Dissertation submitted in partial compliance with the requirements for the Master’s Degree in Technology: Homoeopathy , Durban University of Technology, 2011. / The treatment with homoeopathic remedies, namely Phosphoricum acidum 200CH has proved to be effective in many clinical situations; however confirmation of its use within the sporting industry is limited. This study aimed to test Phosphoricum acidum 200CH efficacy in treating highly trained cyclists. This study was a superiority trial, in which the homoeopathic intervention was hypothesized to be superior to the placebo control group, in a statistically significant way (p < 0.05). Aim This study aimed to test the use of Phosphoricum acidum 200CH in treating the adverse physiological and psychological symptoms induced by exercise on cyclists. The aim for treating these adverse symptoms, induced by exercise, is to enhance performance and recovery of cyclists. Performance and recovery were tracked by assessing blood lactate concentration, oxygen consumption rate, heart rate, peak power output and emotional status. Methodology The study was a randomized controlled clinical trial, parallel group design. Participants were selected using convenience sampling of male road and mountain bike cyclists in the Western Cape. This study followed an explanatory Randomized Controlled Trial test, where the efficacy of the homoeopathic remedy (Phosphoricum acidum 200CH) was under investigation. The participants were selected with great care and testing was completed under highly controlled conditions. Thirty competitive male cyclists volunteered for this study. All participants were required to complete two cycling power to exhaustion interval tests, consisting of a ten minute warm-up at 100 Watts, followed by a five minute constant load at 150 Watts. The purpose of the constant load was to test cycling economy (CE). Thereafter the workload was increased to 200 W for 30 seconds and then the workload was increased by 20 Watts every 30 seconds. The test continued until the participant could no longer maintain the set repetitions per minute for that workload. The peak power output attained iv was recorded. The first test served as a baseline, after which a single dose of Phosphoricum acidum 200CH or identical placebo was administered, a 30 minute recovery period allowed for the remedy to take effect. The test interval was then repeated, the second test results were then compared to the first baseline test to determine the effect of treatment. The study took many different aspects of the remedies action on the cyclists into account, including mental and physiological effects. Breath-by-breath gases were continuously recorded. Expired gases, volumes and air flow were sampled through a flow meter and gas sampling line and heart rate was measured through telemetry (Polar®, Polar Electro, Oy, Finland) and analyzed by a cardio-pulmonary metabolic system (Quark CPET ® Cosmed, Rome, Italy, 2009). Data recorded was filtered for values outside the normal ranges and averaged for every five seconds. Oxygen consumption (vO2 mL.min-1) and heart rate (bpm) at different stages of each test interval were recorded. More specifically oxygen consumption and heart rates were averaged over the 5 minute section following the warm-up to determine cycling economy, directly after exhaustion for one minute and two minutes following exhaustion for another one minute period. Maximum attained heart rate was recorded for each interval test. Maximum oxygen consumption (vO2Max Absolute) was calculated as the mean of the highest three values attained, this mean was then divided by the participants body mass to determine maximum aerobic capacity (vO2Max Relative). Blood lactate levels were tested before, 15 minutes into and directly after each interval, to assess resting lactate status, cycling economy and to determine maximum lactate accumulation. Psychological testing included mood analysis, using a Stellenbosch mood scale (STEMS) questionnaire and further symptoms were analyzed using a Numerical rating scale (NRS) with symptomatic questions. Results From the results, it was clearly apparent that the control group proved to be of a higher calibre when comparing performance variables of the two groups namely:  Higher peak power output  Higher vO2Max  Lower economy lactate v  Lower resting lactate Despite the treatment group being the weaker of the two groups, they showed improvement in performance after administration of the remedy. This improvement was manifest through physiological alteration in the second test. More exclusively is the acceptance of the hypothesis concerning heart rate and oxygen consumption, whereby results showed that the administration of Phosphoricum acidum 200CH decreased heart rate and submaximal oxygen consumption rates during performance and recovery. There was no observable psychological effect during this study. The results suggest that Phosphoricum acidum 200CH primary demonstrated physiological effects on the cyclists. The researcher believes that this is due to insufficient time given for psychological alterations. Conclusion The Phosphoricum acidum 200CH has proven to be effective in enhancing cycling economy, reducing maximum heart rate and enhancing recovery to a large degree for the first minute following exhaustion. These positive effects are of great importance as the treatment group was the weaker of the two groups. Resulting in the possibility of even larger results being observable in repeated studies where both groups have similar performance abilities.

Dissertation submitted in fulfilment for the requirements of the Degree of Master of Technology in Biomedical Technology, Durban University of Technology, 2012. / Introduction Medicinal plants have been used for centuries as remedies for human diseases because they contain components of therapeutic value. Recently, the acceptance of traditional medicine as an alternative form of health care and the development of microbial resistance to the available antibiotics have led scientists to investigate the antimicrobial activity of medicinal plants Aim The aim of this study was to investigate the antimicrobial activity of extracts obtained from medicinal plants used in traditional medicine. A comparative study was carried out on the antimicrobial properties of extracts obtained by two different methods in order to choose that which extracts the most effective antimicrobial compounds. Methodology The plants used in this study Artemisia afra, Erythrina lysistemon and Psidium guajava were harvested from the Silverglen Nature Reserve (Chatsworth) early in the morning (8 a.m.). The leaves of A. afra and P. guajava extracts and the bark of E. Lysistemon were used to prepare the extracts. All plant extracts were prepared according to modified method of the German Homeopathic Pharmacopoea. Two solvents, water and 60 percent ethanol were used to extract the antibacterial compounds from plant material. The extracts were then assessed for their antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. The effect of the plant extracts on these bacteria was determined by the disk diffusion test, which was used as the screening test. Positive results were further subjected to the minimum inhibitory concentration and minimum bactericidal concentration assays. Tubes that showed no turbidity were then sub-cultured onto non-selective plates. Bacterial sensitivity testing was carried out in accordance with modified Kirby-Bauer Antimicrobial Sensitivity Test. An attempt was made to identify some antibacterial compounds using Thin Layer Chromatography and High Pressure Liquid Chromatography. Results None of the gram negative organisms were inhibited by Artemisia afra, Erythrina cafra and Psidium guajava. Only the ethanol extracts of all three plants were able to inhibit Staphylococcus aureus but not Escherichia coli and Pseudomonas auruginosa. None of the test organisms were inhibited by the aqueous extracts of all three plants used in this study. In the screening test, the zones of inhibition for ethanol extracts against Staphylococcus aureus ranged from 3mm – 7mm. The minimum inhibitory concentration ranged from 16.67 percent – 83.3 percent inhibition depending on the dilution of the extract. Quercetin and Catechin were identified as some of the antibacterial compounds present in the leaves of Psidium guajava. These two compounds were not identified on Erythrina lysistemon and Artemisia afra. Conclusion The results obtained in this study have proven that Artemisia afra, Erythrina cafra and Psidium guajava ethanol extracts contain antibacterial substances. The ethanol extracts of all plants in this study inhibited the growth of Staphylococcus aureus but had no effect on the gram negative bacteria. Aqueous plant did not inhibit the growth of any bacteria in this study. This study has also shown that antibacterial effect of these extracts may be considerably enhanced in traditional treatment if traditional healers can include ethanol as one of the extraction solvents. The results obtained in this study might be considered sufficient for further studies aimed at isolating and identifying the active compounds and evaluating possible synergism of antimicrobial activity among these extracts. Investigations on toxicity of these extracts should also be carried out.

Submitted in partial compliance with the requirements for the Master’s Degree in Technology, Department of Homoeopathy, Durban University of Technology, Durban, South Africa, 2015. / INTRODUCTION Homoeopathy is based on the law of similars meaning the medicine that produces symptoms in a healthy individual will cure the same symptoms in a sick individual (Sankaran, 1991:5). AIM Conducting a proving on Ubiquinone 30CH will lead to an establishment of its therapeutic potential through the application of the law of similars thus adding to the Materia Medica and advancing Homoeopathy (Vithoulkas, 2002). It was hypothesised that the 30CH potency of Ubiquinone would clearly produce observable signs and symptoms in healthy prover’s. It was further hypothesised that a comparison of Ubiquinone to those remedies yielding the highest numerical value and total number of rubrics on repertorisation of the proving symptoms would elucidate differences and similarities between Ubiquinone and other Homoeopathic remedies to clarify its therapeutic indications. It was hypothesised that in this manner a better understanding of Ubiquinone and its relationship to other Homoeopathic remedies would be gained. Methodology The proving of Ubiquinone 30CH was a randomised, double blind placebo controlled study, using the 30th centesimal potency and a total of 26 participants who met the inclusion criteria. Each prover was provided with a journal to record their symptoms daily. The data extracted from the journals were added to the case histories and physical examinations to compile a proving profile. The identity of the substance was revealed and the information was correlated after completion of the proving. The symptoms found were translated into Materia Medica and repertory language. Once the proving was concluded, a comparison to the remedies yielding the highest numerical value and total number of rubrics on repertorisation - which is the technique of using a repertory to identify the Homoeopathic medicines whose Materia Medica corresponds most closely to the clinical picture of the patient and from amongst which a simillimum may be chosen (Swayne, 2000:183) - was compared to the proving symptoms. Results The remedy’s main influence was on the mental and physical state. The most prominent symptoms seen in the mental sphere were extreme irritability and exhaustion. There was a sense of emotional fragility with a desire to be alone. On the physical side, headaches were common and weakening pains of the extremities were experienced. It can be concluded that the 30CH potency of Ubiquinone, if used precisely according to Homoeopathic principles, can be applied to a clinical setting, as the extensive range of symptoms produced during the proving suggests an equally wide array of application of the remedy Ubiquinone. Conclusion One of the downfalls of Homoeopathy is the limited number of provings being done, (Vithoulkas, 2002). Vithoulkas (2002:143) maintains that in order for Homoeopathy to advance, it is necessary to perform provings on new substances to expand the Homoeopathic armamentarium. Increasing the number of remedies in the Materia Medica facilitates greater accuracy and individualisation when treating patients (Wright, 1999). According to Herrick (1998) numerous cases cannot be solved because many of the most important remedies have not yet been developed. The purpose of this study was to increase the knowledge of drug substances due to the limited amount of information in our current Materia Medicas, by investigating the therapeutic potential of Ubiquinone 30CH. The investigation supported the hypothesis that Ubiquinone would produce clearly observable signs and symptoms in healthy volunteers. It is essential that the proving symptoms be verified and expanded through clinical use and with further proving of Ubiquinone in various potencies so that it becomes a well utilised remedy in the future.

Growing attention has been given to the potential of the respiratory tract for systemic delivery of macromolecules, particularly proteins and peptides. However, limitations such as short transit time and loss of activity of some proteins and peptides in the respiratory tract need to be overcome. Consequently, the utility of controlled drug delivery systems such as liposomes as protein carriers appear promising. Unfortunately, liposomes are unstable in aqueous dispersions. Additionally, conventional liposome preparation methods such as the thin film hydration are difficult to scale-up, and also demonstrate low entrapment efficiencies for hydrophilic materials. The aim of this work was to develop novel submicron mucoadhesive liposomes entrapping the protein immunoglobulin g (IgG) using the proliposome method. Additionally, this work explored the potential of the generated liposomes for respiratory tract delivery via medical nebulisers and nasal sprays with different operating principles. Liposomes generated from the proliposome technology were multilamellar as cryo-TEM studies revealed. The generated liposomes were capable of entrapping considerable concentrations of salbutamol sulfate (59.1%), ovalbumin (43.3 %) and IgG (29.9 %). Also, the generated liposomes demonstrated superior entrapment efficiency of IgG to other liposome preparation methods (thin film and particulate- based proliposome technology). Reduction of liposome size to 400 nm and the incorporation of the mucoadhesive agent sodium alginate markedly enhanced the entrapment of IgG in liposomes (up to 50 %). The secondary structure and immunological reactivity of IgG were also maintained following its incorporation in liposomes as demonstrated by circular dichroism and microagglutination assay, respectively. Nebulisation was found to fragment liposomes as well as reduce the activity of the entrapped IgG. The degree of liposome fragmentation and loss of activity of IgG varied markedly among different medical nebulisers. Liposome size distribution and IgG immune reactivity studies elucidated that vibrating-mesh nebuliser was least disruptive to liposome structure and the immunoreactivity of the incorporated IgG was least affected following its use (retained activity of 83% versus 24% and 39% for the ultrasonic and air-jet nebulisers, respectively). Contrary to medical nebulizers, this work illustrated that all studied nasal devices preserved both the integrity of liposomes and the incorporated IgG. In conclusion, the findings of this study demonstrate potential benefits in drug delivery employing both intranasal administration and proliposome technology offer with great promise in using proliposome formulations for intranasal protein delivery.

Block copolymers of polylactide and poly( ethylene glycol) (PLA-PEG) were investigated as biodegradable drug delivery systems. They are defined by the differing molecular weight ratios of polylactide to poly( ethylene glycol). Copolymers containing more hydrophilic PEG than hydrophobic PLA per molecule self-dispersed in water giving spherical nonionic micelles. Purification by gel permeation chromatography gave two peaks. The first peak only formed micelles (the second was PLA-depleted). Analysis by dynamic light scattering (DLS) and transmission electron microscopy (TEM) gave diameters of IS.6nm and 18.9nm for 1.5:2 and 2:5 PLA-PEG micelles respectively. PLA-PEG copolymers with more PLA than PEG per molecule (4:2 and 6:2 PLA-PEG) formed "solid particles" by the solvent-precipitation method. GPC purification again gave two peaks, but smaller second peaks. DLS analysis gave diameters of 15.1 nm and 20.8nm for 4:2 and 6:2 PLA-PEG particles respectively (confirmed by TEM and atomic force microscopy (AFM)). Static secondary ion mass spectrometry and X-ray photoelectron spectroscopy showed PEG at the surface of 4:2 and 6:2 PLA-PEG in water and acetone. Stability testing to salt suggested sterically stability. Rheological measurements determined PEG chain layer thicknesses, with the thickest chain for 2:5 PLA-PEG (where PEG chain length is 5000gmol-1 compared with 2000gmol-1). Testosterone and sudan black B (SBB), were used as "model" drugs with different hydrophobicities. Ultracentrifugation sedimentation velocity studies confirmed drug incorporation. Aromatic SBB loaded readily (≥59.0%w/w) compared with steroidal testosterone <2%w/w). Loading of testosterone esters of varying hydrophobicity into PLA-PEG particles showed little difference compared to between testosterone and SBB, suggesting that aromaticity is more significant. In vitro release studies (4:2 PLA-PEG particles/SBB) showed a small burst release, then linear release to twenty eight days. In vivo studies in the rat, using a radioactive marker, demonstrated extended blood circulation times for PLA-PEG micelles during the three-hour study, with increased blood levels and lower liver uptake for 1.5:2 over 2:5 PLA-PEG micelles. PLA-PEG particles were directed to the liver.

Thesis (M.Tech.: Homoeopathy)--Durban University of Technology, 2008 / The proving substance Hemachatus haemachatus commonly known as the Rinkhals belongs to the family of Elapidae. This spitting-cobra is a local snake found only in Southern Africa. This proving tested the effects of the thirtieth centesimal (30CH) potency of venom from Hemachatus haemachatus on healthy provers. OBJECTIVES It was hypothesised that Hemachatus haemachatus 30CH would produce clearly observable signs and symptoms in healthy provers, and that the comparison of Hemachatus haemachatus to those yielding the highest numerical value and total number of rubrics on repertorisation of the proving symptoms would highlight differences and similarities between the remedy symptoms so that confusion as to the indication is eliminated. It was hypothesised that a fuller understanding of Hemachatus haemachatus and its relationship to other remedies would be gained following this comparison. METHODOLOGY A double blind, placebo controlled proving of Hemachatus haemachatus 30CH was conducted on thirty healthy volunteers who met the inclusion criteria. Six of these thirty provers randomly received placebo, with neither prover nor researcher knowing whom received placebo. Provers had a homoeopathic case history taken and a physical examination performed on them prior to commencement of the proving. The provers recorded their signs and symptoms 6 by means of a journal before, during and after administration of the remedy. On completion of the proving, the information obtained was correlated and assessed by the two researchers, De la Rouviere and Cahill. The symptoms elicited during the proving were translated into materia medica and repertory language, and a homoeopathic picture of the remedy was subsequently formulated. Data from the case histories, physical examinations and group discussions were also considered in the assessment. RESULTS During the period of investigation, provers experienced a variety of symptoms on the mental, emotional and physical spheres. On the mental emotional sphere there was a marked degree of irritability and changeability in moods as is commonly seen in many of the snake remedies. Along with this, it was noted that there were feelings of anxiety for reasons unknown, a sense of having lost something or someone close, and a desire to be left alone. There were also a great number of feelings regarding the home, where there were feelings of the home being a place of safety and wanting order in the home. On a physical level, many of the provers noted varying degrees of abdominal discomfort and headaches. Along with anxiety, provers experienced palpitations and sensations of chest restriction or constriction with shortness of breath. There were a variety of musculoskeletal symptoms ranging from painful joints in the fingers to stiffness and tightness in the neck and back. Provers noted flushes of heat and alterations of their internal thermostat. Provers experienced marked dryness of the mucus membranes and the skin, and there was also a general feeling of weakness and heaviness as well as a marked aggravation in the mornings on waking. 7 CONCLUSIONS Symptoms obtained from the proving of Hemachatus haemachatus 30CH were studied and evaluated. Those symptoms that appeared to represent the remedy picture of Hemachatus haemachatus most accurately in the researchers‟ opinion were used in the repertorisation of the remedy. The investigation supported the hypothesis that Hemachatus haemachatus 30CH would produce clearly observable signs and symptoms in healthy provers. The subsequent comparison of the proving symptoms of Hemachatus haemachatus to Lycopodium (Club moss), Sulphur, Alumina (Aluminium oxide), Sepia (Cuttle fish) and Calcarea carbonica (Carbonate of Lime) highlighted differences and similarities between these remedies and Hemachatus haemachatus. The further comparison of remedies that came up on repertorisation restricted to the plant, mineral and animal kingdoms respectively provided a further comparison of remedies, which aimed at enhancing the differentiation of Hemachatus haemachatus to other similar remedies. / M

Mini-dissertation submitted in partial compliance with the requirements for the Master’s Degree in Technology: Homoeopathy in the Department of Homoeopathy at the Durban University of Technology, 2008. / The aim of this study was to determine the effect of Gymnura natalensis 30CH on healthy volunteers, and to record the signs and symptoms produced, so that it may be prescribed to those requiring it according to the Law of Similars. The other aims of this study were to compare the proving symptoms of Gymnura natalensis 30CH to the toxicology of stingray venom and to analyze the remedy picture in terms of the Doctrine of Signatures. It was hypothesised that the thirtieth centesimal potency of the remedy would produce clearly observable signs and symptoms in healthy volunteers (provers). It was further hypothesised that the above signs and symptoms would show a correlation to the toxicology of stingray venom and to the Doctrine of Signatures. The homoeopathic proving of Gymnura natalensis took the form a double-blind, randomised, placebo-controlled trial. The selected proving potency was the thirtieth centesimal potency. A total population of 30 suitable and consenting volunteers participated in this trial. Twenty percent of this population was randomly administered a placebo-control substance. The collection of data from the provers took the form of a journal which was kept by each prover in which their proving signs and symptoms were recorded over a period of five weeks after the administration of the remedy or placebo. On completion of the proving, each journal was assessed by the researcher to determine the suitability of the recorded symptoms for inclusion in the materia medica of Gymnura natalensis. These symptoms were then translated into the language of the materia medica and repertory and a remedy picture was then formulated. Data from case histories, physical examinations (Appendix D) and group discussions were also taken into account during the analysis of the proving the symptoms. A concurrent proving study of Gymnura natalensis, conducted by Naidoo (2008), focused on comparing the symptoms of this remedy to those of other existing remedies that were derived from the sea. A variety of mental, emotional and physical symptoms were extracted from the proving study of Gymnura natalensis. The main mental and emotional symptoms of the remedy included anxiety, irritability, depression, a feeling of disconnection, spaciness of the mind and dreams of events and incidents of the past. The characteristic physical symptoms obtained from the proving included headaches, a reduction of pre-menstrual symptoms, heart palpitations, skin eruptions on the back, low energy levels, tiredness and sleep abnormalities. Symptoms that showed a correlation to the toxicological symptoms of stingray envenomation included frequent urination, muscular cramps, heart palpitations, laboured breathing, fever and copious night sweats. The symptoms that characterised the remedy in terms of the Doctrine of Signatures included anxiety, instinctive behaviour, the desire to be alone, feelings of disconnection, detachment and isolation and skin eruptions on the back. The investigation confirmed the hypothesis that Gymnura natalensis would produce clearly observable sings and symptoms in healthy volunteers. The correlation of the proving symptoms to the toxicology of stingray venom and the Doctrine of Signatures provided a clarification of the remedy picture to assist in the understanding and prescription of this remedy. / M

Inhalation represents a potentially attractive delivery route for biologics, especially those designed to treat pulmonary diseases such as asthma, cystic fibrosis or lung cancer. Delivery directly to the site of action should increase local concentrations of drug, whilst reducing systemic side effects. However, there is limited knowledge regarding the mechanisms of pulmonary clearance, with gaps in understanding; where molecules are absorbed, the mechanisms involved, regional variability throughout the lung, and how to control pulmonary retention and/or facilitate cellular uptake. The work presented in this thesis details the development of a SPECT/CT imaging protocol to determine the pulmonary retention and tissue redistribution of technetium labelled antibodies and their fragments in vivo, in mice, to begin to address these knowledge gaps. The SPECT/CT imaging method was applied to a whole monoclonal murine immunoglobulin G1 (mlgG1), as well as its Fab and scFv fragments and a small protein (FN3) in order to determine whether diffusion controlled pathways were important in pulmonary antibody clearance. Additionally, the pulmonary retention of mutant mlgG1 with differing binding affinities to the murine neonatal Fc receptor (mFcRn) were assessed in order to determine whether antibody transport across the epithelium occurred via active transcytosis. It was determined that 54.4 ± 0.63 % of the total instilled dose of a whole monoclonal antibody remains in the lung over 24 hrs, with Fab and scFv fragments cleared significantly quicker with 28.7 ± 0.73 % and 34.9 ± 0.85 % respectively of the total instilled dose remaining in the lung at 24hrs. The pulmonary retention of the 11 kDa FN3 protein was also assessed with 21.0 ± 0.65 % remaining in the lungs after 24 hrs. No evidence of build up of any protein was detected in the oesophagus/stomach, suggesting little contribution by mucociliary clearance. Very little build up of whole antibody, Fab or scFv was observed in the liver or kidneys. However, very clear evidence of renal filtration of the 11 kDa fragment was observed. There was no difference in the pulmonary retention of wild type IgG and any of the mFcRn binding mutants. Additional investigation of antibody retention rates in the murine house dustmite (HOM) model of asthma, although not making use of the SPECT/CT method, showed that antibody is cleared more rapidly from the diseased lung than the normal lung. It was also shown that the expression pattern of the mFcRn receptor is the same in the normal and HDM exposed lung and so this increase in clearance rate occurs via passive diffusion controlled processes. This is most likely a result of increased paracellular transport due to disrupted mucosal barrier function. The SPECT/CT imaging method developed has proven to be a simple and reliable method to assess, non-invasively, pulmonary antibody retention in vivo. Overall it appears that antibody transport across the pulmonary epithelium occurs predominantly via diffusion controlled mechanisms, which include both paracellular transport and nonspecific transcytosis by pinocytotic routes. Additionally, in the mouse, neither receptor mediated transport by mFcRn nor mucocillary clearance is important in the pulmonary clearance of antibodies.

A multitude of plants have been used extensively for the treatment of cancers throughout the world. In many parts of the world, especially in poor countries, this may be the only form of cancer therapy. Much research has been focused on the scientific evaluation of traditional anti-cancer drugs from the tropical plant; Momordica charantia (MC) is one of them and it has been used frequently as an anti-cancer agent. The green leaves, fruits, seeds and stems of M. charantia composed of many different proteins and steroids that are chemically active. These proteins are α and β momorcharins which possess anti-cancer and anti-HIV properties similar to crude water and methanol soluble extracts of M. charantia. This study investigated the anti cancer effect of either the crude water and methanol soluble extract of M. charantia, α and β and α, β momorcharins based on dose-dependent, time-dependent on the viability of 1321N1, Gos-3, U87-MG, Sk Mel, Corl -23, Weri Rb-1 and L6 cell lines employing different concentrations of each extract or drug. In addition, the study measured the effect of either temozolomide or vinblastine alone or combining each with either the crude water soluble extract of M. charantia or α β momorcharin measuring cell viability in the different cell lines. Furthermore, the present study investigated the cellular mechanism(s) via which the different anti-cancer agents were able to induce cell death measuring the activities of caspase - 3 and caspase - 9, the release of cytochrome c and intracellular free calcium concentrations [Ca 2+ ]i. The results have shown that the crude water soluble extract of M. charantia can evoke both time-course at (800 µg) and dose-dependent (200 µg - 800 µg) decreases in cell viability with maximal increases with 800 µg over a period of 24 hrs following incubation. Either the crude methanol soluble of M. charantia (200 µg - 800 µg), alpha or beta momorcharin (200 µM - 800 µM) had little or no effect on the viability of the different cell lines. In contrast, either alpha, beta momorcharin (200 µM - 800 µM), temozolomide (80 µM - 320 µM) or vinblastine (10 μg - 40 μg) can evoke significant (p < 0.05) decrease in cell viability, similar to the crude water soluble extract of M. charantia. The results also show that combining either temozolomide (240 µM) or vinblastine (40 μg) with either (800 µg) of the crude water- soluble extract of M. charantia or (800 µM) of alpha, beta momorcharin can result in significant decreases in cell viability for each cell line but these effects were neither additive or synergetic compared to the individual effect of temozolomide or vinblastine. The result of this study have also shown that either the crude water-soluble extract of M. charantia (800 µg) or (800 µM) of alpha, beta momorcharin can elicit marked and significant (p < 0.050) increases in the activities of caspase - 3 and caspase - 9 in all the cell lines. Similarly, both the crude water soluble extract of M. charantia and alpha, beta momorcharin can stimulate the release of cytochrome-c and elevated [Ca2+ ]i in the different cancer cell lines compared to untreated cell lines. Together, the results of the study have shown that either the crude water soluble extract of M. charantia or alpha, beta momorcharin can exert their anti-cancer effects (cell death) on cancer cell lines by increasing the activities of caspase - 3 and caspase - 9 and by releasing cytochrome-c and elevating [Ca2+ ]i in the cancer cells. These findings implicate the role of apoptosis and cellular Ca 2+ homeostasis in cancer cell death. Moreover, they confirm the beneficial use of extracts of M. charantia to treat cancers.

Natural products are an important source for drug discovery. At present there is a resurgent interest in pharmacognosy as a platform for new combinations of active principles to provide highly potent and low-cost medications to treat a growing population with an increasing longevity. This product studied phytochemical interactions in Artemisia annua plant extracts using anti-plasmodium and anti-proliferation assays to identify interactions with potential therapeutic implications. To enable the study a rapid tandem quadrupole mass spectrometry (TQD) method was developed for metabolites in the plant and the validation indices showed the method to be robust, quick, sensitive and adequate for a range of applications.