Chang liver cell line as a model for Type II Diabetes in the liver and possible reversal of this condition by an indigenous medicinal plant
- Authors: Williams, Saralene Iona
- Date: 2009
- Subjects: Diabetes -- Alternative treatment , Medicinal plants , Traditional medicine , Liver -- Diseases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10339 , http://hdl.handle.net/10948/d1016179
- Description: The incidence of Type 2 Diabetes Mellittus (T2DM) is increasing world wide. In Africa the limited access to health care and the insidious course of the disease lead to more severe illness and diabetic complications. There is a need to find alternative approaches to treatment and prevention that address the problems and needs of Africa. Sutherlandia frutescens (S.frutescens) is a traditional herbal plant with known anti-diabetic properties, the precise mechanism of action of S.frutescens is not known. In order to develop new approaches for treatment and prevention of T2DM the pathophysiology of T2DM must be understood. T2DM is the final outcome of a multi-organ disease characterized by early defects in muscle, adipocytes, hepatocytes and pancreatic β-cells. In this study the role of the liver was investigated because of its central role in glucose and lipid metabolism. It is hard to differentiate between all the influences in an in vivo model, so the aim of this study was to develop an in vitro model of T2DM in Chang liver cells and to determine if S.frutescens can reverse the state of insulin resistance in this model. Different culture media conditions were screened to identify a method that can be used as the T2DM model in Chang liver cells. Serum free medium (MCBD-201) supplemented with human diabetic serum, (2.5%-10%), high insulin concentrations (0.1μM-1μM), high fructose concentrations (1-10mM). and a combination of high insulin and high fructose was used for this screening. Chang liver cells cultured in MCBD-201 medium supplemented with 1mM fructose and 0.1μM insulin showed reduced glucose uptake and increased lipid accumulation. The effect of two S.frutescens extracts, two anti-diabetic drugs, metformin and ciglitazone, and a hypolipidemic drug ciprofibrate were determined and shown to increase glucose uptake and reduce lipid accumulation. It was postulated that exposing the cells to excess nutrients in the form of high fructose would stimulate the cells to become adipogenic and accumulate lipids, which would interfere with the glucose uptake and induce insulin resistance. Gene expression of PPARγ, PPARα, and SREBP-1 transcription factors regulating lipid metabolism was determined in Chang liver cells cultured in insulin resistance inducing medium over a 48 hour time course. The expression of PPARγ, known to stimulate adipogenesis was increased after 6, 24 and 48 hours of exposure (P(H1)<0.0001). The expression of PPARα, known to stimulate β-oxidation expression, was significantly decreased after 24 hours of exposure (P(H1)<0.0001). The presence of the plant extracts in the insulin resistance inducing media protect against this increase in adipogenesis and decrease in β-oxidation after 48 hours of exposure by increasing PPARα expression and decreasing PPARγ expression. A PCR Array was performed which identified 32 more potential molecular targets of S.frutescens. Five of the 32 targets identified with the PCR Array were validated using qRT-PCR. These genes play a role in lipid and glucose metabolism and protection against oxidative stress and inflammation. In summary a cellular model of insulin resistace in hepatocytes has been established and the capacity of S.frutescens to reverse this process has been demonstrated by acting as a dual PPARγ/α agonist. New genes have been identified in the development of insulin resistance and as targets of S.frutescens.
- Full Text:
- Date Issued: 2009
- Authors: Williams, Saralene Iona
- Date: 2009
- Subjects: Diabetes -- Alternative treatment , Medicinal plants , Traditional medicine , Liver -- Diseases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10339 , http://hdl.handle.net/10948/d1016179
- Description: The incidence of Type 2 Diabetes Mellittus (T2DM) is increasing world wide. In Africa the limited access to health care and the insidious course of the disease lead to more severe illness and diabetic complications. There is a need to find alternative approaches to treatment and prevention that address the problems and needs of Africa. Sutherlandia frutescens (S.frutescens) is a traditional herbal plant with known anti-diabetic properties, the precise mechanism of action of S.frutescens is not known. In order to develop new approaches for treatment and prevention of T2DM the pathophysiology of T2DM must be understood. T2DM is the final outcome of a multi-organ disease characterized by early defects in muscle, adipocytes, hepatocytes and pancreatic β-cells. In this study the role of the liver was investigated because of its central role in glucose and lipid metabolism. It is hard to differentiate between all the influences in an in vivo model, so the aim of this study was to develop an in vitro model of T2DM in Chang liver cells and to determine if S.frutescens can reverse the state of insulin resistance in this model. Different culture media conditions were screened to identify a method that can be used as the T2DM model in Chang liver cells. Serum free medium (MCBD-201) supplemented with human diabetic serum, (2.5%-10%), high insulin concentrations (0.1μM-1μM), high fructose concentrations (1-10mM). and a combination of high insulin and high fructose was used for this screening. Chang liver cells cultured in MCBD-201 medium supplemented with 1mM fructose and 0.1μM insulin showed reduced glucose uptake and increased lipid accumulation. The effect of two S.frutescens extracts, two anti-diabetic drugs, metformin and ciglitazone, and a hypolipidemic drug ciprofibrate were determined and shown to increase glucose uptake and reduce lipid accumulation. It was postulated that exposing the cells to excess nutrients in the form of high fructose would stimulate the cells to become adipogenic and accumulate lipids, which would interfere with the glucose uptake and induce insulin resistance. Gene expression of PPARγ, PPARα, and SREBP-1 transcription factors regulating lipid metabolism was determined in Chang liver cells cultured in insulin resistance inducing medium over a 48 hour time course. The expression of PPARγ, known to stimulate adipogenesis was increased after 6, 24 and 48 hours of exposure (P(H1)<0.0001). The expression of PPARα, known to stimulate β-oxidation expression, was significantly decreased after 24 hours of exposure (P(H1)<0.0001). The presence of the plant extracts in the insulin resistance inducing media protect against this increase in adipogenesis and decrease in β-oxidation after 48 hours of exposure by increasing PPARα expression and decreasing PPARγ expression. A PCR Array was performed which identified 32 more potential molecular targets of S.frutescens. Five of the 32 targets identified with the PCR Array were validated using qRT-PCR. These genes play a role in lipid and glucose metabolism and protection against oxidative stress and inflammation. In summary a cellular model of insulin resistace in hepatocytes has been established and the capacity of S.frutescens to reverse this process has been demonstrated by acting as a dual PPARγ/α agonist. New genes have been identified in the development of insulin resistance and as targets of S.frutescens.
- Full Text:
- Date Issued: 2009
The effect of tulbaghia violacea plant extract on the growth of aspergillus species
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2009
- Subjects: Plant products , Plant extracts , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10315 , http://hdl.handle.net/10948/d1008186 , Plant products , Plant extracts , Traditional medicine
- Description: Traditional medicine has become an important part of healthcare worldwide. It is estimated that about 25 percent of prescribed medicines contain plant products or active compounds derived from plants. In South Africa, traditional medicine forms part of the culture and tradition of most communities. Garlic compounds have been shown to have a variety of antimicrobial properties. Amongst these are antifungal, antibacterial, antiviral and anti protozoal activities. Allicin and its breakdown products have been shown to be the main active compounds which possess these properties. Tulbaghia violacea has been used for the treatment of a variety of illnesses including asthma, fever, oesophageal cancer, constipation and hypertension. This study investigated the antifungal nature of T.violacea on the morphology, spore germination and lipid synthesis of Aspergillus flavus and Aspergillus parasiticus. The results of this study showed that the plant extract inhibited A. flavus growth at a minimal inhibitory concentration of 15mg/ml and was fungicidal at 20mg/ml and above. A. parasiticus was not inhibited at 25mg/ml indicating resistance to the inhibitory component of the plant extract. A measure of metabolic activity using the XTT assay showed reduced metabolic activity in the presence of increasing concentrations of the plant extract. Higher extract concentrations resulted in higher percentage inhibition of fungal growth for both fungal species with up to 98 percent inhibition being observed for the highest extract concentrations for both fungi. Germination was also delayed in the presence of 15mg/ml plant extract concentration by up to 60hr for A. flavus and 48hr for A. parasititcus. The TEM results showed increased thickening of the cell wall with higher extract concentrations. The thickening was greater for A. flavus than for A. parasiticus. Cell wall thickening may be the reason for the delay in germination in both species. Lipid production was reduced in the presence of plant extracts when compared to the control. The plant extracts inhibited triglyceride production at 15mg/ml for both A. flavus and A. parasiticus. The results therefore indicate that T. violacea extracts are antifungal and probably affect germination through interactions with the cell wall. It is possible that the extract affects lipid production in Aspergillus species.
- Full Text:
- Date Issued: 2009
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2009
- Subjects: Plant products , Plant extracts , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10315 , http://hdl.handle.net/10948/d1008186 , Plant products , Plant extracts , Traditional medicine
- Description: Traditional medicine has become an important part of healthcare worldwide. It is estimated that about 25 percent of prescribed medicines contain plant products or active compounds derived from plants. In South Africa, traditional medicine forms part of the culture and tradition of most communities. Garlic compounds have been shown to have a variety of antimicrobial properties. Amongst these are antifungal, antibacterial, antiviral and anti protozoal activities. Allicin and its breakdown products have been shown to be the main active compounds which possess these properties. Tulbaghia violacea has been used for the treatment of a variety of illnesses including asthma, fever, oesophageal cancer, constipation and hypertension. This study investigated the antifungal nature of T.violacea on the morphology, spore germination and lipid synthesis of Aspergillus flavus and Aspergillus parasiticus. The results of this study showed that the plant extract inhibited A. flavus growth at a minimal inhibitory concentration of 15mg/ml and was fungicidal at 20mg/ml and above. A. parasiticus was not inhibited at 25mg/ml indicating resistance to the inhibitory component of the plant extract. A measure of metabolic activity using the XTT assay showed reduced metabolic activity in the presence of increasing concentrations of the plant extract. Higher extract concentrations resulted in higher percentage inhibition of fungal growth for both fungal species with up to 98 percent inhibition being observed for the highest extract concentrations for both fungi. Germination was also delayed in the presence of 15mg/ml plant extract concentration by up to 60hr for A. flavus and 48hr for A. parasititcus. The TEM results showed increased thickening of the cell wall with higher extract concentrations. The thickening was greater for A. flavus than for A. parasiticus. Cell wall thickening may be the reason for the delay in germination in both species. Lipid production was reduced in the presence of plant extracts when compared to the control. The plant extracts inhibited triglyceride production at 15mg/ml for both A. flavus and A. parasiticus. The results therefore indicate that T. violacea extracts are antifungal and probably affect germination through interactions with the cell wall. It is possible that the extract affects lipid production in Aspergillus species.
- Full Text:
- Date Issued: 2009
African traditional medicine-antiretroviral interactions : effects of Sutherlandia frutescens on the pharmacokinetics of Atazanavir
- Authors: Müller, Adrienne Carmel
- Date: 2011 , 2011-03-28
- Subjects: Antiretroviral agents , Medicinal plants , Traditional medicine , AIDS (Disease) -- Treatment , HIV infections -- Drug therapy , Drug interactions , Pharmacokinetics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3859 , http://hdl.handle.net/10962/d1013373
- Description: In response to the urgent call for investigations into antiretroviral (ARV)-African traditional medicine (ATM) interactions, this research was undertaken to ascertain whether chronic administration of the ATM, Sutherlandia frutescens (SF) may alter the bioavailability of the protease inhibitor (PI), atazanavir (ATV), which may impact on the safety or efficacy of the ARV. Prior to investigating a potential interaction between ATV and SF in vitro and in vivo, a high performance liquid chromatography method with ultraviolet detection (HPLC-UV) was developed and validated for the bioanalysis of ATV in human plasma and liver microsomes. An improved and efficient analytical method with minimal use of solvents and short run time was achieved in comparison to methods published in the literature. In addition, the method was selective, linear, accurate and precise for quantitative analysis of ATV in these studies. Molecular docking studies were conducted to compare the binding modes and affinities of ATV and two major SF constituents, Sutherlandioside B and Sutherlandin C, with the efflux transporter, P-glycoprotein (P-gp) and the CYP450 isoenzyme, CYP3A4 to determine the potential for these phytochemicals to competitively inhibit the binding of ATV to these two proteins, which are mediators of absorption and metabolism. These studies revealed that modulation of P-gp transport of ATV by Sutherlandioside B and Sutherlandin C was not likely to occur via competitive inhibition. The results further indicated that weak competitive inhibition of CYP3A4 may possibly occur in the presence of either of these two SF constituents. The Caco-2 cell line was used as an in vitro model of human intestinal absorption. Accumulation studies in these cells were conducted to ascertain whether extracts and constituents of SF have the ability to alter the absorption of ATV. The results showed that the aqueous extract of SF significantly reduced ATV accumulation, suggesting decreased ATV absorption, whilst a triterpenoid glycoside fraction isolated from SF exhibited an opposing effect. Analogous responses were elicited by the aqueous extract and a triterpenoid glycoside fraction in similar accumulation studies in P-gp overexpressing Madin–Darby Canine Kidney Strain II cells (MDCKII-MDR1), which signified that the effects of this extract and component on ATV transport in the Caco-2 cells were P-gp-mediated. The quantitative analysis of ATV in human liver microsomes after co-incubation with extracts and components of SF was conducted to determine the effects of SF on the metabolism of ATV. The aqueous and methanolic extracts of SF inhibited ATV metabolism, whilst the triterpenoid glycoside fraction had a converse effect. Analogous effects by the extracts were demonstrated in experiments conducted in CYP3A4-transfected microsomes, suggesting that the inhibition of ATV metabolism in the liver microsomes by these SF extracts was CYP3A4-mediated. A combination of Sutherlandiosides C and D also inhibited CYP3A4-mediated ATV metabolism, which was in contrast to the response elicited by the triterpenoid fraction in the liver microsomes, where other unidentified compounds, shown to be present therein, may have contributed to the activation of ATV metabolism. The in vitro studies revealed the potential for SF to alter the bioavailability of ATV, therefore a clinical study in which the effect of a multiple dose regimen of SF on the pharmacokinetics (PK) of a single dose of ATV was conducted in healthy male volunteers. The statistical analysis showed that the 90 % confidence intervals around the geometric mean ratios (ATV + SF/ATV alone) for both Cmax and AUC0-24 hours, fell well below the lower limit of the "no-effect" boundary of 0.8 – 1.25, implying that the bioavailability of ATV was significantly reduced in this cohort of subjects. It may thus be concluded that if the reduction in bioavailability observed in this clinical study is found to be clinically relevant, co-administration of SF commercial dosage forms and ATV in HIV/AIDS patients may potentially result in subtherapeutic ATV levels, which may in turn contribute to ATV resistance and/or treatment failure. This research has therefore highlighted the potential risk for toxicity or lack of efficacy of ARV regimens which may result when ATMs and PIs are used concurrently and that patients and health care practitioners alike should be aware of these perils.
- Full Text:
- Date Issued: 2011
- Authors: Müller, Adrienne Carmel
- Date: 2011 , 2011-03-28
- Subjects: Antiretroviral agents , Medicinal plants , Traditional medicine , AIDS (Disease) -- Treatment , HIV infections -- Drug therapy , Drug interactions , Pharmacokinetics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3859 , http://hdl.handle.net/10962/d1013373
- Description: In response to the urgent call for investigations into antiretroviral (ARV)-African traditional medicine (ATM) interactions, this research was undertaken to ascertain whether chronic administration of the ATM, Sutherlandia frutescens (SF) may alter the bioavailability of the protease inhibitor (PI), atazanavir (ATV), which may impact on the safety or efficacy of the ARV. Prior to investigating a potential interaction between ATV and SF in vitro and in vivo, a high performance liquid chromatography method with ultraviolet detection (HPLC-UV) was developed and validated for the bioanalysis of ATV in human plasma and liver microsomes. An improved and efficient analytical method with minimal use of solvents and short run time was achieved in comparison to methods published in the literature. In addition, the method was selective, linear, accurate and precise for quantitative analysis of ATV in these studies. Molecular docking studies were conducted to compare the binding modes and affinities of ATV and two major SF constituents, Sutherlandioside B and Sutherlandin C, with the efflux transporter, P-glycoprotein (P-gp) and the CYP450 isoenzyme, CYP3A4 to determine the potential for these phytochemicals to competitively inhibit the binding of ATV to these two proteins, which are mediators of absorption and metabolism. These studies revealed that modulation of P-gp transport of ATV by Sutherlandioside B and Sutherlandin C was not likely to occur via competitive inhibition. The results further indicated that weak competitive inhibition of CYP3A4 may possibly occur in the presence of either of these two SF constituents. The Caco-2 cell line was used as an in vitro model of human intestinal absorption. Accumulation studies in these cells were conducted to ascertain whether extracts and constituents of SF have the ability to alter the absorption of ATV. The results showed that the aqueous extract of SF significantly reduced ATV accumulation, suggesting decreased ATV absorption, whilst a triterpenoid glycoside fraction isolated from SF exhibited an opposing effect. Analogous responses were elicited by the aqueous extract and a triterpenoid glycoside fraction in similar accumulation studies in P-gp overexpressing Madin–Darby Canine Kidney Strain II cells (MDCKII-MDR1), which signified that the effects of this extract and component on ATV transport in the Caco-2 cells were P-gp-mediated. The quantitative analysis of ATV in human liver microsomes after co-incubation with extracts and components of SF was conducted to determine the effects of SF on the metabolism of ATV. The aqueous and methanolic extracts of SF inhibited ATV metabolism, whilst the triterpenoid glycoside fraction had a converse effect. Analogous effects by the extracts were demonstrated in experiments conducted in CYP3A4-transfected microsomes, suggesting that the inhibition of ATV metabolism in the liver microsomes by these SF extracts was CYP3A4-mediated. A combination of Sutherlandiosides C and D also inhibited CYP3A4-mediated ATV metabolism, which was in contrast to the response elicited by the triterpenoid fraction in the liver microsomes, where other unidentified compounds, shown to be present therein, may have contributed to the activation of ATV metabolism. The in vitro studies revealed the potential for SF to alter the bioavailability of ATV, therefore a clinical study in which the effect of a multiple dose regimen of SF on the pharmacokinetics (PK) of a single dose of ATV was conducted in healthy male volunteers. The statistical analysis showed that the 90 % confidence intervals around the geometric mean ratios (ATV + SF/ATV alone) for both Cmax and AUC0-24 hours, fell well below the lower limit of the "no-effect" boundary of 0.8 – 1.25, implying that the bioavailability of ATV was significantly reduced in this cohort of subjects. It may thus be concluded that if the reduction in bioavailability observed in this clinical study is found to be clinically relevant, co-administration of SF commercial dosage forms and ATV in HIV/AIDS patients may potentially result in subtherapeutic ATV levels, which may in turn contribute to ATV resistance and/or treatment failure. This research has therefore highlighted the potential risk for toxicity or lack of efficacy of ARV regimens which may result when ATMs and PIs are used concurrently and that patients and health care practitioners alike should be aware of these perils.
- Full Text:
- Date Issued: 2011
In vitro cytotoxic effects of selected Nigerian medicinal plant extracts on cancer cell lines
- Authors: Baatjies, Lucinda
- Date: 2012
- Subjects: Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10316 , http://hdl.handle.net/10948/d1008191 , Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Description: Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.
- Full Text:
- Date Issued: 2012
- Authors: Baatjies, Lucinda
- Date: 2012
- Subjects: Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10316 , http://hdl.handle.net/10948/d1008191 , Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Description: Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.
- Full Text:
- Date Issued: 2012
Inhibitory potential of honey on the enzymatic activity of Helicobacter pylori urease
- Authors: Matongo, Fredrick
- Date: 2012
- Subjects: Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11253 , http://hdl.handle.net/10353/431 , Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Description: Urease of Helicobacter pylori is an important virulence factor implicated in the pathogenesis of many clinical conditions, such as chronic gastritis, peptic ulceration, and gastric cancer. Many urease inhibitors have been discovered, like phosphorodiamidates, hydroxamic acid derivatives, and imidazoles. Despite good activities at the enzyme level and excellent kinetic properties most of them have not been used as therapeutic agents in vivo because of their side effects, toxicity and instability. This has led to much attention to focus on exploring the novel urease inhibitory activities of natural products because of their low toxicity and good bioavailability. Honey, a natural product has been used in folk medicine due to its antitumor, antioxidant, antimicrobial and anti-inflammatory properties. The aims of this study were to isolate, characterise, purify urease produced by H. pylori and investigate the inhibitory effects of solvent honey extracts on its enzymatic activity. Urease was found to be both surface-associated and cytoplasmic. Maximum cytoplasmic urease activity was found to occur after 72 hr whereas maximum extracellular urease activities were found to occur after 96 hr. Characterization of the crude cytoplasmic urease revealed optimal activity at a pH of 7.5 and temperature of 40°C. The kinetic parameters Vmax and Km were 45.32 U ml-1 and 61.11 mM respectively.The honey extracts inhibited the activity of the crude urease in a concentration dependent manner. The Lineweaver-Burk plots indicated a non-competitive type of inhibition against H. pylori urease. The two honey extracts gave promising inhibitory activities against urease of H. pylori. Thus the results of this study delineates that inhibition of urease can ease development in therapeutic and preventative approaches based on the enzymatic activity of this Helicobacter protein.
- Full Text:
- Date Issued: 2012
- Authors: Matongo, Fredrick
- Date: 2012
- Subjects: Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11253 , http://hdl.handle.net/10353/431 , Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Description: Urease of Helicobacter pylori is an important virulence factor implicated in the pathogenesis of many clinical conditions, such as chronic gastritis, peptic ulceration, and gastric cancer. Many urease inhibitors have been discovered, like phosphorodiamidates, hydroxamic acid derivatives, and imidazoles. Despite good activities at the enzyme level and excellent kinetic properties most of them have not been used as therapeutic agents in vivo because of their side effects, toxicity and instability. This has led to much attention to focus on exploring the novel urease inhibitory activities of natural products because of their low toxicity and good bioavailability. Honey, a natural product has been used in folk medicine due to its antitumor, antioxidant, antimicrobial and anti-inflammatory properties. The aims of this study were to isolate, characterise, purify urease produced by H. pylori and investigate the inhibitory effects of solvent honey extracts on its enzymatic activity. Urease was found to be both surface-associated and cytoplasmic. Maximum cytoplasmic urease activity was found to occur after 72 hr whereas maximum extracellular urease activities were found to occur after 96 hr. Characterization of the crude cytoplasmic urease revealed optimal activity at a pH of 7.5 and temperature of 40°C. The kinetic parameters Vmax and Km were 45.32 U ml-1 and 61.11 mM respectively.The honey extracts inhibited the activity of the crude urease in a concentration dependent manner. The Lineweaver-Burk plots indicated a non-competitive type of inhibition against H. pylori urease. The two honey extracts gave promising inhibitory activities against urease of H. pylori. Thus the results of this study delineates that inhibition of urease can ease development in therapeutic and preventative approaches based on the enzymatic activity of this Helicobacter protein.
- Full Text:
- Date Issued: 2012
Phytochemical analysis and bioactivity of Garcinia Kola (Heckel) seeds on selected bacterial pathogens
- Seanego, Christinah Tshephisho
- Authors: Seanego, Christinah Tshephisho
- Date: 2012
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11259 , http://hdl.handle.net/10353/420 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Description: Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
- Full Text:
- Date Issued: 2012
- Authors: Seanego, Christinah Tshephisho
- Date: 2012
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11259 , http://hdl.handle.net/10353/420 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Description: Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
- Full Text:
- Date Issued: 2012
Incidence of Staphylococcus species in bovine milk: their antimicrobial sensitivity in selected antibiotics and Usnea barbata lichen extracts
- Idamokoro, Emrobowansan Monday
- Authors: Idamokoro, Emrobowansan Monday
- Date: 2013
- Subjects: Staphylococcus , Microbial sensitivity tests , Traditional medicine , Lichens -- Analysis , Mastitis , Antibiotics in agriculture
- Language: English
- Type: Thesis , Masters , MSc Agric (Animal Science)
- Identifier: vital:11829 , http://hdl.handle.net/10353/d1016193 , Staphylococcus , Microbial sensitivity tests , Traditional medicine , Lichens -- Analysis , Mastitis , Antibiotics in agriculture
- Description: This study was done in order to assess the incidence of Staphylococcus species from milk of cows with subclinical mastitis and their antimicrobial sensitivity in some selected antibiotics and Usnea barbata lichen extracts. The study was conducted in two different commercial dairy farms. Staphylococcus species isolates were identified using several biochemical tests which included Gram’s staining test, catalase test and oxidase test. A commercial API® staph kit (bioMerieux, France) was used to confirm the bacterial organisms to their species level. The antimicrobial sensitivity of individual species was determined according to the Clinical Laboratory Standard Institute (CLSI) for the selected antibiotics. Agar well diffusion method and the broth micro-dilution technique were used to determine the sensitivity of Staphylococcus species in U. barbata extracts. A total of 467 milk samples were screened for bacterial identification from the two farms. Fifteen different Staphylococcus species isolates were identified from all milk samples that were examined. The most frequently isolated species included Staphylococcus xylosus (54.34%), Staphylococcus hominis (24.78%), Staphylococcus aureus (16.38%), Staphylococcus saprophyticus (16.12%) and Staphylococcus haemolyticus (11.63%). Most Staphylococcus species were resistant to Penicillin (75.35%), Nalidixic acid (72.55%) and Ampicillin (63%). Furthermore, the bacterial sensitivity evaluation of U. barbata lichen extracted with methanol and ethyl-acetate against selected Staphylococcus species isolates showed 92.31% and 53.85% susceptibility, respectively. The minimum inhibitory concentration (MIC) of the methanol and ethyl-acetate extracts ranged between 0.0390 to 10 mg/ml. There was a relatively high incidence of Staphylococcus species identified in milk of cows with subclinical mastitis from both farms. Conversely, Staphylococcus species isolates were resistant to antibiotics (mostly penicillin and ampicillin) commonly used in the farms. Furthermore, the study investigated the antimicrobial sensitivity of U. barbata extract in-vitro which may validate its use in traditional medicine for treatment of cows with mastitis.
- Full Text:
- Date Issued: 2013
- Authors: Idamokoro, Emrobowansan Monday
- Date: 2013
- Subjects: Staphylococcus , Microbial sensitivity tests , Traditional medicine , Lichens -- Analysis , Mastitis , Antibiotics in agriculture
- Language: English
- Type: Thesis , Masters , MSc Agric (Animal Science)
- Identifier: vital:11829 , http://hdl.handle.net/10353/d1016193 , Staphylococcus , Microbial sensitivity tests , Traditional medicine , Lichens -- Analysis , Mastitis , Antibiotics in agriculture
- Description: This study was done in order to assess the incidence of Staphylococcus species from milk of cows with subclinical mastitis and their antimicrobial sensitivity in some selected antibiotics and Usnea barbata lichen extracts. The study was conducted in two different commercial dairy farms. Staphylococcus species isolates were identified using several biochemical tests which included Gram’s staining test, catalase test and oxidase test. A commercial API® staph kit (bioMerieux, France) was used to confirm the bacterial organisms to their species level. The antimicrobial sensitivity of individual species was determined according to the Clinical Laboratory Standard Institute (CLSI) for the selected antibiotics. Agar well diffusion method and the broth micro-dilution technique were used to determine the sensitivity of Staphylococcus species in U. barbata extracts. A total of 467 milk samples were screened for bacterial identification from the two farms. Fifteen different Staphylococcus species isolates were identified from all milk samples that were examined. The most frequently isolated species included Staphylococcus xylosus (54.34%), Staphylococcus hominis (24.78%), Staphylococcus aureus (16.38%), Staphylococcus saprophyticus (16.12%) and Staphylococcus haemolyticus (11.63%). Most Staphylococcus species were resistant to Penicillin (75.35%), Nalidixic acid (72.55%) and Ampicillin (63%). Furthermore, the bacterial sensitivity evaluation of U. barbata lichen extracted with methanol and ethyl-acetate against selected Staphylococcus species isolates showed 92.31% and 53.85% susceptibility, respectively. The minimum inhibitory concentration (MIC) of the methanol and ethyl-acetate extracts ranged between 0.0390 to 10 mg/ml. There was a relatively high incidence of Staphylococcus species identified in milk of cows with subclinical mastitis from both farms. Conversely, Staphylococcus species isolates were resistant to antibiotics (mostly penicillin and ampicillin) commonly used in the farms. Furthermore, the study investigated the antimicrobial sensitivity of U. barbata extract in-vitro which may validate its use in traditional medicine for treatment of cows with mastitis.
- Full Text:
- Date Issued: 2013
Phytochemical analyses and Brine shrimp (Artemia Salina) lethality studies on Syzygium cordatum
- Authors: Chiguvare, Herbert
- Date: 2013
- Subjects: Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11336 , http://hdl.handle.net/10353/d1004352 , Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Description: Syzygium cordatum Hoscht ex. C Krauss, also known as water berry, is normally used by the people of South Africa for respiratory ailments including tuberculosis, stomach complaints, treatment of wounds and as emetics. An extract of the leaves can be used as a purgative for diarrhoea treatment. The leaves of Syzygium cordatum Myrtaceae were obtained from the Eastern Cape Province of South Africa, air dried and sequential solvent extraction was done to obtain various non volatile crude extracts. The volatile extract, that is the essential oil was extracted from the leaves using hydrodistillation and analysis of compounds was done by GC/MS for composition. 32 compounds were obtained from the fresh leaves and 18 compounds were obtained from the dry leaves. The fresh oil contains caryophyllene (11.8 percent) and caryophyllene oxide (11.1 percent) as the main sesquiterpene component. α-Pinene(5.0 percent) was the only monoterpene compound identified in the fresh oil in substantial amount. The dry leaves oil had copanene (17.0 percent), β-Caryophellene (26.0 percent), cubenol (6.5 percent) and caryophellene oxide (14.2 percent) as the dominant constituent of the oil. Summary of the classes of compounds in the oil revealed that the chemical profile of both oils were dominated by sesquiterpenoid compounds. This is the first time that terpenoids compounds are being identified in both the fresh and dry leaf oil of S. cordatum. Hexane leaf extract was selected due to the interest in the terpenoid compounds. Column chromatography of the hexane crude gave five (5) of which two are fully reported. The isolates were fully elucidated using spectroscopic methods to be β-Sitosterol (HC3) and Friedela-3-one (HC1A/HC1D). Cytotoxicity analysis was carried out on the crude using the Brine shrimps assay. Isolates 1C and1D showed significant lethality using the brine shrimps assay with lethality values (LC50) of 4.105mg/ml for HC1C and 4.11mg/ml for 1D/1A respectively.
- Full Text:
- Date Issued: 2013
- Authors: Chiguvare, Herbert
- Date: 2013
- Subjects: Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11336 , http://hdl.handle.net/10353/d1004352 , Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Description: Syzygium cordatum Hoscht ex. C Krauss, also known as water berry, is normally used by the people of South Africa for respiratory ailments including tuberculosis, stomach complaints, treatment of wounds and as emetics. An extract of the leaves can be used as a purgative for diarrhoea treatment. The leaves of Syzygium cordatum Myrtaceae were obtained from the Eastern Cape Province of South Africa, air dried and sequential solvent extraction was done to obtain various non volatile crude extracts. The volatile extract, that is the essential oil was extracted from the leaves using hydrodistillation and analysis of compounds was done by GC/MS for composition. 32 compounds were obtained from the fresh leaves and 18 compounds were obtained from the dry leaves. The fresh oil contains caryophyllene (11.8 percent) and caryophyllene oxide (11.1 percent) as the main sesquiterpene component. α-Pinene(5.0 percent) was the only monoterpene compound identified in the fresh oil in substantial amount. The dry leaves oil had copanene (17.0 percent), β-Caryophellene (26.0 percent), cubenol (6.5 percent) and caryophellene oxide (14.2 percent) as the dominant constituent of the oil. Summary of the classes of compounds in the oil revealed that the chemical profile of both oils were dominated by sesquiterpenoid compounds. This is the first time that terpenoids compounds are being identified in both the fresh and dry leaf oil of S. cordatum. Hexane leaf extract was selected due to the interest in the terpenoid compounds. Column chromatography of the hexane crude gave five (5) of which two are fully reported. The isolates were fully elucidated using spectroscopic methods to be β-Sitosterol (HC3) and Friedela-3-one (HC1A/HC1D). Cytotoxicity analysis was carried out on the crude using the Brine shrimps assay. Isolates 1C and1D showed significant lethality using the brine shrimps assay with lethality values (LC50) of 4.105mg/ml for HC1C and 4.11mg/ml for 1D/1A respectively.
- Full Text:
- Date Issued: 2013
Assessment of the anti-Listerial properties of Garcinia kola (Heckel) seeds
- Authors: Penduka, Dambudzo
- Date: 2014
- Subjects: Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11278 , http://hdl.handle.net/10353/d1015527 , Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Description: A follow-up of traditional medicinal plants uses is an important tool in highlighting their therapeutic potentials, as they have been found to be a source of a wide range of bioactive compounds that can be used as base compounds for new pharmaceutical drugs. This study therefore focuses on assessing the anti-Listerial properties of the seeds of Garcinia kola (Heckel) plant, which is a traditional medicinal plant of west and central African origin, and was and is still used to traditionally treat several ailments. Four different solvents crude extracts of the seeds were assessed for their anti-Listerial activities in-vitro, against a panel of 42 Listeria bacteria, which included Listeria monocytogenes, Listeria ivanovii and Listeria grayi species. At 10 mg/ml concentration the aqueous extract had activity against 29% of the test isolates while the other three crude extracts namely dichloromethane, n-hexane and the methanol extracts had activity against 45% of the test bacteria. The minimum inhibitory concentration (MIC) ranges of the extracts were 0.079-0.313 mg/ml for the dichloromethane extract; 0.079-0.625 mg/ml for the n-hexane extract; 0.157-0.625 mg/ml for the methanol extract; and 10->10 mg/ml for the aqueous extract. The minimum bactericidal concentration (MBC) ranges of the extracts were 0.625–10 mg/ml for both the n-hexane and the dichloromethane extract; 5-10 mg/ml for the methanol extract; and those for the aqueous extract were above 10 mg/ml against all the susceptible Listeria isolates. The rate of kill analysis was then determined for the three most active crude extracts that is excluding the aqueous extract and it was assessed against four representative Listeria species namely L. monocytogenes (LAL 8), L. grayi (LAL 15), L. ivanovii (LEL 30) and L. ivanovii (LEL 18). All the three extracts showed a general trend of being concentration and time dependent in their rate of kill profiles such that most bacteria cells were killed at the highest test concentration of 4× MIC value after the maximum exposure time of 2 h. The n-hexane, dichloromethane and methanol extracts were bactericidal against 4, 3 and 1 isolates out of the four test Listeria isolates respectively.
- Full Text:
- Date Issued: 2014
- Authors: Penduka, Dambudzo
- Date: 2014
- Subjects: Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11278 , http://hdl.handle.net/10353/d1015527 , Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Description: A follow-up of traditional medicinal plants uses is an important tool in highlighting their therapeutic potentials, as they have been found to be a source of a wide range of bioactive compounds that can be used as base compounds for new pharmaceutical drugs. This study therefore focuses on assessing the anti-Listerial properties of the seeds of Garcinia kola (Heckel) plant, which is a traditional medicinal plant of west and central African origin, and was and is still used to traditionally treat several ailments. Four different solvents crude extracts of the seeds were assessed for their anti-Listerial activities in-vitro, against a panel of 42 Listeria bacteria, which included Listeria monocytogenes, Listeria ivanovii and Listeria grayi species. At 10 mg/ml concentration the aqueous extract had activity against 29% of the test isolates while the other three crude extracts namely dichloromethane, n-hexane and the methanol extracts had activity against 45% of the test bacteria. The minimum inhibitory concentration (MIC) ranges of the extracts were 0.079-0.313 mg/ml for the dichloromethane extract; 0.079-0.625 mg/ml for the n-hexane extract; 0.157-0.625 mg/ml for the methanol extract; and 10->10 mg/ml for the aqueous extract. The minimum bactericidal concentration (MBC) ranges of the extracts were 0.625–10 mg/ml for both the n-hexane and the dichloromethane extract; 5-10 mg/ml for the methanol extract; and those for the aqueous extract were above 10 mg/ml against all the susceptible Listeria isolates. The rate of kill analysis was then determined for the three most active crude extracts that is excluding the aqueous extract and it was assessed against four representative Listeria species namely L. monocytogenes (LAL 8), L. grayi (LAL 15), L. ivanovii (LEL 30) and L. ivanovii (LEL 18). All the three extracts showed a general trend of being concentration and time dependent in their rate of kill profiles such that most bacteria cells were killed at the highest test concentration of 4× MIC value after the maximum exposure time of 2 h. The n-hexane, dichloromethane and methanol extracts were bactericidal against 4, 3 and 1 isolates out of the four test Listeria isolates respectively.
- Full Text:
- Date Issued: 2014
Histopathology induced by a medicinal plant indigenous to South Africa that has shown in vitro anti-microbial activity against drug resistant strains of Mycobacterium tuberculosis
- Authors: Shauli, Mathulo Mathabiso
- Date: 2015
- Subjects: Mycobacterial diseases , Tuberculosis -- Treatment -- South Africa , Medicinal plants -- Microbiology , Traditional medicine
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: http://hdl.handle.net/10948/3990 , vital:20498
- Description: Tuberculosis (TB) still remains a health problem globally with over a million new infections and a mortality rate of 1.5 million individuals annually (Hawn et al., 2014). The emerging multi-drug resistant (MDR) strains that accompany human immune deficiency virus (HIV) infection in high-incidence populations contribute significantly to the health burden of TB (Areeshi et al., 2014). The standard treatment that is advocated by the World Health Organization (WHO) for active tuberculosis includes long-term therapy that incorporates the use of isoniazid, rifampicin, pyrazinimide and ethambutol as front line drugs (WHO, 2013). Drug resistance against established treatment options for TB makes research into new forms of therapy an imperative in health care (Ntulela et al., 2009). South Africa is currently witnessing a high number of cases of drug-resistant TB. In some parts of the country, one in ten cases of TB is resistant to treatment. It is therefore essential to have new anti-tuberculosis agents, which can be readily and simply produced from some local source (Warner et al., 2014). A logical starting point for this research of new agents would be the herbal medicines which have been used for centuries in rural areas by local healers. Western developed countries have harvested ethno botanical knowledge and have produced drug therapies for conventional medicines for other ailments. The activity of extracts of the active plants and their properties still require study in animal models in order to assess their future as new anti-tuberculosis agents (Lall and Meyer, 1999). This study focuses on qualitative and quantitative experimental findings after the administration of a medicinal plant extract to animals. This will include daily observation of animals, recording of feed consumption, recording of animal weights, macroscopic examination of animals at necropsy, tissue harvesting, histological procedures and microscopy.
- Full Text:
- Date Issued: 2015
- Authors: Shauli, Mathulo Mathabiso
- Date: 2015
- Subjects: Mycobacterial diseases , Tuberculosis -- Treatment -- South Africa , Medicinal plants -- Microbiology , Traditional medicine
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: http://hdl.handle.net/10948/3990 , vital:20498
- Description: Tuberculosis (TB) still remains a health problem globally with over a million new infections and a mortality rate of 1.5 million individuals annually (Hawn et al., 2014). The emerging multi-drug resistant (MDR) strains that accompany human immune deficiency virus (HIV) infection in high-incidence populations contribute significantly to the health burden of TB (Areeshi et al., 2014). The standard treatment that is advocated by the World Health Organization (WHO) for active tuberculosis includes long-term therapy that incorporates the use of isoniazid, rifampicin, pyrazinimide and ethambutol as front line drugs (WHO, 2013). Drug resistance against established treatment options for TB makes research into new forms of therapy an imperative in health care (Ntulela et al., 2009). South Africa is currently witnessing a high number of cases of drug-resistant TB. In some parts of the country, one in ten cases of TB is resistant to treatment. It is therefore essential to have new anti-tuberculosis agents, which can be readily and simply produced from some local source (Warner et al., 2014). A logical starting point for this research of new agents would be the herbal medicines which have been used for centuries in rural areas by local healers. Western developed countries have harvested ethno botanical knowledge and have produced drug therapies for conventional medicines for other ailments. The activity of extracts of the active plants and their properties still require study in animal models in order to assess their future as new anti-tuberculosis agents (Lall and Meyer, 1999). This study focuses on qualitative and quantitative experimental findings after the administration of a medicinal plant extract to animals. This will include daily observation of animals, recording of feed consumption, recording of animal weights, macroscopic examination of animals at necropsy, tissue harvesting, histological procedures and microscopy.
- Full Text:
- Date Issued: 2015
Phytochemical analysis and antibacterial properties of aqueous and ethanol extracts of Brachylaena elliptica (Thurb.) dc. and Brachylaena ilicifolia (Lam.) Phill & Schweick
- Authors: Sagbo, Idowu Jonas
- Date: 2015
- Subjects: Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11297 , http://hdl.handle.net/10353/d1021289 , Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Description: Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
- Full Text:
- Date Issued: 2015
- Authors: Sagbo, Idowu Jonas
- Date: 2015
- Subjects: Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11297 , http://hdl.handle.net/10353/d1021289 , Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Description: Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
- Full Text:
- Date Issued: 2015
Pharmacological, toxicological and phytochemical evaluation of helichrysum petiolare hilliard & b.l. burtt - an indigenous plant traditionally used in the treatment of diabetes in the eastern cape province of South Africa
- Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Authors: Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Date: 2022-04
- Subjects: Diabetes -- Alternative treatment , Traditional medicine , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22787 , vital:52755
- Description: Diabetes mellitus is one of the leading causes of death in South Africa, and it has already placed significant stress on the country’s health sector and economy. The orthodox hypoglycaemic drugs are not only ineffective in the management of the disease and its complications, but they also possess unwanted side effects. The need for alternative non-toxic drugs is therefore imperative. Various studies have listed several medicinal plants that can be successfully used in the herbal treatment of diabetes and have investigated them for their anti-diabetic potentials in vivo and/or in vitro. Out of the different potential herbal species, plants belonging to the Asteraceae family possess highly potent hypoglycaemic properties with negligible toxicities. Five Asteraceae plants widely used in different parts of South Africa for the treatment of diabetes were reviewed. The review provided an update of scientific evidence on the hypoglycaemic properties of the plants. However, Helichrysum petiolare was studied extensively in this study for its antidiabetic activity H. petiolare has been listed in many ethnobotanical surveys as a plant with potent hypoglycaemic potential, this, however, has not been properly verified in scientific literature and there has hardly been any study on the essential oil and nutritional composition, and antioxidant, antidiabetic, and cytotoxicity potentials of the plant. The effects of hydro-distillation (HD) and solvent-free microwave extraction (SFME) methods on the chemical constituents of H. petiolare-derived essential oils were evaluated. The SFME method had a higher yield of essential oil than the HD. There were substantial amounts of monoterpenes, monoterpene alcohols, sesquiterpenes, and sesquiterpene alcohols in both essential oils obtained, but these compounds were more profound in the SFME derived essential oil which has 62 compounds compared to the 52 derived through HD. The SFME derived essential oil can therefore be said to be of better quality than the HD method. The compounds obtained in the essential oils have high pharmaceutical and cosmetic value, and as observed in this study, their quantity is dependent on the method of extraction (Ibáñez and Blázquez, 2021; Kaur et al., 2021). The proximate analysis of the whole plant of H. petiolare showed high levels of Acid Detergent Fibre (ADF), vitamins (A, C and E), Neutral Detergent Fibre (NDF), and minerals. The high ADF level is believed to be responsible for the low energy, fat and carbohydrate levels observed in the study. The result showed a high level of oxalate and therefore suggests cooking of the plant before human consumption. Overall nutrition and mineral compositions of the plant showed that H. petiolare is immensely rich in vital nutrients that are of great importance to health and metabolism; these nutrients are suggested to be partly responsible for the plant’s useful medicinal properties. The phytochemical contents of the acetone (ACQ), ethanol (ETQ), and boiled (BAQ) and cold (CAQ) aqueous whole-plant extracts of Helichrysum petiolare were determined using standard phytochemical reaction methods. ABTS, DPPH, NO and TAC assays were used to evaluate their antioxidant properties. The highest total phenolic content (212,963 mg/g) was reported in the BAQ extract, while the ETQ had the highest flavonoid (172.393 mg/g) and proanthocyanidin contents (65.855 mg/g). Alkaloids, flavonols, and saponin were highest in the ACQ extract, while the CAQ had the lowest phytochemical content. Among the extracts, the BAQ had the highest DPPH•+ (IC50 0.02 mg/mL) and ABTS•+ (IC50 0.07) inhibition capacities, while the ETQ exhibited the highest NO• Inhibition (IC50 0.41 mg/mL) and TAC (IC50 0.19 mg/mL). These findings justify the use of H. petiolare in traditional medicine and further recommend the ETQ and BAQ extracts of the plant as more effective extracts for medicinal treatment. The hepatotoxicity (cytotoxicity, mitotoxicity and lipotoxicity) potential of the BAQ, CAQ and ETQ extracts of Helichrysum petiolare was evaluated using standard procedures. The results showed negligible BAQ and CAQ cytotoxicities, which were further, corroborated by stability in the mitochondrial membrane potentials and were congruent with the CAQ and BAQ results for steatosis and phospholipidosis. The data suggested favourable CAQ and BAQ toxicity profiles with limited risks for hepatotoxicity. The ETQ extract, however, showed significantly high levels of cytotoxicity and lipotoxicity, and a low level of mitotoxicity. Our result suggested a potential risk of the ETQ extract for hepatotoxicity but appears partly independent of direct mitochondrial involvement. Glucose uptake assay showed significantly increased glucose uptake in the BAQ and CAQ treated L6 and C3A cell lines. The CAQ extract enhanced glucose uptake more in the L6 myocytes than in the C3A cell-lines hepatocytes. The BAQ extract showed higher levels of inhibition on α–amylase and α-glucosidase activities as compared to CAQ. The BAQ and CAQ extracts of H. petiolare may, therefore, contain pharmacologically active and relatively non-toxic hypoglycaemic chemicals, which may be effective substitutes in the treatment of diabetes mellitus. This study provides up to date scientific information on the use of H. petiolare in the treatment of diabetes mellitus in the Eastern Cape of South Africa. It justifies the use of this plant in herbal medicine and sheds more light on its previously vaguely understood nutritional and medicinal potentials. More studies, however, need to be done to isolate, identify and purify the constituent bioactive compound(s). Their dosage of application and mode of action also needs to be understood. , Thesis (PhD) -- Faculty of Science and Agriculture, 2022
- Full Text:
- Date Issued: 2022-04
- Authors: Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Date: 2022-04
- Subjects: Diabetes -- Alternative treatment , Traditional medicine , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22787 , vital:52755
- Description: Diabetes mellitus is one of the leading causes of death in South Africa, and it has already placed significant stress on the country’s health sector and economy. The orthodox hypoglycaemic drugs are not only ineffective in the management of the disease and its complications, but they also possess unwanted side effects. The need for alternative non-toxic drugs is therefore imperative. Various studies have listed several medicinal plants that can be successfully used in the herbal treatment of diabetes and have investigated them for their anti-diabetic potentials in vivo and/or in vitro. Out of the different potential herbal species, plants belonging to the Asteraceae family possess highly potent hypoglycaemic properties with negligible toxicities. Five Asteraceae plants widely used in different parts of South Africa for the treatment of diabetes were reviewed. The review provided an update of scientific evidence on the hypoglycaemic properties of the plants. However, Helichrysum petiolare was studied extensively in this study for its antidiabetic activity H. petiolare has been listed in many ethnobotanical surveys as a plant with potent hypoglycaemic potential, this, however, has not been properly verified in scientific literature and there has hardly been any study on the essential oil and nutritional composition, and antioxidant, antidiabetic, and cytotoxicity potentials of the plant. The effects of hydro-distillation (HD) and solvent-free microwave extraction (SFME) methods on the chemical constituents of H. petiolare-derived essential oils were evaluated. The SFME method had a higher yield of essential oil than the HD. There were substantial amounts of monoterpenes, monoterpene alcohols, sesquiterpenes, and sesquiterpene alcohols in both essential oils obtained, but these compounds were more profound in the SFME derived essential oil which has 62 compounds compared to the 52 derived through HD. The SFME derived essential oil can therefore be said to be of better quality than the HD method. The compounds obtained in the essential oils have high pharmaceutical and cosmetic value, and as observed in this study, their quantity is dependent on the method of extraction (Ibáñez and Blázquez, 2021; Kaur et al., 2021). The proximate analysis of the whole plant of H. petiolare showed high levels of Acid Detergent Fibre (ADF), vitamins (A, C and E), Neutral Detergent Fibre (NDF), and minerals. The high ADF level is believed to be responsible for the low energy, fat and carbohydrate levels observed in the study. The result showed a high level of oxalate and therefore suggests cooking of the plant before human consumption. Overall nutrition and mineral compositions of the plant showed that H. petiolare is immensely rich in vital nutrients that are of great importance to health and metabolism; these nutrients are suggested to be partly responsible for the plant’s useful medicinal properties. The phytochemical contents of the acetone (ACQ), ethanol (ETQ), and boiled (BAQ) and cold (CAQ) aqueous whole-plant extracts of Helichrysum petiolare were determined using standard phytochemical reaction methods. ABTS, DPPH, NO and TAC assays were used to evaluate their antioxidant properties. The highest total phenolic content (212,963 mg/g) was reported in the BAQ extract, while the ETQ had the highest flavonoid (172.393 mg/g) and proanthocyanidin contents (65.855 mg/g). Alkaloids, flavonols, and saponin were highest in the ACQ extract, while the CAQ had the lowest phytochemical content. Among the extracts, the BAQ had the highest DPPH•+ (IC50 0.02 mg/mL) and ABTS•+ (IC50 0.07) inhibition capacities, while the ETQ exhibited the highest NO• Inhibition (IC50 0.41 mg/mL) and TAC (IC50 0.19 mg/mL). These findings justify the use of H. petiolare in traditional medicine and further recommend the ETQ and BAQ extracts of the plant as more effective extracts for medicinal treatment. The hepatotoxicity (cytotoxicity, mitotoxicity and lipotoxicity) potential of the BAQ, CAQ and ETQ extracts of Helichrysum petiolare was evaluated using standard procedures. The results showed negligible BAQ and CAQ cytotoxicities, which were further, corroborated by stability in the mitochondrial membrane potentials and were congruent with the CAQ and BAQ results for steatosis and phospholipidosis. The data suggested favourable CAQ and BAQ toxicity profiles with limited risks for hepatotoxicity. The ETQ extract, however, showed significantly high levels of cytotoxicity and lipotoxicity, and a low level of mitotoxicity. Our result suggested a potential risk of the ETQ extract for hepatotoxicity but appears partly independent of direct mitochondrial involvement. Glucose uptake assay showed significantly increased glucose uptake in the BAQ and CAQ treated L6 and C3A cell lines. The CAQ extract enhanced glucose uptake more in the L6 myocytes than in the C3A cell-lines hepatocytes. The BAQ extract showed higher levels of inhibition on α–amylase and α-glucosidase activities as compared to CAQ. The BAQ and CAQ extracts of H. petiolare may, therefore, contain pharmacologically active and relatively non-toxic hypoglycaemic chemicals, which may be effective substitutes in the treatment of diabetes mellitus. This study provides up to date scientific information on the use of H. petiolare in the treatment of diabetes mellitus in the Eastern Cape of South Africa. It justifies the use of this plant in herbal medicine and sheds more light on its previously vaguely understood nutritional and medicinal potentials. More studies, however, need to be done to isolate, identify and purify the constituent bioactive compound(s). Their dosage of application and mode of action also needs to be understood. , Thesis (PhD) -- Faculty of Science and Agriculture, 2022
- Full Text:
- Date Issued: 2022-04
An ethnomedical study of the role and impact of cannabidiol (CBD) treatment of women living with endometriosis and Polycystic Ovarian Syndrome (PCOS)
- Authors: Norman, Paige Sarah
- Date: 2022-10-14
- Subjects: Polycystic ovary syndrome Alternative treatment , Endometriosis Alternative treatment , Cannabidiol , Cannabis , Pain management , Traditional medicine , Feminist anthropology , Women's health services
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/408652 , vital:70513
- Description: Over the last few years, there has been a notable increase in popularity in the use of cannabidiol (CBD) as a form of alternative medicinal treatment for various illnesses. CBD, a by-product of the cannabis plant, is an isolate and does not contain the psychoactive agent, tetrahydrocannabinol (THC). Endometriosis and Polycystic Ovarian Syndrome (PCOS) are chronic reproductive health sicknesses that are increasingly experienced by women. In the absence of cures, biomedical treatment for these diseases aim to manage symptoms, for example; heavy bleeding, heightened levels of pain, and insomnia. CBD offers an alternative to women who feel that biomedical interventions are no longer able to maintain their health and well-being. CBD positions itself as a natural remedy claiming to be safe and effective. This research study, mainly through qualitative data collection, focused on experiences of Zimbabwean and South African women living with endometriosis and/ or PCOS, who have turned to CBD to manage their symptoms. The importance of this study was to position itself within patients’ lived experiences. The research study found that CBD indeed has numerous benefits, including pain management, alleviating stress, and anxiety. Through the emergent themes from the data, it became clear that women are marginalised and treated unequally in the biomedical healthcare sphere. Feminist Anthropology and Structural Violence was applied to analyse the data collected to explore the patriarchal nature of the biomedical healthcare system and the experiences that women have, which has led them to turn to alternative treatments. , Thesis (MSocSci) -- Faculty of Humanities, Anthropology, 2022
- Full Text:
- Date Issued: 2022-10-14
- Authors: Norman, Paige Sarah
- Date: 2022-10-14
- Subjects: Polycystic ovary syndrome Alternative treatment , Endometriosis Alternative treatment , Cannabidiol , Cannabis , Pain management , Traditional medicine , Feminist anthropology , Women's health services
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/408652 , vital:70513
- Description: Over the last few years, there has been a notable increase in popularity in the use of cannabidiol (CBD) as a form of alternative medicinal treatment for various illnesses. CBD, a by-product of the cannabis plant, is an isolate and does not contain the psychoactive agent, tetrahydrocannabinol (THC). Endometriosis and Polycystic Ovarian Syndrome (PCOS) are chronic reproductive health sicknesses that are increasingly experienced by women. In the absence of cures, biomedical treatment for these diseases aim to manage symptoms, for example; heavy bleeding, heightened levels of pain, and insomnia. CBD offers an alternative to women who feel that biomedical interventions are no longer able to maintain their health and well-being. CBD positions itself as a natural remedy claiming to be safe and effective. This research study, mainly through qualitative data collection, focused on experiences of Zimbabwean and South African women living with endometriosis and/ or PCOS, who have turned to CBD to manage their symptoms. The importance of this study was to position itself within patients’ lived experiences. The research study found that CBD indeed has numerous benefits, including pain management, alleviating stress, and anxiety. Through the emergent themes from the data, it became clear that women are marginalised and treated unequally in the biomedical healthcare sphere. Feminist Anthropology and Structural Violence was applied to analyse the data collected to explore the patriarchal nature of the biomedical healthcare system and the experiences that women have, which has led them to turn to alternative treatments. , Thesis (MSocSci) -- Faculty of Humanities, Anthropology, 2022
- Full Text:
- Date Issued: 2022-10-14
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