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In Kenya, the prevalence of diabetes is estimated at 3-10 percent of the population. These figures could be higher because most type 2 diabetics are diagnosed many years after onset. Out of this number, 15 percent are people below 30 years of age who need prompt education to avoid complications that are associated with diabetes (DMI centre, 2004). Due to inadequate or lack of proper information, most patients especially those with type 2 diabetes are diagnosed through complications. Untreated or poorly managed diabetes is now the leading cause of eye disease and kidney failure in the world. Diabetes is the largest cause of kidney failure in the developed world, and is the fourth leading cause of global death by disease in the world (IDF, 2007). At the Kenyatta National Hospital in Nairobi, Kenya, it is the leading cause of all non-accident related amputations. It is with such statistics in mind and the grim reality of poor and inadequate health services that this research is based. The wide use of selected medicinal plants for the treatment and management of diabetes warrants the further study of these plants for potential use and commercialization. The data obtained can also be invaluable for use and reference when using these plants for medicinal purposes. The medicinal plant studied in the research is widely used in Kenya by many communities and was chosen based on ethno-pharmacological references using traditional medicinal practitioners as well as patient’s recommendations. Different in vitro and in vivo assays were studied to try and elucidate the mechanisms of action as well as the organs targeted during treatment using this plant.

Thesis (M. Sc. (Biochemistry)) -- University of Limpopo, 2015 / Diabetes mellitus is one of the major diseases worldwide that is life threatening and is reaching an epidemic proportion. The most important approach in reducing the burden of the disease worldwide is to search for effective, low cost hypoglycaemic drugs with fewer side effects. Past experimental evidence confirmed the hypoglycemic activity of many indigenous African medicinal plants. S. italica (Fabaceae family) is widely used by traditional healers to treat a number of diseases such as sexually transmitted diseases and other forms of intestinal complications traditionally. The current study was aimed at evaluating the in vitro effects of root and leaf extracts of S. italica on GLUT4 translocation in NIH-3T3-L1 preadipocytes and C2C12 muscle cells. In order to address the aim of the study various methods were undertaken. The roots and leaves of S. italica collected from Zebediela sub-region of the Limpopo province, South Africa, were ground to fine powder and extracted using acetone, methanol, ethyl acetate and n-hexane. The various extracts of the root and leaf material were subjected to fingerprint profiling using TLC plates and different mobile phases (BEA, CEF, EMW and BAW). The chromatograms were visualized with vanillin-H2SO4 reagent, p-anisaldehyde and iodine vapour. The extracts were assayed for the type of secondary metabolites contained in the studied plant parts using chemical text and by TLC analysis. The total phenolic content of the root and leaf material were also evaluated. Evaluation for antioxidant activity was performed using 0.2% DPPH qualitatively and quantitatively with vitamin C as a positive control. Toxicity study was performed on C2C12 muscle cells using the MTT assay, with Curcumin as a positive control and untreated cells as a negative control. The CC50 values of the acetone root and leaf extracts were determined by linear regression. The effect of acetone root and leaf extracts on glucose uptake by C2C12 muscle cells was evaluated, also on western blot and immunofluorescence for NIH-3T3-L1 preadipocytes. The solvents employed for extraction in this study are commonly used to extract various biological active compounds from plants in research settings. Methanol extracted more compounds followed by acetone, then ethyl acetate and n-hexane the least. The constituents extracted by methanol may be mostly sugars, amino acids and glycosides due to the polarity of this solvent. Hydro-alcoholic solvents extract a variety of compounds that are mostly polar. Acetone extracts xxii mostly alkaloids, aglycones and glycosides while n-hexane in general extracts mostly waxes, fats and fixed oils. High yield was obtained with leaf extracts with all the solvent used for extraction as compared to the root. The TLC finger-print showed that good separation was achieved with the methanol and acetone extracts in CEF mobile phase, ethyl acetate extracts in CEF and EMW and n-hexane extracts in BEA respectively, especially with the leaf extract. Most compounds present in S. italica extracts were UV active. Some compounds that were not reactive with vanillin-H2SO4 reagent were shown to be reactive with p-anisaldehyde reagent and iodine vapour which revealed the presence of sugars or aromatic compounds. Chemical analysis for secondary metabolites of the acetone root and leaf extracts revealed the presence of flavonoids, terpenes, tannins, steroids, reducing sugars and alkaloids while glycosides were detected only in the leaf extract. The results obtained using TLC analyses were consistent with the results obtained in the chemical analysis. Thin layer chromatography revealed the presence of glycoflavones in the acetone root extract, alkaloids in the root and leaf extracts; and phytosterols and flavonoid aglycones in root and leaf extracts. The acetone root and leaf extracts revealed the presence of phenols. The leaf extract was shown to contain high total phenolic content as compared to the root. The methanol and acetone root and leaf extracts were shown to possess antioxidant activity. However, the concentration of the activity was higher in the acetone root than in the leaf extract. The least activity was observed with the ethyl acetate root and leaf extracts as compared to other extracts. The n-hexane extracts however, was not shown to contain any antioxidant compounds. Although activity observed with the methanol extracts was comparable to that of the acetone extracts in the quantitative assay, the acetone extracts were shown to possess more antioxidant activity in the qualitative assay. The concentration of extracts increased with increase in scavenging activity. The root extract exhibited a more potent antioxidant activity compared to leaf extract. These extracts were evaluated for their cytotoxicity on normal cells. The highest cytotoxic concentration (CC50) was obtained with the root extract with a CC50 value of 297 635 μg/ml at 48 hrs, followed by CC50 value of 21 544 μg/ml at 24 hrs. The CC50 value of the leaf extract at 24 hrs was 2 904 μg/ml with the least value at 48 hrs. The root extract at 24 and 48 hrs together with the leaf extract at 24 hrs were not toxic to C2C12 muscle cells at the concentration tested in this study. The acetone extracts were shown to possibly enhance proliferation of C2C12 muscle cells at a concentration of 0.001–1000 μg/ml. The non-cytotoxic concentration of 25 μg/ml of the leaf extract in combination with insulin showed more glucose uptake as compared to other extracts as well as the control. Prolonged incubation time was shown to increase glucose uptake with leaf extract while increase in concentration of root extract decreased glucose uptake at 24 hrs. At incubation time of 3 and 24 hrs, glucose uptake results at concentration of 2.5 μg/ml were comparable with that of the root extract, with a similar trend observed at 25 μg/ml, although with decrease in uptake. The qualitative and quantitative fluorescence results showed GLUT4 to be translocated to the cell membrane. The leaf extract at a concentration of 25 μg/ml had more fold as compared to other extracts, indicative that more GLUT4 was translocated at this concentration of the leaf extract. The acetone root and leaf extracts were shown to increase protein expression of GLUT4 at 3 hrs incubation time as compared to other incubation times in insulin-stimulated C2C12 muscle cells. The plant constituents of S. italica was shown to contain a variety of secondary metabolites that maybe be acting alone or in concert with each other to exert the various activities observed in this study. Different solvents used for extraction may be responsible for the extraction of different constituents with antioxidant activity observed in the study. The acetone extracts enhanced proliferation of C2C12 muscle cells at concentrations used in the study. However, there was no significant reduction on viability of normal cells. In addition, the extracts were shown to enhance the differentiation of NIH-3T3-L1 preadipocytes into adipocytes and C2C12 muscle cells into myocytes. These in turn induced the translocation of GLUT4 to the cell membrane and as a consequence facilitate glucose transport. Hence, the differentiation of adipose cells as well as glucose uptake of muscle cells and GLUT4 expression might have been enhanced by constituents contained in the acetone extracts. In conclusion, the acetone leaf extract may have a beneficial role in glucose metabolism of differentiated C2C12 muscle cells. Therefore, further studies are however required to elucidate the molecular mechanism by which the acetone leaf extract of S. italica influences the translocation of GLUT4.

No description available.

No description available.

研究背景：肩痛是临床的常见症状，常因不正确姿势、受凉、外伤、运动劳损等引发，多被诊断为局关节周围炎、属部的肌臆炎等’带来的疼痛和肢体活动障碍都严重影响正常生活。针灸治疗盾痛效果显著’传统中医在眉痛的诊疗中已有详尽论边，而近年西方医学提出的激痛点疗法，在治疗痛症上也有明确的疗效。总结局痛的激痛点针刺取穴方案，探讨其与传统针灸的失系’对发展当代针灸有十分重要的理论意义和临床价值。研究目的：初步总结盾痛的激痛点针刺取穴方案，探讨激痛点和传统针灸的相关性, 为临床提供新思路。研究方法：文献研究。结论：眉痛的激痛点针刺取穴方案为：盾前痛取胸大肌锁骨部、胸小肌、肱二头肌、喙肱肌、背阔肌、冈下肌、前三角肌、前斜角肌、中斜角肌激痛点；属后痛取大圆肌、小圆肌、肩胛下肌、肱三头肌长头第一、下斜方肌第三、后三角肌激痛点；肩外侧疼痛取中三角肌、冈上肌激痛点。此法优势在于理论系统与诊疗过程明确、操作规范、疗效显著，是属痛的治疗的新思路。对比发现，激痛点理论与经筋理论相似。激痛点针刺取穴与中医整体辨证论治的结合, 势必有助提高临床疗效，值得深入探索。關键字：肩痛激痛点针刺取穴文献研究

Dissertation submitted in fulfilment of the requirements for the Degree of Master of Technology: Biotechnology, Durban University of Technology, 2012. / Apoptosis, or programmed cell death, is a process which is pivotal in eliminating damaged, infected, or unwanted cells from the body. It has been studied in numerous types of cell lines ranging from normal to infected cell lines, and there have been a wide range of studies on apoptosis in laryngeal cancer because this type of cancer has become one of the most common types of head or neck cancer due to the high incidence of alcohol consumption, tobacco smoking and chewing of betel quid amongst populations. Laryngeal cancer is usually treated with radiotherapy or is surgically removed, but due to the loss of the function of the larynx after surgery, it has been suggested that alternative strategies or ways of treating laryngeal cancer are required. This has prompted the use of, and research in the field of, plant medicine to combat laryngeal cancer. Plant medicine has been used for centuries by the Chinese, Indian and Arabian population in Uhani, Ayurveda and Siddha as a form of replacing conventional medicine with complementary and alternative medicine, these include many plants from the family Asphodelaceae, which have become marketable commodities owing to their medicinal values and traditional uses. Amongst these plants, the genus Bulbine has been used as a form of natural medicine in rural Africa and they are also exploited for their aloe vera properties as well as their possession of phytochemical compounds such as isoflavanoids, nor-lignans, naphthalene derivatives, anthracene and poly prenylated flavonoids. There has been a compelling amount of literature on the traditional uses of the Bulbine spp. because these are linked to the Bulbine spp. having secondary metabolites such as pyroles, chromones, coumarins, bianthraceane, benzene as well as alkaloids. However, for Bulbine natalensis and B. frutescens, the plants of interest in this study, the location of anticancer compounds in them are the only amounts of information available. It has been reported, traditionally, that B. natalensis possesses the anticancer potential in the roots, while the anticancer potential for B. frutescens is in the leaves. However, this requires scientific clarification. Therefore, this study was conducted to assess programmed cell death or apoptosis by analysing the responses of the human laryngeal carcinoma cell line (HEp-2) to crude aqueous and organic (50% and 100% ethanol) fractions of B. natalensis and B. frutescens. In order to have achieved this, the HEp-2 cell line was exposed to the above mentioned fractions at three different final concentrations (20, 2 and 1μg/ml) and assessed for cytotoxicity using the 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay as an indicator of cell death after fraction utilisation (3 days) for 5 and 8 days. The differences in the potency of the Bubline spp. fractions were confirmed using the non-parametric ANOVA test. Thereafter, selected fractions were screened for apoptotic potential using reverse transcriptase-polymerase chain reaction (RT-PCR) to determine the expression of bax and caspase-3 biomarkers, which are the biomarkers that participate in mitochondrial, endoplasmic reticulum and receptor-ligand mechanism of apoptosis. The fractions of B. frutescens were selected relative to those of B. natalensis for the RT-PCR procedure (read section 3.4.1. for the selection procedure) and links between the cytotoxicity and gene expression results were analysed. It was found that the B. natalensis fractions had a much greater cytotoxic effect on the HEp-2 cell line compared to fractions of B. frutescens by the fifth day of the MTT assay. On the eight day of incubation, there was an increase in HEp-2 cell line proliferation by the fractions of both plant species administered. The fractions selected for bax and caspase-3 gene expression analysis for B. natalensis were the: 20 μg/ml root and corm aqueous fractions, 20 μg/ml leaf and corm 100% ethanol fractions, 20 μ g/ml corm 50% ethanol fraction, 2 μg/ml root aqueous fraction, 2 μg/ml leaf 100% ethanol fraction and the corm 1 μg/ml aqueous and 50% ethanol fractions. The fractions that were compared to B. natalensis were the 20 μg/ml root and leaf aqueous and 100% ethanol fractions respectively, the 2 μg/ml root aqueous fraction and the 2 μg/ml leaf 100% ethanol fraction. It was found from RT-PCR analysis that all of the B. natalensis fractions tested induced expression of caspase-3, which indicated that those fractions were capable of inducing apoptosis in laryngeal carcinoma in vitro, since caspase-3 is the molecular indicator of apoptosis. The aqueous B. frutescens root fraction, did not induce expression of caspase-3 gene, although it caused expression of bax. This implied that the root aqueous B. frutescens fraction, may be involved in some other form of cell death, other than apoptosis. It was also found that there was variability in the response of the HEp-2 cell line to the Bulbine spp. fractions because of the variation in bax expression among fractions of different concentration. It was difficult, from this study, to classify fractions into categories for their mechanism of action, because not all of the fractions that caused the expression of capase-3, induced bax gene expression. Hence, proper conclusions were unable to be made, more so, because all the mechanisms of apoptosis mentioned, involve bax gene activation in order to proceed to completion. Therefore for those Bulbine spp. fractions to which the HEp-2 cell line exhibited a variable response to, it was postulated that cell death or apoptosis occurred through some other unknown mechanism. Overall, the cytotoxicity result didn’t correlate to the gene expression results because fractions that promoted HEp-2 cell line growth by day five, expressed apoptotic markers, which highlighted the sensitivity and accuracy of the cells-to-cDNATM II kit for detecting a few possibly apoptosed cells. This was confirmed by the fact that the HEp-2 cell line used in the MTT cytotoxicity assay and gene expression study had the same passage number and were viable, the latter being achieved because the MTT assay only measures the cytotoxicity of compounds once they have been taken up by viable cells – measuring mitochondrial activities expressed as absorbances. Therefore, the deduction that HEp-2 cell death may be due to bax/caspase-3 expression was valid because the mRNA was isolated from viable HEp-2 cells that had been killed by Bulbine spp. fractions of different polarity. Furthermore, the lack of correlation between the cytotoxicity and gene expression results indicated the amount of HEp-2 cell line proliferation by the fraction out-competes those that died, thereby producing a negative cytotoxicity result. There was a relationship between the traditional information about the anticancer potential for B. natalensis and B. frutescens. For example, the aqueous root fractions of B. natalensis were found to be non-toxic to the HEp-2 cell line, but did express caspase-3, which indicated the possibility of apoptosis. Similarly, the 100% ethanol leaf B. frutescens fractions were non-toxic to the HEp-2 cell line, but were able to induce apoptosis as well. This emphasised that the MTT cytotoxicity assay should be compared with other methods of measuring cytotoxicity when performing studies like this, because although literature has emphasised many advantages of using the MTT cytotoxicity assay in apoptotic studies, this study proved otherwise. When identical HEp-2 cells were treated with the same extract, only some cells were killed (apoptosis) whereas others proliferated. This was because although the cells were identical phenotypically, they were all probably at different phases of the cell cycle resulting in the HEp-2 cells responding variably to the same fraction at different concentrations. It was also found that the responses were concentration independent. For example, the 1 μg/ml B. natalensis corm fraction exhibited the highest toxicity of the three concentrations administered. The lowest cytotoxicity was achieved for the 20 μg/ml fraction – showing a proliferative effect on the HEp-2 cell line. Similarly, the 2 μg/ml aqueous B. natalensis leaf fraction induced the highest cytotoxicity level in the HEp-2 cell line followed by the 1 μg/ml and then the 20 μg/ml fractions. Apart from the genetic variation in identical HEp-2 cells; this indicated that the HEp-2 cell line was selective to particular fractions of the Bulbine spp. for utilisation. Concentration independence and HEp-2 cell preferential selection has been reported in many other studies involving plant fractions/extracts and natural products. This study demonstrated that although all the tested B. natalensis fractions were capable of inducing HEp-2 cell death possibly via. apoptosis (caspase-3 induction), a lack of any link between apoptosis and the cytotoxicity results (hence the 20 μg/ml corm fraction had a negative cytotoxicity but expressed both apoptotic markers), indicated the need for phytochemical screening of both Bulbine spp. in future, to determine the compounds that are responsible for the cytotoxicity and gene expression result outcomes of both Bulbine spp. fractions. Furthermore, procaspase genes also have to be analysed since genes are expressed to form procaspases, which then form active caspases. Although normal cells also express caspase-3 genes during apoptosis, this study focused exclusively on the effect of Bulbine natalensis and B. frutescens fractions (selected relative to the cytotoxicity results of B. natalensis) on the HEp-2 cell line (read cell culture and cytotoxicity discussion for selection of HEp-2 cell line). The validity of this study is confirmed by similar experimental designs that assayed the cytotoxicity of plant-derived or natural compounds on cancer cell lines only, and the detection of apoptosis through caspase- 3 induction and other unrelated methods. This is the first study to report the induction of apoptosis in cancer cell lines by Bulbine spp. fractions using cytotoxicity and the expression of bax and caspase-3 apoptotic markers. It provides insight into the interaction between the HEp-2 cell line and the aqueous and organic fractions of B. natalensis and B. frutescens by analyzing links between cytotoxicity and bax and caspase-3 gene expression; which could probably contribute to drug design with selected Bulbine spp. fractions. Further investigations are required in future, to confirm the possible drug targets of the studied Bulbine spp. fractions in an attempt of assaying their therapeutic importance. / National Research Foundation

Submitted in partial compliance with the requirements for Master’s degree in Technology: Chiropractic, Department of Chiropractic, Durban University of Technology, Durban, South Africa, 2015. / Background Functional dyspepsia is a chronic pain/discomfort centred in the upper abdomen in the absence of any known structural cause. Epidemiological studies have shown that functional dyspepsia is a common complaint affecting all population groups that over time places considerable financial strain on public and private resources due to frequent doctors’ visits and expensive diagnostic procedures. The development of non-surgical and non-pharmaceutical treatments of functional dyspepsia would not only make economic sense but would also provide a means to improve patients’ quality of life in the least invasive way possible. Although not traditionally seen to be within the chiropractic scope of practice, anecdotal evidence suggests that chiropractic care and management may have the ability to alleviate visceral symptomatology. Objectives The purpose of this placebo controlled pilot study was to evaluate the preliminary effects of chiropractic manipulation versus inactive laser in the treatment of adult patients suffering from functional dyspepsia. Due to the small sample size, time and budgetary constraints it was hypothesised that the dyspepsia symptoms of participants treated with active chiropractic manipulation would not respond more favourably to the treatment, nor would these patients experience a greater improvement in terms of quality of life, compared to those participants receiving placebo treatment. Method Thirty participants with pre-diagnosed functional dyspepsia were selected after being screened according to the inclusion and exclusion criteria identified by the researcher. These participants were then divided into two groups using consecutive sampling. Data was collected at the Chiropractic Day Clinic at the Durban University of Technology. Group A received an active chiropractic manipulation using diversified technique to pre-identified levels in the cervical, thoracic and lumbar spine. Group B received inactive laser to pre-identified levels in the cervical, thoracic and lumbar spine. Both groups received one treatment a week for three weeks. The fourth and final consultation consisted only of data capturing. At each visit both groups of participants filled in three validated questionnaires: The numerical pain rating scale; PAGI-SYM physical symptom assessment; Results Results were statistically analysed using IBM SPSS version 20 and a p value ≤ 0.05 was considered to be statistically significant. Repeated measures ANOVA testing was used to assess the effect of each of the treatments separately and to assess the comparative effects of the spinal manipulation vs the placebo. Conclusion and Recommendations The gathered results and analysis were statistically insignificant. Clinical improvement in their symptomotology was however noted within both groups over the trial period in terms of treatment received and their perceived quality of life, symptomatology and pain levels. Both groups tended to have reduced pain and discomfort over time, improved: emotional distress, sleep disturbance, food problems, vitality, post-prandial fullness and abdominal pain. This study should be repeated with selected outcome measurements, and perhaps objective outcome measurements, and a larger sample size in order to determine any benefit.

Submitted in partial compliance with the requirements of the Master’s Degree in Technology: Homoeopathy, Department of Homeopathy, Durban University of Technology, Durban, South Africa, 2016. / Introduction This research study investigated the efficacy of a homoeopathic protocol in the post-surgical wound management of medical male circumcision (MMC) in the KwaZulu-Natal region. The study had 30 male participants from 10 of the 11 district municipalities of the KwaZulu-Natal province. The study consultations took place at the Durban University of Technology Health Sciences Clinic. Aim of the study The aim of the study was to determine the efficacy of a topical application of a homoeopathic protocol using; Calendula officinalis (mother tincture (M.T.)), Hypericum perforatum (M.T.) and Delphinum staphysagria 30CH (orally) as an adjunct to the standard care of the post-surgical effects of circumcision and to assess the wounds post-surgically in terms of time taken to heal and associated signs and symptoms. Methodology The data for this study was collected from 30 male participants who had undergone MMC from various hospitals within KwaZulu-Natal. The participants were aged between 18 to 30 years. All participants underwent a medical examination and were given a pain rating scale and a pain and sleep diary. They were instructed to keep a daily record using the data collection tools mentioned above for the study period of six weeks. Descriptive statistics were employed in the form of tables and graphs. The data analysis methods used in the study are: independent samples t-test and the non-parametric Spearman’s Correlation test. Results Group statistics in relation to ‘time to heal’ showed a statistically significant reduction in the time taken for the group on the homoeopathic protocol. Analysis of ‘time to heal’ by group shows that on average, Group 1 (active group: homoeopathic protocol + standard care) (M = 31.4, SD = 7.49) healed in a significantly shorter time than Group 2 (control group: standard care) (M = 38.6667, SD = 1.99), t (28) = -2.617, p=.014. There was no other statistical significance noted except for the results of the Spearman’s rho, where table 4.8 shows there is a significant positive correlation between average quality sleep until healing and the proportion of days across healing NOT feeling refreshed (rho = .676, p<.0005). / M

published_or_final_version / Chinese Medicine / Master / Master of Philosophy