Dans les trois anciens pays indochinois, le Vietnam, le Cambodge et la RDP Lao, la médecine traditionnelle rencontre un succès grandissant auprès de la population. Conscients de l’importance de la médecine traditionnelle, les trois gouvernements l’ont reconnue et intégrée, chacun différemment, dans leur système de soins de santé national. L’encadrement juridique de cette médecine millénaire diffère d’un pays à l’autre. La législation de la médecine traditionnelle du Cambodge demeure particulièrement faible. De plus, de nombreux problèmes persistent et menacent la préservation et le développement de cet héritage précieux. Avec l’aide de partenaires internationaux, les trois gouvernements aséaniens tentent de renforcer l’encadrement juridique de leurs médecines et de leurs pharmacopées traditionnelles / In the three former Indochinese countries, Vietnam, Cambodia and PDR Lao, traditional medicine success is growing among the population. Recognizing the importance of traditional medicine, the three governments have recognized and integrated it in a different way into the national health care system. The legal framework of this millennial medicine differs from one country to another. Traditional medicine legislation in Cambodia remains particularly weak. Moreover, many problems persist and threaten the preservation and development of this precious heritage. With the help of international partners, the three ASEAN governments are trying to strengthen the legal framework for their traditional medicines and pharmacopoeias.

African Potato (Hypoxis hemerocallidea), (AP) is an African traditional medicine (TM) that is commonly used for various nutritional/medicinal purposes and also by people infected with the human immuno deficiency virus HIV and AIDS patients as an immune booster. The use of AP has also been recommended by the former Minister of Health of South Africa for use by HIV positive people. The main phytochemical component of AP is a norlignan glucoside, hypoxoside, and other relatively minor components have also been reported. A recent in vitro study reported the effects of AP extracts, hypoxoside and rooperol (the metabolite of hypoxoside) on human metabolic enzymes such as the cytochrome P450 (CYP450) group of enzymes and also on the transporter protein, p-glycoprotein (P-gp). This research focussed on investigating the clinical significance of those in vitro effects on the pharmacokinetics of efavirenz (EFV) in humans. EFV was chosen as the substrate drug because it is in first-line regimen of treatment of HIV/AIDS in South Africa, and also has been reported to be a substrate for the specific CYP isozymes, 3A4 and 2B6, in common with APs metabolic involvement with 3A4. A high performance liquid chromatography method with ultra-violet detection (HPLC-UV) for the quantitative determination of EFV in plasma was developed and successfully validated according to international standards with good reproducibility, accuracy, recovery, linear response and requisite sensitivity. The preparation of the plasma samples for analysis was effected by using a simple and rapid precipitation method, and the mobile phase consisted of readily available solvents. EFV in plasma samples was found to be stable under the relevant storage conditions studied. The oral dose of AP, administered as a freshly prepared traditional decoction, was standardised based on the hypoxoside content, and the quality of all the AP decoctions was analysed immediately prior to administration, using a validated HPLC-UV method. A single dose, two-phase sequential study was conducted over a period of 31 days in 10 healthy volunteers. The clinical study was approved by the Rhodes University Ethical Standards Committee, and all the participants agreed to the conditions of the study by giving their informed consent. On day 1 of the study, human subjects were administered a 600 mg EFV tablet and blood samples were collected before dosing and at various intervals over a period of 48 hr post dosing. From day 16, a traditionally prepared AP decoction was administered daily at a standardized dose of 15 mg/kg/day per subject until day 30. On day 29, volunteers were administered a single 600 mg dose of EFV as was done on day 1. Plasma samples were harvested immediately after blood sample collection and frozen at -80 ºC until assayed. Geometric mean ratios of relevant pharmacokinetic parameters, Cmax (maximum plasma concentration achieved following dosing) and AUC0-48 (area under the curve of a plot of drug plasma concentrations versus time representing the extent of absorption) of EFV before and after co-administration of 14 successive daily doses of AP were compared and evaluated to determine whether an interaction had occurred. All subjects completed the study and the geometric mean ratios of Cmax and AUC0-48 were 97.30 and 102.82 with corresponding 90% confidence intervals (CIs) of 78.81-120.14% and 89.04-118.80%, respectively. Whereas the acceptance criteria for the ratios of the AUCs fell within the preset 90% CIs indicating no interaction, the Cmax ratios fell outside the limits. Although the protocol was developed in accordance with the United States of America Food & Drug Administration’s Guidance for Drug Interactions, a priori stating that both criteria need to fall within the acceptance limits to indicate no interaction, an argument is presented to waive the Cmax requirement for the declaration of an interaction. As a result, the pharmacokinetic data generated during this study indicated that the effect of AP on the pharmacokinetics of EFV is not clinically significant. Hence, co-administration of AP is unlikely to affect the clinical use of EFV. In summary the objectives of this project were: 1. To develop and validate a suitable HPLC-UV method for the quantitative determination of EFV in plasma. 2. To perform a mini-validation of the determination of hypoxoside for use as a marker in the quality control and standardisation of AP decoctions. 3. To conduct a clinical interaction study in order to determine whether AP affects the pharmacokinetics of EFV following concurrent administration. 4. To apply the validated HPLC-UV method to determine plasma concentrations of EFV in plasma of human subjects. 5. To use appropriate statistical methods and treatments such as a non-compartmental pharmacokinetic analysis to determine the occurrence of an interaction.

Magister Scientiae - MSc / This investigation focused on Rhoicissus tomentosa, belonging to the family, Vitaceae in an attempt to assess the phytochemistry of this plant which is widely used by traditional healers in South Africa to ensure the safe delivery during pregnancy and childbirth (Hutchings et al., 1996). / South Africa

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

Collaborations between researchers and African traditional healers on medicinal plants need to go beyond the flow of information from African traditional healers to researchers. Mutual beneficiation wherein African traditional healers reap rewards due to the information they possess is necessary and has been legislated in South Africa. The manner in which such beneficiation occurs and how it will be distributed should be subject to negotiation between (a) the researchers and holders of indigenous knowledge and (b) among holders of indigenous knowledge themselves. Beneficiation can be in the form of access to information, monetary or through shares in commercialised products, amongst others. African traditional healers involved in the collaboration with researchers of the Department of Biochemistry and Microbiology and the Department of Pharmacy at the Nelson Mandela Metropolitan University beneficiated by having access to knowledge and medicinal plants cultivated in the medicinal garden. Beneficiation was expected to extend to monetary gains from the sale of sewn garments and plants from a medicinal plants nursery which was to be developed and from a herbal topical product which was also to be developed. The aim of this research project was to develop a wound healing herbal cream consisting of Bulbine frutescens and Carpobrotus edulis as active ingredients. B. frutescens demonstrated better wound closure properties when compared to C. edulis and the combined extracts. The scratch assay wound was completely closed after 24 hours at B. frutescens concentrations of 5 μg/ml and 10 μg/ml. B. frutescens was more toxic to 3t3-L1 cells than C. edulis, but it was less toxic than the combined extracts. None of the extracts were toxic to Vero cells and the combined extracts significantly promoted their proliferation. Antibacterial activity of all the extracts was low. C. edulis showed antiviral activity against human herpes simplex virus 1 at 62.5, 125 and 250 μg/ml, while the combined extracts were active at 250 μg/ml. The combined extracts exhibited synergistic antioxidant activity. A reverse phase, gradient, high performance liquid chromatography method was developed and validated and used to conduct quality control tests on the extracts and the finished product. It was concluded that the use of whole chromatographic data instead of common peaks data is best for analysis of medicinal plants. The cream that was developed used buffered cream as the base and was stable at 25 °C/65 percent RH for one month with regard to organoleptic and rheological properties and microbial preservation.

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.

Infection with Helicobacter pylori has been associated with the pathogenesis of numerous stomach and gastroduodenal diseases that pose threats to public health. Eradicaftion of this pathogen is a global challenge due to its alarming rate of multidrug resistance. Consequently, to find an alternative treatment, the search is increasingly focused on new antimicrobial product from natural sources including honey. Honey has been used as medicine in several cultures since ancient time due to its enormous biomedical activities. Its beneficial qualities have been endorsed to its antimicrobial, antioxidant, anti-inflammatory properties added to its phytocomponents. In this study, the anti-H. pylori activity of South African honeys and their solvent extracts as well as the phytochemicals present in the two most active honeys were evaluated. Agar well diffusion test was used to investigate the antimicrobial activity of six honey varieties obtained from different locations in the country. Subsequently, the honeys were extracted with four organic solvents viz n-hexane, diethyl ether, chloroform and ethyl acetate employed in order of increasing polarity. The antibacterial activity of the different solvent extracts of each honey was evaluated by agar well diffusion; broth micro dilution and time kill assays. Different chromatographic techniques (Thin layer & column chromatography) were employed to enumerate the phytochemical constituents in the most active solvent extracts of Pure Honey (PH) and Champagne Royal Train (CRT); and were identified by gas-chromatography linked mass-spectrometry. Linalool pure compound was equally evaluated for anti-H. pylori activity in a bid to trace the antibacterial agent among the variety of compounds identified. Data were analyzed by One-way ANOVA test at 95% confidence interval. Crude honeys and their solvent extracts demonstrated potent anti-H. pylori activity with zone diameter that ranged from [16.0mm (crude) to 22.2mm (extract)] and percentage susceptibilities of test isolates between 73.3% (crude) and 93.3% (extract). The chloroform extracts of PH and CRT were most active with MIC50 in the ranges 0.01- viii 10%v/v and 0.625-10%v/v respectively, not significantly different from amoxicillin (P> 0.05); and efficient bactericidal activity (100% bacterial cells killed) at 1/2MIC and 4xMIC over different time intervals, 36-72hrs and 18-72hrs respectively. The appearance of bands on the thin layer chromatography (TLC) chromatogram spotted with the chloroform extracts of PH and CRT; and developed with hexane: ethyl acetate: acetic acid (HEA) and methanol: acetic acid: water (MAAW) solvent systems indicated the presence of compounds. Purification of the compounds contained in these extracts over silica gel column yielded numerous fractions which were evaluated for antibacterial activity and purity. PHF5 was the most active fraction with a mean MIC50 value of 1.25mg/mL. Volatile compounds belonging to different known chemical families in honey were identified in all the active fractions obtained from PH. Conversely, only four compounds were identified in the active fractions obtained from CRT hence the non volatile constituents could be of prime relevance with respect to antibacterial activity of this honey. Of novelty was the presence of thiophene and N-methyl-D3-azirdine compounds, essential precursors used for the synthesis of natural products and pharmaceuticals with vital biomedical properties. Linalool demonstrated potent inhibitory (MIC95, 0.002- 0.0313mg/mL) and bactericidal activity (0.0039-0.313mg/mL) against the test isolates. On the other hand, a significant difference was recorded (P < 0.05) in comparing the activity of linalool compound to the fractions. PH could serve as a good economic source of bioactive compounds which could be employed as template for the synthesis of novel anti-H. pylori drugs. However, further studies are needed to determine the non volatile active ingredients in PH and CRT as well as toxicological testing

The aim of the study was to examine the procedures followed by traditional healers treating people living with HIV and AIDS in the Instika Municipality and the challenges faced by them in this endeavour. Using the qualitative research method, in-depth interviews and focus group discussions were conducted with twenty traditional healers. Among the significant findings of this study were the hardships encountered by traditional healers in terms of finance, the transport needed to collect medicinal plants, the shortage of medicinal plants, the lack of co-operation from the formal health care sector and the discrimination and abuse suffered at the hands of members of the community, with Christians and members of the SAPS being among the chief antagonists. It is also acknowledged in this study that traditional healers have been trained by the Department of Health concerning issues of hygiene and that traditional healers have knowledge of the symptoms of HIV and AIDS. It can therefore be concluded that traditional healers have a significant role to play in preventing the spread of HIV and AIDS if they employ preventative measures such as the use of protective gloves and limiting the use of a blade to one patient only. However, traditional healers are not supplied with safety kits or condoms to distribute to patients who consult them for treatment of STIs.

Herbal medicines (HMs) have been used to supplement, maintain, and treat health conditions, and have inspired the development of many Western pharmaceuticals. Migrant South Asian (SA) communities in the UK have brought with them their own traditional forms of medicine, yet little is known about their current use of HMs in the UK. Consuming HMs alongside conventional Western medicines could affect pharmacological treatment and lead to herb-drug interactions; hence, healthcare professionals (HCPs) should be aware of their patients’ use of HMs. The import of HMs to the UK raises concerns over the quality, safety and regulation of HMs. Deoxyribonucleic acid (DNA) barcoding can be used to discriminate between different species, and identify contaminants and adulterants, thus can be used for the authentication of HMs. The South Asian Traditional Medicines (SATMED) questionnaire explored the knowledge and use of HMs by diasporic SA communities in the UK. It uncovered a vast range of HMs which were used by participants, where ingredients were sourced from, the concurrent use of herbal and Western medicines, and how minor ailments were treated. An online survey designed to investigate UK based practitioners’ views of HMs revealed that HCPs claimed to lack sufficient knowledge of HMs. HCPs said they needed more training on HMs to help them make better informed decisions. Tulsi (Ocimum tenuiflorum L.) was identified as a culturally and commercially valuable plant, which was used for molecular analysis. A variety of tulsi samples were collected for authentication: community samples from SA families in the UK, commercial samples, and referenced specimens. Both ITS and trnH-psbA regions were successfully used to distinguish between several Ocimum species, and identify a potential species substitution. This research represents the first time that DNA based methods have been used to authenticate medicinal plants species used by migrant SA communities living in the UK. The results of this multi-disciplinary study provide a unique contribution to the evolving discipline of ethnopharmacology.

Seventeen Cree anti-diabetic herbal medicines and eight Traditional Chinese Medicines have been examined for their potential to cause interactions with drugs, which is considered as a major reason for adverse drug effects. Specifically, the effect of these natural health products was examined on major Phase I drug metabolism enzymes including cytochrome P450, human carboxylesterase-1 and flavin-containing monooxygenases. Several of these natural health products have the potential to cause adverse drug effect through the inhibition of major drug metabolism enzymes. The results indicated that 7 Cree medicines plant extracts inhibited CYP3A4 activity, and 3 of them have been proven to cause potent mechanism-based inactivation of CYP3A4. Seven of eight Traditional Chinese Medicines have been identified as strong CYP3A4 inhibitors; the ethanol extract of Goji has identified as a potent inhibitor for CYP2C9 and 2C19. Goji juice showed universal inhibitory effects on most of the tested enzymes except flavin-containing monooxygenases 3.