activity against all tested microorganisms on bioautograms. Hexane and acetone extracts also revealed anti-inflammatory activity and have ability to reduce oxidative stress. In cytotoxicity effect of plant extracts, Methanol extracts had lethal concentration for 50% of the cells (Lc50) of 784 μg/ml on Human Caucasian skin fibroblast (Bud-8) cell line while hexane extracts had Lc50 of 629 μg/ml. Plant extracts with high Lc50 are low toxic to normal cell line and preferable to work with for drug development. Bioassay-guided fractionations results in successful isolation of three antioxidant and two antibacterial compounds from R. communis using column chromatography. Isolated compounds were tested for their biological activities using qualitative DPPH assay on TLC plates for antioxidant activity and bioautography for antibacterial activity. Antioxidant compounds showed strong antioxidant activity after spraying with DPPH in methanol and antibacterial compounds showed less activity compared to the crude extracts. The study suggests the use of crude extracts to fight against pathogenic microorganisms compared to pure compounds. Compound 4 was successful identified as the mixture of stigmasterol and β-sitosterol. The present study recommends the use of R. communis leaves as the potential source of antioxidant, antibacterial and anti-inflammatory compounds. The study serves as a scientific proof for use of this plant in traditional medicine for treatment of various ailments.

This study investigated potential apoptotic and anti-proliferative effects of the phytochemicals, genistein and anthocyanin extract, as single and combined treatments in MCF-7 human breast cancer cells. Cells were exposed to single and combined treatments with the phytochemiclas for 48 and 72 hours. Cell viability was assessed using the MTT bioassay. Apoptosis induction was assessed using acridine orange ethidium bromide and rhodamine 123 ethidium bromide fluorescence assays. Both singe and combination treatments induced dose- and time-dependent apoptotic cell death in MCF-7 cells. The percentage of apoptosis was higher in combination treatments than single treatments with either phytochemical, although the difference was not statistically significant. The combination of genistein and anthocyanin extract peaked in efficacy at 48 hours of treatment, to exhibit significantly greater (P<. O5) dose- and time-dependent cell cytotoxicity than single treatments. This study reveals potential chemopreventive implications for the complementary effects of genistein and anthocyanin extract. / by Corine M. Stinson. / Thesis (M.S.)--Florida Atlantic University, 2011. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2011. Mode of access: World Wide Web.

by Zhang Ze Sheng. / "October 2001." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (p. 157-174). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Thesis (M. Tech. (Biomedical Technology)) -- Central University of Technology, 2014 / The rising costs of health care, the outbreak of drug resistant organisms, health depleting lifestyles and the risky side effects of currently used drugs are world-wide problems. This has led to the search for novel drugs and drug leads. Traditional healers and other individuals across the globe possess unlimited knowledge on the healing powers of different plants that has been passed on through generations. This knowledge together with scientific investigations can lead to the eradication of most of the diseases either by treatment or prevention. Aloe ferox Mill. is one of the plants that have gained a lot of interest from the pharmaceutical industry. The plant has over 80 documented medicinal uses. These include treatment of impotence and infertility, sexually transmitted infections, arthritis, hypertension, leukaemia, bacterial and fungal infections. It is also known as a blood purifier, widely used as a laxative and anti- inflammatory agent. More research is required to discover more about A. ferox and its benefits to health as well as to investigate its potential for the development of novel drugs. The current study was focused at investigating the anti- cancer, anti- microbial antidiabetic, cytotoxic activities and phytochemical composition of leaf extracts of A. ferox. Three cancer cell lines namely: breast (MCF7), colon (HCT116) and prostate (PC3) cancer cell lines were used to investigate the anticancer activity of the extracts using the Sulforhodamine B (SRB) method. To determine the anti- diabetic activity of the plant extracts the C2C12 and Chang cell in- vitro models of glucose uptake were used. The micro- dilution technique was IV used to evaluate the antibacterial and antifungal activity of the extract. The safety of these extracts against normal human foetal lung fibroblasts (W138), Chang and C2C12 cells was done by through the SRB and the MTT methods. To determine the phytochemical profile of A. ferox the DPPH radical scavenging and the Folin Ciocalteu methods were used to test the antioxidant activity and the total phenolic content of the different extracts respectively. Different methods were used to determine the presence of phytochemicals such as steroids, saponins, alkaloids, carbohydrates and flavonoids. LCMS was also done to detect the elemental composition of the plant extracts. According to the CSIR criteria A. ferox was inactive against the cancer cell lines used. It however exhibited antioxidant activity even at low concentrations, with an EC50 of 0.865 ± 0.783. The methanol extract showed more phenolic content than the dichloromethane and aqueous extracts at a concentration of 5mg/ml. It is believed that the antioxidant activity correlates with the phenolic content and quality of the phenols present in the plant and more assays have to be done to prove this hypothesis. Other phytochemicals found in the extract included saponins, steroids, alkaloids as well as flavonoids. Both the methanol and aqueous extracts of A. ferox caused a significant increase in glucose uptake by C2C12 cells but caused a slightly decreased uptake by the Chang cells. The plant extracts inhibited the growth of Staphylococcus epidermidis, Streptococcus pneumonia, Escherichia coli and Candida albicans at a concentration of 15mg/ml extract. Candida tropicalis and Escherichia faecalis were resistant to A. ferox extracts. Finally the extracts showed no toxic activity against the normal foetal lung fibroblasts, Chang and C2C12 cells validating the safety of this plant for human use. V The results in conjunction with literature findings show A. ferox to be a promising source of drugs and therapeutic agents. Due to the fact that traditional healers already rely on it as treatment for different ailments, it is important that the safety of the plant for use has been validated though other studies and clinical trial still need to be done to fully confirm this. All the information gathered also showed this plant to be of great benefit against major health problems, responsible for millions of deaths each year such as cancer, cardiovascular and inflammatory diseases, and diabetes. There is however still a great need for more investigation to be done on this plant against a vast majority of organisms and diseases so as to fully benefit from it.

Wang Deqing. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (p. 236-277). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.