M.M. Hala: Memoirs of an Umkhonto WeSizwe Cadre
- Authors: Hala, Mzimasi Mike
- Date: 2022-10-14
- Subjects: African National Congress , Umkhonto we Sizwe (South Africa) , Anti-apartheid movements South Africa , Anti-apartheid activists South Africa , South Africa Politics and government 1948-1994 , Hani, Chris, 1942-1993 , Holomisa, Bantu, 1955- , Bisho massacre
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/406785 , vital:70307
- Description: Born in Komani (Queenstown) in 1959 and detained for Congress of South African Students (COSAS) activities while still at school, Mzimasi Mike Hala departed South Africa via Swaziland in 1981 and joined uMkhonto WeSizwe (MK). Trained in Angola, Cuba and East Germany, he commanded Cacuso camp in Angola, until redeployed to South Africa in 1987 to work underground in Venda and Cape Town. Following the unbanning of the liberation movements in 1990, he was appointed Commander of MK’s Transkei Region, where he was in charge of Chris Hani’s personal security. For reasons of space, the memoir does not proceed beyond his integration into the South African National Defence Force (SANDF) with the rank of Lieutenant-Colonel and second-in-command of SANDF Group 46 in Mthatha. Besides its value as a primary source of previously undocumented information, the thesis seeks to bridge the gap between the academic literature on MK and the lived experience of MK soldiers. Having considered both the academic literature and the published MK memoirs in Chapter One, the thesis refers back to the literature in narrative chapters Two to Five. Consolidating its findings in its conclusion, the final chapter is divided into three sections: the political culture of MK, MK gender dynamics and the consequences of the political merger of the “exiles,” including MK, and the “inziles” who subsequently came to dominate the ANC. , Thesis (MA) -- Faculty of Humanities, Institute of Social and Economic Research, 2022
- Full Text:
- Authors: Hala, Mzimasi Mike
- Date: 2022-10-14
- Subjects: African National Congress , Umkhonto we Sizwe (South Africa) , Anti-apartheid movements South Africa , Anti-apartheid activists South Africa , South Africa Politics and government 1948-1994 , Hani, Chris, 1942-1993 , Holomisa, Bantu, 1955- , Bisho massacre
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/406785 , vital:70307
- Description: Born in Komani (Queenstown) in 1959 and detained for Congress of South African Students (COSAS) activities while still at school, Mzimasi Mike Hala departed South Africa via Swaziland in 1981 and joined uMkhonto WeSizwe (MK). Trained in Angola, Cuba and East Germany, he commanded Cacuso camp in Angola, until redeployed to South Africa in 1987 to work underground in Venda and Cape Town. Following the unbanning of the liberation movements in 1990, he was appointed Commander of MK’s Transkei Region, where he was in charge of Chris Hani’s personal security. For reasons of space, the memoir does not proceed beyond his integration into the South African National Defence Force (SANDF) with the rank of Lieutenant-Colonel and second-in-command of SANDF Group 46 in Mthatha. Besides its value as a primary source of previously undocumented information, the thesis seeks to bridge the gap between the academic literature on MK and the lived experience of MK soldiers. Having considered both the academic literature and the published MK memoirs in Chapter One, the thesis refers back to the literature in narrative chapters Two to Five. Consolidating its findings in its conclusion, the final chapter is divided into three sections: the political culture of MK, MK gender dynamics and the consequences of the political merger of the “exiles,” including MK, and the “inziles” who subsequently came to dominate the ANC. , Thesis (MA) -- Faculty of Humanities, Institute of Social and Economic Research, 2022
- Full Text:
Teachers’ narratives of their experience of teaching Fetal Alcohol Spectrum Disorders (FASD) -affected children in mainstream schooling
- Authors: Makasi-Simukonda, Mihlali
- Date: 2022-10-14
- Subjects: Fetal alcohol spectrum disorders South Africa Buffalo City , Inclusive education South Africa Buffalo City , Teachers of children with disabilities South Africa Buffalo City , Children of prenatal alcohol abuse South Africa Buffalo City Social conditions , Social constructionism
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/406183 , vital:70246
- Description: This study considers the experiences of teachers working with Fetal Alcohol Spectrum Disorders (FASD)-affected children in mainstream schooling. FASD refers to a range of conditions in children resulting from maternal alcohol consumption during pregnancy. Prevalence rates in South Africa are said to be among the highest in the world. There is a paucity of research on FASD-affected children in the Eastern Cape. This study was thus conducted in the Buffalo City Metropolitan Municipality (BCMM). Teachers told stories of their experiences of teaching undiagnosed FASD-affected children in mainstream schooling. The stories reveal the quality of life for children in classrooms and socio-contextual influences of their experiences at school, as well as the responsibilisation of teachers in these settings. This qualitative research is situated within a social constructionism paradigm. The social model of disability was utilised as a theoretical framework for this study to represent the systemic barriers against which FASD-affected learner support in the mainstream school can be conceptualised. A sample of five (5) Black female teachers were interviewed. Data gathering was done by conducting telephonic interviews using the single question inducing narrative, known as SQUIN. A thematic analysis of the data was conducted for the purpose of identifying themes within the data collected. Four superordinate themes emerged from the data, viz. narratives of FASD-affected learners, narratives of parental involvement, narratives of the role of teachers and narratives of stakeholder support. This study reveals a significant level of responsibilisation of teachers. Teachers appear to play a bigger role than just being teachers and they positioned themselves as playing a parental role in relation to FASD-affected learners. FASD-affected learners were positioned as “the problem” in the mainstream classroom whereas the environment is a disabling factor to FASD-affected learners. There is a need for a holistic approach in working with FASD-affected learners from all relevant stakeholders. Key recommendations are made for effective inclusion of FASD-affected learners in mainstream schooling. Future research recommendations include repeating this study in a different geographical area, a focus on senior phase learners and/or a more diverse sample. The findings assist in the development of policies of inclusive education in mainstream schooling and supportive strategies to enhance the developmental trajectory of FASD-affected children. , Thesis (MA) -- Faculty of Humanities, Psychology, 2022
- Full Text:
- Authors: Makasi-Simukonda, Mihlali
- Date: 2022-10-14
- Subjects: Fetal alcohol spectrum disorders South Africa Buffalo City , Inclusive education South Africa Buffalo City , Teachers of children with disabilities South Africa Buffalo City , Children of prenatal alcohol abuse South Africa Buffalo City Social conditions , Social constructionism
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/406183 , vital:70246
- Description: This study considers the experiences of teachers working with Fetal Alcohol Spectrum Disorders (FASD)-affected children in mainstream schooling. FASD refers to a range of conditions in children resulting from maternal alcohol consumption during pregnancy. Prevalence rates in South Africa are said to be among the highest in the world. There is a paucity of research on FASD-affected children in the Eastern Cape. This study was thus conducted in the Buffalo City Metropolitan Municipality (BCMM). Teachers told stories of their experiences of teaching undiagnosed FASD-affected children in mainstream schooling. The stories reveal the quality of life for children in classrooms and socio-contextual influences of their experiences at school, as well as the responsibilisation of teachers in these settings. This qualitative research is situated within a social constructionism paradigm. The social model of disability was utilised as a theoretical framework for this study to represent the systemic barriers against which FASD-affected learner support in the mainstream school can be conceptualised. A sample of five (5) Black female teachers were interviewed. Data gathering was done by conducting telephonic interviews using the single question inducing narrative, known as SQUIN. A thematic analysis of the data was conducted for the purpose of identifying themes within the data collected. Four superordinate themes emerged from the data, viz. narratives of FASD-affected learners, narratives of parental involvement, narratives of the role of teachers and narratives of stakeholder support. This study reveals a significant level of responsibilisation of teachers. Teachers appear to play a bigger role than just being teachers and they positioned themselves as playing a parental role in relation to FASD-affected learners. FASD-affected learners were positioned as “the problem” in the mainstream classroom whereas the environment is a disabling factor to FASD-affected learners. There is a need for a holistic approach in working with FASD-affected learners from all relevant stakeholders. Key recommendations are made for effective inclusion of FASD-affected learners in mainstream schooling. Future research recommendations include repeating this study in a different geographical area, a focus on senior phase learners and/or a more diverse sample. The findings assist in the development of policies of inclusive education in mainstream schooling and supportive strategies to enhance the developmental trajectory of FASD-affected children. , Thesis (MA) -- Faculty of Humanities, Psychology, 2022
- Full Text:
Design, synthesis, manufacture, characterization and evaluation of lipid nanocapsules in chitosan-iota-carrageenan based hydrogel scaffold as a potential anti-Covid-19 drug delivery system
- Authors: Mukubwa, Grady Kathondo
- Date: 2022-10-14
- Subjects: Nanocapsules Design , Hydrogel , COVID-19 (Disease) , Characterization , Drug delivery systems
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/364955 , vital:65665
- Description: Covid-19 is a deadly viral disease that has been rampant around the world since 2019. Although the successful introduction of the vaccine has reduced the spread of covid-19, new cases and deaths are still being recorded. To date, no specific curative antiviral treatment has been approved for covid-19. However, many existing antiviral drugs have been and are still being studied against covid-19 and some of them, such as Remdesivir, have shown promise and could be repurposed to treat this infection. Unfortunately, antiviral drugs are prone to resistance as most of them have poor biopharmaceutical properties, including low solubility, permeability and bioavailability, which could hinder any clinical success. Recent advances in nanotechnology-based delivery systems have made it possible to improve the biopharmaceutical properties of many drugs, especially those of poorly water-soluble drugs, by formulating them as lipid nanoparticles (LNP). Thus, in order to contribute to the fight against covid-19, this work aimed to develop Lipid Nanocapsules (LNC), based on some natural raw materials, which could improve the biopharmaceutical properties of antiviral drugs. In addition, since covid-19 infection is mainly respiratory, this work also aimed to fabricate a targeted delivery system based on a hydrogel capable of entrapping LNC and ensuring their efficient deposition and release in the lungs. The LNC consisted of a mixture of medium-chain triglycerides oil (MCT oil), crude soy lecithin, tween 80, NaCl and water, while the hydrogel consisted of a chitosan-grafted-iota carrageenan-grafted-poly (acrylamide-co-acrylic acid) system (CS-iCar-p (AAm-Co-AA)). Efavirenz (EFV), a drug with very low water solubility that has recently been demonstrated to have the potential to influence sars-cov-2 life cycle through different targets (3CLP, RdRp, Hellicase, 3’to5’exonuclease, 2’-O-ribose methyltransferase and EndoRNAse), was chosen as the model drug to evaluate the developed delivery system. The combination of LNP and hydrogel results in a delivery system known as the LNP-hydrogel composite, an emerging area of research in the field of drug delivery. To date, no research has reported the design and fabrication of an LNC-CS-iCar-p (AAm-Co-AA) hydrogel composite that could effectively deliver an antiviral drug to the lungs in addition to its advantages in terms of biological activities. Prior to the design of experiment, EFV solubility was assessed in water, labrafac lipophile 1349 and MCT oil. After that, the Design Expert Software version 13 was used to design the different experiments performed in this work. The I-optimal mixture design of experiments was performed for both LNC preparation and CS-iCar-p (AAm-Co-AA) hydrogel synthesis to study the impact of raw materials on the characteristics of these delivery systems. LNC were prepared using the phase inversion method while the free radical precipitation graft copolymerization method was used to synthesize hydrogel. In order to build polynomial models that could predict the amount of drug both LNC and CS-iCar-p (AAm-Co-AA) hydrogel can entrap, a D-optimal (custom) randomized design was performed. Moreover, various characterization techniques were used to investigate the physicochemical properties of the developed delivery systems. Thereafter, drug release studies were performed using a 1% sodium lauryl sulfate solution adjusted to either pH 4 or 7. Solubility studies revealed that EFV was more soluble in labrafac lipophile 1349 and in MCT oil than in water; therefore, given its affordability, MCT oil was used for the LNC formulation. The design of experiment carried out allowed the construction of polynomial models that could predict, on the one hand, the droplet size, the polydispersity index and the Zeta potential of LNC, which were respectively around 50nm, below 0.2 and below -33. On the other hand, the model could predict the swelling capacity of the synthesized hydrogel, which was optimised to about 30,000% (300 g of water to 1 g of hydrogel). This turned out to be influenced by the proportion of polymers, the ratio of monomers as well as the concentration of the cross-linking agent. In addition, the characterization techniques further supported the improvement of EFV solubility by highlighting its conversion into its amorphous state after encapsulation in LNC. They also confirmed successful synthesis of CS-iCar-p (AAm-co-AA) hydrogel. LNC were able to encapsulate about 87% of EFV while the synthesized CS-iCar-p (AAm-co-AA) hydrogel entrapped around 53% of EFV encapsulated in LNC. While LNC were able to release 42% and 27% of EFV after 74 hours in a 1% sodium lauryl sulfate solution (SLS) at pH 7 and pH 4 respectively, the LNC-CS-iCar-p (AAm-co-AA) hydrogel composite released about 50% and 40% of the drug after 9 days in the same release medium. Interestingly, the chemical integrity of the drug was preserved throughout the manufacturing process up to after its release, suggesting that the developed LNC-CS-iCar-p (AAm-co-AA) hydrogel composite could be used as a novel potential anticovid-19 drugs delivery system. , Thesis (MSc) -- Faculty of Science, Chemistry, 2022
- Full Text:
- Authors: Mukubwa, Grady Kathondo
- Date: 2022-10-14
- Subjects: Nanocapsules Design , Hydrogel , COVID-19 (Disease) , Characterization , Drug delivery systems
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/364955 , vital:65665
- Description: Covid-19 is a deadly viral disease that has been rampant around the world since 2019. Although the successful introduction of the vaccine has reduced the spread of covid-19, new cases and deaths are still being recorded. To date, no specific curative antiviral treatment has been approved for covid-19. However, many existing antiviral drugs have been and are still being studied against covid-19 and some of them, such as Remdesivir, have shown promise and could be repurposed to treat this infection. Unfortunately, antiviral drugs are prone to resistance as most of them have poor biopharmaceutical properties, including low solubility, permeability and bioavailability, which could hinder any clinical success. Recent advances in nanotechnology-based delivery systems have made it possible to improve the biopharmaceutical properties of many drugs, especially those of poorly water-soluble drugs, by formulating them as lipid nanoparticles (LNP). Thus, in order to contribute to the fight against covid-19, this work aimed to develop Lipid Nanocapsules (LNC), based on some natural raw materials, which could improve the biopharmaceutical properties of antiviral drugs. In addition, since covid-19 infection is mainly respiratory, this work also aimed to fabricate a targeted delivery system based on a hydrogel capable of entrapping LNC and ensuring their efficient deposition and release in the lungs. The LNC consisted of a mixture of medium-chain triglycerides oil (MCT oil), crude soy lecithin, tween 80, NaCl and water, while the hydrogel consisted of a chitosan-grafted-iota carrageenan-grafted-poly (acrylamide-co-acrylic acid) system (CS-iCar-p (AAm-Co-AA)). Efavirenz (EFV), a drug with very low water solubility that has recently been demonstrated to have the potential to influence sars-cov-2 life cycle through different targets (3CLP, RdRp, Hellicase, 3’to5’exonuclease, 2’-O-ribose methyltransferase and EndoRNAse), was chosen as the model drug to evaluate the developed delivery system. The combination of LNP and hydrogel results in a delivery system known as the LNP-hydrogel composite, an emerging area of research in the field of drug delivery. To date, no research has reported the design and fabrication of an LNC-CS-iCar-p (AAm-Co-AA) hydrogel composite that could effectively deliver an antiviral drug to the lungs in addition to its advantages in terms of biological activities. Prior to the design of experiment, EFV solubility was assessed in water, labrafac lipophile 1349 and MCT oil. After that, the Design Expert Software version 13 was used to design the different experiments performed in this work. The I-optimal mixture design of experiments was performed for both LNC preparation and CS-iCar-p (AAm-Co-AA) hydrogel synthesis to study the impact of raw materials on the characteristics of these delivery systems. LNC were prepared using the phase inversion method while the free radical precipitation graft copolymerization method was used to synthesize hydrogel. In order to build polynomial models that could predict the amount of drug both LNC and CS-iCar-p (AAm-Co-AA) hydrogel can entrap, a D-optimal (custom) randomized design was performed. Moreover, various characterization techniques were used to investigate the physicochemical properties of the developed delivery systems. Thereafter, drug release studies were performed using a 1% sodium lauryl sulfate solution adjusted to either pH 4 or 7. Solubility studies revealed that EFV was more soluble in labrafac lipophile 1349 and in MCT oil than in water; therefore, given its affordability, MCT oil was used for the LNC formulation. The design of experiment carried out allowed the construction of polynomial models that could predict, on the one hand, the droplet size, the polydispersity index and the Zeta potential of LNC, which were respectively around 50nm, below 0.2 and below -33. On the other hand, the model could predict the swelling capacity of the synthesized hydrogel, which was optimised to about 30,000% (300 g of water to 1 g of hydrogel). This turned out to be influenced by the proportion of polymers, the ratio of monomers as well as the concentration of the cross-linking agent. In addition, the characterization techniques further supported the improvement of EFV solubility by highlighting its conversion into its amorphous state after encapsulation in LNC. They also confirmed successful synthesis of CS-iCar-p (AAm-co-AA) hydrogel. LNC were able to encapsulate about 87% of EFV while the synthesized CS-iCar-p (AAm-co-AA) hydrogel entrapped around 53% of EFV encapsulated in LNC. While LNC were able to release 42% and 27% of EFV after 74 hours in a 1% sodium lauryl sulfate solution (SLS) at pH 7 and pH 4 respectively, the LNC-CS-iCar-p (AAm-co-AA) hydrogel composite released about 50% and 40% of the drug after 9 days in the same release medium. Interestingly, the chemical integrity of the drug was preserved throughout the manufacturing process up to after its release, suggesting that the developed LNC-CS-iCar-p (AAm-co-AA) hydrogel composite could be used as a novel potential anticovid-19 drugs delivery system. , Thesis (MSc) -- Faculty of Science, Chemistry, 2022
- Full Text:
Rural households’ perceptions of an invasive alien species rosa rubiginosa l. (rosehip) and the role it plays in rural livelihoods in Lesotho
- Authors: Makhorole, Thato Violet
- Date: 2022-10-14
- Subjects: Rose hips Lesotho , Invasive plants Lesotho , Rural poor Lesotho , Probit model , Principal components analysis
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/403069 , vital:69919
- Description: Despite the vast research on the negative impacts of invasive alien species on the environment, these species remain part of the rural communities due to their numerous livelihood uses. Thus, more research is required, focusing mainly on the impacts of invasive alien species on the livelihoods of rural communities. This study investigated the community perceptions of rosehip (Rosa rubiginosa) and its contribution to rural communities as an invasive alien species. Four community councils, Pitseng, Matlameng, Limamarela and Mphorosane in the Leribe District Lesotho, were assessed. The study followed the pragmatism paradigm. The contribution of rosehip to rural livelihoods was analysed by comparing income from rosehip with other income sources. The study used simple random sampling and snowball sampling to select a representative of 160 respondents. The primary data was collected using semi-structured questionnaires. Moreover, SPSS and Stata statistical package programs were used for statistical analyses. The results showed that rosehip's livelihood benefits, its negative impacts, the length of time it has been available in the area, and its abundance highly influence the social, economic and environmental perception of rural communities. Furthermore, the study revealed that although the income from rosehip is extremely low and available for only three months of the year, the income plays an important part to the poorer households who have no other income sources. The study found that the main reason for engagement in rosehip harvesting despite its challenging nature was unemployment. The study also revealed that rosehip is part of the risk-reducing strategy or income diversification. Some households used it to complement other sources of income, such as agricultural production. Rosehip trade, if well-controlled, has the potential to alleviate rural poverty by creating job opportunities, providing a source of household income, and acting as a safety net in the face of shocks such as limited job opportunities and food shortages. In conclusion, households’ perceptions of rosehip have proven that rosehip is a valuable resource that provides a supplementary income that contributes towards alleviating poverty in Lesotho’s rural communities. The study recommends the private sector to establish and manages small agro-processing industries focusing on products used daily. The study also encourages environmental education and indigenous knowledge among community members, which would include knowledge and recognition of invasive alien species and their potential benefits and threats. , Thesis (MEcon) -- Faculty of Commerce, Economics and Economic History, 2022
- Full Text:
- Authors: Makhorole, Thato Violet
- Date: 2022-10-14
- Subjects: Rose hips Lesotho , Invasive plants Lesotho , Rural poor Lesotho , Probit model , Principal components analysis
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/403069 , vital:69919
- Description: Despite the vast research on the negative impacts of invasive alien species on the environment, these species remain part of the rural communities due to their numerous livelihood uses. Thus, more research is required, focusing mainly on the impacts of invasive alien species on the livelihoods of rural communities. This study investigated the community perceptions of rosehip (Rosa rubiginosa) and its contribution to rural communities as an invasive alien species. Four community councils, Pitseng, Matlameng, Limamarela and Mphorosane in the Leribe District Lesotho, were assessed. The study followed the pragmatism paradigm. The contribution of rosehip to rural livelihoods was analysed by comparing income from rosehip with other income sources. The study used simple random sampling and snowball sampling to select a representative of 160 respondents. The primary data was collected using semi-structured questionnaires. Moreover, SPSS and Stata statistical package programs were used for statistical analyses. The results showed that rosehip's livelihood benefits, its negative impacts, the length of time it has been available in the area, and its abundance highly influence the social, economic and environmental perception of rural communities. Furthermore, the study revealed that although the income from rosehip is extremely low and available for only three months of the year, the income plays an important part to the poorer households who have no other income sources. The study found that the main reason for engagement in rosehip harvesting despite its challenging nature was unemployment. The study also revealed that rosehip is part of the risk-reducing strategy or income diversification. Some households used it to complement other sources of income, such as agricultural production. Rosehip trade, if well-controlled, has the potential to alleviate rural poverty by creating job opportunities, providing a source of household income, and acting as a safety net in the face of shocks such as limited job opportunities and food shortages. In conclusion, households’ perceptions of rosehip have proven that rosehip is a valuable resource that provides a supplementary income that contributes towards alleviating poverty in Lesotho’s rural communities. The study recommends the private sector to establish and manages small agro-processing industries focusing on products used daily. The study also encourages environmental education and indigenous knowledge among community members, which would include knowledge and recognition of invasive alien species and their potential benefits and threats. , Thesis (MEcon) -- Faculty of Commerce, Economics and Economic History, 2022
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An investigation of how newly appointed team leaders in an automotive manufacturing organisation experience the role transition from specialists to team leaders
- Authors: Mzelemu, Patrick Sbusiso
- Date: 2022-10-14
- Subjects: Leadership , Automobile industry executives , Role expectation , Identity (Psychology) , Personnel management , Organizational behavior
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/403036 , vital:69916
- Description: The purpose of this study is to investigate and understand how specialists who are newly appointed team leaders in an automotive manufacturing organisation experience the transition from specialists to team leaders. The role of leading others in an organisation is linked to human resource leadership. The human resource is highlighted as a valuable resource as it is the only thinking resource in an organisation. It also influences cost and quality and it is central to the organisation's competitive advantage. Consequently, any interruptions or lack of effective management of the human resource will result in a loss of value that the human resource brings into the organisation since they maximise the use of all other resources. The role transitioning process takes place in three phases. These are the separation, transitioning and incorporation phases. In the separation phase, the transitioning individuals have difficulty detaching from their previous role; the individuals still having the urge to do the work for their subordinates instead of delegating illustrate this. In the transition phase, the individuals in the study experienced the anxiety of wanting to make the new role a success whilst experiencing a sense of lack of belonging due to not being in the familiar old role and not entrenched in the new role. In the incorporation phase, the transitioning individuals experience a sense of stability where they are comfortable acting in the new role. Problem: This study aims to answer the question of how newly appointed team leaders in an automotive manufacturing organisation experience role transition from specialists to team leaders Method: The study will be employing the qualitative thematic deductive method. The target population consists of team leaders who have transitioned into the team leader position in the past two years in different departments. The data was collected by way of face-to-face semistructured interviews with a sample size of seven participants. Data analysis was conducted with a method that is consistent with thematic analysis. 3 Key Findings: Individuals transitioning have urgencies to perform their previous roles in the early phase; during the transition phase, they have anxiety because they are not yet successful in the new role, and in the final phase, they have become comfortable with their new roles and their new identities. Implications: The study's outcome is to identify insights and resultant recommendations for non-leading specialists transitioning into roles where they have to lead others. The study will add to the research in the field of leadership behaviour and organisational behaviour. , Thesis (MBA) -- Faculty of Commerce, Rhodes Business School, 2022
- Full Text:
- Authors: Mzelemu, Patrick Sbusiso
- Date: 2022-10-14
- Subjects: Leadership , Automobile industry executives , Role expectation , Identity (Psychology) , Personnel management , Organizational behavior
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/403036 , vital:69916
- Description: The purpose of this study is to investigate and understand how specialists who are newly appointed team leaders in an automotive manufacturing organisation experience the transition from specialists to team leaders. The role of leading others in an organisation is linked to human resource leadership. The human resource is highlighted as a valuable resource as it is the only thinking resource in an organisation. It also influences cost and quality and it is central to the organisation's competitive advantage. Consequently, any interruptions or lack of effective management of the human resource will result in a loss of value that the human resource brings into the organisation since they maximise the use of all other resources. The role transitioning process takes place in three phases. These are the separation, transitioning and incorporation phases. In the separation phase, the transitioning individuals have difficulty detaching from their previous role; the individuals still having the urge to do the work for their subordinates instead of delegating illustrate this. In the transition phase, the individuals in the study experienced the anxiety of wanting to make the new role a success whilst experiencing a sense of lack of belonging due to not being in the familiar old role and not entrenched in the new role. In the incorporation phase, the transitioning individuals experience a sense of stability where they are comfortable acting in the new role. Problem: This study aims to answer the question of how newly appointed team leaders in an automotive manufacturing organisation experience role transition from specialists to team leaders Method: The study will be employing the qualitative thematic deductive method. The target population consists of team leaders who have transitioned into the team leader position in the past two years in different departments. The data was collected by way of face-to-face semistructured interviews with a sample size of seven participants. Data analysis was conducted with a method that is consistent with thematic analysis. 3 Key Findings: Individuals transitioning have urgencies to perform their previous roles in the early phase; during the transition phase, they have anxiety because they are not yet successful in the new role, and in the final phase, they have become comfortable with their new roles and their new identities. Implications: The study's outcome is to identify insights and resultant recommendations for non-leading specialists transitioning into roles where they have to lead others. The study will add to the research in the field of leadership behaviour and organisational behaviour. , Thesis (MBA) -- Faculty of Commerce, Rhodes Business School, 2022
- Full Text:
Mediating transition to household electricity conservation through co-designing in Makhanda, South Africa
- Authors: Mutumbi, Uzziah
- Date: 2022-04-06
- Subjects: Households Energy consumption South Africa Makhanda , Renewable energy sources South Africa , Consumer behavior South Africa Makhanda , Energy conservation South Africa Makhanda Citizen participation , Low-income consumers South Africa Makhanda
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/291262 , vital:56838
- Description: Households consume up to 20% of total global electricity consumption, hence they are important role players in efforts for promoting sustainable consumption. Research on electricity-use behaviour and effectiveness of interventions is important for informing intervention strategies. However, relative to developed countries, research on this subject is lacking in developing countries, where electricity access is limited, and a substantial proportion of households are energy poor. The main goal of this study was to examine reported electricity-use behaviour and the effects of co-designed interventions on household electricity consumption in low-income households in Makhanda, South Africa, using a field-based experiment. Findings from self-reported electricity-use practices showed mixed results, with households reporting both good electricity-use behaviour (e.g., cooling down hot food before refrigeration) and wasteful actions (e.g., leaving appliances on standby). Results showed that electricity-use behaviour was influenced by socio-psychological values including universalism, benevolence, hedonism, and power. Over the intervention period, households exposed to co-designed electricity-saving interventions showed mean electricity savings of about 5.5%, while households in the Partial Treatment and Control group showed increased electricity consumption. Socio-demographic factors such as age and psychological factors (involvement and perceived behavioural control) were very important determinants of household consumption. Broadly, the findings empirically validate the impact of collective problem formulation and co-designing interventions in promoting sustainable actions. , Thesis (MSc) -- Faculty of Science, Environmental Science, 2022
- Full Text:
- Authors: Mutumbi, Uzziah
- Date: 2022-04-06
- Subjects: Households Energy consumption South Africa Makhanda , Renewable energy sources South Africa , Consumer behavior South Africa Makhanda , Energy conservation South Africa Makhanda Citizen participation , Low-income consumers South Africa Makhanda
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/291262 , vital:56838
- Description: Households consume up to 20% of total global electricity consumption, hence they are important role players in efforts for promoting sustainable consumption. Research on electricity-use behaviour and effectiveness of interventions is important for informing intervention strategies. However, relative to developed countries, research on this subject is lacking in developing countries, where electricity access is limited, and a substantial proportion of households are energy poor. The main goal of this study was to examine reported electricity-use behaviour and the effects of co-designed interventions on household electricity consumption in low-income households in Makhanda, South Africa, using a field-based experiment. Findings from self-reported electricity-use practices showed mixed results, with households reporting both good electricity-use behaviour (e.g., cooling down hot food before refrigeration) and wasteful actions (e.g., leaving appliances on standby). Results showed that electricity-use behaviour was influenced by socio-psychological values including universalism, benevolence, hedonism, and power. Over the intervention period, households exposed to co-designed electricity-saving interventions showed mean electricity savings of about 5.5%, while households in the Partial Treatment and Control group showed increased electricity consumption. Socio-demographic factors such as age and psychological factors (involvement and perceived behavioural control) were very important determinants of household consumption. Broadly, the findings empirically validate the impact of collective problem formulation and co-designing interventions in promoting sustainable actions. , Thesis (MSc) -- Faculty of Science, Environmental Science, 2022
- Full Text:
Exploring extensive reading in cultivating positive attitudes and reading comprehension with Grade 8 Learners: a case study in a selected Namibian secondary school
- Authors: Engelbrecht, Sylviana
- Date: 2022-10-14
- Subjects: Reading comprehension Namibia , High school students Books and reading Namibia , High school students Attitudes , Fluency (Language learning) Namibia , Motivation in education Namibia , Affective education Namibia , Second language acquisition
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/403092 , vital:69921
- Description: Extensive reading, otherwise known as “reading for pleasure”, “recreational reading” or “voluntary reading”, appears to be absent in the Namibian society, especially within public educational institutions. With the reformed Education System, the Namibian Curriculum for Basic Education makes room for a reading period in a 7-day cycle, from grade 1 to 11, in which all learners are expected to engage in a sustained silent reading (SSR) of interesting material of their choice. Nevertheless, such a period seems to be insufficient for engaging learners in an enjoyable reading experience, which could possibly create a positive attitude towards reading as well as enhance their reading skills of comprehension and reading fluency, possibly resulting in a habit of individual SSR. This research intended to explore an out-of-school, extensive reading program, over the period of one year, in the hope of cultivating Grade 8 learners’ positive attitude toward reading, in a selected secondary school in the township of Katutura, Windhoek, Khomas Region, Namibia. This one-year study, in which eight Grade 8 learners participated, focused on English only reading, as it is the Language of Learning and Teaching (LoLT). Operating within the interpretivist paradigm, the research adopted a qualitative approach, using a single case study to explore how participants make meaning of their experiences and attitudes toward ER. Qualitative data was collected through pre- and post-questionnaires and document analysis of learners’ reading journals as well as the researcher’s reflective journal. The research site was purposively and conveniently selected. The study drew on Krashen’s, input and affective filter hypotheses as the driving theory of this study, which according to some scholars, is the main second language acquisition theory applied in ER research. Findings of this study suggest that ER programs have a positive impact on learners’ attitudes towards reading as well as on their reading comprehension, even in poorly resourced schools in previously disadvantaged communities. , Thesis (MEd) -- Faculty of Education, Education, 2022
- Full Text:
- Authors: Engelbrecht, Sylviana
- Date: 2022-10-14
- Subjects: Reading comprehension Namibia , High school students Books and reading Namibia , High school students Attitudes , Fluency (Language learning) Namibia , Motivation in education Namibia , Affective education Namibia , Second language acquisition
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/403092 , vital:69921
- Description: Extensive reading, otherwise known as “reading for pleasure”, “recreational reading” or “voluntary reading”, appears to be absent in the Namibian society, especially within public educational institutions. With the reformed Education System, the Namibian Curriculum for Basic Education makes room for a reading period in a 7-day cycle, from grade 1 to 11, in which all learners are expected to engage in a sustained silent reading (SSR) of interesting material of their choice. Nevertheless, such a period seems to be insufficient for engaging learners in an enjoyable reading experience, which could possibly create a positive attitude towards reading as well as enhance their reading skills of comprehension and reading fluency, possibly resulting in a habit of individual SSR. This research intended to explore an out-of-school, extensive reading program, over the period of one year, in the hope of cultivating Grade 8 learners’ positive attitude toward reading, in a selected secondary school in the township of Katutura, Windhoek, Khomas Region, Namibia. This one-year study, in which eight Grade 8 learners participated, focused on English only reading, as it is the Language of Learning and Teaching (LoLT). Operating within the interpretivist paradigm, the research adopted a qualitative approach, using a single case study to explore how participants make meaning of their experiences and attitudes toward ER. Qualitative data was collected through pre- and post-questionnaires and document analysis of learners’ reading journals as well as the researcher’s reflective journal. The research site was purposively and conveniently selected. The study drew on Krashen’s, input and affective filter hypotheses as the driving theory of this study, which according to some scholars, is the main second language acquisition theory applied in ER research. Findings of this study suggest that ER programs have a positive impact on learners’ attitudes towards reading as well as on their reading comprehension, even in poorly resourced schools in previously disadvantaged communities. , Thesis (MEd) -- Faculty of Education, Education, 2022
- Full Text:
Pathi’s sister is still troubling
- Authors: Naidoo, Savani
- Date: 2023-03-30
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/408942 , vital:70539
- Description: My thesis is a collection of micro fiction, flash fiction, fairy tales, vignettes and short stories which explore the tension of being both an insider and an outsider. I have access to different cultures without belonging to any of them: as a child, my family moved from a South African Indian community to a formerly whites-only suburb; as an adult I have lived in South Korea, the United Arab Emirates and Sudan. My prose draws on my life experiences, family legends, neighbourhood gossip, news reports and historical events to question norms and ideas that I may have taken for granted had I been fully inside a single culture. In my thesis I frequently spell words phonetically to mimic how I hear or remember them. I also borrow words from languages I don’t speak. I want the languages I use and mix to corrupt each other, as Raymond Federman put it, in order to better express the voices and contexts of the communities I draw inspiration from. Kuzhali Manickavel’s Things We Found During the Autopsy showed me that culturally rich imagery can be used without interrupting narrative flow with explanations. I am also influenced by the poetic sense of rhythm and melody of Lydia Davis’s minimalist prose, and by Sandra Cisneros’s The House on Mango Street, where each concise short story stands alone but together creates a broad understanding of people and place. Anthologies such as PP/FF, edited by Peter Conners, and My Mother She Killed Me, My Father He Ate Me, edited by Kate Bernheimer, have inspired me to be bold in finding the form that best allows each narrative to be told. , Thesis (MA) -- Faculty of Humanities, School of Languages and Literatures, 2023
- Full Text:
- Authors: Naidoo, Savani
- Date: 2023-03-30
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/408942 , vital:70539
- Description: My thesis is a collection of micro fiction, flash fiction, fairy tales, vignettes and short stories which explore the tension of being both an insider and an outsider. I have access to different cultures without belonging to any of them: as a child, my family moved from a South African Indian community to a formerly whites-only suburb; as an adult I have lived in South Korea, the United Arab Emirates and Sudan. My prose draws on my life experiences, family legends, neighbourhood gossip, news reports and historical events to question norms and ideas that I may have taken for granted had I been fully inside a single culture. In my thesis I frequently spell words phonetically to mimic how I hear or remember them. I also borrow words from languages I don’t speak. I want the languages I use and mix to corrupt each other, as Raymond Federman put it, in order to better express the voices and contexts of the communities I draw inspiration from. Kuzhali Manickavel’s Things We Found During the Autopsy showed me that culturally rich imagery can be used without interrupting narrative flow with explanations. I am also influenced by the poetic sense of rhythm and melody of Lydia Davis’s minimalist prose, and by Sandra Cisneros’s The House on Mango Street, where each concise short story stands alone but together creates a broad understanding of people and place. Anthologies such as PP/FF, edited by Peter Conners, and My Mother She Killed Me, My Father He Ate Me, edited by Kate Bernheimer, have inspired me to be bold in finding the form that best allows each narrative to be told. , Thesis (MA) -- Faculty of Humanities, School of Languages and Literatures, 2023
- Full Text:
In silico substrate binding profiling for SARS-COV-2 main protease (mpro) using hexapeptide substrates
- Authors: Zabo, Sophakama
- Date: 2022-10-14
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/365566 , vital:65760
- Description: COVID-19, as a disease resulting from SARS-CoV-2 infection, and a pandemic has had a devastating effect on the world. There are limited effective measures that control the spread and treatment of COVID-19 illness. The homodimeric cysteine main protease (Mpro) is crucial to the life cycle of the virus, as it cleaves the large polyproteins 1a and 1ab into matured, functional non-structural proteins. The Mpro exhibits high degrees of conservation in sequence, structure and specificity across coronavirus species, making it an ideal drug target. The Mpro substrate-binding profiles remain, despite the resolution of its recognition sequence and cleavage points (Leu-Gln↓(Ser/Ala/Gly)). In this study, a series of hexapeptide sequences containing the appropriate recognition sequence and cleavage points were generated and screened against the Mpro to study these binding profiles, and to further be the basis for efficiency-driven drug design. A multi-conformer hexapeptide substrate library comprising optimised 81000 models of 810 unique sequences was generated using RDKit within the context of python. Terminal capping with ACE and NMe was effected using SMILES and SMARTS matching. Multiple hexapeptides were complexed with chain B of crystallographic Mpro (PDS ID: 6XHM), following the validation of chain B for this purpose using AutoDock Vina at high levels of exhaustiveness (480). The resulting Vina scores ranged between -8.7 and -7.0 kcal.mol-1, and the reproducibility of best poses was validated through redocking. Ligand efficiency indices were calculated to identify substrate residues with high binding efficiency at their respective positions, revealing Val (P3), Ala (P1′); and Gly and Ala (P2′ and P3′) as leading efficient binders. Binding efficiencies were lowered by molecular weight. Substrate recognition was assessed by mapping of binding subsites, and Mpro specificity was evaluated through the resolution of intermolecular interaction at the binding interface. Molecular dynamics simulations for 20 ns were performed to assess the stability and behaviour of 132 Mpro systems complexed with KLQ*** substrates. Principal component analysis (PCA), was performed to assess II protein motions and conformational changes during the simulations. A strategy was formulated to classify and evaluate relations in the Mpro PCA motions, revealing four main clades of similarity. Similarity within a clade (Group 2) and dissimilarity between clades were confirmed. Trajectory visualisation revealed complex stability, substrate unbinding and dimer dissociation for various Mpro systems. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2022
- Full Text:
- Authors: Zabo, Sophakama
- Date: 2022-10-14
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/365566 , vital:65760
- Description: COVID-19, as a disease resulting from SARS-CoV-2 infection, and a pandemic has had a devastating effect on the world. There are limited effective measures that control the spread and treatment of COVID-19 illness. The homodimeric cysteine main protease (Mpro) is crucial to the life cycle of the virus, as it cleaves the large polyproteins 1a and 1ab into matured, functional non-structural proteins. The Mpro exhibits high degrees of conservation in sequence, structure and specificity across coronavirus species, making it an ideal drug target. The Mpro substrate-binding profiles remain, despite the resolution of its recognition sequence and cleavage points (Leu-Gln↓(Ser/Ala/Gly)). In this study, a series of hexapeptide sequences containing the appropriate recognition sequence and cleavage points were generated and screened against the Mpro to study these binding profiles, and to further be the basis for efficiency-driven drug design. A multi-conformer hexapeptide substrate library comprising optimised 81000 models of 810 unique sequences was generated using RDKit within the context of python. Terminal capping with ACE and NMe was effected using SMILES and SMARTS matching. Multiple hexapeptides were complexed with chain B of crystallographic Mpro (PDS ID: 6XHM), following the validation of chain B for this purpose using AutoDock Vina at high levels of exhaustiveness (480). The resulting Vina scores ranged between -8.7 and -7.0 kcal.mol-1, and the reproducibility of best poses was validated through redocking. Ligand efficiency indices were calculated to identify substrate residues with high binding efficiency at their respective positions, revealing Val (P3), Ala (P1′); and Gly and Ala (P2′ and P3′) as leading efficient binders. Binding efficiencies were lowered by molecular weight. Substrate recognition was assessed by mapping of binding subsites, and Mpro specificity was evaluated through the resolution of intermolecular interaction at the binding interface. Molecular dynamics simulations for 20 ns were performed to assess the stability and behaviour of 132 Mpro systems complexed with KLQ*** substrates. Principal component analysis (PCA), was performed to assess II protein motions and conformational changes during the simulations. A strategy was formulated to classify and evaluate relations in the Mpro PCA motions, revealing four main clades of similarity. Similarity within a clade (Group 2) and dissimilarity between clades were confirmed. Trajectory visualisation revealed complex stability, substrate unbinding and dimer dissociation for various Mpro systems. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2022
- Full Text:
Social Learning and Regenerative Sustainability: Unlocking value created in sustainability projects in higher education
- Authors: Royle, Carlene
- Date: 2023-10-13
- Subjects: Social Learning South Africa Makhanda , Sustainability South Africa Makhanda , Education, Higher Social aspects South Africa Makhanda , Social accounting South Africa Makhanda , Education, Higher Activity programs South Africa Makhanda , Value creation , Agent (Philosophy) , Sustainable Development Goals
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/402981 , vital:69911
- Description: This qualitative study considers two bodies of theory, regenerative sustainability and social learning theory, within a tertiary education context. Universities offer unique opportunities, across both the formal and informal curriculum, that engage with the two-fold socioeconomic and environmental crises. The study explores both the promise of unlocking value in an ex-linear economy; and the healing and regenerative benefits in moving toward greener practices. Through an embedded case study at Rhodes University, in the Eastern Cape of South Africa, this research explores the value that is being unlocked in campus-based sustainability projects, when considered through a social learning lens (Lave and Wenger, 1991). Invitations to participate in this study were extended to project-leads of sustainability initiatives originating from Rhodes University campus. Six projects, which have been active on campus in recent years, were selected and, through semi-structured interviews, the insights of 12 participants were captured and analysed using the associated Value Creation Framework (VCF) developed by Wenger, Trayner and De Laat (Wenger, Trayner, de Laat, 2011) and later Wenger-Trayner and Wenger Trayner (2014, 2015, 2020). The VCF consists of value cycles that interconnect dynamically, including Orientating, Strategic, Enabling, Immediate, Potential, Applied, Realised and Reframed or Transformative value cycles. The study identified instances of all these value cycles, within and across the six embedded case studies (project). Drawing on social learning theorists the analysis further focused on instances of collaboration, agency and boundary crossing. Additional themes that emerged across the majority of the embedded study projects, were a convergence of socio and eco; a whole-systems perspective shared by project leads; the importance of the social commons; language as an enabler; and problem solving for system change. A recommendation is proposed to further unlock the unbounded value created through such projects by reframing the informal curriculum opportunities offered on university campuses as social learning spaces where students can practice agency. To do so, would require formalizing various forms of system support, thus facilitating expanded learning in regenerative sustainability activities, for the common social and planetary good. , Thesis (MEd) -- Faculty of Education, Education, 2023
- Full Text:
- Authors: Royle, Carlene
- Date: 2023-10-13
- Subjects: Social Learning South Africa Makhanda , Sustainability South Africa Makhanda , Education, Higher Social aspects South Africa Makhanda , Social accounting South Africa Makhanda , Education, Higher Activity programs South Africa Makhanda , Value creation , Agent (Philosophy) , Sustainable Development Goals
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/402981 , vital:69911
- Description: This qualitative study considers two bodies of theory, regenerative sustainability and social learning theory, within a tertiary education context. Universities offer unique opportunities, across both the formal and informal curriculum, that engage with the two-fold socioeconomic and environmental crises. The study explores both the promise of unlocking value in an ex-linear economy; and the healing and regenerative benefits in moving toward greener practices. Through an embedded case study at Rhodes University, in the Eastern Cape of South Africa, this research explores the value that is being unlocked in campus-based sustainability projects, when considered through a social learning lens (Lave and Wenger, 1991). Invitations to participate in this study were extended to project-leads of sustainability initiatives originating from Rhodes University campus. Six projects, which have been active on campus in recent years, were selected and, through semi-structured interviews, the insights of 12 participants were captured and analysed using the associated Value Creation Framework (VCF) developed by Wenger, Trayner and De Laat (Wenger, Trayner, de Laat, 2011) and later Wenger-Trayner and Wenger Trayner (2014, 2015, 2020). The VCF consists of value cycles that interconnect dynamically, including Orientating, Strategic, Enabling, Immediate, Potential, Applied, Realised and Reframed or Transformative value cycles. The study identified instances of all these value cycles, within and across the six embedded case studies (project). Drawing on social learning theorists the analysis further focused on instances of collaboration, agency and boundary crossing. Additional themes that emerged across the majority of the embedded study projects, were a convergence of socio and eco; a whole-systems perspective shared by project leads; the importance of the social commons; language as an enabler; and problem solving for system change. A recommendation is proposed to further unlock the unbounded value created through such projects by reframing the informal curriculum opportunities offered on university campuses as social learning spaces where students can practice agency. To do so, would require formalizing various forms of system support, thus facilitating expanded learning in regenerative sustainability activities, for the common social and planetary good. , Thesis (MEd) -- Faculty of Education, Education, 2023
- Full Text:
Expression, partial characterisation and utilization of a GH11 xylanase (Xyn2A) from Trichoderma viride as an additive in monogastric animal feeds
- Mzimkulu-Ncoyi, Nosabatha Happyness
- Authors: Mzimkulu-Ncoyi, Nosabatha Happyness
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422409 , vital:71940
- Description: Endo-xylanases (shortly called xylanases) are a group of glycoside hydrolase enzymes that target β-D-1,4-linkages in the xylan backbone, leading to the production of xylooligosaccharides (XOS) of varying degree of polymerization (DP). Xylan is an indigestible non-starch polysaccharide present in monogastric animal feeds which in high amounts leads to increased digesta viscosity, slow movement of digesta in the intestines, malabsorption of nutrients among other challenges. The aim of this study was to investigate the effect of xylanase 2A (Xyn2A) from Trichoderma viride on broiler chicken feeds, particularly the hydrolysis of the xylan content, reduction of feed viscosity and the effect of produced XOS on eliciting the growth of gut associated probiotic bacteria. Xyn2AE was successfully induced with 0.8 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and produced in Escherichia coli BL21 (DE3) and Xyn2AC was expressed in tobacco mosaic plants. For the purification of Xyn2AE, an immobilized metal affinity chromatography (IMAC) column and diafiltration using a 3kDa cut-off Amicon filter membranes were used. Xyn2AE and Xyn2AC showed a xylanase active band at a relative weight of 21 kDa. Both enzymes showed high specificity towards soluble wheat arabinoxylan (WAX), with specific activities of 7.61 U/mg for Xyn2AE and 536.5 U/mg for Xyn2AC. Xyn2A kinetic parameters (Vmax and Km) were determined by Michaelis-Menten plots on soluble and insoluble WAX. The Vmax and Km values of Xyn2AC were 1003.01 U/mg and 9.25 mg/mL, 302.89 U/mg and 13.54 mg/mL, respectively. The Vmax and Km values of Xyn2AE for soluble and insoluble WAX were 20.45 U/mg and 12.95 mg/mL, and 8.31 U/mg and 13.15 mg/mL. Xyn2A enzymes displayed optimum activity at pH and temperature parameters of 5.0 and 50°C, respectively, and stability in temperatures ranging between 50 and 80°C and pH 4.0-9.0. Broiler chicken feeds were hydrolysed using Xyn2AE over a 24 h period and analysed using the dinitrosalicylic (DNS) assay, thin layer chromatography (TLC), viscometry and visualized using scanning electron microscope (SEM). The results showed a release of release of XOS xylotriose, xylopentose and xylohexose; enzyme’s ability to decrease the viscosity of the feeds and punched holes of feed surface, which was indicative of xylanase action. XOS produced during hydrolysis was used to study prebiotic effect on selected few bacteria and released short chain fatty acids (SCFAs) were measured. Additionally, SCFAs formation was detected in the presence of XOS as a carbon source for S. thermophilus and L. bulgaricus, whereas B. subtilis formed fewer organic acids in the presence of XOS. The results obtained from this study demonstrated that the supplementation of Xyn2A on broiler feeds has ii a positive effect in decreasing feed viscosity. Furthermore, the results of this investigation will assist the South African poultry farming sector to increase profitability in poultry farming and gain stability in the global trade as far as poultry feed is concerned. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
- Authors: Mzimkulu-Ncoyi, Nosabatha Happyness
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422409 , vital:71940
- Description: Endo-xylanases (shortly called xylanases) are a group of glycoside hydrolase enzymes that target β-D-1,4-linkages in the xylan backbone, leading to the production of xylooligosaccharides (XOS) of varying degree of polymerization (DP). Xylan is an indigestible non-starch polysaccharide present in monogastric animal feeds which in high amounts leads to increased digesta viscosity, slow movement of digesta in the intestines, malabsorption of nutrients among other challenges. The aim of this study was to investigate the effect of xylanase 2A (Xyn2A) from Trichoderma viride on broiler chicken feeds, particularly the hydrolysis of the xylan content, reduction of feed viscosity and the effect of produced XOS on eliciting the growth of gut associated probiotic bacteria. Xyn2AE was successfully induced with 0.8 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and produced in Escherichia coli BL21 (DE3) and Xyn2AC was expressed in tobacco mosaic plants. For the purification of Xyn2AE, an immobilized metal affinity chromatography (IMAC) column and diafiltration using a 3kDa cut-off Amicon filter membranes were used. Xyn2AE and Xyn2AC showed a xylanase active band at a relative weight of 21 kDa. Both enzymes showed high specificity towards soluble wheat arabinoxylan (WAX), with specific activities of 7.61 U/mg for Xyn2AE and 536.5 U/mg for Xyn2AC. Xyn2A kinetic parameters (Vmax and Km) were determined by Michaelis-Menten plots on soluble and insoluble WAX. The Vmax and Km values of Xyn2AC were 1003.01 U/mg and 9.25 mg/mL, 302.89 U/mg and 13.54 mg/mL, respectively. The Vmax and Km values of Xyn2AE for soluble and insoluble WAX were 20.45 U/mg and 12.95 mg/mL, and 8.31 U/mg and 13.15 mg/mL. Xyn2A enzymes displayed optimum activity at pH and temperature parameters of 5.0 and 50°C, respectively, and stability in temperatures ranging between 50 and 80°C and pH 4.0-9.0. Broiler chicken feeds were hydrolysed using Xyn2AE over a 24 h period and analysed using the dinitrosalicylic (DNS) assay, thin layer chromatography (TLC), viscometry and visualized using scanning electron microscope (SEM). The results showed a release of release of XOS xylotriose, xylopentose and xylohexose; enzyme’s ability to decrease the viscosity of the feeds and punched holes of feed surface, which was indicative of xylanase action. XOS produced during hydrolysis was used to study prebiotic effect on selected few bacteria and released short chain fatty acids (SCFAs) were measured. Additionally, SCFAs formation was detected in the presence of XOS as a carbon source for S. thermophilus and L. bulgaricus, whereas B. subtilis formed fewer organic acids in the presence of XOS. The results obtained from this study demonstrated that the supplementation of Xyn2A on broiler feeds has ii a positive effect in decreasing feed viscosity. Furthermore, the results of this investigation will assist the South African poultry farming sector to increase profitability in poultry farming and gain stability in the global trade as far as poultry feed is concerned. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
Biochemical and genetic analysis of the Mycobacterium smegmatis CnoX Chaperedoxin
- Authors: Watkins, Ariana Heloise Jo
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422403 , vital:71939
- Description: Mycobacterium (M.) tuberculosis (Mtb) encounters numerous physical and chemical stresses associated with host immunity during infection. These include exposure to reactive oxygen, chlorine and nitrogen species, low pH, hypoxia, nutrient starvation, and metal toxicity. Cellular proteins are particularly susceptible to damage by these stresses, and the ability to prevent their irreversible damage is consequently crucial for bacterial growth and survival. Mtb employs a network of proteins that includes chaperones, disaggregases, and proteases to maintain the integrity of its proteome. The chaperedoxin, CnoX, is a recently identified stress-inducible chaperone that combines redox and holdase activities to prevent the over-oxidation and aggregation of proteins in E. coli and other proteobacterial species. In this study, we identified orthologs of the E. coli CnoX (EcCnoX) in Mtb and M. smegmatis (Msm). Bioinformatics analysis of the Mtb and Msm CnoX orthologs (MtCnoX and MsCnoX, respectively) revealed that they possess similar domains, domain architectures and predicted tertiary structures as previously characterised CnoX enzymes, i.e. an N-terminal thioredoxin (Trx) domain fused to a C-terminal TPR-motif containing domain. The EcCnoX, MsCnoX, and MtCnoX enzymes were expressed as recombinant, His-tagged proteins in E. coli and purified to near homogeneity. Biochemical analysis of the recombinant CnoX enzymes revealed that the MsCnoX and MtCnoX both lack thiol-disulphide oxidoreductase (thioredoxin) activity, as evidenced by their inability to catalyse the reduction of the disulphide bonds of insulin in vitro. Both mycobacterial CnoX enzymes displayed activity as chaperones (holdases) during thermal aggregation assays of the model substrate, malate dehydrogenase (MDH). In contrast to previously reported findings for EcCnoX, the holdase activity of the mycobacterial CnoX enzymes was constitutive and did not require exposure to hypochlorous acid (HOCl) for activation. To establish the physiological role of CnoX in Msm, cnoX knockdown (KD) and knockout (KO) mutants were generated using CRISPRi-mediated gene silencing or homologous recombination, respectively. Consistent with previous findings, CnoX activity was not essential for the growth of Msm under conventional growth conditions. Reducing or eliminating CnoX activity in the Msm KD or KO mutants, respectively, did not confer increased sensitivity to HOCl as has been observed for an E. coli cnoX mutant. Reduced CnoX activity in Msm did, however, confer sensitivity to the superoxide generator, plumbagin, and front-line antitubercular drugs rifampicin and isoniazid. The combination of biochemical and physiological data presented suggests that MsCnoX may function as a holdase for substrates following proteotoxic damage induced by certain types of oxidants, a line of investigation that will be pursued in future studies. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
- Authors: Watkins, Ariana Heloise Jo
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422403 , vital:71939
- Description: Mycobacterium (M.) tuberculosis (Mtb) encounters numerous physical and chemical stresses associated with host immunity during infection. These include exposure to reactive oxygen, chlorine and nitrogen species, low pH, hypoxia, nutrient starvation, and metal toxicity. Cellular proteins are particularly susceptible to damage by these stresses, and the ability to prevent their irreversible damage is consequently crucial for bacterial growth and survival. Mtb employs a network of proteins that includes chaperones, disaggregases, and proteases to maintain the integrity of its proteome. The chaperedoxin, CnoX, is a recently identified stress-inducible chaperone that combines redox and holdase activities to prevent the over-oxidation and aggregation of proteins in E. coli and other proteobacterial species. In this study, we identified orthologs of the E. coli CnoX (EcCnoX) in Mtb and M. smegmatis (Msm). Bioinformatics analysis of the Mtb and Msm CnoX orthologs (MtCnoX and MsCnoX, respectively) revealed that they possess similar domains, domain architectures and predicted tertiary structures as previously characterised CnoX enzymes, i.e. an N-terminal thioredoxin (Trx) domain fused to a C-terminal TPR-motif containing domain. The EcCnoX, MsCnoX, and MtCnoX enzymes were expressed as recombinant, His-tagged proteins in E. coli and purified to near homogeneity. Biochemical analysis of the recombinant CnoX enzymes revealed that the MsCnoX and MtCnoX both lack thiol-disulphide oxidoreductase (thioredoxin) activity, as evidenced by their inability to catalyse the reduction of the disulphide bonds of insulin in vitro. Both mycobacterial CnoX enzymes displayed activity as chaperones (holdases) during thermal aggregation assays of the model substrate, malate dehydrogenase (MDH). In contrast to previously reported findings for EcCnoX, the holdase activity of the mycobacterial CnoX enzymes was constitutive and did not require exposure to hypochlorous acid (HOCl) for activation. To establish the physiological role of CnoX in Msm, cnoX knockdown (KD) and knockout (KO) mutants were generated using CRISPRi-mediated gene silencing or homologous recombination, respectively. Consistent with previous findings, CnoX activity was not essential for the growth of Msm under conventional growth conditions. Reducing or eliminating CnoX activity in the Msm KD or KO mutants, respectively, did not confer increased sensitivity to HOCl as has been observed for an E. coli cnoX mutant. Reduced CnoX activity in Msm did, however, confer sensitivity to the superoxide generator, plumbagin, and front-line antitubercular drugs rifampicin and isoniazid. The combination of biochemical and physiological data presented suggests that MsCnoX may function as a holdase for substrates following proteotoxic damage induced by certain types of oxidants, a line of investigation that will be pursued in future studies. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
Implications of changing land use and introducing extralimital giraffe on vegetation in the subtropical thicket, South Africa
- Authors: Trafford, Bronwyn Kate
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422356 , vital:71935
- Description: Land use change has increased in southern Africa as many livestock farms convert to game reserves, to restore degraded and overgrazed lands. This leads to a change in herbivore communities with potential positive and negative consequences for vegetation and landscape productivity. This study aimed to look at the effect of land use change and subsequent herbivore community changes, on thicket vegetation structure, composition, and landscape productivity. To attain the study objectives, data were collected from four game reserves and their adjacent livestock farms within the Eastern Cape province in South Africa. These sites were selected because they are within the Subtropical Thicket biome (similar vegetation) and were once used for livestock farming. At each study site, vegetation composition and structure were assessed using a unique bush clump sampling technique along fence line contrasts. Six plots were sampled at each site, three plots on the game reserve lands and three plots on the adjacent livestock farmlands during the dry (July/August 2020) and wet (January/February 2021) seasons. The Normalized Difference Vegetation Index (NDVI), in-situ leaf area index (LAI) and the fraction of intercepted photosynthetically active radiation (FIPAR), were used to assess vegetation productivity between the two land use types. Additionally, diet of an extralimital herbivore, giraffe (Giraffa camelopardalis), was assessed during contrasting seasons (dry and wet) to infer the role this species might have on these landscapes. The diet of giraffes was assessed through field observations, recording plant species that giraffes were feeding on. Floristic results showed that there were no differences in vegetation structure, species composition and richness between the two land types. There was no difference in the overall diversity of plant species on the game reserves than in the adjacent livestock farms. The primary productivity was greater on the game reserves compared to the livestock farms. Field observations of giraffe feeding showed that they fed predominantly on Vachellia karroo followed by Searsia longispina. Their diets did not differ between the two seasons. My study provides an insight into the effects of changing land use and this information can be used to infer the extent to which these changes might have on restoration of degraded landscape and how land use changes may be effective in restoring the degraded thicket vegetation caused by overgrazing and overstocking of domestic livestock. , Thesis (MSc) -- Faculty of Science, Zoology and Entomology, 2023
- Full Text:
- Authors: Trafford, Bronwyn Kate
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422356 , vital:71935
- Description: Land use change has increased in southern Africa as many livestock farms convert to game reserves, to restore degraded and overgrazed lands. This leads to a change in herbivore communities with potential positive and negative consequences for vegetation and landscape productivity. This study aimed to look at the effect of land use change and subsequent herbivore community changes, on thicket vegetation structure, composition, and landscape productivity. To attain the study objectives, data were collected from four game reserves and their adjacent livestock farms within the Eastern Cape province in South Africa. These sites were selected because they are within the Subtropical Thicket biome (similar vegetation) and were once used for livestock farming. At each study site, vegetation composition and structure were assessed using a unique bush clump sampling technique along fence line contrasts. Six plots were sampled at each site, three plots on the game reserve lands and three plots on the adjacent livestock farmlands during the dry (July/August 2020) and wet (January/February 2021) seasons. The Normalized Difference Vegetation Index (NDVI), in-situ leaf area index (LAI) and the fraction of intercepted photosynthetically active radiation (FIPAR), were used to assess vegetation productivity between the two land use types. Additionally, diet of an extralimital herbivore, giraffe (Giraffa camelopardalis), was assessed during contrasting seasons (dry and wet) to infer the role this species might have on these landscapes. The diet of giraffes was assessed through field observations, recording plant species that giraffes were feeding on. Floristic results showed that there were no differences in vegetation structure, species composition and richness between the two land types. There was no difference in the overall diversity of plant species on the game reserves than in the adjacent livestock farms. The primary productivity was greater on the game reserves compared to the livestock farms. Field observations of giraffe feeding showed that they fed predominantly on Vachellia karroo followed by Searsia longispina. Their diets did not differ between the two seasons. My study provides an insight into the effects of changing land use and this information can be used to infer the extent to which these changes might have on restoration of degraded landscape and how land use changes may be effective in restoring the degraded thicket vegetation caused by overgrazing and overstocking of domestic livestock. , Thesis (MSc) -- Faculty of Science, Zoology and Entomology, 2023
- Full Text:
Photodynamic therapy using morpholine substituted porphyrins in the presence of cancer specific molecules linked to graphene quantum dots
- Magaela, Ngwanabjala Bridged
- Authors: Magaela, Ngwanabjala Bridged
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422486 , vital:71947
- Description: This thesis reports on the synthesis, characterization, photophysiochemical properties of morpholine substituted symmetrical and asymmetrical porphyrins. The synthesized porphyrins are conjugated to cancer selective biomolecules (folic acid and biotin) which are linked to nitrogen doped graphene quantum dots, as potential photosensitizers for photodynamic therapy (PDT). The symmetrical morpholine porphyrin complexes 2 (Sn(IV) 5,10,15,20 tetra-4-morpholinyl porphyrin) and 3 (Zn 5,10,15,20 tetra-4-morpholinyl porphyrin) had the same substituent but different central metals, and they were both conjugated to biotin decorated nitrogen doped graphene quantum dots (B-NGQDs), however complex 2 (Sn(IV) 5,10,15,20 tetra-4-morpholinyl porphyrin) was conjugated to B-NGQDs through an ester bond and complex 3 (Zinc 5,10,15,20 tetra-4-morpholinyl porphyrin) through 𝜋-𝜋 stacking. The effect of asymmetry was studied by comparing complex 3 (Zn 5,10,15,20 tetra-4-morpholinyl porphyrin) and complex 5 (Zn 5- bromophenyl-10-15-20-(tris-4- morpholinyl) porphyrin). Complex 5 (Zn 5- bromophenyl-10-15-20-(tris-4- morpholinyl) porphyrin) was an asymmetric porphyrin with morpholine and bromine as substituents. It was observed that asymmetry enhances singlet oxygen quantum yield and PDT activity. It was also observed that folic acid is a better targeting biomolecule when compared to biotin, and this was studied by comparing complex 3 conjugated to B-NGQDs and complex 3 conjugated to folic acid decorated nitrogen doped graphene quantum dots (FA-NGQDs). 3-FA-NGQDs had a better cellular uptake and PDT activity. , Thesis (MSc) -- Faculty of Science, Chemistry, 2023
- Full Text:
- Authors: Magaela, Ngwanabjala Bridged
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422486 , vital:71947
- Description: This thesis reports on the synthesis, characterization, photophysiochemical properties of morpholine substituted symmetrical and asymmetrical porphyrins. The synthesized porphyrins are conjugated to cancer selective biomolecules (folic acid and biotin) which are linked to nitrogen doped graphene quantum dots, as potential photosensitizers for photodynamic therapy (PDT). The symmetrical morpholine porphyrin complexes 2 (Sn(IV) 5,10,15,20 tetra-4-morpholinyl porphyrin) and 3 (Zn 5,10,15,20 tetra-4-morpholinyl porphyrin) had the same substituent but different central metals, and they were both conjugated to biotin decorated nitrogen doped graphene quantum dots (B-NGQDs), however complex 2 (Sn(IV) 5,10,15,20 tetra-4-morpholinyl porphyrin) was conjugated to B-NGQDs through an ester bond and complex 3 (Zinc 5,10,15,20 tetra-4-morpholinyl porphyrin) through 𝜋-𝜋 stacking. The effect of asymmetry was studied by comparing complex 3 (Zn 5,10,15,20 tetra-4-morpholinyl porphyrin) and complex 5 (Zn 5- bromophenyl-10-15-20-(tris-4- morpholinyl) porphyrin). Complex 5 (Zn 5- bromophenyl-10-15-20-(tris-4- morpholinyl) porphyrin) was an asymmetric porphyrin with morpholine and bromine as substituents. It was observed that asymmetry enhances singlet oxygen quantum yield and PDT activity. It was also observed that folic acid is a better targeting biomolecule when compared to biotin, and this was studied by comparing complex 3 conjugated to B-NGQDs and complex 3 conjugated to folic acid decorated nitrogen doped graphene quantum dots (FA-NGQDs). 3-FA-NGQDs had a better cellular uptake and PDT activity. , Thesis (MSc) -- Faculty of Science, Chemistry, 2023
- Full Text:
Identification of novel Arf1 GTPase inhibitors for cancer target validation
- Authors: Mqwathi, Nomxolisi Vuyokasi
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424666 , vital:72173
- Description: The key regulators of both anterograde and retrograde vesicular traffic, adenosine diphosphate-ribosylation factors (Arfs), also coordinate various signalling pathways and regulate cellular processes required for cell survival and function. In addition to its role in mediating secretory trafficking in the Golgi apparatus, the involvement of Arf1 in signalling pathways that contribute to the formation and progression of cancer has become apparent, and the overexpression and deregulation of Arf1 activity has been associated with cancer cell invasion, proliferation and metastasis. As with other small GTPases, Arf1 must cycle back and forth between an inactive (GDP-bound) and active (GTP-bound) conformation to carry out its function. However, the cycle of Arf1 inactivation and activation is controlled by Arf GTPase activating proteins (Arf-GAPs) that stimulate Arf1 to hydrolyse the bound GTP to GDP and Arf guanine nucleotide exchange factors (Arf-GEFs) that facilitate GDP for GTP exchange on Arf1, respectively. The identification of Arf1 inhibitors that indirectly disrupt Arf1 function by blocking its interaction with Arf-GAPs or Arf-GEFs has generated interest in their use as possible anti-cancer agents. The suppression of Arf1 activation (by targeting Arf-GEFs) has been investigated as a potential cancer therapeutic target and resulted in inhibitor compounds that have micromolar-range activity against cancer cells and targets and promising results in mouse models, but experience problems with bioavailability when used in vivo. This motivates the search for novel Arf1 inhibitors for validation purposes to question whether Arf1 is a viable target for cancer therapy. The purpose of the study was to employ a recently developed colourimetric screening assay to identify inhibitors of Arf1 activation (Arf-GEF inhibitors) and deactivation (Arf-GAP inhibitors), with a focus on evaluating the potential of Arf1 deactivation as an entirely novel anti-cancer target. The proteins required for the assay (Arf1, Arf-GEF and -GAP domains and a reporter protein, GST-GGA3) were expressed in E. coli. and purified using affinity chromatography. The assay could detect the activation of Arf1 by the catalytic Sec7 domain of the three Arf-GEFs chosen for this study, but reproducibility was compromised by the occasional spontaneous activation of Arf1 in the absence of the Arf-GEFs. By contrast, the assay could reproducibly detect Arf1 deactivation by an Arf-GAP domain (Arf-GAP1GAP) and was subsequently used to screen a library of α-helix mimetics. Thirteen hit compounds with IC50 values ranging from 0.53 to 20.95 μM were found to inhibit Arf-GAP1GAP-mediated stimulation of GTP hydrolysis by Arf1-GTP in this assay format, however, they did not effectively suppress the proliferation of three tested cell lines (HeLa, MCF-7 and MCF-12A). Interestingly, the results obtained from fluorescence microscopy studies suggested that the compounds disrupt Golgi structure and Arf1 localisation, presumably by keeping Arf1 in its active conformation by blocking Arf-GAP1 function. This suggests that the compounds affect Arf1 function in cells, and may be used to explore the feasibility of targeting Arf1 deactivation for anti-cancer purposes in a wider range of cell lines and experiments. It has been reported that Arf-GAP1 inhibition is associated with the suppression of cell migration, and the potential of the compounds as metastasis inhibitors may also be explored. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
- Authors: Mqwathi, Nomxolisi Vuyokasi
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424666 , vital:72173
- Description: The key regulators of both anterograde and retrograde vesicular traffic, adenosine diphosphate-ribosylation factors (Arfs), also coordinate various signalling pathways and regulate cellular processes required for cell survival and function. In addition to its role in mediating secretory trafficking in the Golgi apparatus, the involvement of Arf1 in signalling pathways that contribute to the formation and progression of cancer has become apparent, and the overexpression and deregulation of Arf1 activity has been associated with cancer cell invasion, proliferation and metastasis. As with other small GTPases, Arf1 must cycle back and forth between an inactive (GDP-bound) and active (GTP-bound) conformation to carry out its function. However, the cycle of Arf1 inactivation and activation is controlled by Arf GTPase activating proteins (Arf-GAPs) that stimulate Arf1 to hydrolyse the bound GTP to GDP and Arf guanine nucleotide exchange factors (Arf-GEFs) that facilitate GDP for GTP exchange on Arf1, respectively. The identification of Arf1 inhibitors that indirectly disrupt Arf1 function by blocking its interaction with Arf-GAPs or Arf-GEFs has generated interest in their use as possible anti-cancer agents. The suppression of Arf1 activation (by targeting Arf-GEFs) has been investigated as a potential cancer therapeutic target and resulted in inhibitor compounds that have micromolar-range activity against cancer cells and targets and promising results in mouse models, but experience problems with bioavailability when used in vivo. This motivates the search for novel Arf1 inhibitors for validation purposes to question whether Arf1 is a viable target for cancer therapy. The purpose of the study was to employ a recently developed colourimetric screening assay to identify inhibitors of Arf1 activation (Arf-GEF inhibitors) and deactivation (Arf-GAP inhibitors), with a focus on evaluating the potential of Arf1 deactivation as an entirely novel anti-cancer target. The proteins required for the assay (Arf1, Arf-GEF and -GAP domains and a reporter protein, GST-GGA3) were expressed in E. coli. and purified using affinity chromatography. The assay could detect the activation of Arf1 by the catalytic Sec7 domain of the three Arf-GEFs chosen for this study, but reproducibility was compromised by the occasional spontaneous activation of Arf1 in the absence of the Arf-GEFs. By contrast, the assay could reproducibly detect Arf1 deactivation by an Arf-GAP domain (Arf-GAP1GAP) and was subsequently used to screen a library of α-helix mimetics. Thirteen hit compounds with IC50 values ranging from 0.53 to 20.95 μM were found to inhibit Arf-GAP1GAP-mediated stimulation of GTP hydrolysis by Arf1-GTP in this assay format, however, they did not effectively suppress the proliferation of three tested cell lines (HeLa, MCF-7 and MCF-12A). Interestingly, the results obtained from fluorescence microscopy studies suggested that the compounds disrupt Golgi structure and Arf1 localisation, presumably by keeping Arf1 in its active conformation by blocking Arf-GAP1 function. This suggests that the compounds affect Arf1 function in cells, and may be used to explore the feasibility of targeting Arf1 deactivation for anti-cancer purposes in a wider range of cell lines and experiments. It has been reported that Arf-GAP1 inhibition is associated with the suppression of cell migration, and the potential of the compounds as metastasis inhibitors may also be explored. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
The development of a plate-based assay to detect the activation status of ARF1 GTPase in Plasmodium falciparum parasites
- Authors: Du Toit, Skye Carol
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424654 , vital:72172
- Description: The exponential rise in antimalarial drug resistance in the most infectious malaria species, Plasmodium falciparum, has emphasised the urgency to identify and validate novel drug targets that decrease parasite viability upon inhibition. In addition to several publications indicating that the regulation of human Arf1 GTPase activity (mediated by ArfGEFs and ArfGAPs) serves as a pertinent drug target for cancer research, the identification of Arf1 and its regulatory proteins in Plasmodium falciparum led to the question whether these protein homologs could be exploited as drug targets for anti-malarial drug therapies. To investigate this prospect, the establishment of a novel in vitro colorimetric ELISA-based assay was needed to be able to detect changes in the activation status of P. falciparum Arf1 (PfArf1) in parasite cultures exposed to potential Arf1 inhibitors. By exploiting the selective protein interaction that occurs between active GTP-bound Arf1 and its downstream effector, GGA3, an assay protocol was established that could be used to detect the activation status of purified, truncated PfArf1 obtained from E. coli and endogenous PfArf1 sourced from parasite lysates. The assay relies on the use of anti-Arf1 antibodies to detect the binding of active PfArf1 in the lysates of inhibitor-exposed cultured parasites to GST-GGA3 immobilised in glutathione-coated plates. The results from chemical validation experiments conducted using the novel assay developed in this study, using the known ArfGEF inhibitor brefeldin A (BFA) and ArfGAP inhibitors Chem1099 and Chem3050, yielded the anticipated results: decrease in active PfArf1 after parasite incubation with the ArfGEF inhibitor, and increased active PfArf1 after ArfGAP inhibition. The results confirmed PfArf1 as a potential anti-malarial drug target and encourages the further development of this assay format for the identification of subsequent inhibitors in library screening campaigns. Additional pilot experiments were conducted to further explore whether the assay could detect the activation status of human Arf1 using HeLa cell lysates and to provide further evidence that the assay could be exploited as a tool in the identification of Arf1 GTPase inhibitors with BFA and the known ArfGAP inhibitor, QS11. The results suggested that, while the assay can detect the increase in active cellular Arf1 due to the inhibition of human ArfGEF following BFA treatment, subsequent treatment with QS11 showed no evidence of a reduction in active human Arf1 due to ArfGAP inhibition. Further experimentation is required to investigate the ability the assay to confirm inhibition of human Arf1 deactivation by ArfGAP inhibitors and develop the assay as a useful tool to support cancer drug discovery, in addition to antimalarial drug discovery projects aimed at Arf1. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
- Authors: Du Toit, Skye Carol
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424654 , vital:72172
- Description: The exponential rise in antimalarial drug resistance in the most infectious malaria species, Plasmodium falciparum, has emphasised the urgency to identify and validate novel drug targets that decrease parasite viability upon inhibition. In addition to several publications indicating that the regulation of human Arf1 GTPase activity (mediated by ArfGEFs and ArfGAPs) serves as a pertinent drug target for cancer research, the identification of Arf1 and its regulatory proteins in Plasmodium falciparum led to the question whether these protein homologs could be exploited as drug targets for anti-malarial drug therapies. To investigate this prospect, the establishment of a novel in vitro colorimetric ELISA-based assay was needed to be able to detect changes in the activation status of P. falciparum Arf1 (PfArf1) in parasite cultures exposed to potential Arf1 inhibitors. By exploiting the selective protein interaction that occurs between active GTP-bound Arf1 and its downstream effector, GGA3, an assay protocol was established that could be used to detect the activation status of purified, truncated PfArf1 obtained from E. coli and endogenous PfArf1 sourced from parasite lysates. The assay relies on the use of anti-Arf1 antibodies to detect the binding of active PfArf1 in the lysates of inhibitor-exposed cultured parasites to GST-GGA3 immobilised in glutathione-coated plates. The results from chemical validation experiments conducted using the novel assay developed in this study, using the known ArfGEF inhibitor brefeldin A (BFA) and ArfGAP inhibitors Chem1099 and Chem3050, yielded the anticipated results: decrease in active PfArf1 after parasite incubation with the ArfGEF inhibitor, and increased active PfArf1 after ArfGAP inhibition. The results confirmed PfArf1 as a potential anti-malarial drug target and encourages the further development of this assay format for the identification of subsequent inhibitors in library screening campaigns. Additional pilot experiments were conducted to further explore whether the assay could detect the activation status of human Arf1 using HeLa cell lysates and to provide further evidence that the assay could be exploited as a tool in the identification of Arf1 GTPase inhibitors with BFA and the known ArfGAP inhibitor, QS11. The results suggested that, while the assay can detect the increase in active cellular Arf1 due to the inhibition of human ArfGEF following BFA treatment, subsequent treatment with QS11 showed no evidence of a reduction in active human Arf1 due to ArfGAP inhibition. Further experimentation is required to investigate the ability the assay to confirm inhibition of human Arf1 deactivation by ArfGAP inhibitors and develop the assay as a useful tool to support cancer drug discovery, in addition to antimalarial drug discovery projects aimed at Arf1. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
Application of web design techniques and best practices in implementing web development, maintenance and enhancement of RUBi websites and web application systems
- Authors: Tshabalalala, Thulani
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424688 , vital:72175
- Description: The popularity of the web has seen various fields, such as the sciences taking advantage of this resource to further their scientific endeavours. This has seen science groups moving into developing websites and web applications, and such a group is the Research Unit in Bioinformative (RUBi). With the use of the web, the development and maintenance of whatever web-related tools become inevitable, given the continuous changes in the web space. This continuous evolution of web development and maintenance will come with techniques, principles and standards which will not only enable faster development of web entities but also ensure that modern hardware, fulfilment of the requirements to use such hardware and modern concepts are incorporated into forming web tools that enable such progression. Furthermore, introducing the previously mentioned progress of the web becomes an essential part of its development and maintenance. This paper did implement the processes of progressing the web using the technique of documentation and version control systems. The web development for the COVIDRUG website was done for the Covidrug-Africa Consortium (COVIDRUG) using the Django webdevelopment framework. The RUBi website and the MDM-Task we band the Job Management System (JMS) web applications were maintained for the maintenance aspect. Archives brought value regarding the traceability it provides of the various web-related aspects. The development showed a website’s potential value, particularly for research groups. The maintenance carried out showed how different techniques and approaches could be used in different maintenance prospects to achieve set objectives. The development and maintenance resulted in websites and web applications that have the features stated in their respective maintenance plans. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
- Authors: Tshabalalala, Thulani
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424688 , vital:72175
- Description: The popularity of the web has seen various fields, such as the sciences taking advantage of this resource to further their scientific endeavours. This has seen science groups moving into developing websites and web applications, and such a group is the Research Unit in Bioinformative (RUBi). With the use of the web, the development and maintenance of whatever web-related tools become inevitable, given the continuous changes in the web space. This continuous evolution of web development and maintenance will come with techniques, principles and standards which will not only enable faster development of web entities but also ensure that modern hardware, fulfilment of the requirements to use such hardware and modern concepts are incorporated into forming web tools that enable such progression. Furthermore, introducing the previously mentioned progress of the web becomes an essential part of its development and maintenance. This paper did implement the processes of progressing the web using the technique of documentation and version control systems. The web development for the COVIDRUG website was done for the Covidrug-Africa Consortium (COVIDRUG) using the Django webdevelopment framework. The RUBi website and the MDM-Task we band the Job Management System (JMS) web applications were maintained for the maintenance aspect. Archives brought value regarding the traceability it provides of the various web-related aspects. The development showed a website’s potential value, particularly for research groups. The maintenance carried out showed how different techniques and approaches could be used in different maintenance prospects to achieve set objectives. The development and maintenance resulted in websites and web applications that have the features stated in their respective maintenance plans. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
- Full Text:
Investigation of brewery waste grains and microbial fuel cells as value-additive technologies improving solvent production yields in Clostridium acetobutylicum (ATCC 824) fermentation
- Authors: Du Toit, Ryan Guillaume
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424643 , vital:72171
- Description: The production of the solvent compounds acetone, ethanol and butanol through fermentation of organic feedstocks using Clostridia species could be a promising route for biofuel production. However, the cost of raw materials, low yields and the complexity of anaerobic fermentation continue to hinder this means of generating these compounds. The research presented in this Thesis investigated low-cost interventions that could decrease the costs of production and to direct the synthesis of fuel compounds using microbial fuel cells. Low-cost anaerobic chambers were designed and constructed for the propagation and manipulation of Clostridium acetobutylicum, selected as a low-risk microbial catalyst. Fermentation was monitored using in situ pH measurements and a combination of turbidity measurements, nutrient assays (especially total carbohydrates) and HPLC-RI detection as a means of monitoring the consumption of nutrients (glucose), production of precursor compounds (butyric acid) and the formation of solvent molecules (acetone/ethanol and butanol) during fermentation by this organism. Brewer’s spent grains were tested as a sustainable and low-cost feedstock for solvent production, comparing the effects of sterilising before fermentation, or allowing resident microflora to remain during Clostridium-catalysed solvent production. Sterilised spent grains significantly improved the production of solvent molecules (e.g. 12.97 ± 0.38 g/L of butanol yielded, compared to 0.40 ± 0.33 g/L for defined media sampled during the solventogenic phase); compared to these, the use of non-sterilised brewer’s grain decreased both the reproducibility and yields of fermentation (8.66 ± 1.6 g/L of butanol). Microbial fuel cells were studied as a possible means of altering electron transfer to/from electrode-attached Clostridia to control the metabolic shift in bacteria from acidogenesis to solventogenesis. The base line MFC (11.00 ± 4.69 g/L) fermentation experiment did produce higher acetone/ethanol than the baseline batch experiment MB (5.47 ± 4.48 g/L), indicating an improvement to solvent production in C. acetobutylicum (ATCC 824) in a MFC fermentation. In this study, MFC-1 demonstrated remarkable superiority over MB in terms of butyric acid production, yielding significantly higher concentrations while also improving acetone and ethanol production. However, the enhanced butyric acid production did not correspond to significantly increased butanol yields when compared to batch fermentation of chemically defined media. These findings highlight the potential of MFC-1 as an efficient approach for enhancing the fermentative production of valuable compounds, with a particular focus on butyric acid and acetone/ethanol. , Thesis (MSc) -- Faculty of Science, Biotechnology Innovation Centre, 2023
- Full Text:
- Authors: Du Toit, Ryan Guillaume
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424643 , vital:72171
- Description: The production of the solvent compounds acetone, ethanol and butanol through fermentation of organic feedstocks using Clostridia species could be a promising route for biofuel production. However, the cost of raw materials, low yields and the complexity of anaerobic fermentation continue to hinder this means of generating these compounds. The research presented in this Thesis investigated low-cost interventions that could decrease the costs of production and to direct the synthesis of fuel compounds using microbial fuel cells. Low-cost anaerobic chambers were designed and constructed for the propagation and manipulation of Clostridium acetobutylicum, selected as a low-risk microbial catalyst. Fermentation was monitored using in situ pH measurements and a combination of turbidity measurements, nutrient assays (especially total carbohydrates) and HPLC-RI detection as a means of monitoring the consumption of nutrients (glucose), production of precursor compounds (butyric acid) and the formation of solvent molecules (acetone/ethanol and butanol) during fermentation by this organism. Brewer’s spent grains were tested as a sustainable and low-cost feedstock for solvent production, comparing the effects of sterilising before fermentation, or allowing resident microflora to remain during Clostridium-catalysed solvent production. Sterilised spent grains significantly improved the production of solvent molecules (e.g. 12.97 ± 0.38 g/L of butanol yielded, compared to 0.40 ± 0.33 g/L for defined media sampled during the solventogenic phase); compared to these, the use of non-sterilised brewer’s grain decreased both the reproducibility and yields of fermentation (8.66 ± 1.6 g/L of butanol). Microbial fuel cells were studied as a possible means of altering electron transfer to/from electrode-attached Clostridia to control the metabolic shift in bacteria from acidogenesis to solventogenesis. The base line MFC (11.00 ± 4.69 g/L) fermentation experiment did produce higher acetone/ethanol than the baseline batch experiment MB (5.47 ± 4.48 g/L), indicating an improvement to solvent production in C. acetobutylicum (ATCC 824) in a MFC fermentation. In this study, MFC-1 demonstrated remarkable superiority over MB in terms of butyric acid production, yielding significantly higher concentrations while also improving acetone and ethanol production. However, the enhanced butyric acid production did not correspond to significantly increased butanol yields when compared to batch fermentation of chemically defined media. These findings highlight the potential of MFC-1 as an efficient approach for enhancing the fermentative production of valuable compounds, with a particular focus on butyric acid and acetone/ethanol. , Thesis (MSc) -- Faculty of Science, Biotechnology Innovation Centre, 2023
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Characterizing shift systems and exploring their effects on sleep and fatigue in South African freight railway operators
- Authors: Mona, Chumani
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424300 , vital:72141
- Description: Embargoed. Expected release date 2025. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2023
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- Authors: Mona, Chumani
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424300 , vital:72141
- Description: Embargoed. Expected release date 2025. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2023
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An investigation into fatigue prevalence amongst citrus packhouse sorters in the Eastern Cape province of South Africa
- Authors: Robinson, Harry Jack
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424312 , vital:72142
- Description: Background: South Africa is the second most influential exporter of citrus fruits internationally and holds a pivotal role in economic revenue for the country. Rural areas, such as the Sundays River Valley in the Eastern Cape province of South Africa, contribute to the country’s citrus production. Many women depend on citrus packhouses for employment as citrus sorters. As humans perceive certain defects in citrus fruits more accurately than machines, these sorters must identify and manually remove any fruit that does not conform with export requirements. Citrus sorters are exposed to numerous physical and cognitive stressors during the task while faced with organizational challenges, such as shift work and long working hours. Therefore, the potential for fatigue development is anticipated. Given the multifactorial nature of fatigue and the negative consequences it may have on workers, it also has the potential to impede the accuracy of the sorting performance. Stringent disciplinary action for the entire South African citrus industry may be of consequence if nonconforming or pest-infested fruit is missed by citrus sorters and exported to foreign countries. This study aimed to investigate the prevalence of fatigue among citrus sorters in a citrus packhouse in the Sundays River Valley of the Eastern Cape throughout a citrus harvesting season and to identify factors that may contribute towards fatigue development. Methods: The research design utilized a crosssectional, two-part approach that applied mixed methods. Part one was administered once-off, incorporating demographic, work-, and non-work-related questions. Part two was a self-developed repeated measures assessment comprising close-ended contextual questions, the Modified Fatigue Impact Scale, and the Karolinska Sleepiness Scale. Environmental and work output data were also recorded. Results: Citrus sorters (n= 35) recorded a mean MFIS score of 39.35 throughout the harvest season, which was above the prescribed fatigue threshold (38). However, there was no significant difference in fatigue ratings over time (p= 0.122). Day shift workers exceeded the fatigue threshold for the entire season compared to night shift workers, who only exceeded it in the last two weeks. The physical, cognitive, and psychosocial subscales found no significant difference in fatigue scores, although physical fatigue recorded the highest scores across all weeks and displayed a significant difference over time. Overall, participants, on average, perceived to be “neither sleepy nor alert” over the season. However, eight participants (22%) recorded sleepiness scores ii exceeding the excessive sleepiness threshold of seven. Educational levels, health status, work-pace, and the number of family dependents significantly contributed to fatigue development, albeit a weak correlation. Discussion: Sorters were perceived to be fatigued from week three till the end of the study; however, there was no variation in fatigue scores over time. An accumulation of physical fatigue over time was revealed where prolonged standing, repetitive work, and irregular working postures may have contributed. Night shift workers did not receive the recommended hours of sleep (7-8 hours); hence, they registered greater sleepiness scores over the season than day shift workers. Environmental and work output recordings did not prove to have a significant influence on fatigue development, and neither did work experience or physical exercise. Conclusion: An amalgamation of numerous contributing factors within the work situation, private situation, and the individual influenced the development of fatigue, where there was no primary causal factor. Future studies should consider recording the accuracy of the sorting performance to acquire rich, objective data. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2023
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- Authors: Robinson, Harry Jack
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424312 , vital:72142
- Description: Background: South Africa is the second most influential exporter of citrus fruits internationally and holds a pivotal role in economic revenue for the country. Rural areas, such as the Sundays River Valley in the Eastern Cape province of South Africa, contribute to the country’s citrus production. Many women depend on citrus packhouses for employment as citrus sorters. As humans perceive certain defects in citrus fruits more accurately than machines, these sorters must identify and manually remove any fruit that does not conform with export requirements. Citrus sorters are exposed to numerous physical and cognitive stressors during the task while faced with organizational challenges, such as shift work and long working hours. Therefore, the potential for fatigue development is anticipated. Given the multifactorial nature of fatigue and the negative consequences it may have on workers, it also has the potential to impede the accuracy of the sorting performance. Stringent disciplinary action for the entire South African citrus industry may be of consequence if nonconforming or pest-infested fruit is missed by citrus sorters and exported to foreign countries. This study aimed to investigate the prevalence of fatigue among citrus sorters in a citrus packhouse in the Sundays River Valley of the Eastern Cape throughout a citrus harvesting season and to identify factors that may contribute towards fatigue development. Methods: The research design utilized a crosssectional, two-part approach that applied mixed methods. Part one was administered once-off, incorporating demographic, work-, and non-work-related questions. Part two was a self-developed repeated measures assessment comprising close-ended contextual questions, the Modified Fatigue Impact Scale, and the Karolinska Sleepiness Scale. Environmental and work output data were also recorded. Results: Citrus sorters (n= 35) recorded a mean MFIS score of 39.35 throughout the harvest season, which was above the prescribed fatigue threshold (38). However, there was no significant difference in fatigue ratings over time (p= 0.122). Day shift workers exceeded the fatigue threshold for the entire season compared to night shift workers, who only exceeded it in the last two weeks. The physical, cognitive, and psychosocial subscales found no significant difference in fatigue scores, although physical fatigue recorded the highest scores across all weeks and displayed a significant difference over time. Overall, participants, on average, perceived to be “neither sleepy nor alert” over the season. However, eight participants (22%) recorded sleepiness scores ii exceeding the excessive sleepiness threshold of seven. Educational levels, health status, work-pace, and the number of family dependents significantly contributed to fatigue development, albeit a weak correlation. Discussion: Sorters were perceived to be fatigued from week three till the end of the study; however, there was no variation in fatigue scores over time. An accumulation of physical fatigue over time was revealed where prolonged standing, repetitive work, and irregular working postures may have contributed. Night shift workers did not receive the recommended hours of sleep (7-8 hours); hence, they registered greater sleepiness scores over the season than day shift workers. Environmental and work output recordings did not prove to have a significant influence on fatigue development, and neither did work experience or physical exercise. Conclusion: An amalgamation of numerous contributing factors within the work situation, private situation, and the individual influenced the development of fatigue, where there was no primary causal factor. Future studies should consider recording the accuracy of the sorting performance to acquire rich, objective data. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2023
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