According to the World Health Organization (WHO), M. tuberculosis, the causative agent of TB, accounts for approximately 1.7 million deaths annually. Further contributing causes to the worldwide TB incidence, is the widespread unavailability and ineffectiveness of TB vaccines, time consuming diagnostic methods and unsuccessful treatment approaches. Research for better characterising mycobacteria in general, or other Mycobacterium species, may help us to better understand M. tuberculosis and TB disease mechanisms, which will in turn lower the future TB disease prevalence, as this may lead to the development of better treatments, diagnostics and vaccines. Mycobacteria use various secretion pathways, including the ESX- or type VII secretion (T7S) system, to ensure transport across the complex cell wall. The genome of M. tuberculosis has five copies of a gene cluster known as the ESX gene cluster region, which is associated with virulence and viability of mycobacteria. The ESX-3 gene cluster is thought to be essential for growth of M. tuberculosis and proposed to be involved in iron / zinc homeostasis. Mycobacteria synthesise siderophores, which are proposed to be involved in the uptake of iron over their cell wall. M. tuberculosis are known to produce two types of siderophores, namely: carboxymycobactins and mycobactins. Loots and colleagues, however illustrated, that ESX-3 knockouts, show signs of iron overload, despite the absence of the mycobactins induced by knocking out the ESX-3 gene cluster. It was hypothesised, that this overload occurs due to an increase in exochelin synthesis, another iron uptake protein not associated with ESX-3, overcompensating for the perceived iron depletion in the knockout organism. A Metabolomics research approach was subsequently used in this study, to generate new information in order to better characterise the role of iron on the metabolism of these organisms, and additionally confirm the role of ESX-3 in iron uptake. In this study, we firstly determined the most optimal extraction conditions for this metabolomics investigation. Two extraction methods were subsequently investigated and compared, considering their repeatability and their respective capacities to extract those compounds which best differentiate the M. smegmatis ESX-3 knockouts and wild-type parent strains. Considering the results generated, the total metabolome method was chosen for further analyses, for the following reasons: 1) it is simpler, 2) faster, 3) showed better repeatability, 4) extracts those compounds best differentiating the compared groups and 5) has been previously described for metabolomics analyses characterising ESX-3 gene functionality, hence potentially allowing us to compare results to that previously generated and published data. Subsequently, we used the chosen extraction method, followed by GCxGC-TOFMS analysis of the separately cultured M. smegmatis wild-type sample extracts, cultured in normal, low and high iron conditions, to determine the influence of varying iron concentrations on the metabolome of this organism, by metabolomics comparisons of these groups. Following this, an identical research approach was used to compare the metabolome of a M. smegmatis ESX-3 knock-out strain, to that of a M. smegmatis wild type parent strain, both cultured in normal / standardised iron concentrations. Considering the results generated when comparing the metabolome of a M. smegmatis ESX-3 knock-out strain to that of a M. smegmatis wild type parent strain, the altered metabolome of the M. smegmatis ESX-3 knockouts correlated well to that of the M. smegmatis wild type cultured in elevated iron growth conditions. This suggests ESX-3 is involved in iron uptake, and that knocking out the ESX-3 gene cluster of M. smegmatis does in fact result in a metabolome profile suggesting iron overload, as was proposed by Loots et al (2012), most probably due the exochelins overcompensating for the absence of mycobactins, in M. smegmatis ESX-3 knockouts. / MSc (Biochemistry) North-West University, Potchefstroom Campus 2013.

According to the World Health Organization (WHO), M. tuberculosis, the causative agent of TB, accounts for approximately 1.7 million deaths annually. Further contributing causes to the worldwide TB incidence, is the widespread unavailability and ineffectiveness of TB vaccines, time consuming diagnostic methods and unsuccessful treatment approaches. Research for better characterising mycobacteria in general, or other Mycobacterium species, may help us to better understand M. tuberculosis and TB disease mechanisms, which will in turn lower the future TB disease prevalence, as this may lead to the development of better treatments, diagnostics and vaccines. Mycobacteria use various secretion pathways, including the ESX- or type VII secretion (T7S) system, to ensure transport across the complex cell wall. The genome of M. tuberculosis has five copies of a gene cluster known as the ESX gene cluster region, which is associated with virulence and viability of mycobacteria. The ESX-3 gene cluster is thought to be essential for growth of M. tuberculosis and proposed to be involved in iron / zinc homeostasis. Mycobacteria synthesise siderophores, which are proposed to be involved in the uptake of iron over their cell wall. M. tuberculosis are known to produce two types of siderophores, namely: carboxymycobactins and mycobactins. Loots and colleagues, however illustrated, that ESX-3 knockouts, show signs of iron overload, despite the absence of the mycobactins induced by knocking out the ESX-3 gene cluster. It was hypothesised, that this overload occurs due to an increase in exochelin synthesis, another iron uptake protein not associated with ESX-3, overcompensating for the perceived iron depletion in the knockout organism. A Metabolomics research approach was subsequently used in this study, to generate new information in order to better characterise the role of iron on the metabolism of these organisms, and additionally confirm the role of ESX-3 in iron uptake. In this study, we firstly determined the most optimal extraction conditions for this metabolomics investigation. Two extraction methods were subsequently investigated and compared, considering their repeatability and their respective capacities to extract those compounds which best differentiate the M. smegmatis ESX-3 knockouts and wild-type parent strains. Considering the results generated, the total metabolome method was chosen for further analyses, for the following reasons: 1) it is simpler, 2) faster, 3) showed better repeatability, 4) extracts those compounds best differentiating the compared groups and 5) has been previously described for metabolomics analyses characterising ESX-3 gene functionality, hence potentially allowing us to compare results to that previously generated and published data. Subsequently, we used the chosen extraction method, followed by GCxGC-TOFMS analysis of the separately cultured M. smegmatis wild-type sample extracts, cultured in normal, low and high iron conditions, to determine the influence of varying iron concentrations on the metabolome of this organism, by metabolomics comparisons of these groups. Following this, an identical research approach was used to compare the metabolome of a M. smegmatis ESX-3 knock-out strain, to that of a M. smegmatis wild type parent strain, both cultured in normal / standardised iron concentrations. Considering the results generated when comparing the metabolome of a M. smegmatis ESX-3 knock-out strain to that of a M. smegmatis wild type parent strain, the altered metabolome of the M. smegmatis ESX-3 knockouts correlated well to that of the M. smegmatis wild type cultured in elevated iron growth conditions. This suggests ESX-3 is involved in iron uptake, and that knocking out the ESX-3 gene cluster of M. smegmatis does in fact result in a metabolome profile suggesting iron overload, as was proposed by Loots et al (2012), most probably due the exochelins overcompensating for the absence of mycobactins, in M. smegmatis ESX-3 knockouts. / MSc (Biochemistry) North-West University, Potchefstroom Campus 2013.

Background: Children in South Africa are still affected by micronutrient deficiencies and children living in farm communities are especially vulnerable. African Leafy Vegetables (ALVs) are well endowed with micronutrients such as iron, zinc and vitamin A and might contribute to the nutritional status of children. However, these vegetables have been perceived as “poor people’s food” and over the years knowledge of and use of ALVs has decreased. Aim: This study assessed the potential contribution of ALVs to the nutritional status of children in a semi-rural farm community. Method: In this cross-sectional study, anthropometric indices, serum iron, zinc and retinol concentrations were determined in school children aged 5−13 years (n=155). Dietary intake of iron, zinc and vitamin A was evaluated by three 24-hour diet recalls of children (n=154). The iron, zinc and β-carotene content of selected ALVs was determined. Knowledge of and use of ALVs by primary caregivers was established using focus group discussions (FGDs). Descriptive statistics, independent t-tests, the Pearson Chi-Square Test and Mann-Whitney U Test were used. Anthropometric data were analysed using the World Health Organization Reference 2007 data. Dietary data were analysed using FoodFinder (version 3). Qualitative data from FGDs were translated, transcribed and color-coded to generate emerging themes. Results: Stunting (11%) was the most prevalent anthropometric indicator of malnutrition. This was supported by the low socio-economic status of households. Deficiency prevalence in iron (serum ferritin <15 μg/L; 15.5%) and vitamin A (serum retinol <20 μg/dL; 3.2%) was low. Zinc deficiency was the most prevalent (serum zinc <65 μg/dL; 74.8%) deficiency. Median dietary intake of iron, zinc and vitamin A was generally above the Estimated Average Requirement. ALVs were potentially good sources of iron, zinc and β-carotene and could contribute substantially to the Recommended Dietary Allowance for these nutrients in children, without taking into account inhibiting factors that might affect the bioavailability. Iron content of the ALVs studied ranged from 1.4−3.2 mg/100 g edible portion. Amaranthus cruentus was the best source of iron. Zinc content of the ALVs ranged from 0.7−1.4 mg/100g edible portions, with Cleome gynandra having the highest zinc composition. The β-carotene content of the ALVs ranged from 182−314 μg RAE/100 g edible portion, with both Amaranthus cruentus and Cleome gynandra being the best sources. Knowledge of ALVs and their use was indigenous and was transferred between generations. Caregivers had positive attitudes towards the use of ALVs. Conclusion: Although the prevalence of deficiencies was not severe (with exception of zinc deficiency), micronutrient deficiencies exist in the rural farm community studied. ALVs are potentially good sources of iron, zinc and β-carotene and might contribute to the nutritional status of school children. Knowledge of ALVs and the positive attitude and perceptions regarding their use by primary caregivers implied a potentially positive future response to interventions promoting consumption of ALVs in order to contribute to the alleviation of micronutrient deficiencies. / Thesis (MSc (Nutrition))--North-West University, Potchefstroom Campus, 2013.

Background: Children in South Africa are still affected by micronutrient deficiencies and children living in farm communities are especially vulnerable. African Leafy Vegetables (ALVs) are well endowed with micronutrients such as iron, zinc and vitamin A and might contribute to the nutritional status of children. However, these vegetables have been perceived as “poor people’s food” and over the years knowledge of and use of ALVs has decreased. Aim: This study assessed the potential contribution of ALVs to the nutritional status of children in a semi-rural farm community. Method: In this cross-sectional study, anthropometric indices, serum iron, zinc and retinol concentrations were determined in school children aged 5−13 years (n=155). Dietary intake of iron, zinc and vitamin A was evaluated by three 24-hour diet recalls of children (n=154). The iron, zinc and β-carotene content of selected ALVs was determined. Knowledge of and use of ALVs by primary caregivers was established using focus group discussions (FGDs). Descriptive statistics, independent t-tests, the Pearson Chi-Square Test and Mann-Whitney U Test were used. Anthropometric data were analysed using the World Health Organization Reference 2007 data. Dietary data were analysed using FoodFinder (version 3). Qualitative data from FGDs were translated, transcribed and color-coded to generate emerging themes. Results: Stunting (11%) was the most prevalent anthropometric indicator of malnutrition. This was supported by the low socio-economic status of households. Deficiency prevalence in iron (serum ferritin <15 μg/L; 15.5%) and vitamin A (serum retinol <20 μg/dL; 3.2%) was low. Zinc deficiency was the most prevalent (serum zinc <65 μg/dL; 74.8%) deficiency. Median dietary intake of iron, zinc and vitamin A was generally above the Estimated Average Requirement. ALVs were potentially good sources of iron, zinc and β-carotene and could contribute substantially to the Recommended Dietary Allowance for these nutrients in children, without taking into account inhibiting factors that might affect the bioavailability. Iron content of the ALVs studied ranged from 1.4−3.2 mg/100 g edible portion. Amaranthus cruentus was the best source of iron. Zinc content of the ALVs ranged from 0.7−1.4 mg/100g edible portions, with Cleome gynandra having the highest zinc composition. The β-carotene content of the ALVs ranged from 182−314 μg RAE/100 g edible portion, with both Amaranthus cruentus and Cleome gynandra being the best sources. Knowledge of ALVs and their use was indigenous and was transferred between generations. Caregivers had positive attitudes towards the use of ALVs. Conclusion: Although the prevalence of deficiencies was not severe (with exception of zinc deficiency), micronutrient deficiencies exist in the rural farm community studied. ALVs are potentially good sources of iron, zinc and β-carotene and might contribute to the nutritional status of school children. Knowledge of ALVs and the positive attitude and perceptions regarding their use by primary caregivers implied a potentially positive future response to interventions promoting consumption of ALVs in order to contribute to the alleviation of micronutrient deficiencies. / Thesis (MSc (Nutrition))--North-West University, Potchefstroom Campus, 2013.

Background: In South Africa, as in many other developing countries, iron deficiency (the most common micronutrient deficiency) still remains unresolved; while obesity has emerged as a public health challenge causing increases in the incidence and prevalence of cardiovascular diseases (CVDs). Research has shown that certain iron indices are associated with both anthropometric and biological markers of CVDs. Adiposity is thought to modulate the pathway linking iron status to CVDs. Objective: To examine the associations between iron indices, anthropometric and biological markers of CVDs in an African population undergoing transition. Methods: This thesis was based on secondary analysis of data generated during the Transition and Health during Urbanisation of South Africans (THUSA) study; and primary and secondary analysis of the baseline Prospective Urban and Rural Epidemiological (PURE) study. Both studies were cross–sectional in design and were conducted between 1996–1998 and in 2005 respectively in the North West Province of South Africa. The 1854 men and women participants in the THUSA study (>15years) and 1262 women participants in the PURE study (>35years) were included in the analysis. The relationship between iron and anthropometric indicators of CVD risk was examined in the THUSA study while that of iron status, anthropometric and biological markers of CVD risk was examined in the PURE study. Results: In the THUSA study, ferritin was positively associated with body mass index (BMI), waist circumference (WC), waist to hip ratio (WHR), body fat and subscapular skinfold (r=0.141, 0.359, 0.396, 0.308, 0.141 respectively for men and 0.126, 0.232, 0.319, 0.126, 0.105 respectively for women; p<0.01). Only the women showed decreased serum iron concentration with increasing BMI (p<0.05). WC and WHR increased with increasing serum ferritin concentration for both genders (p<0.05). As for the PURE study, associations between iron status parameters and CVD risk factors were generally weak (r<0.3, p<0.01) and were not retained after adjusting for valid confounders. WC and WHR increased with increasing ferritin concentration (p<0.05). Conclusion: Although these results do not indicate any significant association between iron indices and biological markers of CVD, its association with anthropometric indices gives an indication of the possible contribution of iron in the aetiology of CVDs. Thus, it may be necessary to exercise caution on the emphasis placed on iron as a nutrient and iron intervention programmes because of the suggestive role of iron in CVD development. / Thesis (Ph.D. (Nutrition))--North-West University, Potchefstroom Campus, 2011.

Background: In South Africa, as in many other developing countries, iron deficiency (the most common micronutrient deficiency) still remains unresolved; while obesity has emerged as a public health challenge causing increases in the incidence and prevalence of cardiovascular diseases (CVDs). Research has shown that certain iron indices are associated with both anthropometric and biological markers of CVDs. Adiposity is thought to modulate the pathway linking iron status to CVDs. Objective: To examine the associations between iron indices, anthropometric and biological markers of CVDs in an African population undergoing transition. Methods: This thesis was based on secondary analysis of data generated during the Transition and Health during Urbanisation of South Africans (THUSA) study; and primary and secondary analysis of the baseline Prospective Urban and Rural Epidemiological (PURE) study. Both studies were cross–sectional in design and were conducted between 1996–1998 and in 2005 respectively in the North West Province of South Africa. The 1854 men and women participants in the THUSA study (>15years) and 1262 women participants in the PURE study (>35years) were included in the analysis. The relationship between iron and anthropometric indicators of CVD risk was examined in the THUSA study while that of iron status, anthropometric and biological markers of CVD risk was examined in the PURE study. Results: In the THUSA study, ferritin was positively associated with body mass index (BMI), waist circumference (WC), waist to hip ratio (WHR), body fat and subscapular skinfold (r=0.141, 0.359, 0.396, 0.308, 0.141 respectively for men and 0.126, 0.232, 0.319, 0.126, 0.105 respectively for women; p<0.01). Only the women showed decreased serum iron concentration with increasing BMI (p<0.05). WC and WHR increased with increasing serum ferritin concentration for both genders (p<0.05). As for the PURE study, associations between iron status parameters and CVD risk factors were generally weak (r<0.3, p<0.01) and were not retained after adjusting for valid confounders. WC and WHR increased with increasing ferritin concentration (p<0.05). Conclusion: Although these results do not indicate any significant association between iron indices and biological markers of CVD, its association with anthropometric indices gives an indication of the possible contribution of iron in the aetiology of CVDs. Thus, it may be necessary to exercise caution on the emphasis placed on iron as a nutrient and iron intervention programmes because of the suggestive role of iron in CVD development. / Thesis (Ph.D. (Nutrition))--North-West University, Potchefstroom Campus, 2011.

The evolution of aerobic metabolic processes unavoidably led to the production of reactive oxygen species (ROS). ROS have the ability to cause harmful oxidative damage to biomolecules. Increased ROS generation and subsequent oxidative stress have been associated with aging and neurodegenerative disorders such as Parkinson’s and Alzheimer’s diseases as a result of the extreme sensitivity of the central nervous system to damage from ROS. Antioxidant defence systems have co–evolved with aerobic metabolic processes to counteract oxidative damage inflicted by ROS. The impact of neurodegenerative disorders on society is increasing rapidly as the life expectancy of the global population increases. In this day and age, a much younger group of the population is also experiencing neurodegenerative symptoms as a result of the harmful effect of the human immunodeficiency virus (HIV) on the central nervous system. Plants are an invaluable source of medicinal compounds. The use of plants for their healing properties is rooted in ancient times. The aim of this study was to select from twenty one plants, the plant with the most promising antioxidant activity and to determine whether extracts of this plant could act as free radical scavengers, comparing the results to Trolox, a known free radical scavenger. The next step was to isolate and characterize a compound from an extract exhibiting promising antioxidant activity. Bioassay–guided fractionation was followed to achieve this. During screening trials, twenty one plants, namely Berula erecta, Heteromorpha arborescens, Tarchonanthus camphoratus, Vernonia oligocephala, Gymnosporia buxifolia, Acacia karroo, Elephantorrhiza elephantina, Erythrina zeyheri, Leonotis leonurus, Plectranthus ecklonii, P. rehmanii, P. venteri, Salvia auretia, S. runcinata, Solenostemon latifolius, S. rotundifolius, Plumbago auriculata, Clematis brachiata, Vangueria infausta, Physalis peruviana and Lippia javanica were selected from literature, based on reported antioxidant activity within the plant families, for screening of their antioxidant activity. One hundred and ten extracts were prepared from the leaves, using Soxhlet extraction and the solvents petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc) and ethanol (EtOH), consecutively. The focus during initial screening trials was on chemistry–based assays. The oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays were employed for the primary screening of the one hundred and ten leaf extracts. The ORAC assay was used to determine whether the plant extracts were able to scavenge peroxyl radicals and the FRAP assay was used to determine the reducing abilities of the extracts. Quantification of the peroxyl radical scavenging activity by the ORAC assay revealed that activity was observed for most of the extracts, with the ethyl acetate and ethanol extracts of L. javanica exhibiting the most promising activity. This pattern of activity was also found with the reducing capacity evaluated by the FRAP assay in which the EtOAc and EtOH extracts of L. javanica also exhibited the most promising activity. L. javanica was selected for further study by screening for biological activity, employing the nitro–blue tetrazolium (NBT) assay and thiobarbituric acid reactive substances (TBARS) assay. Using a cyanide model to induce neurotoxic effects in rat brain homogenate, the neuroprotective properties of the extracts of L. javanica leaves were examined using the NBT assay and compared to that of Trolox. The NBT assay determines the level of superoxide anions. All the extracts of L. javanica significantly reduced superoxide anion generation at all concentrations used. The petroleum ether and ethyl acetate extracts, at all concentrations, reduced superoxide anion generation to values lower than that of the control, suggesting that these extracts may be able to attenuate normal free radical processes in the brain. The petroleum ether extract exhibited the most promising activity at a concentration of 1.25 and 2.5 mg/ml and also exhibited similar results as the ethyl acetate extract at a lower concentration than the ethyl acetate extract (2.5 mg/ml compared to 5 mg/ml). A toxin–solution consisting of hydrogen peroxide (H2O2), iron(III)chloride (FeCl3) and ascorbic acid was used to induce lipid peroxidation and the ability of the extracts of the leaves of L. javanica to attenuate lipid peroxidation was investigated in rat brain homogenate and compared to that of Trolox. All of the extracts of L. javanica significantly attenuated toxininduced lipid peroxidation at all concentrations used. All of the extracts were also able to significantly attenuate toxin–induced lipid peroxidation to values lower than that of the control. These results suggest that all of the extracts of L. javanica possess the ability to attenuate not only toxin–induced lipid peroxidation, but also lipid peroxidation that occurs during normal processes in the brain. The petroleum ether extract was subjected to bioassay–guided fractionation using column and thin–layer chromatography and the NBT and TBARS assays. Fraction DD1 was investigated by means of nuclear magnetic resonance, infrared and mass spectrometry. The exact structure of fraction DD1 was not elucidated. Considering all the results, it is clear that L. javanica shows great potential as a medicinal plant with antioxidant activity and may therefore be beneficial in diminishing the destructive oxidative effects inflicted by free radicals. There are however still many compounds to be isolated from L. javanica. Key words: Verbenaceae, Lippia javanica, antioxidant, neurodegeneration, oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), nitro–blue tetrazolium assay (NBT), thiobarbituric acid reactive substances assay (TBARS). / Thesis (M.Sc. (Pharmaceutical Chemistry))--North-West University, Potchefstroom Campus, 2011.

The evolution of aerobic metabolic processes unavoidably led to the production of reactive oxygen species (ROS). ROS have the ability to cause harmful oxidative damage to biomolecules. Increased ROS generation and subsequent oxidative stress have been associated with aging and neurodegenerative disorders such as Parkinson’s and Alzheimer’s diseases as a result of the extreme sensitivity of the central nervous system to damage from ROS. Antioxidant defence systems have co–evolved with aerobic metabolic processes to counteract oxidative damage inflicted by ROS. The impact of neurodegenerative disorders on society is increasing rapidly as the life expectancy of the global population increases. In this day and age, a much younger group of the population is also experiencing neurodegenerative symptoms as a result of the harmful effect of the human immunodeficiency virus (HIV) on the central nervous system. Plants are an invaluable source of medicinal compounds. The use of plants for their healing properties is rooted in ancient times. The aim of this study was to select from twenty one plants, the plant with the most promising antioxidant activity and to determine whether extracts of this plant could act as free radical scavengers, comparing the results to Trolox, a known free radical scavenger. The next step was to isolate and characterize a compound from an extract exhibiting promising antioxidant activity. Bioassay–guided fractionation was followed to achieve this. During screening trials, twenty one plants, namely Berula erecta, Heteromorpha arborescens, Tarchonanthus camphoratus, Vernonia oligocephala, Gymnosporia buxifolia, Acacia karroo, Elephantorrhiza elephantina, Erythrina zeyheri, Leonotis leonurus, Plectranthus ecklonii, P. rehmanii, P. venteri, Salvia auretia, S. runcinata, Solenostemon latifolius, S. rotundifolius, Plumbago auriculata, Clematis brachiata, Vangueria infausta, Physalis peruviana and Lippia javanica were selected from literature, based on reported antioxidant activity within the plant families, for screening of their antioxidant activity. One hundred and ten extracts were prepared from the leaves, using Soxhlet extraction and the solvents petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc) and ethanol (EtOH), consecutively. The focus during initial screening trials was on chemistry–based assays. The oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays were employed for the primary screening of the one hundred and ten leaf extracts. The ORAC assay was used to determine whether the plant extracts were able to scavenge peroxyl radicals and the FRAP assay was used to determine the reducing abilities of the extracts. Quantification of the peroxyl radical scavenging activity by the ORAC assay revealed that activity was observed for most of the extracts, with the ethyl acetate and ethanol extracts of L. javanica exhibiting the most promising activity. This pattern of activity was also found with the reducing capacity evaluated by the FRAP assay in which the EtOAc and EtOH extracts of L. javanica also exhibited the most promising activity. L. javanica was selected for further study by screening for biological activity, employing the nitro–blue tetrazolium (NBT) assay and thiobarbituric acid reactive substances (TBARS) assay. Using a cyanide model to induce neurotoxic effects in rat brain homogenate, the neuroprotective properties of the extracts of L. javanica leaves were examined using the NBT assay and compared to that of Trolox. The NBT assay determines the level of superoxide anions. All the extracts of L. javanica significantly reduced superoxide anion generation at all concentrations used. The petroleum ether and ethyl acetate extracts, at all concentrations, reduced superoxide anion generation to values lower than that of the control, suggesting that these extracts may be able to attenuate normal free radical processes in the brain. The petroleum ether extract exhibited the most promising activity at a concentration of 1.25 and 2.5 mg/ml and also exhibited similar results as the ethyl acetate extract at a lower concentration than the ethyl acetate extract (2.5 mg/ml compared to 5 mg/ml). A toxin–solution consisting of hydrogen peroxide (H2O2), iron(III)chloride (FeCl3) and ascorbic acid was used to induce lipid peroxidation and the ability of the extracts of the leaves of L. javanica to attenuate lipid peroxidation was investigated in rat brain homogenate and compared to that of Trolox. All of the extracts of L. javanica significantly attenuated toxininduced lipid peroxidation at all concentrations used. All of the extracts were also able to significantly attenuate toxin–induced lipid peroxidation to values lower than that of the control. These results suggest that all of the extracts of L. javanica possess the ability to attenuate not only toxin–induced lipid peroxidation, but also lipid peroxidation that occurs during normal processes in the brain. The petroleum ether extract was subjected to bioassay–guided fractionation using column and thin–layer chromatography and the NBT and TBARS assays. Fraction DD1 was investigated by means of nuclear magnetic resonance, infrared and mass spectrometry. The exact structure of fraction DD1 was not elucidated. Considering all the results, it is clear that L. javanica shows great potential as a medicinal plant with antioxidant activity and may therefore be beneficial in diminishing the destructive oxidative effects inflicted by free radicals. There are however still many compounds to be isolated from L. javanica. Key words: Verbenaceae, Lippia javanica, antioxidant, neurodegeneration, oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), nitro–blue tetrazolium assay (NBT), thiobarbituric acid reactive substances assay (TBARS). / Thesis (M.Sc. (Pharmaceutical Chemistry))--North-West University, Potchefstroom Campus, 2011.