Thesis (M.S.)--University of Hawaii at Manoa, 2005. / Includes bibliographical references (leaves 106-109). / xxi, 109 leaves, bound ill. (some col.) 29 cm

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2001. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 5 uppsatser.

M.Tech. / Haemostasis is an internal mechanism to stop bleeding from a damaged blood vessel. Conceptually this process occurs in a number of essential steps following tissue injury. Although the herbal preparation of Arnica montana has been well documented for its tendency to prolong bleeding, according to the Law of Similars, homeopathically prepared Arnica montana 6C is well indicated for traumatic injuries and post surgical bruising. Arnica montana 6C can be used when there is mechanical trauma that causes wounds, haemorrhages, haematomas, sore-bruised bone and muscular pains, inflammations, fractures, muscular strains and sprains. The remedy is often prescribed before and immediately after surgery to reduce post-operative pain and to speed up recuperation. Three in vitro studies conducted at the Technikon Witwatersrand (now the University of Johannesburg) on various potencies of homeopathically prepared Arnica montana showed lowered overall coagubility of blood, but no significant difference between the experimental and control groups. Bengsch (2000), Hohl (2005), Vermeulen (2000) and van Tonder (2005) recommended that studies on the effect of homeopathically prepared Arnica montana on blood coagulability be repeated in vivo. This study formed part of a three part in vivo study to determine the effect of Arnica montana homeopathic preparations on blood coagulation by measuring the Bleeding Time (BT), activated Partial Thromboplastin Time (aP'TT) and Prothrombin Time (PT). This study investigated the effect of Arnica montana 6C on these measurements. Eighty participants were allocated a participant number and randomised by the research supervisor into four groups of twenty participants. Twenty participants were in the placebo group that was shared by all three studies. Twenty participants were allocated to the experimental group for this study. The study was conducted over a period of two weeks at the University of Johannesburg (UJ) Doomfontein Campus Homeopathy Health Centre. Consenting participants were screened by means of a questionnaire (Appendix D) regarding relevant medical history and other background information. A case history was taken and a physical examination was performed. Any prospective participants that were diagnosed with and/or suffer from hypertension, hypotension, heart disease, a iii bleeding disorder, anaemia, iron or any vitamin deficiency, liver disease, malaria or are currently on aspirin or anticoagulants (Appendix D) were excluded from the study. The bleeding time was measured by a trained medical technologist using a standardised bleeding time technique. Blood samples drawn by a phlebotomist went for coagulation tests comprising of aPTT and PT at the NHLS Main Haematology laboratory of the Johannesburg Hospital. Twenty participants were given a 25mL bottle of Arnica montana 6C in 20% ethanol. Twenty participants received an identical bottle containing only 20% ethanol. All participants were requested to take ten drops twice a day for two weeks. All three coagulation test measurements were performed again at the end of the second week. The BT, PT and aPTT results were analysed by using ordinary descriptive statistics such as mean and standard deviation. Changes over time in blood coagulation were ascertained utilising ANOVA (analysis of variance). The results showed that there is no statistically significant difference between the experimental and control group in BT, aPTT and PT. There was also no statistically significant difference between the first BT, PT and aPTT before medication and the second BT, PT and aPTT after two weeks of medication. The results of the study support the hypothesis that Arnica montana 6C would have no effect on the bleeding or coagulation times in vivo. These results support the view that prescribing the remedy before surgery is not likely to increase the post surgical risk of haemorrhage

Melanoma is the leading cause of skin cancer-related death. The STAT3 (signal transducer and activator of transcription 3) and TLR4 (toll-like receptor 4) signaling pathways have been shown to be activated in melanoma. Activation of each of the two pathways can promote melanoma growth, angiogenesis and metastasis. Suppressing TLR4 signaling or STAT3 signaling has been proposed as an approach for melanoma management although the TLR4/STAT3 pathway has not yet been established in melanoma. Atractylodis Macrocephalae Rhizoma (Baizhu in Chinese), a Qi-tonifying Chinese medicinal herb, is commonly prescribed by Chinese medicine doctors for treating melanoma. Our previous studies demonstrated that atractylenolide II (AT-II), isolated from Atractylodis Macrocephalae Rhizoma, could induce apoptosis, and inhibit proliferation and migration in B16 melanoma cells. However, the antimelanoma properties of AT-II and the underlying molecular mechanisms have not been fully understood. In this study, we further investigated the antimelanoma effects of AT-II in vivo and in vitro, and explored the TLR4/STAT3 signaling-related mechanism of action of AT-II. In conclusion, we established the TLR4/STAT3 pathway in melanoma, which provides novel insight into melanoma pathophysiology. We demonstrated that AT-II exerted antimelanoma effects in vivo and in vitro, and inhibition of TLR4/STAT3 signaling contributed to these effects. These findings advanced our understanding of the antimelanoma properties and the underlying mechanism of action of AT-II, and provided a chemical and pharmacological justification for the clinical application of Atractylodis Macrocephalae Rhizoma in melanoma management. This contribution is significant because it is one step in a continuum of research that is expected to lead to future clinical trials of AT-II as a novel antimelanoma agent. Results showed that AT-II induced apoptosis, and inhibited proliferation, migration and invasion in multiple melanoma cells, and significantly inhibited melanoma growth, angiogenesis and metastasis in mice. AT-II suppressed the activation of STAT3 and Src (a STAT3 upstream tyrosine kinase) in mouse melanoma tissues and inhibited the EGFR/Src/STAT3 signaling in cultured melanoma cells. The free binding energy of AT-II with EGFR (an upstream receptor tyrosine kinase of STAT3) was relatively low in molecular docking assays, suggesting that AT-II might inhibit EGFR activation via other molecules. We found that activation of TLR4 enhanced EGFR/Src/STAT3 signaling in melanoma cells, and activation of the TLR4/STAT3 pathway contributed to melanoma progression in vivo and in vitro. These observations suggested that the TLR4/STAT3 pathway was established in melanoma. Molecular docking showed that AT-II could bind to the TLR4/MD-2 receptor complex. AT-II reduced the binding of LPS (a TLR4 ligand) to TLR4, and inhibited LPS-triggered activation of EGFR/Src/STAT3 signaling as well as LPS or MPLAs (synthetic monophosphoryl lipid A, a TLR4 agonist) induced invasion in melanoma cells. Overexpression of a constitutively active variant of STAT3 (STAT3C) in A375 cells diminished anti-proliferative, apoptotic and anti-invasive effects of AT-II; and overexpression of an active form of TLR4 in A375 cells diminished AT-II-exerted anti-invasive effects in cultured cells, and attenuated the inhibitory effects of AT-II on tumor growth and angiogenesis in mice. These suggested that suppression of TLR4/STAT3 signaling contributed to the antimelanoma effects of AT-II.

M. Tech. / Phototobiomodulation (PBM), also known as low level laser therapy (LLLT) or photobiostimulation, is a non-invasive form of therapy that utilizes low intensity laser light or irradiation to provide healing. However, in order for healing to be successful certain laser parameters need to be taken into consideration i.e. fluence (dosage), wavelength and power density. Laser therapy has been used for various medical applications and fields. Multiple cytokines and growth factors are involved in wound healing including Interleukin (IL)-1, IL-6 and Tumour Necrosis Factor alpha (TNF- a). In diseased state(s) such as diabetes mellitus (DM) or psoriasis, these growth factors or cytokines are either found elevated or decreased depending on various factors and for abnormally prolonged periods. However, inflammatory cytokines are usually elevated. Phototherapy has been reported to accelerate wound healing, attenuate pain and cease inflammation. However, the effect of phototherapy on cytokine modulation has not been explored extensively, especially under various stress mechanisms. Furthermore, the pathway that laser irradiation induces on modulated pro-inflammatory cytokines has not been clearly elucidated as scientists typically report on the up- or down-regulated expression of cytokines. Numerous authors have reported on the efficacy of laser irradiation to enhance the rate of wound healing and proliferation in normal and diabetic cells or tissue; however, literature that has demonstrated the latter on hypoxic insulted cells is inadequate. In this study hypoxic insult was induced as it is one of the factors that usually prolong the healing process in diabetic wounds. Prior to commencing with the main study, a pilot study was done to exclude the effect of osmotic pressure on cells grown in media containing additional glucose, and thus simulating a diabetic model iv in vitro. Mannitol was used as a control since it is not absorbed by the cells. The study involved four groups namely: normal, normal wounded, mannitol wounded and diabetic wounded cells with each group having a non-irradiated control. Mannitol wounded and diabetic wounded cells had a final concentration of 30 mM mannitol and glucose respectively. A wavelength of 636 nm at a fluence of 5 J/cm2 was used on day 1; experiments were repeated four times and all tests were done in duplicate. Cellular responses (Trypan Blue, adenosine triphosphate (ATP) and lactate dehydrogenase (LDH)) and morphological changes were assessed after 1 h incubation post-irradiation in both irradiated and non-irradiated cultures.

M.Tech. (Homoeopathy) / Haemostasis is defined as the arrest of bleeding by formation of a haemostatic plug or clot. The herb Arnica montana interferes with this process thus resulting in increased bleeding. Homoeopathic physicians use Arnica montana in a potentised form for the treatment of post-operative swelling, pain and ecchymosis but little is known on what effect this potentised form of Arnica montana has on blood coagulation and bleeding time. This study forms part of a three part in vivo study to determine the effects of various homoeopathic potencies of Arnica montana on blood coagulation. This was done by measuring the Bleeding Time (BT), activated Partial Thromboplastin Time (aPTT) and the International Normalised Ratio (INR) of Prothrombin Time (PT). The aim of this particular study is to investigate the in vivo effect of Arnica montana 30C on blood coagulation and Bleeding Time. This study is a double blind, placebo controlled study that took place over a period of two weeks. A total sample group for the three part study consisted of eighty healthy participants between the ages of eighteen to thirty five. Consenting participants that met the criteria were randomised into four groups of twenty each. One group for each part of the three part study were the experimental group and one group was allocated to the placebo group that was shared by all three studies. BT was taken as well as blood samples which underwent coagulation tests (aPTT and INR). Twenty participants received Arnica montana 30C in 20% ethanol and twenty participants received an identical bottle containing 20% ethanol. After two weeks another blood sample was taken where all three coagulation test measurements were repeated. The results of the BT, INR and aPTT were analysed using Statkon Statistical Package for Social Sciences. This showed no statistical difference between the experimental or control group with regard to BT, INR and aPTT. The results indicate that Arnica montana 30C appears to have no effect on Bleeding Time..

There has been an upsurge in the use of Complementary and Alternate Medicines (CAMs) in both developed and developing countries. Although herbal medicines have been in use for many centuries, their quality, safety and efficacy are still of major concern. Many countries are in the process of integrating CAMs into conventional health care systems based on the knowledge and use of traditional medicines. The quality control (QC) of herbal products usually presents a formidable analytical challenge in view of the complexity of the constituents in plant material and the commercial non-availability of appropriate qualified reference standards. Sceletium, a genus belonging to the family Aizoaceae, has been reported to contain psychoactive alkaloids, specifically mesembrine, mesembrenone, mesembrenol and some other related alkaloids. Sceletium is marketed as dried plant powder and as phyto-pharmaceutical dosage forms. Sceletium products and plant material marketed through health shops and on the internet are associated with unjustified claims of specific therapeutic efficacy and may be of dubious quality. Validated analytical methods to estimate Sceletium alkaloids have not previously been reported in the scientific literature and the available methods have focused only on qualitative estimation. Furthermore, since appropriate markers were not commercially available for use as reference standards, a primary objective of this study was to isolate relevant compounds, qualify them as reference standards which could be applied to develop appropriate validated qualitative and quantitative analytical methods for fingerprinting and assay of Sceletium plant material and dosage forms. The alkaloidal markers mesembrine, mesembrenone and ∆⁷ mesembrenone were isolated by solvent extraction and chromatography from dried plant material. Mesembranol and epimesembranol were synthesised by hydrogenation of the isolated mesembrine using the catalyst platinum (IV) oxide and then further purified by semi-preparative column chromatography. All compounds were subjected to analysis by ¹H, ¹³C, 2-D nuclear magnetic resonance and liquid chromatography-tandem mass spectroscopy. Mesembrine was converted to hydrochloride crystals and mesembranol was isolated as crystals from the hydrogenation reaction mass. These compounds were analysed and characterised by X-ray crystallography. A relatively simple HPLC method for the separation and quantitative analysis of five relevant alkaloidal components in Sceletium was developed and validated. The method was applied to determine the alkaloids in plant material and dosage forms containing Sceletium. An LCMS method developed during the study provided accurate identification of the five relevant Sceletium alkaloids. The method was applied for the quantitative analysis and QC of Sceletium plant material and its dosage forms. This LCMS method was found to efficiently ionize the relevant alkaloidal markers in order to facilitate their detection, identification and quantification in Sceletium plant material as well as for the assay and QC of dosage forms containing Sceletium. The chemotaxonomy of some Sceletium species and commercially available Sceletium dosage forms were successfully studied by the LCMS method. The HPLC and LCMS methods were also used to monitor the bio-conversion of some of the alkaloids while processing the plant material as per traditional method of fermentation. Additionally a high resolution CZE method was developed for the separation of several Sceletium alkaloids in relatively short analysis times. This analytical method was used successfully to fingerprint the alkaloids and quantify mesembrine in Sceletium and its products. Sceletium species grown under varying conditions at different locations, when analyzed, showed major differences in their composition of alkaloids and an enormous difference was found to exist between the various species with respect to the presence and content of alkaloids. Sceletium and its products marketed through health shops and the internet may thus have problems with respect to the quality and related therapeutic efficacy. The QC of Sceletium presents a formidable challenge as Sceletium plants and products contain a complex mixture of compounds. The work presented herein contributes to a growing body of scientific knowledge to improve the QC standards of herbal medicines and also to provide vital information regarding the selection of plant species and information on the specific alkaloidal constituents to the cultivators of Sceletium and the manufacturers of its products.

In South Africa part of the cultural and religious beliefs of the African people is the use of traditional remedies to treat diseases. These remedies are obtained from medicinal plants (Steenkamp, 2003). One of the most frequently traded plants in the Eastern Cape is Hypoxis, commonly known as Afrika patat, or African potato. South African traditional healers instruct patients to brew the fresh Hypoxis corm as a tea and then ingest it (Steenkamp, 2006a). This prompted an investigation into the digestive stability of a traditionally prepared Hypoxis extract. The H. sobolifera extracts were digested using a simulated gastric/small intestinal digestion and their biological activity determined. The hot water H. sobolifera extract before digestion only showed cytotoxic activity against cancer cell lines at very high concentrations which are not likely to be achieved under normal ingestion circumstances. In Chang liver cells on the other hand, chronic exposure to the hot water H. sobolifera extract increased glucose uptake in amounts similar to that of metformin. On the negative side, the glucose utilization stimulation was lost due to the simulated digestion process. The significant inhibition of AGEs by hot water H. sobolifera extract (IC50 of 6.3 Ig/ml) is a very encouraging result as treatment in the management of diabetes. This activity was only slightly reduced by the in vitro digestion process. Also observed was enzyme inhibition activity by traditionally prepared H. sobolifera, with ∝-amylase being inhibited (IC50 of approximately 250 Ig/ml) and therefore preventing or limiting starch breakdown. From the DPPH results it was clear that H. sobolifera, even when digested, is a potent anti-oxidant (IC50 of 134.4 Ig/ml when undigested compared to 162.9 when digested with β-glucosidase added to stomach digestive step). HPLC and TLC experiments revealed that rooperol which has previously been thought to be the compound responsible for the anti-oxidant activity in Hypoxis extracts, was absent from the traditional extract of H. sobolifera and therefore cannot be the sole compound exhibiting anti-oxidant activity; other compounds such as phenolics may be contributing. The phenolic and flavonoid content results revealed very highconcentrations of these compounds in the traditionally prepared H. sobolifera extract. These compounds may therefore play major roles in all of the biological activities observed from treatment with Hypoxis spp. The ROS results yielded interesting and promising results. Using standard or traditionally prepared H. sobolifera extracts, activation of differentiated U937 cells with PMA was greatly enhanced by cotreatment with the extracts, while extracts on their own did not cause significant activation. Future studies should investigate this property of the extracts as a promising immune boosterThe HPLC results showed that hypoxoside was undetectable in the hot water traditional extract and the TLC anti-oxidant experiment proved that rooperol is not present in the hot water traditional extract after treatment with β-glucosidase. This indicates that neither one of the Hypoxis compounds previously believed to be responsible for the biological activities observed are present in the extract when prepared the traditional way. Therefore, the biological activities observed in this study can be attributed to other phytochemical compounds.

The intent of this research is to demonstrate the effectiveness of hypnosis as a treatment for school refusal. The research design is a single-case study employing an A-B Follow-up format. The 10 year old male subject completed measures of personality (The Children's Personality Questionnaire), self-concept (The Piers-Harris Children's Self-concept Scale), identified stressors, and anxiety. The baseline period was two weeks and therapy lasted four weeks. Follow-up data was collected on the same measures ten months later. All post-therapy results indicate change in a more adaptive direction. The subject showed increased self-concept, lessened anxiety, greater ability to cope and he returned to school with little or no of the previous psychosomatic complaints evident. The follow-up results show that the subject has maintained his gains. Hypnosis is seen as an effective, fast method of treatment for school refusal, a syndrome which needs to be dealt with quickly since consequences can be severe for the child. / Education, Faculty of / Educational and Counselling Psychology, and Special Education (ECPS), Department of / Graduate