The African potato is used as an African traditional medicine for its nutritional and medicinal properties. Most research has been carried out on H. hemerocallidea, with very little or nothing on other Hypoxis spp. The main aim of this project was to provide scientific data on the anticancer, anti-inflammatory and antioxidant properties of H. hemerocallidea, H. stellipilis and H. sobolifera chloroform extracts and their active compounds. The hypoxoside and phytosterol contents of the three Hypoxis spp. were determined using TLC, HPLC and GC. H. hemerocallidea and H. sobolifera chloroform extracts contained the highest amounts of hypoxoside and β-sitosterol, respectively. For the anticancer properties, cytotoxicity of the Hypoxis extracts and its purified compounds were determined against the HeLa, HT-29 and MCF-7 cancer cell lines (using MTT), and PBMCs (using CellTiter-Blue®). H. sobolifera had the best cytotoxicity against the three cancer cell lines, whereas H. stellipilis stimulated HeLa and HT-29 cancer cell growth. IC50 values of hypoxoside and rooperol were determined. DNA cell cycle arrest (using PI staining) occurred in the late G1/early S (confirmed by increased p21Waf1/Cip1 expression) and G2/M phases after 15 and 48 hrs, respectively, when treated with Hypoxis extracts and rooperol. H. sobolifera and rooperol activated caspase-3 and -7 (using fluorescently labelled antibodies) in HeLa and HT-29 cancer cells, and caspase-7 in MCF-7 cancer cells after 48 hrs. Annexin V binding to phosphatidylserines in rooperol treated U937 cells confirmed early apoptosis after 15 hrs. The TUNEL assay showed DNA fragmentation in the three cancer cell lines when treated with H. sobolifera and rooperol for 48 hrs. A shift pass the G2/M phase has led to the investigation of endoreduplication, which was confirmed by cell/nucleus size, and anti-apoptotic proteins (Akt, phospho-Akt, phospho-Bcl-2 and p21Waf1/Cip1). U937 cell differentiation to monocyte-macrophages was optimized using PMA and 1,25(OH)2D3, which was confirmed by morphological and biochemical changes. For the anti-inflammatory properties, Hypoxis extracts and rooperol significantly increased NO production in monocyte-macrophages (pre-loaded with DAF-2 DA) and phagocytosis of pHrodoTM E. coli BioParticles®. The treatments had no effect on COX-2 expression in monocyte-macrophages. The phytosterols significantly increased IL-1β and IL-6 secretion xv (using the FlowCytomix Multiplex human Th1/Th2 10plex Kit I) in the PBMCs of one donor. For the antioxidant properties, Hypoxis extracts and rooperol significantly increased ROS production in undifferentiated and differentiated U937 cells, which were pre-loaded with DCFH-DA. Hypoxis extracts and purified compounds had ferric reducing activities, but only rooperol had ferric reducing activities significantly greater than ascorbic acid. β-sitosterol, campesterol and cholesterol significantly increased SOD activity in Chang liver cells, while H. stellipilis, H. sobolifera and rooperol decreased SOD activity. Anticancer, anti-inflammatory and antioxidant properties of the Hypoxis extracts may be attributed to the β-sitosterol content, because Hypoxis chloroform extracts contained very little or no hypoxoside. Unidentified compounds, and synergistic and additive effects of the compounds may have contributed to the biological effects. This study confirms previous reports that rooperol is the active compound. Results provide scientific data on the medicinal properties of one of the most frequently used medicinal plants in South Africa.

Cyclic dipeptides (CDPs) are amino acid-based compounds, some of which possess antibacterial activity. The encapsulation of certain drugs into liposomes has been found to improve their activity in terms of bioavailability and duration of action. Liposomes are small vesicles that are under investigation as drug carriers for the delivery of therapeutic agents. A number of liposome formulations are currently under clinical trial review, whilst some have already been approved for clinical use. The aim of this study was to optimize a liposomal cyclo(Tyr-Pro) formulation and to assess its antibacterial activity against various Gram-positive and Gram-negative bacteria. Response surface methodology (RSM) using the central composite design (CCD) model was used to optimize liposomal formulations of cyclo(Tyr-Pro) for each of the four bacteria, namely Bacillus cereus, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Percent drug encapsulated and bacterial inhibition were investigated with respect to two independent variables, i.e. lipid composition and cholesterol content. Design Expert 8 was used for the purpose of finding the combination of independent variables that would yield an optimal formulation for each bacterium. The model selected by the software failed to adequately correlate the predicted models to the experimental data. The in vitro experiments showed that the antibacterial activity of liposome-encapsulated cyclo(Tyr-Pro) was superior to that of its free counterpart. Binding maximum or Bmax for the encapsulated compound at concentrations as low as 0.412 mg/ml, was significantly higher than that obtained for free cyclo(Tyr-Pro) which was tested at a concentration of 20 mg/ml. Furthermore, encapsulation of cyclo(Tyr-Pro) into a liposome formulation enhanced its potency. This was evident in the lower IC50 values for the liposomal compound when compared to free cyclo(Tyr-Pro).

During the 1800’s cannabis use was described as a treatment for a variety of metabolic disorders but its recreational use in the twentieth century resulted in laws which made the usage of cannabis illegal despite its medicinal properties. Cannabis usage has been reported to be useful in the treatment of Type 2 diabetes but unfortunately conflicting results are often published and its mechanism of action is still unknown. The aim of this project was to investigate the effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells, to unravel their mechanism of action and also to test for potential anti-diabetic properties. The studies showed that phytocannabinoid treatment promoted higher glucose uptake and significantly less fat accumulation when compared to Rosiglitazone. Rosiglitazone is an anti-diabetic drug that has recently been withdrawn from the market since its usage has been associated with severe side effects. It was also found that during the 1800’s cannabis use was described as a treatment for a variety of metabolic disorders but its recreational use in the twentieth century resulted in laws which made the usage of cannabis illegal despite its medicinal properties. Cannabis usage has been reported to be useful in the treatment of Type 2 diabetes but unfortunately conflicting results are often published and its mechanism of action is still unknown. The aim of this project was to investigate the effect of phytocannabinoid treatment on adipogenesis and lipolysis in 3T3-L1 cells, to unravel their mechanism of action and also to test for potential anti-diabetic properties. The studies showed that phytocannabinoid treatment promoted higher glucose uptake and significantly less fat accumulation when compared to Rosiglitazone. Rosiglitazone is an anti-diabetic drug that has recently been withdrawn from the market since its usage has been associated with severe side effects. It was also found that phytocannabinoid treatment was able to reverse the insulin-resistant state of 3T3-L1 cells. The study indicates that the mechanism of action occurs at the mitochondrial level where enzymes such as succinate dehydrogenase and glycerol-3-phosphate dehydrogenase are modulated thereby affecting oxidative phosphorylation involved in the respiratory chain. In addition the effect observed with phytocannabinoid treatment is time dependent and affects the cells differently at different developmental stages. Therefore it can be concluded that phytocannabinoid treatment not only helps to maintain the balance between adipogenesis and lipolysis in 3T3-L1 cells but its use may also be helpful in the treatment of Type 2 diabetes and/or obesity-related insulin resistance.phytocannabinoid treatment was able to reverse the insulin-resistant state of 3T3-L1 cells. The study indicates that the mechanism of action occurs at the mitochondrial level where enzymes such as succinate dehydrogenase and glycerol-3-phosphate dehydrogenase are modulated thereby affecting oxidative phosphorylation involved in the respiratory chain. In addition the effect observed with phytocannabinoid treatment is time dependent and affects the cells differently at different developmental stages. Therefore it can be concluded that phytocannabinoid treatment not only helps to maintain the balance between adipogenesis and lipolysis in 3T3-L1 cells but its use may also be helpful in the treatment of Type 2 diabetes and/or obesity-related insulin resistance.

Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.

Two studies were conducted to determine whether controlled, abdominally-predominant breathing could be accurately implemented during periods of acute anxiety by speech anxious/phobic individuals, and what effect breathing control has on autonomic and subjective indices of anxiety. Twenty-two moderately speech anxious young adults took part in Study 1. The results of this study indicated that after two weeks of training, only 50% of trainees were able to implement the controlled breathing technique with any degree of accuracy while waiting to deliver an impromptu speech before a small audience. No one were successful at reliably implementing the technique during the speech itself. As in previous research, training had little impact on autonomic arousal but was associated with improvements in self-reported anxiety. Similar findings emerged for Study 2, which differed from Study 1 in that it involved a larger (N = 48) and more highly speech anxious sample who participated in a longer (4-week), more intensive training program. Although training had little effect on subjective or autonomic arousal during speech anticipation and speech delivery, it did result in significantly higher predictions of speech aptitude and emotional control relative to no treatment. Such findings suggest that breathing control is not a useful emotion-focused coping strategy on its own, but may add to the effectiveness of exposure-based therapies by enhancing patients' self-efficacy and willingness to expose themselves to feared situations. / Arts, Faculty of / Psychology, Department of / Graduate

The purpose of this study was to understand the meaning of change through therapeutic enactment in psychodrama. Existential and hermeneutic phenomenology conducted from the perspective of a dialectic between storied narrative and thematic analysis was used to investigate the essential meaning of the experience. Eight co-researchers who had experienced significant change through therapeutic enactment in psychodrama were interviewed in depth. Transcripts from these interviews were transposed into narrative form in order to straighten the story of change through enactment in a before, during, and after sequence. These eight individual narratives were validated by the co-researchers. An independent reviewer checked each narrative against the original transcript, video tapes of the enactments, and comments of each co-researcher for trustworthiness. Each validated narrative provided a rich description of the lived experience of change through therapeutic enactment. In addition, fifty-nine (59) essential themes were formulated from the individual narratives: Fourteen (14) in the planning stage, twenty-four (24) in the enactive stage, and twenty-one (21) in the reflective, or integrative stage, of the enactment process. These themes were then woven into a common story representing the pattern and meaning of change through therapeutic enactment for this group of co-researchers. Finally, notations made during the transposing of the transcripts into personal narratives, formulation of the essential themes, and construction of the common story were used to develop a theoretical story of change through therapeutic enactment, as a final level of hermeneutic interpretation. This theoretical story was then presented in summary form as a thematic sequence of multi-modal change processes representing a model of change through therapeutic enactment. The results of this study suggested numerous theoretical and technical implications. Foremost among theoretical implications was the suggestion that Tomkins (1992) script theory of affect may best illuminate the effects and processes of psychodrama and enactment. This study also had implications for interactional theories of development, contemporary psychoanalytic theories of interpersonal functioning, theories of moral development, theories of dream functioning, and ethological theories of myth and ritual. The results of this study also suggested a number of additional qualitative and comparative outcome studies for future research. / Education, Faculty of / Educational and Counselling Psychology, and Special Education (ECPS), Department of / Graduate

M.Tech. / It is estimated that 59% of South African adult women and 29% of South African adult men are overweight (Department of Health, 2004). Significant risks arise from being overweight including: elevated cholesterol and the development of cardiovascular disease which increases with a greater gain in weight (Duyff, 2006). There is a lack of sufficient evidence supporting the safety and efficacy of many of the herbal weight-loss products currently available thus indicating that more research on herbal products and their efficacy in weight-loss is required (Lenz and Hamilton, 2004). The aim of this study is to determine the efficacy of a herbal complex (Aloe ferox, Fucus vesiculosis, Taraxacum officinale and Trigonella foenum– graecum) as an aid in weight loss in females utilising comparative measurements of the participants’ weight, Body Mass Index (BMI), body fat percentage and circumferential measurements of their hips, waist, thighs, upper arms and abdomen. The study was a quantitative, double blind placebo controlled study. The study involved thirty overweight female participants (BMI 25.5 - 30 kg/m²) between the ages of twenty and thirty five. The participants were recruited by means of advertisement posters placed at the University of Johannesburg, Homoeopathy Health Centre. The participants were randomly divided into two groups of fifteen. One group received the herbal complex (Aloe ferox, Fucus vesiculosis, Taraxacum officinale and Trigonella foenum– graecum) and the other group received the placebo. Participants from both groups attended an initial interview where they were screened by means of a questionnaire and physical examination, including the measurement of their height and weight, calculation of their Body Mass Index (BMI) and body fat percentage, as well as the circumferential measurement of their hips, waist, thighs, upper arms and abdomen. Each participant was given a weekly diary and instructed to take fifteen drops of the issued medication three times daily, after meals for the duration of the full eight week study. Participants were examined, weighed, and the measurement of their body circumference and fat percentage were recorded every second week for the duration of the eight week study. Data from each participant was collected and analysed using repeated measures analysis of variance (ANOVA). From statistical evaluation, it was determined that the herbal complex (Aloe ferox, Fucus vesiculosis, Taraxacum officinale and Trigonella foenum– graecum) was ineffective as an aid in weight loss in females.

No description available.

Human gut harbors 100 trillion microbial organisms that is intrinsically linked to individual’s health and diseases, including cancer. Food fiber and phytochemicals such as polyphenols are considered as prebiotic-like dietary modifiers. They can influence the gut microbial communities, and in turn to modulate disease outcome and drug responses of the host. Saponins belong to a family of phytochemicals commonly found in many medicinal and edible plants. Herbal saponins have raised keen interest among scientists for their health-promoting effects, but have not been investigated for their potential as prebiotics. Gynostemma pentaphyllum (Gp) is riched in triterpenoid saponins and has been consumed in China and other part of the world as an herbal tea and as a folk medicine. In our lab, we have demonstrated that Gp possesses strong anticancer and anti-inflammatory effects. Whether Gp possesses prebiotic property and whether gut microbiota plays any part of the anticancer effect of Gp are the questions addressed in the present study. Thus, we hypothesized that Gp saponins (GpS) might modulate the gut microbiota, which in turn enhance its anticancer activities. In the study, the gut microbiome analysis were carried out using two main techniques, neamly the enterobacterial repetitive intergenic consensus (ERIC-PCR) and 16S pyrosequencing approaches. Both xenograft nude mice and Apcmin/+ mice were employed as the animal models to investigate the interaction between the herbal saponins and the gut microbiota in the host. Athymic nude mice have been employed for tumorigenic research for decades, however, the relationships between the gut microbiome and host’s response to the grafted tumors and drug treatments are unexplored. For the first part of the thesis, we investigated the relationship between the gut microbiota and grafted tumor in the nude mice under the treatment of Gp saponins. Partial least squared discriminant analysis (PLS-DA) of ERIC-PCR data showed that the microbiota profile of xenograft nude mice departed from that of the nonxenograft mice. However, prolonged treatment of GpS seems to realign the fecal microbiota with the pretreatment control. Pyrosequencing data reiterated the differences in fecal microbiome between the nonxenograft and xenograft animals. GpS treatment had a much stronger impact on the phylotypes of the xenograft than the nonxenograft mice. In addition, GpS treatment markedly induced the relative abundance of Clostridium cocleatum and Bacteroides acidifaciens, for which the beneficial effects on the host have been well documented. ApcMin/+ colorectal cancer mouse model was further employed for the investigation of the association of the gut microbiota and cancer occurred inside the gut, which was a more direct site to interact with the gut microbiota. In the ApcMin/+ mouse model, we found distinct difference of fecal microbiome between the ApcMin/+ and the wild-type littermates. GpS treatment significantly reduced the number of intestinal polyps. GpS also increased the ratio of Bacteroidetes/Firmicutes and reduced the sulfate- and sulfur-reducing bacteria lineage and potential opportunistic pathogens, which might cause certain deleterious effects to the host. The impact of GpS on the gut mucosal environment was also examined. We found GpS treatment improved the gut barrier function by increasing the numbers of Paneth cells, goblet cells, up-regulating the expression of E-cadherin and down-regulating the expression of N-cadherin in the intestine. In addition, GpS treatment down-regulated the protein expression of beta-catenin and p-STAT3. Furthermore, higher levels of anti-inflammatory and tissue repair-related cytokines as well as Arginase I, but lower level of iNOS expression were found in GpS-treated ApcMin/+ mice, indicating increased anti-inflammatory macrophage phenotype M2 (associated with tissue repair) and reduced proinflammatory phenotype M1. Furthermore, in addition to GpS, other herbal saponins also showed prebiotic-like effects in C57BL/6 mice. In summary, this study provides first hand evidence for the impact of herbal saponins on the gut microbial ecosystem and new insight into mechanisms responsible, at least in part, for the activities of GpS. We demonstrate that tumor growth induce intestinal dysbiosis. GpS treatment can inhibit tumor progression and concurrently alter the microbiome by increasing symbionts and/or decreasing pathobionts, which may contribute to its chemopreventive effect against tumorigenesis. Herbal saponins showing prebiotic-like effects may be used for improving the health of the host by manipulation of the gut microbiota.

No description available.