Dissertation submitted in partial compliance with the requirements for the Master's Degree in Technology: Homoeopathy at Technikon Natal, 1999. / The purpose of this placebo-controlled study was to determine the morbid symptomcomplex, in healthy people, produced by the administration of Sceletium tortuosum (a South African herb) in the 6 CH potency, so that it may be prescribed according to the Law of Similars, as required by homoeopathic principles. / M

Thesis submitted to the Faculty of Science under the school of Molecular and Cell Biology, University of the Witwatersrand, in partial fulfilment of the requirements for PhD. Gauteng, Johannesburg, 2015 / Cancer is an enormous burden of disease, accounting for millions of deaths annually worldwide. Today, more people are dying from cancer than HIV/AIDS, malaria and tuberculosis combined. According to the American Cancer Society, it is expected that the global cancer burden will double by 2030 if preventative measures are not applied. Breast and gynaecological cancers remains a big scourge in developing countries, with breast cancer being the most common cancer and gynaecological cancers accounting for approximately 25% of all cancers in women in developing countries. Currently, the standard cancer treatments include surgery, radiation and chemotherapy. Adverse toxicities have been associated with these therapies and their effectiveness is also limited to drug resistance. The cost of treatment is another major burden. Limitations associated with these conventional cancer treatments have made discoveries of novel therapeutics which exhibit less toxicity and at a lowered cost of paramount importance. Medicinal plant extracts have recently attracted attention to modern medical science research with their non-lethal activity. Currently, up to 50% of the world drugs including chemotherapeutic drugs such as taxol and camptothecin are made from natural products or their derivatives. In this study we aimed to investigate the anti-cancer properties of the medicinal plant Euphorbia tirucalli. The crude extracts of E. tirucalli extracted using butanol; hexane and methanol solvents were screened for antiproliferative activity in breast (MCF-7 and MDA-MB231), ovarian (RMG-1) and cervical (SiHa) cancer cell lines. MTT assay and Real-Time Cell Analyzer (RTCA), xCELLigence were used to determine cytotoxicity of the extracts and calculate IC50. From MTT and xCELLigence results, we observed that E. tirucalli extracts exhibited dose-dependent inhibition of cell proliferation with RMG-1 and MCF-7 cells being more sensitive than MDA-MB231 and SiHa cells to all three extracts for an unclear reason. The butanol extract appeared to exhibit iv the most cytotoxicity with all cell lines reaching IC50 at low extract concentrations. Most therapies in anticancer treatment, such as chemotherapy, mainly induce cell death by causing either G0/G1 or G2/M cell cycle arrest and then inducing an apoptotic pathway. Therefore, cell cycle arrest and the induction of apoptosis in cancer cells become the major indicators of anticancer effects. Cells were stained with propidium iodide dye to determine if cells were arrested at G0/G1 or G2/M cell cycle stages while annexin V and PI staining were used to determine the type of cell death induced by the extracts. Cell cycle analyses revealed MCF-7, MDA-MB231 and SiHa cancer cells underwent arrest at G0/G1 following treatment with the plant extracts. Annexin V and PI staining revealed different proportions of apoptotic and necrotic populations. The extracts mainly induced apoptosis on MCF-7 and MDA-MB 231 cells, with the butanolic extracts inducing the most apoptosis. RMG-1 and SiHa cells had a high proportion of cells undergoing both late apoptosis and necrosis. The molecular mechanism of cell death induction was investigated using real time PCR and western blot. From the gene expression studies, p21 was observed to be over expressed in all cells following all treatments, in line with the observed cell cycle arrest at G0/G1. The extrinsic pathway of apoptosis was identified as the type of cell death induced with caspase 8 being overexpressed in MDA-MB 231 cells treated with butanol and hexane extracts. Upon further fractionation, flavonoids and especially isorhamnetin were identified as the active compounds in these extracts. Overall, the plant contains compounds that have some activity against cell proliferation and can be a promising tool to treat cancer cells. However, more work needs to be done to verify which compounds are mainly involved.

A thesis submitted in compliance with the requirements for the Masters Degree in Technology: Mechanical Engineering at Technikon Natal, 2002. / The experiments and their results contained herein will form the basis for the development of a portable non-destructive testing device for composite structures. This device is to be capable of detecting any of a variety of defects and assessing their severity within a short time / M

In this research we aimed to investigate the anti-proliferative properties of three South African plants: Kedrostis foetidissima, Euphorbia mauritanica and Elytropappus rhinocerotis against breast cancer cells. This was done on the basis of their documented ethno-medicinal use against cancer and other ailments. The plant extracts were screened for cytotoxicity and pro-apoptotic activity against two breast cancer cell lines MCF-7 and YMB-1. With an IC50 ~ 100 μg/ml, K. foetidissima was the only extract that exhibited significant cytotoxicity on both cell lines, whilst E. mauritanica was cytotoxic to MCF-7 cells only. The cytotoxicity assay was followed by the Annexin-V detection assay to evaluate the occurrence of apoptosis. The results observed suggested that K. foetidissima was inducing significant apoptosis on both YMB-1 and MCF-7 cells, whilst E. mauritanica was inducing significant apoptosis on MCF-7 cells. Since both K. foetidissima and E. mauritanica crude extracts induced apoptosis to MCF-7 cells, they were selected for gene expression studies on MCF-7 using real-time PCR. This was done with the aim of investigating if these extracts were having an effect on the tumour suppressors p53 and RBBP6, which were shown in previous studies to be deregulated in up to 50% of cancers. From the real-time PCR data we observed no changes in the expression levels of these genes following treatment with the herbal extracts. This may suggest that these plants have an effect on other components of the apoptotic pathway other than the tumour suppressors p53 and RBBP6. The antiproliferative activity observed whilst treating these particular cell lines with K. foetidissima and E. mauritanica suggests that these South African herbal plants present themselves as potential future cancer therapeutic agents; however, further studies on these herbal plants need to be performed to validate these results. KEYWORDS: Apoptosis Breast cancer Euphorbia mauritanica Kedrostis foetidissima p53

M. Tech. Entrepreneurship / The purpose of this study is to develop a consumer profile of traditional Chinese medicine consumption in South Africa. The research was based upon a nomethetic survey, a total sample size of 321 respondents was selected according to the convenience sampling technique to participate in the research. Descriptive data analysis was performed indicating that behaviour control and social impact norms primarily influencing the purchase of traditional Chinese medicine.

Herbal formulations claimed to cure several medical conditions including skin eruptions, chest pains, wounds, gout, menstrual pains, stress, nervous disorders, microbial and viral infections as well as stomach ailments have recently appeared as part of South African traditional medicine. The formulations consist of mixtures of extracts of different plant parts from several different plant species packaged in labelled bottles or boxes. The mixtures are available for sale in herbal shops and public places. While there has been widespread use of these herbal mixtures, there has been no scientific evidence to support their use. This project was aimed at documenting, validating claimed efficacy and testing the safety of fourteen unregistered commercial herbal preparations commonly sold in Pietermaritzburg. A detailed investigation of the pharmacological effects and safety of the plant components of one of the mixtures, Imbiza ephuzwato was also carried out. / Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2009.

Please read the abstract in the section 07chapter7 of this document / Thesis (DPhil (Plant Physiology))--University of Pretoria, 2007. / Plant Science / unrestricted

Over the past 25 years, there has been a resurgence of worldwide scientific research in the fields of ethnopharmacology. The Western world has acknowledged the continued use of traditional medicines by the majority of third world countries, and the need for novel drug development. Hence, much of the pharmaceutical research in recent years has focused on the ethnobotanical approach to drug discovery (Light et al., 2005). In South Africa, as in most developing parts of the world, traditional herbal medicine still forms the backbone of rural healthcare. The government health services in South Africa provide only western medical care although the majority of the population consult traditional healers for some or all of their healthcare needs (McGaw et al., 2005). Medicinal plants like Harpephyllum caffrum are used as blood purifiers or emetics (Watt and Breyer-Brandwijk, 1962), and also for treating acne and eczema. The antimicrobial activity of this plant can be used to treat septicaemia, which is ranked the sixth leading cause of death among neonates and the eighth leading cause of death for infants through the first year of life (Heron, 2007). In this study, the plants investigated for antimicrobial activity were Harpephyllum caffrum, Hermannia cuneifolia, Chironia baccifera, Rhigozum obovatum, Felicia muricata and Pentzia incana. These plants were tested against ATTC (American Type Culture Collection) strains and microorganisms isolated from clinical isolates of patients suffering from septicaemia. The assay methods used included the agar diffusion method using the Mast multipoint inoculator, the microtitre dilution method were used to determine the minimum inhibitory concentration, thin layer chromatography fingerprints accompanied by bioautographic assay were used to detect the inhibition of bacterial growth by active compounds separated from plant extracts and the Ames test was required to assess the possibility of bacterial mutagenesis upon the exposure to plant extracts which can lead to carcinogenicity. In agar diffusion method, extracts of Harpephyllum caffrum inhibited nine strains of Candida albicans, three species of Acinetobacter and four strains of E.faecalis. Extracts of Hermannia cuneifolia inhibited four strains of B.cereus and three strains of Staphylococcus aureus. Extracts of Chironia baccifera inhibited one strain of Acinetobacter and five strains of E.faecalis. Extracts of plants Rhigozum obovatum, Felicia muricata, and Pentzia incana showed no antimicrobial activity. In the microtitre dilution method used to determine the minimum inhibitory concentration (MIC), the results were different from the agar diffusion method. More activity was observed. Extracts of Harpephyllum caffrum inhibited three strains of E.coli, six strains of S.aureus, three species of Acinetobacter and one strain of Klebsiella pneumonia. Extracts of Hermannia cuneifolia inhibited four strains of B.cereus, three strains of S.aureus, two strains of K.oxytoca and one species of Acinetobacter. Extracts of Chironia baccifera inhibited three strains of S.aureus, one strain of MRSA, one species of Acinetobacter and one strain of S.haemolyticus. The MIC values ranged from 0.049 to 6.25mg/ml. Using the thin layer chromatography fingerprints, bioautography showed the presence of various inhibitory chemical compounds. Methanol and acetone extracts of Harpephyllum caffrum, separated very well and showed various inhibition zones on exposure to Candida albicans, Enterococcus faecalis and Staphylococcus aureus. The different inhibition zones were recorded as Rf In the Ames test (Maron and Ames, 1983) the methanol and acetone extracts of Harpephyllum caffrum and Hermannia cuneifolia were negative which means they were devoid of any mutagenic properties. Methanol extracts of Harpephyllum caffrum showed similar results in the Ames assay as reported by Verschaeve and Van Staden (2008). values ranging from 0.25 to 0.95. The zones indicate the different inhibiting chemical compounds present in the plant. Petroleum ether, ethyl acetate, chloroform and formic acid were the solvents used in the assay in the ratio 8:7:5:1, respectively. Establishing the antimicrobial activity of these plants contribute to the systematic scientific investigation of indigenous South African medicinal plants.

M.Sc. (Botany) / A few poorly studied Cape herbal medicinal plants of historic and potential commercial interest which form part of the traditional medicine system of the Cape Region have been selected for this study. These are Centella glabrata L., Olea europaea L. subsp. africana (Mill.), Rafnia amplexicaulis (L.) Thunb. and Tulbaghia alliacea L.f. Centella C. asiatica is mainly used to treat skin conditions and wounds, tuberculosis and coughs and cooked and eaten as a vegetable in South Africa. The treatment of heartburn in the Wild Coast is a recent record. Decoctions of the leaves and stalks of C. glabrata were used for treating diarrhea and dysentery by the early Cape settlers. C. stenophylla or “klipdagga” is used in the treatment of high blood pressure and diabetes in the Still Bay area. This is also a recent new record. The anatomy of the few Centella species studied differed dramatically both in the leaf lamina and the petiole and could be used to differentiate between the species studied. A wider study would be very interesting and could be useful to distinguish between the species. This could have great potential as the ca. 45 species of Centella are often very difficult to distinguish from one another.

Herbal remedies are commonly used in developing countries for the treatment of various diseases, including constipation. The rationale for utilizing medicinal plants for the treatment of diseases rested largely on the belief that they are safe and free of side effects. However, there is limited scientific evidence on the safety and efficacy of these herbal medicines to back up their continued therapeutic application. Aloe ferox Mill. (Aspodelaceae), known as Cape aloe, locally called ikhala is a medicinal plant used by the people of the Eastern Cape Province for the treatment of gastrointestinal problems and constipation. The plant is a perennial shrub with thick succulent leaves bearing brown thorns on the margin and bright orange flowers arranged in oval lanceolate. It occurs in all weather in bush veld, road side, gardens and undisturbed places. According to the ethnomedicinal information, A. ferox is used as purgative. This research project was therefore designed to evaluate its ability in the treatment of constipation and to investigate its possible toxicological property. At the beginning of this programme, a survey of plants used for the treatment of constipation in Nkonkobe Municipality of the Eastern Cape Province was carried out using a questionnaire, which was administered to herbalists, traditional healers and rural dwellers. The study revealed 10 plant species from 8 families that are used for the treatment of constipation in the province. Four plants, Aloe ferox Mill, Boophone distischa L.f Herb, Alepidea amatybica Eckl and Artemisia afra Jacq, were repeatedly mentioned. Based on the frequency of usage, perceived efficacy and availability to the rural dwellers and the traditional healers, Aloe ferox was the most commonly used of the plants for the treatment of constipation. The plant was thus, chosen for the study. The invivo laxative effect of the aqueous extract of Aloe ferox in the treatment of loperamide-induced constipation in Wistar rats was investigated at varying concentrations. The leaf extract at all the dosages investigated (50, 100 and 200 mg/kg body weight) improved intestinal motility, increased fecal volume and normalized body weight in the constipated rats. This was an indication of its laxative properties. However, the laxative property of the herb at 200 mg/kg body weight of the extract showed best efficacy and compares favourably well with senokot, a standard laxative drug. These findings have therefore, lent scientific credence to the folkloric use of the herb by the people of the Eastern Cape of South Africa as a laxative agent. Toxicological evaluation of aqueous leaf extract of Aloe ferox in loperamide-induced constipation was studied at 50, 100, and 200 mg/kg body weight. The oral administration of the extracts did not show any significant effect on the liver and kidney body weight ratios as well as the kidney and liver function indices. The extracts, at all the dosages investigated, did not alter the levels of creatinine, uric acid, urea, calcium and potassium ions. Similarly, the levels of total protein, albumin, bilirubin and gamma glutamyl transferase (GGT) were not significantly different from the control. The plant extract appreciably normalized the elevated activities of alkaline phosphatase (ALP), alanine transaminase (ALT) and aspartate transaminase (AST) in the untreated constipated rats following treatment with the extract. The extract did not show a significant effect on the hematological parameters except for the increase in the lymphocyte count in the untreated constipated rats, which was attenuated after administering the herb. ThThe available evidence in this study suggests that A. ferox may be safe as an oral remedy for constipation. Generally, the effect of the extract compared favourably well with senokot, a recommended drug for the treatment of constipation. The antioxidant activities against 1, 1 diphenyl- 2 picrylhydrazl (DPPH), 2,2’ – azinobis [3- ethylbenzothiazoline -6- sulfonic acid] diammonium salt (ABTS), hydrogen peroxide (H2O2), Nitric oxide (NO), lipid peroxidation and the ferric reducing agents were investigated spectrophotometrically. Alkaloids, saponins, tannins, total phenols, flavonoids, flavonols and proanthocyanidin were also determined to assess their effects on the antioxidants activity of this plant. The phytochemical content of the ethanol and acetone extracts were consistently high compared to other solvents extracts. The level of tannins was not significant (P > 0.05) as compared with other solvent extracts. The free radical scavenging activity of the extracts was high even at lower concentrations (0.025 mg/ml) except in DPPH and lipid peroxidation. The ferric reducing potential of the extracts was concentration dependent and significantly different from Vitamin C and butylated hydroxytoluene (BHT) that were used as standard drugs. The present study showed a high level of scavenging activity of the leaf extracts of Aloe ferox in all the solvent extracts. Both ethanol and methanolic extract showed potent antioxidant activities than acetone and aqueous extracts. The study indicated that the leaf extracts of Aloe ferox might be a valuable source of natural antioxidant for both medicine and food industries. A. ferox leaf consists of the gel, latex and mesophyll layer; however, the main active constituents of the latex and the leaf exudate of Aloe ferox are anthraquinones which are believed to be responsible for the laxative property. The laxative compound in Aloe ferox leaf extract was isolated and characterized by extracting the plant material in methanol and extract suspended in distilled water. Partitioning was done with n-hexane, ethyl acetate and butanol respectively and was co-spotted with the over-the-counter (OTC) laxative drugs. This led to the successive column chromatography and thin layer chromatography (TLC) of the most active ethyl acetate fraction on silica gel with benzene/ethanol/ammonia hydroxide (BEA: 90:10:1), ethyl acetate/methanol/water (EMW: 40:5.4:5) and chloroform/ethyl acetate/formic acid (CEF: 50:40:10) as the mobile phase. The successive chromatograph and TLC afforded two compounds of Rf 0.420 (blue) and 0.831 (yellow) with the over-the-counter (OTC) drugs. These compounds were not totally elucidated due to their small quantity and instability. However, hydroxyl (OH) and carboxyl groups (COOH) was established as common to the extracted compounds, which might be responsible for the biological activity recorded for the plant extract.