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An assessment of Hypoxis hemerocallidea extracts, and actives as natural antibiotic, and immune modulation phytotherapies.Muwanga, Catherine January 2006 (has links)
<p>In South Africa, the crude aqueous extract from Hypoxis hemerocallidea is used by AIDS patients to treat opportunistic infections, such as tuberculosis. The rapid emergence of multidrug-resistant tuberculosis, and extreme drug resistant tuberculosis, in recent years, is a major threat to human health. The treatment of TB, nosocomial bacterial infections, and fungal infections is now a clinical challenge, especially in the immuno-compromised individual. There is a dire need for novel antibiotic alternatives with phytotherapies and plant-derived compounds as potentially promising alternatives. The main objective of this study was to investigate the antimycobacterial activity of Hypoxis hemerocallidea, a South African medicinal plant, using Mycobacterium smegmatis.</p>
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An assessment of Hypoxis hemerocallidea extracts, and actives as natural antibiotic, and immune modulation phytotherapies.Muwanga, Catherine January 2006 (has links)
<p>In South Africa, the crude aqueous extract from Hypoxis hemerocallidea is used by AIDS patients to treat opportunistic infections, such as tuberculosis. The rapid emergence of multidrug-resistant tuberculosis, and extreme drug resistant tuberculosis, in recent years, is a major threat to human health. The treatment of TB, nosocomial bacterial infections, and fungal infections is now a clinical challenge, especially in the immuno-compromised individual. There is a dire need for novel antibiotic alternatives with phytotherapies and plant-derived compounds as potentially promising alternatives. The main objective of this study was to investigate the antimycobacterial activity of Hypoxis hemerocallidea, a South African medicinal plant, using Mycobacterium smegmatis.</p>
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In vitro anti-HIV activities of Sutherlandia frutescens and Lobostemon trigonum extractsHarnett, Siobhán Margaret January 2004 (has links)
Currently, the approved anti-HIV drugs on the market only target the three HIV enzymes: reverse transcriptase, protease and more recently, integrase. Due to the limited nature of the current therapy, it is possible that a multi-drug resistant virus can emerge. The main concerns in developing countries however, are the expense and availability of the drugs and because of this, it is essential to investigate all alternatives. Traditional medicine offers many advantages as compared to allopathic treatment in so far as being relatively cheaper, accessible and it is broadly accepted in the population groups of the developing countries. Little is known though, of the exact efficacy and toxicity of these remedies so it is vital that these possible leads be investigated thoroughly. For the purpose of this study, two plants, Sutherlandia frutescens and Lobostemon trigonum were studied to ascertain their potential anti-HIV activity. Sutherlandia has received international attention as a possible cheap herbal remedy to improve the health of AIDS sufferers. Anecdotal evidence from health workers claim that HIV- infected patients on Sutherlandia treatment have shown improved CD4 counts, decreased viral loads and a general improvement in well-being. Extracts were prepared from dried leaves and flowers in methanol, ethanol, acetone, methylene dichloride or distilled water. Sulphated polysaccharides have been described extensively in literature with regards to their anti-HIV activity, so as a form of dereplication; an ethanol precipitation was performed on the aqueous extracts to remove sulphated polysaccharides. A toxicity study was performed on all crude extracts using uninfected peripheral mononuclear blood cells (PBMCs) isolated from whole blood. To measure anti-HIV activity, HIV-infected PBMCs were cultured with each of the crude extracts and cell viability measured using the tetrazolium salt, XTT. HIV-infected CEM-NKR-CCR5 cells were also used and supernatant from the viral studies was tested for the HIV antigen p24. xii Results varied greatly between assays but with the inclusion of a point-scale system to evaluate the extracts it was clear that overall the organic extracts of the Sutherlandia flowers, especially the acetone extract (SFA), showed great anti-HIV potential. SFA in every case decreased p24 levels and in the toxicity study did not decrease cell proliferation. With the HIV-infected PBMCs SFA actually helped improve cell proliferation despite the infection. To determine the specific anti- HIV activity, all crude extracts were tested for inhibition of HIV-I reverse transcriptase, the glycohydrolase enzymes: a-glucosidase, ß-glucosidase, ßglucuronidase, HIV-I integrase and HIV-II protease. No significant inhibition was seen with these experiments except for the HIV-I RT assay. The aqueous extract of the Lobostemon leaves produced an inhibitor of HIV-RT with a very low IC50 value of 0.049mg/ml. Some inhibitory effect was lost with the removal of the sulphated polysaccharides and the addition of BSA to the assay, but still 64% inhibition of the HIVRT remained, which confirmed that the inhibitor could be something novel, and not of the polysaccharide or tannin compounds.
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An assessment of Hypoxis hemerocallidea extracts, and actives as natural antibiotic, and immune modulation phytotherapiesMuwanga, Catherine January 2006 (has links)
Magister Scientiae - MSc / In South Africa, the crude aqueous extract from Hypoxis hemerocallidea is used by AIDS patients to treat opportunistic infections, such as tuberculosis. The rapid emergence of multidrug-resistant tuberculosis, and extreme drug resistant tuberculosis, in recent years, is a major threat to human health. The treatment of TB, nosocomial bacterial infections, and fungal infections is now a clinical challenge, especially in the immuno-compromised individual. There is a dire need for novel antibiotic alternatives with phytotherapies and plant-derived compounds as potentially promising alternatives. The main objective of this study was to investigate the antimycobacterial activity of Hypoxis hemerocallidea, a South African medicinal plant, using Mycobacterium smegmatis. / South Africa
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An investigation into the antibacterial activities of medicinial plants traditionally used in the Eastern Cape to treat secondary skin infections associated with burn woundsWeideman, Liezel January 2005 (has links)
Traditional medicine has a long history of being used for treating various ailments ranging in severity. Although traditional medicine has typically been the health care for the poorest levels of society, there is a worldwide growth in popularity. The growing popularity of traditional medicine, termed the green boom, may be ascribed to people taking a more holistic approach to maintain their health. Traditional medicine is widely used on a regular basis by 70% of South Africans. Various indigenous medicinal plants are used for the preparation of traditional herbal medicine. These plants are mostly indigenous to the regions were it is used. In this study four medicinal plants (Bulbine frutescens, Leonotis leounurus, Melianthus major & Zantedecshia aethiopica) that are traditionally used in the Eastern Cape region for treating burn wound infections, were collected for investigation. The in vitro antibacterial activity of these plants was tested against different bacterial strains of eight different bacteria. The bacteria used in this investigation included bacterial strains of four Gram-positive bacteria, S. aureus, methicillin-resistant S. aureus (MRSA), E. feacalis, S. pyogenes and four Gramnegative bacteria, P. aeruginosa, A. baumanii, K. pneumoniae and P. mirabilis. Traditional preparations as well as three different extracts (methanol, aqueous & acetone) of the plants were used for in vitro antibacterial activity testing. The microtitre plate assay and agar dilution assay were used for determining the antibacterial activity of the traditional preparations and plant extracts against the different bacterial strains. In the microtitre plate assay the antibacterial activity was tested using the bacterial growth indicator, INT and a microtitre plate spectrophotometer to determine the minimal inhibitory concentrations of the plant extracts and traditional preparations. The microtitre plate assay was used for testing the antibacterial activity of the plants against the bacterial strains of five bacteria, S. aureus, MRSA, P. aeruginosa, A. baumanii and K. pneumoniae. The bacterial strains of the three bacteria, S. pyogenes, E. feacalis and P. mirabilis were not compatible with the microtitre plate assay using INT and spectrophotometric readings to determine bacterial inhibition. Therefore the agar dilution assay were used as an alternative method for determining the MIC’s of the plant extracts against the bacterial strains of these bacteria. The initial plant extract concentration in the microtitre plate assay differed with the different plant extracts in the microtitre plate assay. Acetone followed by methanol extracted the highest plant extract concentrations with the different medicinal plants. M. major followed by L. leonurus produced the highest plant extract concentrations following extraction with the different extraction solvents. Consequently the acetone extract of M. major had the highest plant extract concentration before serial dilution in the microtitre plate assay. Uniform plant extract concentrations were tested in the agar dilution assay. The methanol extract followed by the acetone extract of the plants gave the highest antibacterial activity against the different bacterial strains. The extracts of M. major followed by L. leonurus inhibited the highest number of bacterial strains in the microtitre plate assay and the extracts of B. frutescens inhibited the lowest number of bacterial strains. The acetone and methanol extracts of M. major were the only extracts that displayed antibacterial activity in the agar dilution assay. The bacterial strains of P. mirabilis were the only bacteria that were inhibited using this method. The bacterial strains of S. pyogenes and E. feacalis were not inhibited at any of the plant extract concentrations in the agar dilution assay.
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Antimicrobial activity of selected Eastern Cape medical plantsMohlakoana, Keneuoe January 2010 (has links)
Bacterial resistance to antibiotics has been a great problem for many years. The degree of resistance and the speed with which resistance develops varies with different organisms and different drugs. Enzymes called β-lactamases are produced by bacteria and are one mechanism in which bacteria develop antimicrobial resistance. Gram-negative bacteria producing enzymes called ESBLs because of their wide substrate range are of a particular concern in nosocomial infections. In many countries people still use traditional medicine derived from plants as an alternative to the Western medicine due to increased cost of Western medicine and microbial resistance of antibiotic treatments. Biologically active compounds isolated from plants species are used in herbal medicine. Because of the high prevalence of the ESBLs and their increasing resistance to the antibiotics, this research study was done to test the antimicrobial activities of selected medicinal plants of the Eastern Cape; G. incanum, D. angustifolia and E. autumnalis which were traditionally used to treat various infections. The in vitro antimicrobial activity of three different extracts (acetone, methanol & distilled water) and the traditional preparations of the three plants were tested against the selected strains of ESBL-producing bacteria, non β-lactamase producers and the different fungal species. The extracts were screened against 26 Gram-positive bacterial strains, 53 Gram-negative bacterial strains and 15 fungal strains. The Gram-positive bacteria included strains from S. aureus, B. cereus and E. faecalis. The Gram-negative bacteria included strains from E. ii coli, E. cloacae, K. pneumoniae, P. aeruginosa and Acinetobacter spp. The fungal strains included 9 strains of Candida albicans and a single strain of each of the following opportunistic fungi, Mucor sp, Geotrichium sp, Penicillium sp, Fusarium sp and Rhizopus sp. The agar dilution assay was used for the antimicrobial screening of the plants extracts and for the determination of the MICs. The Ames test was performed for the determination of probable carcinogenicity of the extracts of G. incanum and D. angustifolia. The distilled water extracts followed by acetone extracts of the plants revealed the highest antimicrobial activity against the different microbial strains. The extracts of G. incanum followed by the extracts of D. angustifolia inhibited the highest number of microbial strains. The extracts of E. autumnalis did not show any antimicrobial activity against all the pathogens in this study. More of the Gram-positive bacteria were inhibited by the plant extracts. The lowest MIC was obtained with Gram-positive bacteria. The bacterial strains of E. faecalis and P. aeruginosa were not inhibited by any of the plants extracts in the agar dilution assay yet Acinetobacter species which are MDR were inhibited by the distilled water and methanol extracts of G. incanum. A single strain of Mucor sp was the only spore forming fungi that was inhibited by the distilled water extracts of G. incanum. None of the plants extracts showed any mutagenic effects on the TA100 S. typhimurium strains incorporated on the Ames test. Apart from revealing of new antimicrobial agents that may be used against resistant organisms, the proper use of antimicrobial agents should be recommended. The study has highlighted a need for further investigations on the properties of the medicinal plants used in this study.
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Optimisation of an in vitro model for anti-diabetic screeningWilson, Gayle Pamela January 2006 (has links)
The need for alternative strategies for the prevention and treatment of diabetes is growing rapidly as type II diabetes is reaching epidemic status in our society. This need was the basis for the creation of this study, as it was necessary to start looking towards medicinal plants as potential antidiabetic treatment and no comprehensive in vitro model existed. In creating a model for determining the effects of alternative traditional medicines as antidiabetic potentiates, it was necessary that two metabolic pathways, namely glucose uptake and insulin secretion, which play a significant role in glucose homeostasis, be at the centre of our investigations. The objective of this project was to optimize the methodology required to screen and ultimately determine the effectiveness of the plant extracts Kankerbos and MRC2003, as antidiabetic potentiates, through observing their effects on glucose utilisation and insulin secretion. If these medicinal plants are going to make a positive contribution to the health of type II diabetic South Africans, then the determination of their efficacy is essential. The cell lines used in this study included 3T3-L1 preadipocytes, Chang liver, C2C12 muscle and INS-1 rat pancreatic cells. Each cell line represents a different in vivo organ that is known to have an influence on glucose homeostasis in our bodies, each with its own unique metabolic pathways and mechanisms of activity, thereby making each one a vital component in the study. The positive controls for the two models were insulin and metformin (glucose utilisation) and glibenclamide (insulin secretion). Insulin was shown to provide a significant increase in the amount of glucose taken up in C2C12 muscle and Chang liver cells for acute conditions. Chronic treatments with metformin provided a significant increase in glucose utilised by Chang liver cells. Glibenclamide was an effective positive control for stimulating insulin secretion by INS-1 cells under acute conditions as there was a significant increase in the amount of insulin secreted. MRC2003 did not show any significant antidiabetic activity. Sutherlandia frutescens (Kankerbos) showed biological activities comparable to some of the more recognized antidiabetic compounds throughout the study. With regards to the glucose utilisation model, Kankerbos was seen to have both acute and chronic effects in different cell lines. In the C2C12 muscle cell line, Kankerbos significantly increased glucose uptake when they were exposed to acute conditions. Kankerbos also had a significant effect on the Chang liver cells as it was observed that under both acute and chronic conditions, this plant extract induced the uptake of glucose into these cells. With respect to the insulin secretion model involving INS-1 cells, no significant effect was seen during acute exposure with Kankerbos treatment. However during chronic exposure, an increase in insulin secretion was initiated by this plant extract. Overall, the results of this study suggest that Kankerbos has a twofold mechanism of action for its glucose-lowering effects. Given that Kankerbos is widely available in South Africa, this study was valuable as it provided an indication that Kankerbos has antidiabetic activities and could possibly be used as an alternative antidiabetic medication.
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Production of biologically active recombinant HIV-1 protease and intehrase for the purpose of screening medicianl plant extractsBosch, Janine January 2009 (has links)
Human immunodeficiency virus (HIV) and its gradual weakening of the immune system is an ever growing threat. Acquired immune deficiency syndrome (AIDS), the final stage of HIV, renders a person vulnerable to various opportunistic infections, which in the end lead to death. Apart from intensive vaccine studies, treatment research mainly focuses on preventing the individual HIV enzymes (reverse transcriptase, integrase and protease) from performing their functions. Entry inhibitors, however, block viral entry into the cell, while antisense drugs lock onto the viral genome to keep it from functioning. In this study production of active recombinant HIV-1 protease and integrase was attempted for future drug screening programs. HIV-1 protease was cloned into a pET28b(+) vector and expressed in ROSETTA(DE3)pLysS cells. The protein was purified using a nickel-affinity column utilizing the hexa-histidine tag encoded by the vector. Gel filtration chromatography was attempted after refolding of the protease, but protease yield seemed to decrease with the additional purification step. Partially purified protease was characterized with kinetic studies. Kinetic parameters of HIV-1 protease were determined to be Km = 592 μM, Vmax = 0.59 μM/min and kcat = 31 s-1. HIV-1 integrase, which was cloned into a pET15b vector, was expressed in E. coli BL21(DE3) cells. The coding sequence had been mutated to introduce the amino acid substitutions F185K and C280S, increasing solubility of the protein. The first step in purification of this protein was nickel-affinity chromatography, after which cation exchange chromatography was attempted. HIV-1 integrase concentration was low throughout experiments and no clear elution from the cation exchange column could be observed. A non-radioactive enzyme linked HIV-1 integrase assay failed to detect integrase activity. Modifications to future studies of the integrase are suggested in the chapter involved.
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In vitro testing to investigate the anticoagulant/antithrombotic and antidiabetic biological activity of Leonotis LeonurusMnonopi, Nandipha Olivia January 2007 (has links)
The rising costs of prescription drugs in the maintenance of personal health and wellbeing have increased the interest in medicinal plants. The World Health Organization estimates that 65 percent-80 percent of the world’s population use traditional medicine as their primary form of health care. In this project the focus has been on the use of Leonotis leonurus extracts as a traditional medicine. The major chemical constituent of this plant is marrubiin, which is a diterpenoid labdane lactone formed from a precursor called premarrubiin. Aqueous and acetone extract (AL and OL extract, respectively) of this plant has been found to have an antithrombotic effect, with IC50 values of 3mg/ml and 6mg/ml, respectively. The extracts also have an effect on fibrinolysis, where the lysis time was decreased by more than 50 percent by the organic extract and standard marrubiin. In whole blood ADP-induced platelet aggregation, the organic extract inhibited aggregation by 68 percent at a final concentration of 138μg/ml (equivalent to 7.2μg/ml marrubiin). Marrubiin has also been screened for antithrombotic/anticoagulant activity; no antithrombotic activity has been observed but it increased the rate of fibrinolysis, by decreasing lysis time by 64 percent and also decreasing fibrin formation. From these findings it can be concluded that marrubiin has a fibrinolytic effect and antiplatelet aggregation effect. In the diabetic studies, in hyperglycemic condition, the OL (10μg/ml) extract and standard marrubiin significantly increased insulin secretion by 200 percent (2-fold) and 400 percent (4-fold), respectively, with respect to the control. The OL extract and standard marrubiin stimulated the release of insulin, the stimulatory index was significantly increased by 450 percent (4.5-fold) and 500 percent (5-fold), respectively, with respect to the control. In the apoptotic studies, in the normoglycemic and hyperglycemic conditions, the OL extract decreased the occurrence of apoptosis, in a dose-dependent manner, with the lower concentrations inducing apoptosis significantly higher than the relevant controls. Standard marrubiin did not have an effect on apoptosis in hyperglycemic condition, but it decreased the occurrence of apoptosis by 200 percent (2-fold) under normoglycemic conditions. The OL extract increased proliferation by 148 percent (1.48- fold) and 155 percent (1.55-fold) in normoglycemic and hyperglycemic conditions, respectively. The same effect was observed for standard marrubiin, where, proliferation was increased by 180 percent (1.8-fold) and 200 percent (2.0-fold) in normoglycemic and hyperglycemic conditions, respectively. RT-PCR displayed that standard marrubiin inhibited the expression of insulin by 50 percent under normoglycemic conditions.
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The extraction, purification and evaluation of compounds from the leaves of Leonotis Leonorus for anticonvulsant activity.Muhizi, Thèoneste January 2002 (has links)
The aim of this study is to isolate and evaluate the anticonvulsant components from the leaves of Leonotis leonorus (L) R.aR. and to see if there is any change in activity with the origin of the plant material and I or the season in which plant material is collected. Therefore, in this study, two sites were chosen for collection of plant material and the collection was made in summer and in winter. Chemical, physical and pharmacological methods were used to isolate, identify and to evaluate compounds isolated from the leaves of Leonotis leonorus for anticonvulsant activity.
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