An unsung dialogue: music, society and the history of The Flames
- Authors: Park, Duncan Keith
- Date: 2020
- Subjects: The Flames (Musical group) , Fataar, Steve , Durban (South Africa) -- History , Durban (South Africa) -- Race relations , Musical groups -- South Africa , Music -- Social aspects -- South Africa , Music -- Social aspects -- South Africa -- Durban , South Africa -- Politics and government -- 1961-1978
- Language: English
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/169632 , vital:41780
- Description: This research aims to examine the relationship between music and social conditions within the context of apartheid . The focus area is the city of Durban, specifically 1960-1970. A case study of the multi-racial Durban-based band, The Flames 1963–1972) will be used to rigorously assess the extent to which social conditions and music can affect each other in both directions. This will also be the first coherent and historical narrative of the band, The Flames. As such, the thesis aims to make an original contribution to this field of music history, while it will provide the very first academic discussion of The Flames. South Africa during apartheid, with its racialised legislation provides an ideal context to scrutinize the effects (both potential and real) of music on social conditions. The Flames provide a particularly relevant case point; being a collective of “mixed-race” individuals classified as Coloured performing for a multi-racial fan- base. This means that at their concerts, audience members would have frequently been from a variety of different racial backgrounds. This would have been in direct contravention to the National Party‟s (NP) Separate Amenities Act as well as the policy of “separate development”, the cornerstone of apartheid. The thesis will firstly contextualise the research within the broader, global historical context, as well as in the existing theoretical literature which focuses on the relationship between music, politics and society. Firstly, this will entail a brief historical account of popular music around the world during the twentieth century. Through this we will observe both the ways in which the development and popularisation of various forms of popular music came to be shaped in a particular way, as well as the ways in which the public received this music. This will include an examination of positive responses to certain kinds of popular music which resonated and became popular with various sectors of society. Conversely, it will also closely examine the backlashes against these musical forms, and attempt to identify why certain sectors of the public were deeply opposed to particular kinds of popular music which may have been perceived to embody certain values and meanings. This will be conducted by studying various forms of popular music from the turn of the twentieth century into the early 1970s with a cross-cultural, global perspective, examining particular historical instances and existing theories relating to these instances. Chapter one will then shift from a global perspective and will situate the research within the South African musical context specifically. This will include an account of the development of South African popular music through the twentieth century in relation to the country's politics. Through examining this relationship, the dialogue between historical case studies and theoretical literature will continue, in which existing theories relating to the relationship between music, politics and society will be discussed. This theoretical literature will be made use of in the final section of the paper in order to make sense of The Flames and their role in South African history. The second section of this thesis will focus on the socio-historical context of the city of Durban under apartheid. The author will contextualise the research through a narrative historical retelling of Durban‟s social history, focusing on both political and social public mobilisation and the role of cultural spaces and practices within these his torical moments and their relevant structures. Attention will be paid to the development of segregation and apartheid within Durban specifically, while racial relations will also be fo cused upon. This will provide the reader with the necessary background required in order to meaningfully understand how the band The Flames came into existence, became popular, and whether or not they had any meaningful effect on their historical context. The section on Durban's history will be followed by a brief account of Durban's Coloured community's history, as well as a discussion around Coloured identity more broadly within the South African context. This will be vital to the research due to the fact that the musicians of The Flames were classified as Coloured under apartheid, and such an historical background will be critical to understanding the social, political and economic context of the band. Finally, the thesis will end with the written history of The Flame . This final section will provide an account of The Flames' history in which the band will be made sense of within the context of Durban during apartheid, as well as within the context of South African music, and the globalised context of popular music. By situating the band within these three contexts, we can truly assess both how they were shaped by their contexts, and whether they had any effect on these contexts of which they were a part. In this section those existing theories around the relationship between music, politics and society will be discussed and assessed in order to determine how useful a framework they may be for understanding popular music in certain historical contexts. In this way, the research aims to make a meaningful contribution to the study of music and its socio-political role throughout history, as well as to the broader understanding of Durban's history. Specifically, the role of music in Durban's history is being examined, and the researcher hopes that this work can begin to open up new discussions around the importance of studying music in Durban, and South Africa's history. Additionally, this thesis will open up a new area of research into a band which, until this point, has not been engaged with in the field of music history whatsoever. .
- Full Text:
- Date Issued: 2020
- Authors: Park, Duncan Keith
- Date: 2020
- Subjects: The Flames (Musical group) , Fataar, Steve , Durban (South Africa) -- History , Durban (South Africa) -- Race relations , Musical groups -- South Africa , Music -- Social aspects -- South Africa , Music -- Social aspects -- South Africa -- Durban , South Africa -- Politics and government -- 1961-1978
- Language: English
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/169632 , vital:41780
- Description: This research aims to examine the relationship between music and social conditions within the context of apartheid . The focus area is the city of Durban, specifically 1960-1970. A case study of the multi-racial Durban-based band, The Flames 1963–1972) will be used to rigorously assess the extent to which social conditions and music can affect each other in both directions. This will also be the first coherent and historical narrative of the band, The Flames. As such, the thesis aims to make an original contribution to this field of music history, while it will provide the very first academic discussion of The Flames. South Africa during apartheid, with its racialised legislation provides an ideal context to scrutinize the effects (both potential and real) of music on social conditions. The Flames provide a particularly relevant case point; being a collective of “mixed-race” individuals classified as Coloured performing for a multi-racial fan- base. This means that at their concerts, audience members would have frequently been from a variety of different racial backgrounds. This would have been in direct contravention to the National Party‟s (NP) Separate Amenities Act as well as the policy of “separate development”, the cornerstone of apartheid. The thesis will firstly contextualise the research within the broader, global historical context, as well as in the existing theoretical literature which focuses on the relationship between music, politics and society. Firstly, this will entail a brief historical account of popular music around the world during the twentieth century. Through this we will observe both the ways in which the development and popularisation of various forms of popular music came to be shaped in a particular way, as well as the ways in which the public received this music. This will include an examination of positive responses to certain kinds of popular music which resonated and became popular with various sectors of society. Conversely, it will also closely examine the backlashes against these musical forms, and attempt to identify why certain sectors of the public were deeply opposed to particular kinds of popular music which may have been perceived to embody certain values and meanings. This will be conducted by studying various forms of popular music from the turn of the twentieth century into the early 1970s with a cross-cultural, global perspective, examining particular historical instances and existing theories relating to these instances. Chapter one will then shift from a global perspective and will situate the research within the South African musical context specifically. This will include an account of the development of South African popular music through the twentieth century in relation to the country's politics. Through examining this relationship, the dialogue between historical case studies and theoretical literature will continue, in which existing theories relating to the relationship between music, politics and society will be discussed. This theoretical literature will be made use of in the final section of the paper in order to make sense of The Flames and their role in South African history. The second section of this thesis will focus on the socio-historical context of the city of Durban under apartheid. The author will contextualise the research through a narrative historical retelling of Durban‟s social history, focusing on both political and social public mobilisation and the role of cultural spaces and practices within these his torical moments and their relevant structures. Attention will be paid to the development of segregation and apartheid within Durban specifically, while racial relations will also be fo cused upon. This will provide the reader with the necessary background required in order to meaningfully understand how the band The Flames came into existence, became popular, and whether or not they had any meaningful effect on their historical context. The section on Durban's history will be followed by a brief account of Durban's Coloured community's history, as well as a discussion around Coloured identity more broadly within the South African context. This will be vital to the research due to the fact that the musicians of The Flames were classified as Coloured under apartheid, and such an historical background will be critical to understanding the social, political and economic context of the band. Finally, the thesis will end with the written history of The Flame . This final section will provide an account of The Flames' history in which the band will be made sense of within the context of Durban during apartheid, as well as within the context of South African music, and the globalised context of popular music. By situating the band within these three contexts, we can truly assess both how they were shaped by their contexts, and whether they had any effect on these contexts of which they were a part. In this section those existing theories around the relationship between music, politics and society will be discussed and assessed in order to determine how useful a framework they may be for understanding popular music in certain historical contexts. In this way, the research aims to make a meaningful contribution to the study of music and its socio-political role throughout history, as well as to the broader understanding of Durban's history. Specifically, the role of music in Durban's history is being examined, and the researcher hopes that this work can begin to open up new discussions around the importance of studying music in Durban, and South Africa's history. Additionally, this thesis will open up a new area of research into a band which, until this point, has not been engaged with in the field of music history whatsoever. .
- Full Text:
- Date Issued: 2020
Shoutfighting and other fiction
- Authors: Rasmenike, Nonqubela Evelyn
- Date: 2020
- Subjects: South African fiction (English) -- 21st century , Xhosa fiction -- 21st century
- Language: English , Xhosa
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/144500 , vital:38351
- Description: This document consists of three parts: Part A: English Half Thesis (Creative Work) ; Part B: IsiXhosa Half Thesis (Creative Work) ; Part C: Portfolio.
- Full Text:
- Date Issued: 2020
- Authors: Rasmenike, Nonqubela Evelyn
- Date: 2020
- Subjects: South African fiction (English) -- 21st century , Xhosa fiction -- 21st century
- Language: English , Xhosa
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/144500 , vital:38351
- Description: This document consists of three parts: Part A: English Half Thesis (Creative Work) ; Part B: IsiXhosa Half Thesis (Creative Work) ; Part C: Portfolio.
- Full Text:
- Date Issued: 2020
Synthesis, photophysicochemical properties and photodynamic therapy activities of indium and zinc phthalocyanines when incorporated into Pluronic polymer micelles
- Authors: Motloung, Banele Mike
- Date: 2020
- Subjects: Indium , Zinc , Phthalocyanines , Polymers , Photochemotherapy , Micelles
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167529 , vital:41489
- Description: This thesis reports on the syntheses, photophysicochemical properties and photodynamic therapy activities of symmetrical metallophthalocyanines (MPcs) when alone or when incorporated into Pluronic polymer micelles. The Pcs contain either zinc or indium as central metals and have phenyldiazenylphenoxy, pyridine-2-yloxy and benzo[d]thiazol-2-ylthio as ring substituents. Spectroscopic and microscopic techniques were used to confirm the formation MPcs with micelles. The photophysics and photochemistry of the Pcs were assessed when alone and with micelles. All the studied Pcs showed good photophysicochemical behavior with relatively high triplet and singlet oxygen quantum yields corresponding to their low fluorescence quantum yields. The Pcs with indium in their central cavity exhibited higher triplet and singlet oxygen quantum yields in comparison to their zinc counterparts due to the heavy atom effect obtained from the former. The in vitro dark cytotoxicity and photodynamic therapy of the Pc complexes and conjugates against MCF7 cells was tested. All studied Pc complexes alone and with micelles showed minimum dark toxicity making them applicable for PDT. All complexes displayed good phototoxicity < 50% cell viability (except for complex 2 > 50% cell viability) at concentrations ≤100 μg/mL, however the conjugates showed < 45% cell viability at concentrations ≤ 100 μg/mL, probably due to the small micellar size and EPR effect. The findings from this work show the importance of incorporating photosensitizers such as phthalocyanines into Pluronic polymers micelles and making them water soluble and ultimately improving their photodynamic effect.
- Full Text:
- Date Issued: 2020
- Authors: Motloung, Banele Mike
- Date: 2020
- Subjects: Indium , Zinc , Phthalocyanines , Polymers , Photochemotherapy , Micelles
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167529 , vital:41489
- Description: This thesis reports on the syntheses, photophysicochemical properties and photodynamic therapy activities of symmetrical metallophthalocyanines (MPcs) when alone or when incorporated into Pluronic polymer micelles. The Pcs contain either zinc or indium as central metals and have phenyldiazenylphenoxy, pyridine-2-yloxy and benzo[d]thiazol-2-ylthio as ring substituents. Spectroscopic and microscopic techniques were used to confirm the formation MPcs with micelles. The photophysics and photochemistry of the Pcs were assessed when alone and with micelles. All the studied Pcs showed good photophysicochemical behavior with relatively high triplet and singlet oxygen quantum yields corresponding to their low fluorescence quantum yields. The Pcs with indium in their central cavity exhibited higher triplet and singlet oxygen quantum yields in comparison to their zinc counterparts due to the heavy atom effect obtained from the former. The in vitro dark cytotoxicity and photodynamic therapy of the Pc complexes and conjugates against MCF7 cells was tested. All studied Pc complexes alone and with micelles showed minimum dark toxicity making them applicable for PDT. All complexes displayed good phototoxicity < 50% cell viability (except for complex 2 > 50% cell viability) at concentrations ≤100 μg/mL, however the conjugates showed < 45% cell viability at concentrations ≤ 100 μg/mL, probably due to the small micellar size and EPR effect. The findings from this work show the importance of incorporating photosensitizers such as phthalocyanines into Pluronic polymers micelles and making them water soluble and ultimately improving their photodynamic effect.
- Full Text:
- Date Issued: 2020
‘Regulated Flexibility’ and labour market regulation: a case Study of Twizza Soft Drinks in the Eastern Cape, South Africa
- Authors: Flatau, Scott
- Date: 2020
- Subjects: Labor market -- South Africa , Labor laws and legislation -- South Africa , Industrial relations -- South Africa -- Case studies
- Language: English
- Type: text , Thesis , Masters , MSocSc
- Identifier: http://hdl.handle.net/10962/141288 , vital:37959
- Description: Following the negotiated settlement, which led to the ANC assuming power in South Africa in1994, debates concerning the nature of the South African labour market ensued between policy makers and economists alike. Central to understanding the South African labour market was the policy objective of regulated flexibility that has guided the formation of labour legislation in the post-1994 period, including the Labour Relations Act of 1995, the Basic Conditions of Employment Act of 1997, the Employment Equity Act of 1998 and the Skills Development Act of 1998. Regulated flexibility attempts to accommodate the interests of the employer for flexibility and the interests of the employee in regulation or security. These four Acts and the relevant provisions contained within them are the central focus of this research paper, in particular how they affect the case study firm Twizza Soft Drinks. An interpretivist approach was utilised as the preferred research methodology with in-depth, semi-structured interviews being the primary source of data collection. This research paper attempts to situate more clearly the impact of South Africa’s macro-economic policies since 1994 on labour market policy and undertakes an exploration of internal dynamics of firms in response to exogenous factors, such as government regulation. The key finding suggest that some Acts (BCEA, LRA) do not impose a significant burden on the firm and some provisions can lead to beneficial outcomes such as business modernisation and the adoption of formal Human Resource Practices. Conversely, some provisions contained in the EEA increase the administrative burden and therefore increase the indirect cost on the firm.
- Full Text:
- Date Issued: 2020
- Authors: Flatau, Scott
- Date: 2020
- Subjects: Labor market -- South Africa , Labor laws and legislation -- South Africa , Industrial relations -- South Africa -- Case studies
- Language: English
- Type: text , Thesis , Masters , MSocSc
- Identifier: http://hdl.handle.net/10962/141288 , vital:37959
- Description: Following the negotiated settlement, which led to the ANC assuming power in South Africa in1994, debates concerning the nature of the South African labour market ensued between policy makers and economists alike. Central to understanding the South African labour market was the policy objective of regulated flexibility that has guided the formation of labour legislation in the post-1994 period, including the Labour Relations Act of 1995, the Basic Conditions of Employment Act of 1997, the Employment Equity Act of 1998 and the Skills Development Act of 1998. Regulated flexibility attempts to accommodate the interests of the employer for flexibility and the interests of the employee in regulation or security. These four Acts and the relevant provisions contained within them are the central focus of this research paper, in particular how they affect the case study firm Twizza Soft Drinks. An interpretivist approach was utilised as the preferred research methodology with in-depth, semi-structured interviews being the primary source of data collection. This research paper attempts to situate more clearly the impact of South Africa’s macro-economic policies since 1994 on labour market policy and undertakes an exploration of internal dynamics of firms in response to exogenous factors, such as government regulation. The key finding suggest that some Acts (BCEA, LRA) do not impose a significant burden on the firm and some provisions can lead to beneficial outcomes such as business modernisation and the adoption of formal Human Resource Practices. Conversely, some provisions contained in the EEA increase the administrative burden and therefore increase the indirect cost on the firm.
- Full Text:
- Date Issued: 2020
Demographic change in the Upper Tsitsa Catchment: the integration of census and land cover data for 2001 and 2011
- Authors: Hodgson, Danuta Lorina
- Date: 2018
- Subjects: Land use -- South Africa -- Eastern Cape , Watersheds -- South Africa -- Eastern Cape , Thematic maps -- South Africa -- Eastern Cape , Population density -- South Africa -- Eastern Cape , Mzimvubu River Watershed (South Africa) -- Census , Upper Tsitsa Catchment (South Africa)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/57296 , vital:26870
- Description: The purpose of this research was to determine if the integration of census and land cover data could provide evidence of spatial patterns and temporal change for the Upper Tsitsa Catchment. This thesis contributed to academic literature with regards to dasymetric mapping and provided a database for the Ntabelanga and Laleni Ecological Infrastructure Project. The study took place in the Upper Tsitsa Catchment which is located in the north-eastern region of the Eastern Cape and falls within the uMzimvubu Catchment. South Africa National Population Censuses for 2001 and 2011 and the National Land Cover data sets for 2000 and 2013/14 were used to create dasymetric maps depicting demographic changes over time for the catchment area. Spatial statistics were performed on the dasymetric and choropleth map to determine the accuracy of the data that was created. From the results, it was found that although the statistics were skewed, the method was more accurate in displaying the population densities, which was noted during the sampling process of the spatial analysis. It was found that there had been a decrease in the population density within the catchment. This affected the density of several other variables such as population race group, language and employment status statistics as decreases in these values could be due to individuals migrating out of the catchment as well as socio-economic upliftment, such as having better access to services. The use of dasymetric mapping allowed an accurate representation of the population density from the census data to be created. The results of the dasymetric mapping were more accurate as they depicted where the population within the enumeration areas were located, and recognised that some areas were populated while some areas were not. To conclude, it was found that using dasymetric mapping provided reliable and useful data about population density and enables comparison over time.
- Full Text:
- Date Issued: 2018
- Authors: Hodgson, Danuta Lorina
- Date: 2018
- Subjects: Land use -- South Africa -- Eastern Cape , Watersheds -- South Africa -- Eastern Cape , Thematic maps -- South Africa -- Eastern Cape , Population density -- South Africa -- Eastern Cape , Mzimvubu River Watershed (South Africa) -- Census , Upper Tsitsa Catchment (South Africa)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/57296 , vital:26870
- Description: The purpose of this research was to determine if the integration of census and land cover data could provide evidence of spatial patterns and temporal change for the Upper Tsitsa Catchment. This thesis contributed to academic literature with regards to dasymetric mapping and provided a database for the Ntabelanga and Laleni Ecological Infrastructure Project. The study took place in the Upper Tsitsa Catchment which is located in the north-eastern region of the Eastern Cape and falls within the uMzimvubu Catchment. South Africa National Population Censuses for 2001 and 2011 and the National Land Cover data sets for 2000 and 2013/14 were used to create dasymetric maps depicting demographic changes over time for the catchment area. Spatial statistics were performed on the dasymetric and choropleth map to determine the accuracy of the data that was created. From the results, it was found that although the statistics were skewed, the method was more accurate in displaying the population densities, which was noted during the sampling process of the spatial analysis. It was found that there had been a decrease in the population density within the catchment. This affected the density of several other variables such as population race group, language and employment status statistics as decreases in these values could be due to individuals migrating out of the catchment as well as socio-economic upliftment, such as having better access to services. The use of dasymetric mapping allowed an accurate representation of the population density from the census data to be created. The results of the dasymetric mapping were more accurate as they depicted where the population within the enumeration areas were located, and recognised that some areas were populated while some areas were not. To conclude, it was found that using dasymetric mapping provided reliable and useful data about population density and enables comparison over time.
- Full Text:
- Date Issued: 2018
Development and optimisation of a novel Plasmodium falciparum Hsp90-Hop interaction assay
- Authors: Wambua, Lynn
- Date: 2018
- Subjects: Plasmodium falciparum , Molecular chaperones , Heat shock proteins , Protein-protein interactions , Antimalarials
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/62626 , vital:28216
- Description: Protein-protein interactions are involved in a range of disease processes and thus have become the focus of many drug discovery programs. Widespread drug resistance to all currently used antimalarial drugs drives the search for alternative drug targets with novel mechanisms of action that offer new therapeutic options. Molecular chaperones such as heat shock proteins facilitate protein folding, play a role in protein trafficking and prevent protein misfolding in cells under stress. Heat shock protein 90 (Hsp90) is a well-studied chaperone that has been the focus of cancer drug development with moderate success. In Plasmodium falciparum (P. falciparum), heat shock proteins are thought to play a vital role in parasite survival of the physiologically diverse habitats of the parasite lifecycle and because Hsp90 is prominently expressed in P. falciparum, the chaperone is considered a potentially ideal drug target. Hsp90 function in cells is regulated by interactions with co-chaperones, which includes Heat shock protein 70-Heat shock protein 90 organising protein (Hop). As opposed to directly inhibiting Hsp90 activity, targeting Hsp90 interaction with Hop has recently been suggested as an alternative method of Hsp90 inhibition that has not been explored in P. falciparum. The aim of this research project was to demonstrate PfHsp90 and PfHop robustly interact in vitro and to facilitate high-throughput screening of PfHsp90-PfHop inhibitors by developing and optimising a novel plate capture Hsp90-Hop interaction assay. To establish the assay, the respective domains of the proteins that mediate Hsp90-Hop interaction were used (Hsp90 C- terminal domain and Hop TPR2A domain). The human Hsp90 C-terminal domain and glutathione-S-transferase (GST) coding sequences were cloned into pET-28a(+) and murine and P. falciparum TPR2A sequences into pGEX-4T-1 plasmids to enable expression of histidine-tagged and GST fusion proteins, respectively, in Escherichia coli. The P. falciparum Hsp90 C-terminal domain sequence cloned into pET-28a(+) was supplied by GenScript. The constructs were transformed into T7 Express lysYcompetent E. coli cells and subsequent small- scale expression studies showed the recombinant proteins were expressed in a soluble form allowing for subsequent protein purification. Purification of the recombinant proteins was achieved using nickel-NTA and glutathione affinity chromatography for the His-tagged (Hsp90 C-terminal domains and GST) and GST fusion proteins (TPR2A domains), respectively. The purified proteins were used to establish and optimise mammalian and P. falciparum Hsp90- Hop interaction assays on nickel-coated plates by immobilising the His-tagged C-terminal domains on the plates and detecting the binding of the GST-TPR2A domains using a colorimetric GST enzyme assay. Z’-factor values above 0.5 were observed for both assays indicating good separation between the protein interaction signals and negative control background signals, although relatively high background signals were observed for the mammalian interaction due to non-specific binding of murine TPR2A to the plate. Designed human and P. falciparum TPR peptides were observed to be effective inhibitors of the mammalian and P. falciparum interactions, demonstrating the assay’s ability to respond to inhibitor compounds. Comparison of assay performance using GST assay kit reagents and lab- prepared reagents showed the assay was more efficient using lab-prepared reagents, however, lower GST signals were observed when comparing assay performance using a custom prepared Ni-NTA plate to a purchased Ni-NTA plate. The Hsp90-Hop interaction assays were also performed using an alternative assay format in which the GST-TPR2A fusion proteins were immobilised on glutathione-coated plates and binding of the His-tagged C-terminal domains detected with a nickel-horseradish peroxidase (HRP) conjugate and a colorimetric HRP substrate. The assay showed higher interaction signals for the P. falciparum proteins but comparatively low signals for the mammalian proteins. Z’-factor values for the assay were above 0.8 for both protein sets, suggesting this assay format is superior to the GST assay. However, further optimisation of this assay format is required. This study demonstrated direct binding of PfHsp90-PfHop in vitro and established a novel and robust PfHsp90-PfHop interaction assay format that can be used in future screening campaigns.
- Full Text:
- Date Issued: 2018
- Authors: Wambua, Lynn
- Date: 2018
- Subjects: Plasmodium falciparum , Molecular chaperones , Heat shock proteins , Protein-protein interactions , Antimalarials
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/62626 , vital:28216
- Description: Protein-protein interactions are involved in a range of disease processes and thus have become the focus of many drug discovery programs. Widespread drug resistance to all currently used antimalarial drugs drives the search for alternative drug targets with novel mechanisms of action that offer new therapeutic options. Molecular chaperones such as heat shock proteins facilitate protein folding, play a role in protein trafficking and prevent protein misfolding in cells under stress. Heat shock protein 90 (Hsp90) is a well-studied chaperone that has been the focus of cancer drug development with moderate success. In Plasmodium falciparum (P. falciparum), heat shock proteins are thought to play a vital role in parasite survival of the physiologically diverse habitats of the parasite lifecycle and because Hsp90 is prominently expressed in P. falciparum, the chaperone is considered a potentially ideal drug target. Hsp90 function in cells is regulated by interactions with co-chaperones, which includes Heat shock protein 70-Heat shock protein 90 organising protein (Hop). As opposed to directly inhibiting Hsp90 activity, targeting Hsp90 interaction with Hop has recently been suggested as an alternative method of Hsp90 inhibition that has not been explored in P. falciparum. The aim of this research project was to demonstrate PfHsp90 and PfHop robustly interact in vitro and to facilitate high-throughput screening of PfHsp90-PfHop inhibitors by developing and optimising a novel plate capture Hsp90-Hop interaction assay. To establish the assay, the respective domains of the proteins that mediate Hsp90-Hop interaction were used (Hsp90 C- terminal domain and Hop TPR2A domain). The human Hsp90 C-terminal domain and glutathione-S-transferase (GST) coding sequences were cloned into pET-28a(+) and murine and P. falciparum TPR2A sequences into pGEX-4T-1 plasmids to enable expression of histidine-tagged and GST fusion proteins, respectively, in Escherichia coli. The P. falciparum Hsp90 C-terminal domain sequence cloned into pET-28a(+) was supplied by GenScript. The constructs were transformed into T7 Express lysYcompetent E. coli cells and subsequent small- scale expression studies showed the recombinant proteins were expressed in a soluble form allowing for subsequent protein purification. Purification of the recombinant proteins was achieved using nickel-NTA and glutathione affinity chromatography for the His-tagged (Hsp90 C-terminal domains and GST) and GST fusion proteins (TPR2A domains), respectively. The purified proteins were used to establish and optimise mammalian and P. falciparum Hsp90- Hop interaction assays on nickel-coated plates by immobilising the His-tagged C-terminal domains on the plates and detecting the binding of the GST-TPR2A domains using a colorimetric GST enzyme assay. Z’-factor values above 0.5 were observed for both assays indicating good separation between the protein interaction signals and negative control background signals, although relatively high background signals were observed for the mammalian interaction due to non-specific binding of murine TPR2A to the plate. Designed human and P. falciparum TPR peptides were observed to be effective inhibitors of the mammalian and P. falciparum interactions, demonstrating the assay’s ability to respond to inhibitor compounds. Comparison of assay performance using GST assay kit reagents and lab- prepared reagents showed the assay was more efficient using lab-prepared reagents, however, lower GST signals were observed when comparing assay performance using a custom prepared Ni-NTA plate to a purchased Ni-NTA plate. The Hsp90-Hop interaction assays were also performed using an alternative assay format in which the GST-TPR2A fusion proteins were immobilised on glutathione-coated plates and binding of the His-tagged C-terminal domains detected with a nickel-horseradish peroxidase (HRP) conjugate and a colorimetric HRP substrate. The assay showed higher interaction signals for the P. falciparum proteins but comparatively low signals for the mammalian proteins. Z’-factor values for the assay were above 0.8 for both protein sets, suggesting this assay format is superior to the GST assay. However, further optimisation of this assay format is required. This study demonstrated direct binding of PfHsp90-PfHop in vitro and established a novel and robust PfHsp90-PfHop interaction assay format that can be used in future screening campaigns.
- Full Text:
- Date Issued: 2018
The economic rationale and modalities for rural infrastructure development: developmental local government in rural service delivery
- Stilwell, Ted, Atkinson, Doreen
- Authors: Stilwell, Ted , Atkinson, Doreen
- Date: 1999-09
- Subjects: Rural development -- South Africa , South Africa -- Economic conditions , Infrastructure (Economics) -- South Africa , Sustainable development -- South Africa , Local government -- South Africa
- Language: English
- Type: text , book
- Identifier: http://hdl.handle.net/10962/73841 , vital:30234 , 1919692444
- Description: The Policy Unit of the Development Bank of Southern Africa (DBSA) has developed a number of strategic themes embracing many cross-cutting issues. The main thrust of the Unit’s work, however, focuses on the core mandate of the Bank - infrastructure. In order to provide for a coherent whole, rural infrastructure must fit into a broader policy framework. Globally, the goals set for the first two decades of the next millennium are to address poverty and achieve food security. The DBSA can contribute to South Africa’s position on these global themes by addressing the issue of rural infrastructure delivery, taking cognisance of the South African government’s Growth, Employment and Redistribution (GEAR) programme. This discussion document will specifically address the local economic development activities of farm production and rural livelihoods in order to achieve food security, address poverty and foster economic growth in the marginalised, infrastructure-deprived rural areas of the country. It is assumed that additional entrepreneurial economic opportunities will arise at village and town level as producers become nett surplus producers, eg in small grain milling, cottage industry, village markets, processing (value adding), etc. In contrast to urban development, where economic activity is assumed by investing in infrastructure, the DBSA will actively have to support the transformation of economic activity in the rural scenario. One of the past successes of the DBSA has been the introduction of the Farmer Support Programme (FSP). The FSP’s provision of services and support, based on the needs of existing smallholders, is embedded in participatory planning and action. Without local initiatives, rural infrastructure cannot be delivered effectively to bolster economic growth, create jobs and redistribute income. Most of the country’s poor live in rural areas, and without rural development there can be no GEAR. Life in these areas needs to be made liveable by encouraging entrepreneurial development, which includes facilitating agriculture, creating jobs and increasing rural income through appropriate government actions for improving the welfare of rural households. One of the ways in which national, provincial and local government could have a broad-based impact is by providing rural economic infrastructure and, to this end, local government, including regional and district councils, has been mandated to develop integrated development plans. These plans have to include economic development plans, and entrepreneurial development has been singled out as important in this regard. From this it follows that district councils and municipalities have a prime responsibility for developing entrepreneurs, including farmers in their rural constituency.
- Full Text:
- Date Issued: 1999-09
- Authors: Stilwell, Ted , Atkinson, Doreen
- Date: 1999-09
- Subjects: Rural development -- South Africa , South Africa -- Economic conditions , Infrastructure (Economics) -- South Africa , Sustainable development -- South Africa , Local government -- South Africa
- Language: English
- Type: text , book
- Identifier: http://hdl.handle.net/10962/73841 , vital:30234 , 1919692444
- Description: The Policy Unit of the Development Bank of Southern Africa (DBSA) has developed a number of strategic themes embracing many cross-cutting issues. The main thrust of the Unit’s work, however, focuses on the core mandate of the Bank - infrastructure. In order to provide for a coherent whole, rural infrastructure must fit into a broader policy framework. Globally, the goals set for the first two decades of the next millennium are to address poverty and achieve food security. The DBSA can contribute to South Africa’s position on these global themes by addressing the issue of rural infrastructure delivery, taking cognisance of the South African government’s Growth, Employment and Redistribution (GEAR) programme. This discussion document will specifically address the local economic development activities of farm production and rural livelihoods in order to achieve food security, address poverty and foster economic growth in the marginalised, infrastructure-deprived rural areas of the country. It is assumed that additional entrepreneurial economic opportunities will arise at village and town level as producers become nett surplus producers, eg in small grain milling, cottage industry, village markets, processing (value adding), etc. In contrast to urban development, where economic activity is assumed by investing in infrastructure, the DBSA will actively have to support the transformation of economic activity in the rural scenario. One of the past successes of the DBSA has been the introduction of the Farmer Support Programme (FSP). The FSP’s provision of services and support, based on the needs of existing smallholders, is embedded in participatory planning and action. Without local initiatives, rural infrastructure cannot be delivered effectively to bolster economic growth, create jobs and redistribute income. Most of the country’s poor live in rural areas, and without rural development there can be no GEAR. Life in these areas needs to be made liveable by encouraging entrepreneurial development, which includes facilitating agriculture, creating jobs and increasing rural income through appropriate government actions for improving the welfare of rural households. One of the ways in which national, provincial and local government could have a broad-based impact is by providing rural economic infrastructure and, to this end, local government, including regional and district councils, has been mandated to develop integrated development plans. These plans have to include economic development plans, and entrepreneurial development has been singled out as important in this regard. From this it follows that district councils and municipalities have a prime responsibility for developing entrepreneurs, including farmers in their rural constituency.
- Full Text:
- Date Issued: 1999-09
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