The influence of indigenous languages on Ugandan English as used in the media
- Authors: Tukwasibwe, Constance
- Date: 2014
- Subjects: Mass media and language -- Uganda , Languages in contact -- Uganda , English language -- Uganda , Language transfer (Language learning) -- Uganda , Communication and culture -- Uganda , Bilingualism -- Uganda , Linguistic demography -- Uganda , Uganda -- Languages
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3646 , http://hdl.handle.net/10962/d1015637
- Description: When two or more languages come in contact, they influence each other in various ways, for example through word borrowing, transfer of sounds, morphology and syntax taken from one language system and imported to another. In this study, the primary concern is on the indigenous communities of Uganda learning the English language, plus the influence that this interaction brings into the linguistic space. Bringing the Ugandan multilingual situation into perspective, the study looks at how the English language has interacted with the local languages and the local speech habits, customs and traditions of the indigenous people, to the extent that it has been indigenized. Some word usage results in miscommunication due to the socio-cultural uniqueness of Ugandan cultural expressions. As an example, because of the practice of polygamy in most Ugandan cultures, words like co-wife are coined to mean 'a woman who shares a husband, or a husband's other wife', a word that is absent in both the language and culture of native English speakers. Furthermore some words are formed by calquing some indigenous language expressions, e.g. 'to eat money' or 'to eat cash', an expression that is calqued from the Luganda phrase, kulya sente. Such word coinages are meant to fill the 'shortfall' where the English language fails to provide adequate equivalents. Understanding the context of this kind of English usage and the influence from the indigenous languages is helpful in handling inter-cultural discourses, as the same expression may convey different senses to different people in different contexts. So then, this study deals with some peculiarities of Ugandan English, namely; the features of Ugandan English grammar which are influenced by the indigenous languages. Evidence from the Corpus of Ugandan English is explored to establish that indigenous languages in Uganda have a significant influence on the English language variety spoken in the country, and that a large part of English bilingual speakers cannot speak English without transferring the features from their mother tongue or indeed, switching and mixing codes. A British corpus was used for the purposes of comparison with Ugandan English. The research was conducted in Uganda, drawing data from English newspapers, radio and television talk -shows that were recorded to provide a structural analysis of the contact situations. The result of the study points to the fact that, indeed, the phonological, morpho-syntactic and semantic characteristics of Ugandan English have a considerable amount of influence from indigenous local languages. This study is hinged on the assumption that when indigenous languages and the target language come together in a linguistic contact situation, the resulting variety would exhibit distinct phonological, lexical, grammatical and semantic/pragmatic features ( cf. Sankoff, 2001; Thomason, 1995; Thomason & Kaufman, 1988; Winford, 2005). However, some of these innovations have attracted criticism from 'prescriptivists' such as Quirk (1985, 1988, 1990); Gaudio (2011); and Abbot (1991) who perceive them as 'nonstandard', 'incorrect English language usage' and a 'direct translation from the language user's mother tongue into English'. Yet, indigenous languages continue to play important roles in shaping the kind of English language usage in Uganda.
- Full Text:
- Date Issued: 2014
- Authors: Tukwasibwe, Constance
- Date: 2014
- Subjects: Mass media and language -- Uganda , Languages in contact -- Uganda , English language -- Uganda , Language transfer (Language learning) -- Uganda , Communication and culture -- Uganda , Bilingualism -- Uganda , Linguistic demography -- Uganda , Uganda -- Languages
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3646 , http://hdl.handle.net/10962/d1015637
- Description: When two or more languages come in contact, they influence each other in various ways, for example through word borrowing, transfer of sounds, morphology and syntax taken from one language system and imported to another. In this study, the primary concern is on the indigenous communities of Uganda learning the English language, plus the influence that this interaction brings into the linguistic space. Bringing the Ugandan multilingual situation into perspective, the study looks at how the English language has interacted with the local languages and the local speech habits, customs and traditions of the indigenous people, to the extent that it has been indigenized. Some word usage results in miscommunication due to the socio-cultural uniqueness of Ugandan cultural expressions. As an example, because of the practice of polygamy in most Ugandan cultures, words like co-wife are coined to mean 'a woman who shares a husband, or a husband's other wife', a word that is absent in both the language and culture of native English speakers. Furthermore some words are formed by calquing some indigenous language expressions, e.g. 'to eat money' or 'to eat cash', an expression that is calqued from the Luganda phrase, kulya sente. Such word coinages are meant to fill the 'shortfall' where the English language fails to provide adequate equivalents. Understanding the context of this kind of English usage and the influence from the indigenous languages is helpful in handling inter-cultural discourses, as the same expression may convey different senses to different people in different contexts. So then, this study deals with some peculiarities of Ugandan English, namely; the features of Ugandan English grammar which are influenced by the indigenous languages. Evidence from the Corpus of Ugandan English is explored to establish that indigenous languages in Uganda have a significant influence on the English language variety spoken in the country, and that a large part of English bilingual speakers cannot speak English without transferring the features from their mother tongue or indeed, switching and mixing codes. A British corpus was used for the purposes of comparison with Ugandan English. The research was conducted in Uganda, drawing data from English newspapers, radio and television talk -shows that were recorded to provide a structural analysis of the contact situations. The result of the study points to the fact that, indeed, the phonological, morpho-syntactic and semantic characteristics of Ugandan English have a considerable amount of influence from indigenous local languages. This study is hinged on the assumption that when indigenous languages and the target language come together in a linguistic contact situation, the resulting variety would exhibit distinct phonological, lexical, grammatical and semantic/pragmatic features ( cf. Sankoff, 2001; Thomason, 1995; Thomason & Kaufman, 1988; Winford, 2005). However, some of these innovations have attracted criticism from 'prescriptivists' such as Quirk (1985, 1988, 1990); Gaudio (2011); and Abbot (1991) who perceive them as 'nonstandard', 'incorrect English language usage' and a 'direct translation from the language user's mother tongue into English'. Yet, indigenous languages continue to play important roles in shaping the kind of English language usage in Uganda.
- Full Text:
- Date Issued: 2014
The dramatic theory of William Hazlitt : "Imagination in criticism"
- Authors: De Villiers, André
- Date: 1964
- Subjects: Hazlitt, William, 1778-1830 , Drama -- History and criticism -- 19th century , English drama -- History and criticism -- 19th century
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2320 , http://hdl.handle.net/10962/d1013383
- Full Text:
- Date Issued: 1964
- Authors: De Villiers, André
- Date: 1964
- Subjects: Hazlitt, William, 1778-1830 , Drama -- History and criticism -- 19th century , English drama -- History and criticism -- 19th century
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2320 , http://hdl.handle.net/10962/d1013383
- Full Text:
- Date Issued: 1964
A model for privacy-aware presence management in mobile communications
- Authors: Ophoff, Jacobus Albertus
- Date: 2011
- Subjects: Mobile communications systems , Mobile communication systems -- Management
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9757 , http://hdl.handle.net/10948/1354 , Mobile communications systems , Mobile communication systems -- Management
- Description: As humans we find communicating natural and compelling. Over the centuries we have created many innovations which enable and improve communication between people; during the past decade mobile phone networks have brought about a technological revolution in this area. Never before have people been so connected to one another. Today we have the ability to communicate with almost anyone, anytime, anywhere. Our increased connectivity and reachability also leads to new issues and challenges that we need to deal with. When we phone someone we expect an instant connection, and when this does not occur it can be frustrating. On the other hand it is equally disruptive to receive a call when one is busy with an important task or in a situation where communication is inappropriate. Social protocol dictates that we try to minimize such situations for the benefit of others nearby and for ourselves. This management of communications is a constant and difficult task. Using presence – which signals a person’s availability and willingness to communicate – is a solution to this problem. Such information can benefit communication partners by increasing the likelihood of a successful connection and decreasing disruptions. This research addresses the problem of staying connected while keeping control over mobile communications. It adopts a design-science research paradigm, with the primary research artifact being a model for privacy-aware presence management in mobile communications. As part of the model development knowledge contributions are made in several ways. Existing knowledge about the problem area is extended through a quantitative analysis of mobile communications management. This analysis uses a novel survey, collecting useful empirical data for future research. This includes how people currently manage their communications and what features they expect from a potential “call management” system. The examination and use of presence standards, as a foundation for the model, provides a comparison of the main presence technologies available today. A focus on privacy features identifies several shortcomings in standards which, if addressed, can help to improve and make these standards more complete. The model stresses the privacy of potentially sensitive presence information. A unique perspective based on social relationship theories is adopted. The use of relationship groups not only makes logical sense but also assists in the management of presence information and extends existing standards. Finally, the evaluation of the model demonstrates the feasibility of a practical implementation as well the ability to extend the model in next generation mobile networks. Thus the model presents a solid foundation for the development of future services. In these ways the proposed model contributes positively towards balancing efficient mobile communications with the need for privacy-awareness.
- Full Text:
- Date Issued: 2011
- Authors: Ophoff, Jacobus Albertus
- Date: 2011
- Subjects: Mobile communications systems , Mobile communication systems -- Management
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9757 , http://hdl.handle.net/10948/1354 , Mobile communications systems , Mobile communication systems -- Management
- Description: As humans we find communicating natural and compelling. Over the centuries we have created many innovations which enable and improve communication between people; during the past decade mobile phone networks have brought about a technological revolution in this area. Never before have people been so connected to one another. Today we have the ability to communicate with almost anyone, anytime, anywhere. Our increased connectivity and reachability also leads to new issues and challenges that we need to deal with. When we phone someone we expect an instant connection, and when this does not occur it can be frustrating. On the other hand it is equally disruptive to receive a call when one is busy with an important task or in a situation where communication is inappropriate. Social protocol dictates that we try to minimize such situations for the benefit of others nearby and for ourselves. This management of communications is a constant and difficult task. Using presence – which signals a person’s availability and willingness to communicate – is a solution to this problem. Such information can benefit communication partners by increasing the likelihood of a successful connection and decreasing disruptions. This research addresses the problem of staying connected while keeping control over mobile communications. It adopts a design-science research paradigm, with the primary research artifact being a model for privacy-aware presence management in mobile communications. As part of the model development knowledge contributions are made in several ways. Existing knowledge about the problem area is extended through a quantitative analysis of mobile communications management. This analysis uses a novel survey, collecting useful empirical data for future research. This includes how people currently manage their communications and what features they expect from a potential “call management” system. The examination and use of presence standards, as a foundation for the model, provides a comparison of the main presence technologies available today. A focus on privacy features identifies several shortcomings in standards which, if addressed, can help to improve and make these standards more complete. The model stresses the privacy of potentially sensitive presence information. A unique perspective based on social relationship theories is adopted. The use of relationship groups not only makes logical sense but also assists in the management of presence information and extends existing standards. Finally, the evaluation of the model demonstrates the feasibility of a practical implementation as well the ability to extend the model in next generation mobile networks. Thus the model presents a solid foundation for the development of future services. In these ways the proposed model contributes positively towards balancing efficient mobile communications with the need for privacy-awareness.
- Full Text:
- Date Issued: 2011
Effect of adding a probiotic to an aquaponic system on plant and fish growth, water quality, and microbial diversity
- Authors: Kasozi, Nasser
- Date: 2022-04-08
- Subjects: To be added
- Language: English
- Type: thesis , text , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/294596 , vital:57236
- Description: Thesis embargoed. Release date April 2023. , Thesis (PhD) -- Faculty of Science, Ichthyology and Fisheries Science, 2022
- Full Text:
- Date Issued: 2022-04-08
- Authors: Kasozi, Nasser
- Date: 2022-04-08
- Subjects: To be added
- Language: English
- Type: thesis , text , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/294596 , vital:57236
- Description: Thesis embargoed. Release date April 2023. , Thesis (PhD) -- Faculty of Science, Ichthyology and Fisheries Science, 2022
- Full Text:
- Date Issued: 2022-04-08
Movement patterns, stock delineation and conservation of an overexploited fishery species, Lithognathus Lithognathus (Pisces: Sparidae)
- Authors: Bennett, Rhett Hamilton
- Date: 2012
- Subjects: Reef fishes -- Behavior , Endangered species -- South Africa , Fish stock assessment -- South Africa , Fishery management -- South Africa , Fish communities -- South Africa , Sparidae , Lithognathus , Lithognathus -- Growth
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5374 , http://hdl.handle.net/10962/d1015709
- Description: White steenbras Lithognathus lithognathus (Pisces: Sparidae) has been a major target species of numerous fisheries in South Africa, since the late 19th century. Historically, it contributed substantially to annual catches in commercial net fisheries, and became dominant in recreational shore catches in the latter half of the 20th century. However, overexploitation in both sectors resulted in severe declines in abundance. The ultimate collapse of the stock by the end of the last century, and the failure of traditional management measures to protect the species indicate that a new management approach for this species is necessary. The species was identified as a priority for research, management and conservation in a National Linefish Status Report. Despite knowledge on aspects of its biology and life history, little is known about juvenile habitat use patterns, home range dynamics and movement behaviour in estuaries. Similarly, the movement and migration of larger juveniles and adults in the marine environment are poorly understood. Furthermore, there is a complete lack of information on its genetic stock structure. Such information is essential for effective management of a fishery species. This thesis aimed to address the gaps in the understanding of white steenbras movement patterns and genetic stock structure, and provide an assessment of its current conservation status. The study adopted a multidisciplinary approach, incorporating a range of methods and drawing on available information, including published literature, unpublished reports and data from long-term monitoring programmes. Acoustic telemetry, conducted in a range of estuaries, showed high site fidelity, restricted area use, small home ranges relative to the size of the estuary, and a high level of residency within estuaries at the early juvenile life stage. Behaviour within estuaries was dominated by station-keeping, superimposed by a strong diel behaviour, presumably based on feeding and/or predator avoidance, with individuals entering the shallow littoral zone at night to feed, and seeking refuge in the deeper channel areas during the daytime. Conventional dart tagging and recapture data from four ongoing, long-term coastal fish tagging projects, spread throughout the distribution of this species, indicated high levels of residency in the surf zone at the late juvenile and sub-adult life stages. Consequently, juvenile and sub-adult white steenbras are vulnerable to localised depletion, although they can be effectively protected by suitably positioned estuarine protected areas (EPAs) and marine protected areas (MPAs), respectively. It has been hypothesized that adult white steenbras undertake large-scale coastal migrations between summer aggregation areas and winter spawning grounds. The scale of observed coastal movements was correlated with fish size (and age), with larger fish undertaking considerably longer-distance coastal movements than smaller individuals, supporting this hypothesis. Given the migratory behaviour of adults, and indications that limited spawning habitat exists, MPAs designed to protect white steenbras during the adult life stage should encompass all known spawning aggregation sites. The fishery is plagued by problems such as low compliance and low enforcement capacity, and alternative management measures, such as seasonal closure, need to be evaluated. Despite considerable conventional dart tagging effort around the coastline (5 782 fish tagged) with 292 recaptures there remains a lack of empirical evidence of fish migrating long distances (> 600 km) between aggregation and spawning areas. This uncertainty in the level of connectivity among coastal regions was addressed using mitochondrial DNA sequencing and genotyping of microsatellite repeat loci in the nuclear genome, which showed no evidence of major geographic barriers to gene flow in this species. Samples collected throughout the white steenbras core distribution showed high genetic diversity, low genetic differentiation and no evidence of isolation by distance or localised spawning. Although historically dominant in several fisheries, analysis of long-term commercial and recreational catch data for white steenbras indicated considerable declines and ultimately stock collapse. Improved catch-per-unit-effort in two large MPAs subsequent to closure confirmed that MPAs can be effective for the protection of white steenbras. However, the current MPA network encompasses a low proportion of sandy shoreline, for which white steenbras exhibits an affinity. Many MPAs do not prohibit recreational shore angling, which currently accounts for the greatest proportion of the total annual catch. Furthermore, EPAs within the juvenile distribution protect a negligible proportion of the total available surface area of estuaries – habitat on which white steenbras is wholly dependent. Despite some evidence of recent increases in abundance in estuaries and the surf zone in certain areas, white steenbras meets the criteria for “Endangered” on the IUCN Red List of Threatened Species, and for “Protected species” status on the National Environmental Management: Biodiversity Act of South Africa. The species requires improved management, with consideration for its life-history style, estuarine dependency, surf zone residency, predictable spawning migrations and its poor conservation status. The multidisciplinary approach provides valuable information towards an improved scientific basis for the management of white steenbras and a framework for research that can be adopted for other overexploited, estuarine-associated coastal fishery species.
- Full Text:
- Date Issued: 2012
- Authors: Bennett, Rhett Hamilton
- Date: 2012
- Subjects: Reef fishes -- Behavior , Endangered species -- South Africa , Fish stock assessment -- South Africa , Fishery management -- South Africa , Fish communities -- South Africa , Sparidae , Lithognathus , Lithognathus -- Growth
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5374 , http://hdl.handle.net/10962/d1015709
- Description: White steenbras Lithognathus lithognathus (Pisces: Sparidae) has been a major target species of numerous fisheries in South Africa, since the late 19th century. Historically, it contributed substantially to annual catches in commercial net fisheries, and became dominant in recreational shore catches in the latter half of the 20th century. However, overexploitation in both sectors resulted in severe declines in abundance. The ultimate collapse of the stock by the end of the last century, and the failure of traditional management measures to protect the species indicate that a new management approach for this species is necessary. The species was identified as a priority for research, management and conservation in a National Linefish Status Report. Despite knowledge on aspects of its biology and life history, little is known about juvenile habitat use patterns, home range dynamics and movement behaviour in estuaries. Similarly, the movement and migration of larger juveniles and adults in the marine environment are poorly understood. Furthermore, there is a complete lack of information on its genetic stock structure. Such information is essential for effective management of a fishery species. This thesis aimed to address the gaps in the understanding of white steenbras movement patterns and genetic stock structure, and provide an assessment of its current conservation status. The study adopted a multidisciplinary approach, incorporating a range of methods and drawing on available information, including published literature, unpublished reports and data from long-term monitoring programmes. Acoustic telemetry, conducted in a range of estuaries, showed high site fidelity, restricted area use, small home ranges relative to the size of the estuary, and a high level of residency within estuaries at the early juvenile life stage. Behaviour within estuaries was dominated by station-keeping, superimposed by a strong diel behaviour, presumably based on feeding and/or predator avoidance, with individuals entering the shallow littoral zone at night to feed, and seeking refuge in the deeper channel areas during the daytime. Conventional dart tagging and recapture data from four ongoing, long-term coastal fish tagging projects, spread throughout the distribution of this species, indicated high levels of residency in the surf zone at the late juvenile and sub-adult life stages. Consequently, juvenile and sub-adult white steenbras are vulnerable to localised depletion, although they can be effectively protected by suitably positioned estuarine protected areas (EPAs) and marine protected areas (MPAs), respectively. It has been hypothesized that adult white steenbras undertake large-scale coastal migrations between summer aggregation areas and winter spawning grounds. The scale of observed coastal movements was correlated with fish size (and age), with larger fish undertaking considerably longer-distance coastal movements than smaller individuals, supporting this hypothesis. Given the migratory behaviour of adults, and indications that limited spawning habitat exists, MPAs designed to protect white steenbras during the adult life stage should encompass all known spawning aggregation sites. The fishery is plagued by problems such as low compliance and low enforcement capacity, and alternative management measures, such as seasonal closure, need to be evaluated. Despite considerable conventional dart tagging effort around the coastline (5 782 fish tagged) with 292 recaptures there remains a lack of empirical evidence of fish migrating long distances (> 600 km) between aggregation and spawning areas. This uncertainty in the level of connectivity among coastal regions was addressed using mitochondrial DNA sequencing and genotyping of microsatellite repeat loci in the nuclear genome, which showed no evidence of major geographic barriers to gene flow in this species. Samples collected throughout the white steenbras core distribution showed high genetic diversity, low genetic differentiation and no evidence of isolation by distance or localised spawning. Although historically dominant in several fisheries, analysis of long-term commercial and recreational catch data for white steenbras indicated considerable declines and ultimately stock collapse. Improved catch-per-unit-effort in two large MPAs subsequent to closure confirmed that MPAs can be effective for the protection of white steenbras. However, the current MPA network encompasses a low proportion of sandy shoreline, for which white steenbras exhibits an affinity. Many MPAs do not prohibit recreational shore angling, which currently accounts for the greatest proportion of the total annual catch. Furthermore, EPAs within the juvenile distribution protect a negligible proportion of the total available surface area of estuaries – habitat on which white steenbras is wholly dependent. Despite some evidence of recent increases in abundance in estuaries and the surf zone in certain areas, white steenbras meets the criteria for “Endangered” on the IUCN Red List of Threatened Species, and for “Protected species” status on the National Environmental Management: Biodiversity Act of South Africa. The species requires improved management, with consideration for its life-history style, estuarine dependency, surf zone residency, predictable spawning migrations and its poor conservation status. The multidisciplinary approach provides valuable information towards an improved scientific basis for the management of white steenbras and a framework for research that can be adopted for other overexploited, estuarine-associated coastal fishery species.
- Full Text:
- Date Issued: 2012
Understanding current teacher implementation of Zimbabwe's primary school AIDS curriculum: a case study
- Authors: Musingarabwi, Starlin
- Date: 2013
- Subjects: Teacher participation in curriculum planning -- Zimbabwe , Primary school teachers -- Zimbabwe , Education -- Zimbabwe
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9584 , http://hdl.handle.net/10948/d1020912
- Description: Zimbabwe’s Ministry of Education Sport, Arts and Culture offers as one of the primary school curricula, an AIDS curriculum which all Grades 4 to 7 teachers in Zimbabwe’s primary schools mandatorily implement with a view to contributing towards the prevention of the spread of HIV/AIDS among the young primary school learners. The purpose of this research was to explore and describe teachers’ understanding and implementation of Zimbabwe’s primary school AIDS curriculum regarding the ways in which they articulated teaching practices and processes in their classrooms. The study also aimed to elicit the teachers’ views on how personal and contextual factors impact their adaptation and enactment of the curriculum. The study also sought to establish teachers’ perceptions of their practical experiences with the implementation of Zimbabwe’s primary school AIDS curriculum and their suggestions for improving practice. The study follows a qualitative case study design with minimal quantitative results. It involved three purposively selected primary school grade six teachers (n=3) each of whom was asked to teach five lessons while being observed over a period of three months. Each teacher availed his or her teaching scheme/plan to the researcher who conducted document analysis to glean their symbolic conceptualisation of actual classroom practice of the curriculum. This was followed by three semi-structured interviews with each participating teacher to elicit their perceptions. A content analysis using ideas borrowed from the grounded theory approach was employed resulting in thematic findings. The findings of the study confirm and enhance the theoretical significance of the phenomenological-adaptive perspective of educational change and Honig’s (people, policy, places) and cognition model for describing teacher implementation of the mandatory AIDS curriculum. The findings also confirm the complex ways in which human-generated personal and contextual factors played out in framing and shaping teachers’ personal adaptation of the mandatory AIDS curriculum. The study confirms the adaptation claim that as cognitive sense-makers, teachers mutate and enact a curriculum according to their personal subjective interpretations in the context of unique use-setting implementation realities. Although one of the participants’ understanding and practice displayed considerable comprehension of the requirements of the curriculum, the other teachers displayed an understanding of this curriculum in a superficial way, and experienced few positive experiences and several conceptual and operational constraints in its implementation. Drawing on their practical experiences with the implementation of the curriculum, teachers offered suggestions for transforming the implementation proficiency of this curriculum, which formed part of the conceptual strategy I developed for improving practice. Thus the resultant achievement of the study was a conceptual strategy that was constructed from the key findings of the study to provide educational change leaders with nuanced ideas and insights for improving practice.
- Full Text:
- Date Issued: 2013
- Authors: Musingarabwi, Starlin
- Date: 2013
- Subjects: Teacher participation in curriculum planning -- Zimbabwe , Primary school teachers -- Zimbabwe , Education -- Zimbabwe
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9584 , http://hdl.handle.net/10948/d1020912
- Description: Zimbabwe’s Ministry of Education Sport, Arts and Culture offers as one of the primary school curricula, an AIDS curriculum which all Grades 4 to 7 teachers in Zimbabwe’s primary schools mandatorily implement with a view to contributing towards the prevention of the spread of HIV/AIDS among the young primary school learners. The purpose of this research was to explore and describe teachers’ understanding and implementation of Zimbabwe’s primary school AIDS curriculum regarding the ways in which they articulated teaching practices and processes in their classrooms. The study also aimed to elicit the teachers’ views on how personal and contextual factors impact their adaptation and enactment of the curriculum. The study also sought to establish teachers’ perceptions of their practical experiences with the implementation of Zimbabwe’s primary school AIDS curriculum and their suggestions for improving practice. The study follows a qualitative case study design with minimal quantitative results. It involved three purposively selected primary school grade six teachers (n=3) each of whom was asked to teach five lessons while being observed over a period of three months. Each teacher availed his or her teaching scheme/plan to the researcher who conducted document analysis to glean their symbolic conceptualisation of actual classroom practice of the curriculum. This was followed by three semi-structured interviews with each participating teacher to elicit their perceptions. A content analysis using ideas borrowed from the grounded theory approach was employed resulting in thematic findings. The findings of the study confirm and enhance the theoretical significance of the phenomenological-adaptive perspective of educational change and Honig’s (people, policy, places) and cognition model for describing teacher implementation of the mandatory AIDS curriculum. The findings also confirm the complex ways in which human-generated personal and contextual factors played out in framing and shaping teachers’ personal adaptation of the mandatory AIDS curriculum. The study confirms the adaptation claim that as cognitive sense-makers, teachers mutate and enact a curriculum according to their personal subjective interpretations in the context of unique use-setting implementation realities. Although one of the participants’ understanding and practice displayed considerable comprehension of the requirements of the curriculum, the other teachers displayed an understanding of this curriculum in a superficial way, and experienced few positive experiences and several conceptual and operational constraints in its implementation. Drawing on their practical experiences with the implementation of the curriculum, teachers offered suggestions for transforming the implementation proficiency of this curriculum, which formed part of the conceptual strategy I developed for improving practice. Thus the resultant achievement of the study was a conceptual strategy that was constructed from the key findings of the study to provide educational change leaders with nuanced ideas and insights for improving practice.
- Full Text:
- Date Issued: 2013
Synthesis and biological evaluation of anti-HIV-I integrase agents
- Jesumoroti, Omobolanle Janet
- Authors: Jesumoroti, Omobolanle Janet
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59215 , vital:27479
- Description: Expected release date-April 2019
- Full Text:
- Date Issued: 2017
- Authors: Jesumoroti, Omobolanle Janet
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59215 , vital:27479
- Description: Expected release date-April 2019
- Full Text:
- Date Issued: 2017
Investigation into the technical feasibility of biological treatment of precious metal refining wastewater
- Authors: Moore, Bronwyn Ann
- Date: 2013
- Subjects: Sewage -- Purification -- Biological treatment -- South Africa Sewage -- Purification -- Activated sludge process -- South Africa Water reuse -- South Africa Flotation -- South Africa Platinum mines and mining -- Waste disposal -- South Africa Platinum mines and mining -- Economic aspects -- South Africa Mine water -- Environmental aspects -- South Africa Platinum mines and mining -- Waste minimization -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3888 , http://hdl.handle.net/10962/d1002013
- Description: The hydrometallurgical refining of platinum group metals results in large volumes of liquid waste that requires suitable treatment before any disposal can be contemplated. The wastewater streams are characterized by extremes of pH, high inorganic ion content (such as chloride), significant residual metal loads and small amounts of entrained organic compounds. Historically these effluents were housed in evaporation reservoirs, however lack of space and growing water demands have led Anglo Platinum to consider treatment of these effluents. The aim of this study was to investigate whether biological wastewater treatment could produce water suitable for onsite reuse. Bench-scale activated sludge and anaerobic digestion for co-treatment of an acidic refinery waste stream with domestic wastewater were used to give preliminary data. Activated sludge showed better water treatment at lab scale in terms of removal efficiencies of ammonia (approximately 25%, cf. 20% in anaerobic digestion) and COD (70% cf. 43% in digestion) and greater robustness when biomass health was compared. Activated sludge was consequently selected for a pilot plant trial. The pilot plant was operated on-site and performed comparably with the bench-scale system, however challenges in the clarifier design led to losses of biomass and poor effluent quality (suspended solids washout). The pilot plant was unable to alter the pH of the feed, but a two week maturation period resulted in the pH increasing from 5.3 to 7.0. Tests on algal treatment as an alternative or follow-on unit operation to activated sludge showed it not to be a viable process. The activated sludge effluent was assessed for onsite reuse in flotation and it was found that there was no significant difference between its flotation performance and that of the process water currently used, indicating the effluent generated by the biological treatment system can be used successfully for flotation. Flotation is the method whereby minerals refining operations recover minerals of interest from ore through the addition of chemicals and aeration of the ore slurry. Target minerals adhere to the bubbles and can be removed from the process.
- Full Text:
- Date Issued: 2013
- Authors: Moore, Bronwyn Ann
- Date: 2013
- Subjects: Sewage -- Purification -- Biological treatment -- South Africa Sewage -- Purification -- Activated sludge process -- South Africa Water reuse -- South Africa Flotation -- South Africa Platinum mines and mining -- Waste disposal -- South Africa Platinum mines and mining -- Economic aspects -- South Africa Mine water -- Environmental aspects -- South Africa Platinum mines and mining -- Waste minimization -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3888 , http://hdl.handle.net/10962/d1002013
- Description: The hydrometallurgical refining of platinum group metals results in large volumes of liquid waste that requires suitable treatment before any disposal can be contemplated. The wastewater streams are characterized by extremes of pH, high inorganic ion content (such as chloride), significant residual metal loads and small amounts of entrained organic compounds. Historically these effluents were housed in evaporation reservoirs, however lack of space and growing water demands have led Anglo Platinum to consider treatment of these effluents. The aim of this study was to investigate whether biological wastewater treatment could produce water suitable for onsite reuse. Bench-scale activated sludge and anaerobic digestion for co-treatment of an acidic refinery waste stream with domestic wastewater were used to give preliminary data. Activated sludge showed better water treatment at lab scale in terms of removal efficiencies of ammonia (approximately 25%, cf. 20% in anaerobic digestion) and COD (70% cf. 43% in digestion) and greater robustness when biomass health was compared. Activated sludge was consequently selected for a pilot plant trial. The pilot plant was operated on-site and performed comparably with the bench-scale system, however challenges in the clarifier design led to losses of biomass and poor effluent quality (suspended solids washout). The pilot plant was unable to alter the pH of the feed, but a two week maturation period resulted in the pH increasing from 5.3 to 7.0. Tests on algal treatment as an alternative or follow-on unit operation to activated sludge showed it not to be a viable process. The activated sludge effluent was assessed for onsite reuse in flotation and it was found that there was no significant difference between its flotation performance and that of the process water currently used, indicating the effluent generated by the biological treatment system can be used successfully for flotation. Flotation is the method whereby minerals refining operations recover minerals of interest from ore through the addition of chemicals and aeration of the ore slurry. Target minerals adhere to the bubbles and can be removed from the process.
- Full Text:
- Date Issued: 2013
Colour vision of the citrus psylla Trioza erytreae (Del Guercio) (Homoptera: Psyllidae) in relation to alightment colour preferences
- Authors: Urban, Alan Joseph
- Date: 1977
- Subjects: Trioza , Homoptera , Jumping plant-lice , Color vision
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5885 , http://hdl.handle.net/10962/d1013286
- Description: The colour vision of adult citrus psylla, Trioza erytreae, was investigated in the laboratory using the behavioural parameters: alightment and walking. Light green flushing leaves (under which the nymphs develop) were significantly preferred, visually, to dark green mature leaves for alightment. Diffuse reflectance spectroscopy showed (when expressed in the parameters of human colour vision) that flush has a very slightly longer dominant wavelength, and roughly double the reflectance and purity. Alightrnent frequency correlated almost equally well with "purity" (as noted by Moericke, 1952 et seq., in "yellow-sensitive" aphids) as with the aphidological colour parameter "long/short ratio" developed by Kennedy et al. (1961). Elucidation of the mechanism underlying the citrus psylla's alightment colour preference was initially attempted with a printed spectrum and several paint series of measured spectral characteristics. It was clear that T.erytreae belongs to the "yellow-sensitive" group of Homoptera, but it was impossible to distinguish which pararneter(s) of colour the psyllids were responding to. Phototactic (walking) response to the individual parameters of colour was therefore measured using a monochromator. The phototactic action spectrum (against wavelength) was tri-modal, with peaks in the yellow-green (YG), blue (B), and ultra= violet (UV). Rate of phototaxis was not influenced by bandwidth (roughly equivalent to purity), but was proportional to intensity (roughly equivalent to reflectance). To investigate the influence of the above three wavelength regions on alightment, use was made of a very simple flight chamber incorporating a target of coloured light. Yellow-green and UV light both independently stimulated alightment . Their effect was additive. Different thresholds indicated distinct YG and UV receptor systems. Blue light alone did not stimulate alightment, and was strongly alightment-inhibitory in combination both with YG and with UV light. On the basis of the above physiological/behavioural findings, a new alightment formula was drawn up for describing the hamopteran's apparent manner of alightment determining integration of surface reflectance. The flush preference and alightment distributions on the series of artificial surfaces were found to correlate slightly more accurately, on average, as well as more consistently, with the new formula than with previously-available colour parameters. These findings are placed in perspective to the literature, and their possible economic relevance is discussed.
- Full Text:
- Date Issued: 1977
- Authors: Urban, Alan Joseph
- Date: 1977
- Subjects: Trioza , Homoptera , Jumping plant-lice , Color vision
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5885 , http://hdl.handle.net/10962/d1013286
- Description: The colour vision of adult citrus psylla, Trioza erytreae, was investigated in the laboratory using the behavioural parameters: alightment and walking. Light green flushing leaves (under which the nymphs develop) were significantly preferred, visually, to dark green mature leaves for alightment. Diffuse reflectance spectroscopy showed (when expressed in the parameters of human colour vision) that flush has a very slightly longer dominant wavelength, and roughly double the reflectance and purity. Alightrnent frequency correlated almost equally well with "purity" (as noted by Moericke, 1952 et seq., in "yellow-sensitive" aphids) as with the aphidological colour parameter "long/short ratio" developed by Kennedy et al. (1961). Elucidation of the mechanism underlying the citrus psylla's alightment colour preference was initially attempted with a printed spectrum and several paint series of measured spectral characteristics. It was clear that T.erytreae belongs to the "yellow-sensitive" group of Homoptera, but it was impossible to distinguish which pararneter(s) of colour the psyllids were responding to. Phototactic (walking) response to the individual parameters of colour was therefore measured using a monochromator. The phototactic action spectrum (against wavelength) was tri-modal, with peaks in the yellow-green (YG), blue (B), and ultra= violet (UV). Rate of phototaxis was not influenced by bandwidth (roughly equivalent to purity), but was proportional to intensity (roughly equivalent to reflectance). To investigate the influence of the above three wavelength regions on alightment, use was made of a very simple flight chamber incorporating a target of coloured light. Yellow-green and UV light both independently stimulated alightment . Their effect was additive. Different thresholds indicated distinct YG and UV receptor systems. Blue light alone did not stimulate alightment, and was strongly alightment-inhibitory in combination both with YG and with UV light. On the basis of the above physiological/behavioural findings, a new alightment formula was drawn up for describing the hamopteran's apparent manner of alightment determining integration of surface reflectance. The flush preference and alightment distributions on the series of artificial surfaces were found to correlate slightly more accurately, on average, as well as more consistently, with the new formula than with previously-available colour parameters. These findings are placed in perspective to the literature, and their possible economic relevance is discussed.
- Full Text:
- Date Issued: 1977
Management perceptions regarding skills shortages in gold mines
- Authors: Xingwana, Lumkwana
- Date: 2012
- Subjects: Labor supply--South Africa , Professional employees--South Africa , Occupational training--South Africa , Gold mines and mining--South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9279 , http://hdl.handle.net/10948/d1007959 , Labor supply--South Africa , Professional employees--South Africa , Occupational training--South Africa , Gold mines and mining--South Africa
- Description: The skills shortages in mining and mineral sector had existed for a decade and had a widespread effect on South Africa economy. It affects the level of economic productivity and reduces the country’s capacity to develop a knowledge society. This, in turn, affects the country’s functioning in the global economy. Despite the sector’s best efforts, the shortages continue to grow and threaten the delivery of projects and growth plans. Some researchers contend with the view that the persistence of skills shortages in mining and mineral sector is largely due to entrenched attitudes among both the industry and the community. They claim that employers have the means to change the educational profile of the subsectors by appointing recruits with higher levels of schooling. However, owing to the limited number of higher educated people living in the communities surrounding the mining operations and lack of interest in mining of people with higher levels of education, to name but few, employers are perceived to have a habit of employing people with little skills. The current study was aimed at investigating the impact of skills shortage on organisational performance, propensity to leave, competitive advantage and sustainability, from the management perceptive. The main objective of this study was to incorporate and embed previous research findings and theories into a comprehensive hypothetical model. A hypothetical model showed various factors that may influence skills shortage. Four independent variables (working environment, employment conditions, resources and education and training) were identified as variables that may influence skills shortage; and mediating variable (skills shortage) was also identified as a variable that have potential to affect dependent variables (organisational performance, propensity to leave, competitive advantage and sustainability) of gold mining sector. Furthermore, eight hypotheses were developed to test the relationship between independent, mediating and dependent variables. All these variables were clearly defined and operationalized with various items obtained from measuring instruments used in other similar studies. A purposive sample of 343 respondents was drawn from the population. A seven-point Likert scale and structured questionnaire were administered in person to the respondents and of which 300 were usable and subjected further to several statistical analyses. The validity and reliability of the measuring instrument was evaluated using significant effect p< = 0.001 and Pearson’s correlation test (α = 0.05). Data gathered were fed to and analysed by STATISTICA (version 10) and factor analysis and regression analyses were the statistical procedures used to test the significance of the relationships between the various independent and dependent variables. Consequently, working environment, resources and education and training were three independent variables that were identified as having ability to predict propensity to leave, competitive advantage and sustainability. An attempt was made to establish whether various demographic variables have an influence on mediating and dependent variables through the introduction of gender and position in the organisation while conducting an Analysis of Variance and Multiple linear regressions, but they obtained negative values. The conclusion is that demographic variables do not have over mediating and depended variables. The findings of this study states that with conducive working environment, availability of resources, the high levels of education and training, the country could produce skills that would reduce propensity to leave, drive competitive advantage and sustainability, innovation and entrepreneurship, create competitive advantages and boost employment sustainability.
- Full Text:
- Date Issued: 2012
- Authors: Xingwana, Lumkwana
- Date: 2012
- Subjects: Labor supply--South Africa , Professional employees--South Africa , Occupational training--South Africa , Gold mines and mining--South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9279 , http://hdl.handle.net/10948/d1007959 , Labor supply--South Africa , Professional employees--South Africa , Occupational training--South Africa , Gold mines and mining--South Africa
- Description: The skills shortages in mining and mineral sector had existed for a decade and had a widespread effect on South Africa economy. It affects the level of economic productivity and reduces the country’s capacity to develop a knowledge society. This, in turn, affects the country’s functioning in the global economy. Despite the sector’s best efforts, the shortages continue to grow and threaten the delivery of projects and growth plans. Some researchers contend with the view that the persistence of skills shortages in mining and mineral sector is largely due to entrenched attitudes among both the industry and the community. They claim that employers have the means to change the educational profile of the subsectors by appointing recruits with higher levels of schooling. However, owing to the limited number of higher educated people living in the communities surrounding the mining operations and lack of interest in mining of people with higher levels of education, to name but few, employers are perceived to have a habit of employing people with little skills. The current study was aimed at investigating the impact of skills shortage on organisational performance, propensity to leave, competitive advantage and sustainability, from the management perceptive. The main objective of this study was to incorporate and embed previous research findings and theories into a comprehensive hypothetical model. A hypothetical model showed various factors that may influence skills shortage. Four independent variables (working environment, employment conditions, resources and education and training) were identified as variables that may influence skills shortage; and mediating variable (skills shortage) was also identified as a variable that have potential to affect dependent variables (organisational performance, propensity to leave, competitive advantage and sustainability) of gold mining sector. Furthermore, eight hypotheses were developed to test the relationship between independent, mediating and dependent variables. All these variables were clearly defined and operationalized with various items obtained from measuring instruments used in other similar studies. A purposive sample of 343 respondents was drawn from the population. A seven-point Likert scale and structured questionnaire were administered in person to the respondents and of which 300 were usable and subjected further to several statistical analyses. The validity and reliability of the measuring instrument was evaluated using significant effect p< = 0.001 and Pearson’s correlation test (α = 0.05). Data gathered were fed to and analysed by STATISTICA (version 10) and factor analysis and regression analyses were the statistical procedures used to test the significance of the relationships between the various independent and dependent variables. Consequently, working environment, resources and education and training were three independent variables that were identified as having ability to predict propensity to leave, competitive advantage and sustainability. An attempt was made to establish whether various demographic variables have an influence on mediating and dependent variables through the introduction of gender and position in the organisation while conducting an Analysis of Variance and Multiple linear regressions, but they obtained negative values. The conclusion is that demographic variables do not have over mediating and depended variables. The findings of this study states that with conducive working environment, availability of resources, the high levels of education and training, the country could produce skills that would reduce propensity to leave, drive competitive advantage and sustainability, innovation and entrepreneurship, create competitive advantages and boost employment sustainability.
- Full Text:
- Date Issued: 2012
The role of African theology and spirituality in bereavement among Xhosa children
- Authors: Diniso Patrick Mncedisi
- Date: 2017
- Subjects: Bereavement in children -- Religious aspects Black theology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10353/17839 , vital:41374
- Description: The background to this study highlights the fact that Xhosa children are not being taken care of during bereavement. There is no theological guidance applied to them, to say the least. There are devotions that are usually conducted by lay preachers (Preachers that are not trained as Pastors/Therapists) who do their utmost best to support bereaved families, and this is done out of love. The challenge, however, is that the language mostly used by these preachers does not seem to accommodate children, as there are expressions that do not make any sense to children such as: tutwini, imela igobele esandleni. These are good and relevant concepts as they underpin the reality of death and the need for comfort during bereavement. However, these need to be communicated in a language that can be understood by children. Due to this identified problem, the researcher deemed it necessary to undertake this study. This study comprises five (5) chapters. In chapter one (1), the researcher focused on the research background and introduction. Chapter two (2) dealt with the literature review of this study. The chapter comprises three sections: generic literature review section, theory base section and primary literature review section. In chapter three (3), the focus is on research methodology and its components. Concepts such as sampling procedures, research instrument and research design are dealt with. In chapter four (4), the researcher dealt with concepts such as data analysis, interpretation and presentation of findings. Chapter five dealt with the research recommendations as per the role of the church (Free Church in Southern Africa [FSA]). The researcher is convinced that this work has done justice in addressing the identified gap in care giving for bereaved Xhosa children. The need for theological guidance for bereaved Xhosa children has been highlighted; findings and practical recommendations have also been presented to meet such a need. These confirmed the need for theological guidance for bereaved Xhosa children. The next section contains the appendix list, namely: appendix one (1), two (2) and three (3).
- Full Text:
- Date Issued: 2017
- Authors: Diniso Patrick Mncedisi
- Date: 2017
- Subjects: Bereavement in children -- Religious aspects Black theology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10353/17839 , vital:41374
- Description: The background to this study highlights the fact that Xhosa children are not being taken care of during bereavement. There is no theological guidance applied to them, to say the least. There are devotions that are usually conducted by lay preachers (Preachers that are not trained as Pastors/Therapists) who do their utmost best to support bereaved families, and this is done out of love. The challenge, however, is that the language mostly used by these preachers does not seem to accommodate children, as there are expressions that do not make any sense to children such as: tutwini, imela igobele esandleni. These are good and relevant concepts as they underpin the reality of death and the need for comfort during bereavement. However, these need to be communicated in a language that can be understood by children. Due to this identified problem, the researcher deemed it necessary to undertake this study. This study comprises five (5) chapters. In chapter one (1), the researcher focused on the research background and introduction. Chapter two (2) dealt with the literature review of this study. The chapter comprises three sections: generic literature review section, theory base section and primary literature review section. In chapter three (3), the focus is on research methodology and its components. Concepts such as sampling procedures, research instrument and research design are dealt with. In chapter four (4), the researcher dealt with concepts such as data analysis, interpretation and presentation of findings. Chapter five dealt with the research recommendations as per the role of the church (Free Church in Southern Africa [FSA]). The researcher is convinced that this work has done justice in addressing the identified gap in care giving for bereaved Xhosa children. The need for theological guidance for bereaved Xhosa children has been highlighted; findings and practical recommendations have also been presented to meet such a need. These confirmed the need for theological guidance for bereaved Xhosa children. The next section contains the appendix list, namely: appendix one (1), two (2) and three (3).
- Full Text:
- Date Issued: 2017
Nutrient supplementation and secondary metaolites in melanoma cells
- Authors: Stoll, Karin Elisabeth
- Date: 1994
- Subjects: Vitamin C -- Therapeutic use Cancer -- Research
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4049 , http://hdl.handle.net/10962/d1004110
- Description: Considerable interest exists with regard to the putative therapeutic role of ascorbic acid in various conditions. A condition which has received much attention is cancer, as it is reported that ascorbic acid may be a prophylactic against cancer development. However, the actual involvement of ascorbic acid, an oxidizing/reducing agent, in the development and progression of tumours is presently a subject of much speculation. This study initially addressed the effect of ascorbic acid supplementation over a nutritional concentration range (0 - 100 μg/ml) on the in vitro growth of non-malignant LLCMK and malignant B16 cells. Ascorbic acid supplementation of these two cell types resulted in an overall decrease in the growth of both types of cells. The actual inhibitory mechanism of ascorbic acid on cell growth was not clear. Further study attempted to define and explain a mechanism responsible for this effect. Ascorbic acid has a role in the maintenance of tissue integrity and host defences, thus providing a rational basis for examining its relationship to cancer. Ascorbic acid is lcnown to be essential for the structural integrity of the intercellular matrix of the cells, the latter being a complex aqueous gel containing, amongst other compounds, fats and prostaglandins. Fats and prostaglandins have diverse effects on. membrane stability, enzyme activity and secondary messengers within cells. Hence, this study investigated the effect of ascorbic acid supplementation on certain enzymes and secondary metabolites within the cells, which had the potential to be involved in the control of cell growth. Throughout this study, emphasis was placed on the Bl6 melanoma cells as ascorbic acid supplementation did not significantly affect levels of secondary metabolites within the non-malignant LLCMK cells. Ascorbic acid supplementation of the B16 cells resulted in significant increases in adenylate cyclase activity and cyclic adenosine monophosphate levels, witb a significant decrease in Bl6 cell growth in that particular experiment. As cyclic adenosine monophosphate has a regulatory role in the cell cycle, this study suggested that the inhibitory effect of ascorbic acid supplementation on cell growth was mediated tbrough a final effect provided by the second messenger, cyclic adenosine monophosphate. However, clarification of tbe mechanism of tbe effect of ascorbic acid on adenylate cyclase activity was required. Hence, a further study investigated prostaglandin E₂ levels, as tbese affect adenylate cyclase activity. Prostaglandin E₂ levels were also found to be inversely related to Bl6 cell growth with ascorbic acid supplementation. It thus appeared tbat adenylate cyclase activity was dependent on prostaglandin E₂ levels in the B16 cells, and further study showed that tbis was indeed the case. Here, higher levels of prostaglandin E₂ supplementation of the Bl6 cells inhibited cell growth significantly and also significantly increased adenylate cyclase activity. Arachidonic acid is the precursor of prostaglandin E₂. In the presence of ascorbic acid supplementation, the percentage arachidonic acid composition of the Bl6 cells was inversely correlated with cell growth. Hence, prostaglandin E₂ levels in ascorbic acid supplemented B16 cells appeared dependent on tbe amount of precursor present. This was confirmed when Bl6 cells were supplemented with arachidonic acid. The latter had an inhibitory effect on Bl6 cell growth and also stimulated prostaglandin E₂ production. The cause of tbe inverse relationship between B16 cell growth and arachidonic acid composition with ascorbic acid supplementation was furtber investigated and found to be dependent on tbe uptake of arachidonic acid and other essential fatty acids from tbe medium. The enzymes phospholipase A₂ delta-5 and delta-6-desaturase, and elongase which could influence arachidonic acid levels were not affected to any extent by ascorbic acid supplementation and therefore did not influence the inverse relationship between B16 cell growth and arachidonic acid. Hence, it can be concluded that the effect of ascorbic acid supplementation on the BI6 cells is mediated, in part at least, by cyclic adenosine monophosphate. However, this is not the result of a direct effect of ascorbic acid supplementation. The initial effect of ascorbic acid supplementation concerns fatty acid - in particular arachidonic acid - uptake from the medium, with subsequent cascade effects On secondary metabolites, ultimately affecting the cellular levels of cyclic adenosine monophosphate.
- Full Text:
- Date Issued: 1994
- Authors: Stoll, Karin Elisabeth
- Date: 1994
- Subjects: Vitamin C -- Therapeutic use Cancer -- Research
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4049 , http://hdl.handle.net/10962/d1004110
- Description: Considerable interest exists with regard to the putative therapeutic role of ascorbic acid in various conditions. A condition which has received much attention is cancer, as it is reported that ascorbic acid may be a prophylactic against cancer development. However, the actual involvement of ascorbic acid, an oxidizing/reducing agent, in the development and progression of tumours is presently a subject of much speculation. This study initially addressed the effect of ascorbic acid supplementation over a nutritional concentration range (0 - 100 μg/ml) on the in vitro growth of non-malignant LLCMK and malignant B16 cells. Ascorbic acid supplementation of these two cell types resulted in an overall decrease in the growth of both types of cells. The actual inhibitory mechanism of ascorbic acid on cell growth was not clear. Further study attempted to define and explain a mechanism responsible for this effect. Ascorbic acid has a role in the maintenance of tissue integrity and host defences, thus providing a rational basis for examining its relationship to cancer. Ascorbic acid is lcnown to be essential for the structural integrity of the intercellular matrix of the cells, the latter being a complex aqueous gel containing, amongst other compounds, fats and prostaglandins. Fats and prostaglandins have diverse effects on. membrane stability, enzyme activity and secondary messengers within cells. Hence, this study investigated the effect of ascorbic acid supplementation on certain enzymes and secondary metabolites within the cells, which had the potential to be involved in the control of cell growth. Throughout this study, emphasis was placed on the Bl6 melanoma cells as ascorbic acid supplementation did not significantly affect levels of secondary metabolites within the non-malignant LLCMK cells. Ascorbic acid supplementation of the B16 cells resulted in significant increases in adenylate cyclase activity and cyclic adenosine monophosphate levels, witb a significant decrease in Bl6 cell growth in that particular experiment. As cyclic adenosine monophosphate has a regulatory role in the cell cycle, this study suggested that the inhibitory effect of ascorbic acid supplementation on cell growth was mediated tbrough a final effect provided by the second messenger, cyclic adenosine monophosphate. However, clarification of tbe mechanism of tbe effect of ascorbic acid on adenylate cyclase activity was required. Hence, a further study investigated prostaglandin E₂ levels, as tbese affect adenylate cyclase activity. Prostaglandin E₂ levels were also found to be inversely related to Bl6 cell growth with ascorbic acid supplementation. It thus appeared tbat adenylate cyclase activity was dependent on prostaglandin E₂ levels in the B16 cells, and further study showed that tbis was indeed the case. Here, higher levels of prostaglandin E₂ supplementation of the Bl6 cells inhibited cell growth significantly and also significantly increased adenylate cyclase activity. Arachidonic acid is the precursor of prostaglandin E₂. In the presence of ascorbic acid supplementation, the percentage arachidonic acid composition of the Bl6 cells was inversely correlated with cell growth. Hence, prostaglandin E₂ levels in ascorbic acid supplemented B16 cells appeared dependent on tbe amount of precursor present. This was confirmed when Bl6 cells were supplemented with arachidonic acid. The latter had an inhibitory effect on Bl6 cell growth and also stimulated prostaglandin E₂ production. The cause of tbe inverse relationship between B16 cell growth and arachidonic acid composition with ascorbic acid supplementation was furtber investigated and found to be dependent on tbe uptake of arachidonic acid and other essential fatty acids from tbe medium. The enzymes phospholipase A₂ delta-5 and delta-6-desaturase, and elongase which could influence arachidonic acid levels were not affected to any extent by ascorbic acid supplementation and therefore did not influence the inverse relationship between B16 cell growth and arachidonic acid. Hence, it can be concluded that the effect of ascorbic acid supplementation on the BI6 cells is mediated, in part at least, by cyclic adenosine monophosphate. However, this is not the result of a direct effect of ascorbic acid supplementation. The initial effect of ascorbic acid supplementation concerns fatty acid - in particular arachidonic acid - uptake from the medium, with subsequent cascade effects On secondary metabolites, ultimately affecting the cellular levels of cyclic adenosine monophosphate.
- Full Text:
- Date Issued: 1994
A constructivist deconstruction of post-apartheid South Africa’s trade negotiation strategies: the politics of development and global value chains
- Authors: Pillay, Morgenie
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64731 , vital:28596
- Description: Expected release date-May 2020
- Full Text:
- Date Issued: 2015
- Authors: Pillay, Morgenie
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64731 , vital:28596
- Description: Expected release date-May 2020
- Full Text:
- Date Issued: 2015
Thermal degradation of diamond compacts: a TEM investigation
- Authors: Westraadt, Johan Ewald
- Date: 2011
- Subjects: Materials -- Thermal properties Chemical weathering
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/10829 , vital:26827
- Description: Diamond compacts consist of fine diamond grains bonded together by using high pressure and high temperature. In this study transmission electron microscopy (TEM)was used to study thermal degradation of diamond compacts. Three different types of diamond compacts – namely cobalt polycrystalline diamond (PCD), calcium carbonate PCD, and diamond-SiC composites – were investigated with TEM to understand the processes that occur during synthesis. These compacts were then heated in inert atmospheres and the chemical changes studied with TEM. It was found that PCD, using cobalt as a bonding agent, will degrade after exposure to temperatures above 750ºC. The cobalt pools contain tungsten in solid solution. During heat treatment above 700ºC the solid solution tungsten combines with cobalt and dissolved carbon to form η-phase particles at the cobalt/diamond interface. At higher temperatures or insufficient tungsten levels the rate of dissolved carbon, into the cobalt pool, is too high and the excess carbon will form as graphite in the cobalt pool. Increased levels of solid solution tungsten, in the cobalt, is expected to delay the onset of graphitization in the diamond compact, thereby increasing the thermal stability of the diamond compact. Non-metallic PCD using calcium carbonate as a bonding agent was successfully sintered in this study. TEM revealed similar micro-structural features as in cobalt based PCD. No signs of thermal degradation were found after exposure to 1200ºC in vacuum for this PCD. Contaminants introduced during processing prevented a detailed study of the binder in this system. The effect of substitutional metal atoms and plastic deformation of diamond on the thermal stability of diamond-SiC composites were investigated. A piston cylinder press was developed and used to synthesize diamond-SiC composites with different levels of plastically deformed diamond. It was concluded that substitutional metal atoms and plastic deformation of diamond grains play no role in the thermal degradation of diamond compacts at 750ºC. The thermal degradation of cobalt PCD is therefore completely determined by the cobalt/diamond interaction at 750ºC.
- Full Text:
- Date Issued: 2011
- Authors: Westraadt, Johan Ewald
- Date: 2011
- Subjects: Materials -- Thermal properties Chemical weathering
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/10829 , vital:26827
- Description: Diamond compacts consist of fine diamond grains bonded together by using high pressure and high temperature. In this study transmission electron microscopy (TEM)was used to study thermal degradation of diamond compacts. Three different types of diamond compacts – namely cobalt polycrystalline diamond (PCD), calcium carbonate PCD, and diamond-SiC composites – were investigated with TEM to understand the processes that occur during synthesis. These compacts were then heated in inert atmospheres and the chemical changes studied with TEM. It was found that PCD, using cobalt as a bonding agent, will degrade after exposure to temperatures above 750ºC. The cobalt pools contain tungsten in solid solution. During heat treatment above 700ºC the solid solution tungsten combines with cobalt and dissolved carbon to form η-phase particles at the cobalt/diamond interface. At higher temperatures or insufficient tungsten levels the rate of dissolved carbon, into the cobalt pool, is too high and the excess carbon will form as graphite in the cobalt pool. Increased levels of solid solution tungsten, in the cobalt, is expected to delay the onset of graphitization in the diamond compact, thereby increasing the thermal stability of the diamond compact. Non-metallic PCD using calcium carbonate as a bonding agent was successfully sintered in this study. TEM revealed similar micro-structural features as in cobalt based PCD. No signs of thermal degradation were found after exposure to 1200ºC in vacuum for this PCD. Contaminants introduced during processing prevented a detailed study of the binder in this system. The effect of substitutional metal atoms and plastic deformation of diamond on the thermal stability of diamond-SiC composites were investigated. A piston cylinder press was developed and used to synthesize diamond-SiC composites with different levels of plastically deformed diamond. It was concluded that substitutional metal atoms and plastic deformation of diamond grains play no role in the thermal degradation of diamond compacts at 750ºC. The thermal degradation of cobalt PCD is therefore completely determined by the cobalt/diamond interaction at 750ºC.
- Full Text:
- Date Issued: 2011
In-betweenness: a postcolonial exploration of sociocultural intergenerational learning through cattle as a medium of cultural expression in Mpembeni, KwaZulu-Natal
- Authors: Masuku, Lynette Sibongile
- Date: 2019
- Subjects: Postcolonialism , Environmental education -- South Africa , Community education -- South Africa -- KwaZulu Natal , Non-formal education -- South Africa -- KwaZulu Natal , Agricultural education -- South Africa -- KwaZulu Natal , Livestock -- Handling -- South Africa -- KwaZulu Natal , Cattle -- Handling -- South Africa -- KwaZulu Natal , Cattle herding -- South Africa -- KwaZulu Natal , Life skills -- Study and teaching -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/68181 , vital:29213
- Description: This case study was conducted in a small rural community called Mpembeni, in South Africa’s KwaZulu-Natal province. It was motivated by my observation of high levels of competence in ‘cattle knowledge’ amongst children coupled with a simultaneous failure at school. I view schools as integral parts of the community and consider them as being influenced by the community, which they in turn influence. This study set out to understand that which embodied informal learning in home/pasture-based contexts as well as formalised learning processes in schools. I used Sociocultural theory as the most congruent of educational theories to surface and illuminate the intergenerational learning processes that were taking place in the area. This warranted my use of research investigation methods that could, in non-intrusive ways, expose the everyday community practices that related to cattle as a particular medium of cultural expression. Ethnography, sourced from anthropology, aided by ethnomethods, was not only compatible with my study and the way in which I wanted to write out the research report, but also with my educational theory and its counterhegemonic intents. To understand the colonialities that framed the discord that embodied home and school as learning contexts, I used postcolonial theory, not only as a lens but as a counterhegemonic response. This theory also informed my research methodology as well as afforded me the reflexivity tools for an examination of my own intergenerational learning and the relational identities of myself as ‘Other’ in the lives of the research participants. It further facilitated the exploration of the potential for potential hybrid third spaces within the bubbling meeting nodes of the socio-cultural context of school and home/pasture based settings of learning. I observed cattle herding related practices, interviewed children, their parents and/or carers, dipping tank managers, livestock inspectors, community elders and members. I also analysed some of the written and unwritten content that made up the formal and informal based learning processes and reviewed some of the most recent South African Curriculum Statements and related texts on the representations of cattle. I sought views from teachers on their interactions with the people of Mpembeni, whose children they taught. I also explored axes of tension, silences and presences on anything related to cattle in schools. I argue and make a case for the development of thought by African scholars to advance Africa’s education rather than aid mimicry and the importation of theories of little congruence and relevance to the African context and Africa’s future. The study has made some contributions to new knowledge. This is in its exploration of sociocultural intergenerational methods and techniques that are employed for learning in community contexts, highlighting the importance of surfacing and understanding of children’s knowledge and experiences. The study has gone further to deliberate the in-betweenness of school and home learning environments, highlighting and unsilencing silenced, peripherised, new, old, considered irrelevant in the past, context and time congruent and liberatory knowledges. I propose that the knowledges located in these cleavages of difference be utilised to transform and create learning bridges between home and school environments. I propose that those ways of knowing that see others as nothings, be exposed and unlearned. Methods of learning that naturally unfold at home could be replicated at school with a recognition of the intergenerational methods, techniques, practices and the learning values in a critically constructive manner that narrows difference and othering.
- Full Text:
- Date Issued: 2019
- Authors: Masuku, Lynette Sibongile
- Date: 2019
- Subjects: Postcolonialism , Environmental education -- South Africa , Community education -- South Africa -- KwaZulu Natal , Non-formal education -- South Africa -- KwaZulu Natal , Agricultural education -- South Africa -- KwaZulu Natal , Livestock -- Handling -- South Africa -- KwaZulu Natal , Cattle -- Handling -- South Africa -- KwaZulu Natal , Cattle herding -- South Africa -- KwaZulu Natal , Life skills -- Study and teaching -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/68181 , vital:29213
- Description: This case study was conducted in a small rural community called Mpembeni, in South Africa’s KwaZulu-Natal province. It was motivated by my observation of high levels of competence in ‘cattle knowledge’ amongst children coupled with a simultaneous failure at school. I view schools as integral parts of the community and consider them as being influenced by the community, which they in turn influence. This study set out to understand that which embodied informal learning in home/pasture-based contexts as well as formalised learning processes in schools. I used Sociocultural theory as the most congruent of educational theories to surface and illuminate the intergenerational learning processes that were taking place in the area. This warranted my use of research investigation methods that could, in non-intrusive ways, expose the everyday community practices that related to cattle as a particular medium of cultural expression. Ethnography, sourced from anthropology, aided by ethnomethods, was not only compatible with my study and the way in which I wanted to write out the research report, but also with my educational theory and its counterhegemonic intents. To understand the colonialities that framed the discord that embodied home and school as learning contexts, I used postcolonial theory, not only as a lens but as a counterhegemonic response. This theory also informed my research methodology as well as afforded me the reflexivity tools for an examination of my own intergenerational learning and the relational identities of myself as ‘Other’ in the lives of the research participants. It further facilitated the exploration of the potential for potential hybrid third spaces within the bubbling meeting nodes of the socio-cultural context of school and home/pasture based settings of learning. I observed cattle herding related practices, interviewed children, their parents and/or carers, dipping tank managers, livestock inspectors, community elders and members. I also analysed some of the written and unwritten content that made up the formal and informal based learning processes and reviewed some of the most recent South African Curriculum Statements and related texts on the representations of cattle. I sought views from teachers on their interactions with the people of Mpembeni, whose children they taught. I also explored axes of tension, silences and presences on anything related to cattle in schools. I argue and make a case for the development of thought by African scholars to advance Africa’s education rather than aid mimicry and the importation of theories of little congruence and relevance to the African context and Africa’s future. The study has made some contributions to new knowledge. This is in its exploration of sociocultural intergenerational methods and techniques that are employed for learning in community contexts, highlighting the importance of surfacing and understanding of children’s knowledge and experiences. The study has gone further to deliberate the in-betweenness of school and home learning environments, highlighting and unsilencing silenced, peripherised, new, old, considered irrelevant in the past, context and time congruent and liberatory knowledges. I propose that the knowledges located in these cleavages of difference be utilised to transform and create learning bridges between home and school environments. I propose that those ways of knowing that see others as nothings, be exposed and unlearned. Methods of learning that naturally unfold at home could be replicated at school with a recognition of the intergenerational methods, techniques, practices and the learning values in a critically constructive manner that narrows difference and othering.
- Full Text:
- Date Issued: 2019
The relationship between health and safety and human risk taking behaviour in the South African electrical construction industry
- Authors: Du Toit, Willem Johannes
- Date: 2012
- Subjects: Electrical engineering -- Safety measures , Electrical engineering -- Health aspects , Construction industry -- Safety measures , Psychology, industrial
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9691 , http://hdl.handle.net/10948/d1009529 , Electrical engineering -- Safety measures , Electrical engineering -- Health aspects , Construction industry -- Safety measures , Psychology, industrial
- Description: Mankind, and the development of people, exists due to risk-taking behaviour. It is not that humans should not take risks, but rather the ability to identify the magnitude of risk exposure in order that mankind‟s actions would be so selected as to mitigate exposed risk factors, that no harm should befall them. The approach to health and safety (H&S) has always been to manage H&S environmental factors that could have a negative impact on people, capital, and organisational systems. However, the critical component of human risk-taking behaviour that would have a far greater impact has rarely been acknowledged as part of the drivers that increase risk exposure. Human behaviour is a major contributing factor in accident causation. Although human error cannot be completely eliminated, it should be identified and correctly managed according to each individual‟s risk-taking profile. The reason people decide to take certain risks under certain conditions and the effect it has on H&S management systems is a key component to managing organisational risk exposure. To quantify the value of individual risk-taking behaviour could provide management with better opportunities of lowering the organisational risk exposure. Human risk-taking behaviour is influenced by each individual‟s perception of risk. Such perception of risk will influence decisions on risk-taking behaviour, which in turn is influenced by the individual‟s psychological profile and environmental factors, including character and the impact of a cultural environment. The electrical construction and maintenance industry differs from other similar industries in that the physical entity of electricity requires not only sensory perception for the identification and evaluation of risk factors, but also requires specialised knowledge and testing equipment to evaluate the parameters of electrical installation, plant or equipment. Without such competence, direct exposure to most electrical installations could be fatal. The optimum human resource (HR) solution for managing the risk potential of high risk-taking behaviour is the rating and allocation of specific job tasks that can match and limit the individual potential for risk-taking behaviour and the impact on organisational incident statistics. Maintaining and optimising employee job performance enables organisations to better achieve pre-set goals and missions. Such improvements being a catalyst for better job performance by setting limitations on high risk-taking behaviour, that will improve H&S performance by lowering incident rates.
- Full Text:
- Date Issued: 2012
- Authors: Du Toit, Willem Johannes
- Date: 2012
- Subjects: Electrical engineering -- Safety measures , Electrical engineering -- Health aspects , Construction industry -- Safety measures , Psychology, industrial
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:9691 , http://hdl.handle.net/10948/d1009529 , Electrical engineering -- Safety measures , Electrical engineering -- Health aspects , Construction industry -- Safety measures , Psychology, industrial
- Description: Mankind, and the development of people, exists due to risk-taking behaviour. It is not that humans should not take risks, but rather the ability to identify the magnitude of risk exposure in order that mankind‟s actions would be so selected as to mitigate exposed risk factors, that no harm should befall them. The approach to health and safety (H&S) has always been to manage H&S environmental factors that could have a negative impact on people, capital, and organisational systems. However, the critical component of human risk-taking behaviour that would have a far greater impact has rarely been acknowledged as part of the drivers that increase risk exposure. Human behaviour is a major contributing factor in accident causation. Although human error cannot be completely eliminated, it should be identified and correctly managed according to each individual‟s risk-taking profile. The reason people decide to take certain risks under certain conditions and the effect it has on H&S management systems is a key component to managing organisational risk exposure. To quantify the value of individual risk-taking behaviour could provide management with better opportunities of lowering the organisational risk exposure. Human risk-taking behaviour is influenced by each individual‟s perception of risk. Such perception of risk will influence decisions on risk-taking behaviour, which in turn is influenced by the individual‟s psychological profile and environmental factors, including character and the impact of a cultural environment. The electrical construction and maintenance industry differs from other similar industries in that the physical entity of electricity requires not only sensory perception for the identification and evaluation of risk factors, but also requires specialised knowledge and testing equipment to evaluate the parameters of electrical installation, plant or equipment. Without such competence, direct exposure to most electrical installations could be fatal. The optimum human resource (HR) solution for managing the risk potential of high risk-taking behaviour is the rating and allocation of specific job tasks that can match and limit the individual potential for risk-taking behaviour and the impact on organisational incident statistics. Maintaining and optimising employee job performance enables organisations to better achieve pre-set goals and missions. Such improvements being a catalyst for better job performance by setting limitations on high risk-taking behaviour, that will improve H&S performance by lowering incident rates.
- Full Text:
- Date Issued: 2012
Ionospheric studies of the solar eclipse 25 December, 1954
- Authors: McElhinny, M W
- Date: 1959
- Subjects: Ionosphere -- Research , Solar eclipses
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5531 , http://hdl.handle.net/10962/d1012869
- Description: Since the Kennelly- Heaviside hypothesis in 1902 of the existence of a partially conducting layer in the upper atmosphere was proved to be true by the experiments of APPLETON and BARNETT (1925) and BREIT and TUVE (1926), this region has become known as the ionosphere. The ionosphere was soon discovered to consist of, not one but several layers (Fig. 1) (i) A layer at a height of just over 100 km. called the E layer. (ii) A layer at a height of approximately 300km. called the F₂ layer. (iii) A layer at a height of approximately 200 km. called the F₁ layer; this layer differs from the other two in that it is only present during the day time in Summer. (iv) Occasional intense reflections from a height of about 100 km. are found - these cannot be attributed to the normal E layer and have received the name "Sporadic E". The presence of two E layers (E₁ and E₂) has been suggested by HALLIDAY (1936) and BEST and RATCLIFFE (l978) but until recently most workers still seem to attribute these reflections to Sporadic E. Recent measurement by rockets of the electron density at E layer heights still do not confirm whether such bifurcation exists in the E region. The diurnal and seasonal variations of the first three layers indicate that the sun is the chief agent in their production. It is generally agreed that these layers consist of ionised molecules or atoms and free electrons produced by radiation from the sun. The origin of Sporadic E ionisation is still obscure, but it is thought that these sudden increases in ionisation which occur in E layer heights are due to passing meteors. Recently it has also been suggested by SEDDON, PICKAR and JACKSON (1954) from rocket measurements that Sporadic E might be due to a steep electron density gradient above the B layer.
- Full Text:
- Date Issued: 1959
- Authors: McElhinny, M W
- Date: 1959
- Subjects: Ionosphere -- Research , Solar eclipses
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5531 , http://hdl.handle.net/10962/d1012869
- Description: Since the Kennelly- Heaviside hypothesis in 1902 of the existence of a partially conducting layer in the upper atmosphere was proved to be true by the experiments of APPLETON and BARNETT (1925) and BREIT and TUVE (1926), this region has become known as the ionosphere. The ionosphere was soon discovered to consist of, not one but several layers (Fig. 1) (i) A layer at a height of just over 100 km. called the E layer. (ii) A layer at a height of approximately 300km. called the F₂ layer. (iii) A layer at a height of approximately 200 km. called the F₁ layer; this layer differs from the other two in that it is only present during the day time in Summer. (iv) Occasional intense reflections from a height of about 100 km. are found - these cannot be attributed to the normal E layer and have received the name "Sporadic E". The presence of two E layers (E₁ and E₂) has been suggested by HALLIDAY (1936) and BEST and RATCLIFFE (l978) but until recently most workers still seem to attribute these reflections to Sporadic E. Recent measurement by rockets of the electron density at E layer heights still do not confirm whether such bifurcation exists in the E region. The diurnal and seasonal variations of the first three layers indicate that the sun is the chief agent in their production. It is generally agreed that these layers consist of ionised molecules or atoms and free electrons produced by radiation from the sun. The origin of Sporadic E ionisation is still obscure, but it is thought that these sudden increases in ionisation which occur in E layer heights are due to passing meteors. Recently it has also been suggested by SEDDON, PICKAR and JACKSON (1954) from rocket measurements that Sporadic E might be due to a steep electron density gradient above the B layer.
- Full Text:
- Date Issued: 1959
Evaluation of Gratiana spadicea (Klug, 1829) and Metriona elatior (Klug, 1829) (Chrysomelidae: Cassidinae) for the biological control of sticky nightshade Solanum sisymbriifolium Lamarck (Solanaceae) in South Africa
- Authors: Hill, Martin Patrick
- Date: 1995
- Subjects: Solanaceae Solanaceae -- Biological control Chrysomelidae Chrysomelidae -- Host plants Cassida
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5688 , http://hdl.handle.net/10962/d1005374 , https://orcid.org/0000-0003-0579-5298
- Description: Solanum sisymbriifolium (sticky nightshade) is a shrubby weed of South American origin that was introduced to South Africa at the turn of the century. Despite being indicative of disturbed habitats, the weed was found to be invasive in conservation, agricultural recreational and suburban areas; this, coupled with the failure of both chemical and mechanical control attempts suggested that the weed was a good candidate for biological control. A biological control programme which followed a standard protocol was initiated. Observations suggested that S. sisymbriifolium dispersed primarily by seeds. Plants produced large quantities of fleshy fruit, favoured by frugivorous birds, which facilitated the rapid spread of the weed into new habitats. The seeds germinated quickly, especially in disturbed soil, often below the parent plant where they dropped from burst fruit, and along fences where birds roost. The pre-introductory survey of the weed revealed that S. sisymbriifolium was attacked by a relatively small number of, mainly polyphagous, herbivorous insects. These were localised and sporadic in incidence and inflicted very little observable damage. The herbivore fauna of S. sisymbriifolium was depauperate even in relation to two other exotic weeds, S. elaeaglllfolium and S. mauritianum, in South Africa. The paucity of native herbivores on S. sisymbriifoliwn was ascribed to a combination of the weed's taxonomic distinctness from South African Solanum species, and the dense covering of glandular trichomes on its leaves. Although it was shown that the exudate produced by these glandular trichomes of S. sisymbriifolium seriously impeded the movement and feeding of native herbivores, there was not enough evidence to suggest that the glandular trichomes, alone could have been responsible the lack of herbivores on the weed. Two leaf-feeding Cassidinae Gratiana spadicea and MetJ-iona elatior were screened as agents for the biological control of S. sisymbrilfolium. Favourable biological characteristics for both species included a high rate of increase, long-lived adults, many generations per year, and a high per capita feeding rate. Host range was investigated in larval survival tests and adult choice tests. The larvae of both species were reared through to the adult stage on several of the native Solanum species tested, and also on eggplant (S. melongena). However, the survival of G. spadicea on the majority of these species was very low, suggesting that the beetles would be unlikely to attack them in the field. This was supported by the adult choice tests, where G. spadicea females displayed, a strong oviposition preference for their natural host. In contrast, M. elatior larvae survived well on non-host plants, and the females selected several non-host species, including eggplant for oviposition. It was argued that the conflict of interests involving eggplant was overrated because eggplant is subjected to a stringent insecticide spray regime. Based on this evidence, permission for release was granted for G. spadicea. The impact of native parasitoid host range extensions to weed biological control agents in South Africa was investigated. Native parasitoids were recorded from nearly half of the agent species that had established on their target weed. The level of concealment and taxon influenced susceptibility of the agents to parasitoid attack Poorly concealed endophagous agents were most susceptible to attack, while exposed feeders were fairly free from attack. However, native parasitoids were reported not to strongly influence weed biological control agent populations and it was concluded that no agent should be rejected based only on its susceptibility to native parasitoid attack. Finally, several predictions are made as to the potential success of G. spadicea on S. sisymbriifolium and some of the challenges facing the biological control of weeds are discussed.
- Full Text:
- Date Issued: 1995
- Authors: Hill, Martin Patrick
- Date: 1995
- Subjects: Solanaceae Solanaceae -- Biological control Chrysomelidae Chrysomelidae -- Host plants Cassida
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5688 , http://hdl.handle.net/10962/d1005374 , https://orcid.org/0000-0003-0579-5298
- Description: Solanum sisymbriifolium (sticky nightshade) is a shrubby weed of South American origin that was introduced to South Africa at the turn of the century. Despite being indicative of disturbed habitats, the weed was found to be invasive in conservation, agricultural recreational and suburban areas; this, coupled with the failure of both chemical and mechanical control attempts suggested that the weed was a good candidate for biological control. A biological control programme which followed a standard protocol was initiated. Observations suggested that S. sisymbriifolium dispersed primarily by seeds. Plants produced large quantities of fleshy fruit, favoured by frugivorous birds, which facilitated the rapid spread of the weed into new habitats. The seeds germinated quickly, especially in disturbed soil, often below the parent plant where they dropped from burst fruit, and along fences where birds roost. The pre-introductory survey of the weed revealed that S. sisymbriifolium was attacked by a relatively small number of, mainly polyphagous, herbivorous insects. These were localised and sporadic in incidence and inflicted very little observable damage. The herbivore fauna of S. sisymbriifolium was depauperate even in relation to two other exotic weeds, S. elaeaglllfolium and S. mauritianum, in South Africa. The paucity of native herbivores on S. sisymbriifoliwn was ascribed to a combination of the weed's taxonomic distinctness from South African Solanum species, and the dense covering of glandular trichomes on its leaves. Although it was shown that the exudate produced by these glandular trichomes of S. sisymbriifolium seriously impeded the movement and feeding of native herbivores, there was not enough evidence to suggest that the glandular trichomes, alone could have been responsible the lack of herbivores on the weed. Two leaf-feeding Cassidinae Gratiana spadicea and MetJ-iona elatior were screened as agents for the biological control of S. sisymbrilfolium. Favourable biological characteristics for both species included a high rate of increase, long-lived adults, many generations per year, and a high per capita feeding rate. Host range was investigated in larval survival tests and adult choice tests. The larvae of both species were reared through to the adult stage on several of the native Solanum species tested, and also on eggplant (S. melongena). However, the survival of G. spadicea on the majority of these species was very low, suggesting that the beetles would be unlikely to attack them in the field. This was supported by the adult choice tests, where G. spadicea females displayed, a strong oviposition preference for their natural host. In contrast, M. elatior larvae survived well on non-host plants, and the females selected several non-host species, including eggplant for oviposition. It was argued that the conflict of interests involving eggplant was overrated because eggplant is subjected to a stringent insecticide spray regime. Based on this evidence, permission for release was granted for G. spadicea. The impact of native parasitoid host range extensions to weed biological control agents in South Africa was investigated. Native parasitoids were recorded from nearly half of the agent species that had established on their target weed. The level of concealment and taxon influenced susceptibility of the agents to parasitoid attack Poorly concealed endophagous agents were most susceptible to attack, while exposed feeders were fairly free from attack. However, native parasitoids were reported not to strongly influence weed biological control agent populations and it was concluded that no agent should be rejected based only on its susceptibility to native parasitoid attack. Finally, several predictions are made as to the potential success of G. spadicea on S. sisymbriifolium and some of the challenges facing the biological control of weeds are discussed.
- Full Text:
- Date Issued: 1995
Synthetic and bioactivity studies of antiplasmodial and antibacterial marine natural products
- Authors: Young, Ryan Mark
- Date: 2012
- Subjects: Antibacterial agents Marine natural products Marine pharmacology Plasmodium falciparum Staphylococcus aureus Isocyanides Imidazoles
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4378 , http://hdl.handle.net/10962/d1005043
- Description: This thesis is divided into two parts, assessing marine and synthetic compounds active firstly against Plasmodium falciparum (Chapter 3 and 4) and secondly active against methicillin resistant Staphylococcus aureus (MRSA, Chapter 5). In Chapter 3 the synthesis of nine new tricyclic podocarpanes (3.203-3.207 and 3.209-3.212) from the diterpene (+)-manool is described. Initial SAR study of synthetic podocarpanes concluded that the most active compound was a C-13 phenyl substituted podocarpane (3.204, IC₅₀ 6.6 μM). By preparing analogues with varying halogenated substituents on the phenyl ring (3.209-3.212) the antiplasmodial activity was improved (IC₅₀ 1.4 μM), while simultaneously decreasing the haemolysis previously reported for this class of compounds. Inspired by the antiplasmodial activity of Wright and Wattanapiromsakul’s tricycle marine isonitriles (2.16-2.21 and 2.24-2.27) an unsuccessfully attempt was made to convert tertiary alcohol moieties to isonitrile functionalities in compounds 3.188, 3.204-3.207 and 3.209-3.212. Over a decade ago Wright et al. proposed a putative antiplasmodial mechanism of action for marine isonitriles (2.4, 2.9, 2.15, 2.19 and 2.35) and isothiocyanate (2.34) which involved interference in haem detoxification by P. falciparum thus inhibiting the growth of the parasite. In Chapter 4 we describe how we successfully managed to scale down Egan’s β-haematin inhibition assay for the analyses of small quantities of marine natural products as potential β-haematin inhibitors. Our modified assay revealed that the most active antiplasmodial marine isonitrile 2.9 (IC₅₀ 13 nM) showed total β-haematin inhibition while 2.15 (IC₅₀ 81 nM) and 2.19 (IC₅₀ 31 nM) showed partial inhibition at three equivalents relative to haem. Using contempary molecular modelling techniques the charge on the isonitrile functionality was more accurately describe and the modified charge data sets was used to explore docking of marine isonitriles to haem using AutoDock. In Chapter 5 we describe how a lead South African marine bisindole MRSA pyruvate kinase inhibitor (5.8) was discovered in collaboration with colleagues at the University of British Columbia (UBC) and how this discovery inspired us to design a synthetic route to the dibrominated bisindole, isobromotopsentin (5.20) in an attempt to increase the bioactivity displayed by 5.8. We devised a fast and high yielding synthetic route using microwave assited organic synthesis. We first tested this synthesis using simple aryl glyoxals (5.27-5.32) as precursors to synthesize biphenylimidazoles (5.21-5.26), which later allowed us to synthesize the ascidian natural product 5.111. This method was sucessfully extended to the synthesis of deoxytopsentin (5.33) from an N-Boc protected indole methyl ketone (5.89). We subsequently were able to effectively remove the carbamate protection via thermal decomposition by heating the protected bisindole imidazole (5.90) in a microwave reactor for 5 min under argon. The synthesis of 5.20 resulted in an inseparable mixture of monoprotected and totally deprotected topsentin products, and due to time constraints we were not able to optimise this synthesis. Nonetheless our synthesis of the marine natural product 5.33 which was faster and higher yielding than previously reported routes could be extended to the synthesis of other topsentin bisindoles (5.138-5.140). Work towards this goal continues in our laboratory.
- Full Text:
- Date Issued: 2012
- Authors: Young, Ryan Mark
- Date: 2012
- Subjects: Antibacterial agents Marine natural products Marine pharmacology Plasmodium falciparum Staphylococcus aureus Isocyanides Imidazoles
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4378 , http://hdl.handle.net/10962/d1005043
- Description: This thesis is divided into two parts, assessing marine and synthetic compounds active firstly against Plasmodium falciparum (Chapter 3 and 4) and secondly active against methicillin resistant Staphylococcus aureus (MRSA, Chapter 5). In Chapter 3 the synthesis of nine new tricyclic podocarpanes (3.203-3.207 and 3.209-3.212) from the diterpene (+)-manool is described. Initial SAR study of synthetic podocarpanes concluded that the most active compound was a C-13 phenyl substituted podocarpane (3.204, IC₅₀ 6.6 μM). By preparing analogues with varying halogenated substituents on the phenyl ring (3.209-3.212) the antiplasmodial activity was improved (IC₅₀ 1.4 μM), while simultaneously decreasing the haemolysis previously reported for this class of compounds. Inspired by the antiplasmodial activity of Wright and Wattanapiromsakul’s tricycle marine isonitriles (2.16-2.21 and 2.24-2.27) an unsuccessfully attempt was made to convert tertiary alcohol moieties to isonitrile functionalities in compounds 3.188, 3.204-3.207 and 3.209-3.212. Over a decade ago Wright et al. proposed a putative antiplasmodial mechanism of action for marine isonitriles (2.4, 2.9, 2.15, 2.19 and 2.35) and isothiocyanate (2.34) which involved interference in haem detoxification by P. falciparum thus inhibiting the growth of the parasite. In Chapter 4 we describe how we successfully managed to scale down Egan’s β-haematin inhibition assay for the analyses of small quantities of marine natural products as potential β-haematin inhibitors. Our modified assay revealed that the most active antiplasmodial marine isonitrile 2.9 (IC₅₀ 13 nM) showed total β-haematin inhibition while 2.15 (IC₅₀ 81 nM) and 2.19 (IC₅₀ 31 nM) showed partial inhibition at three equivalents relative to haem. Using contempary molecular modelling techniques the charge on the isonitrile functionality was more accurately describe and the modified charge data sets was used to explore docking of marine isonitriles to haem using AutoDock. In Chapter 5 we describe how a lead South African marine bisindole MRSA pyruvate kinase inhibitor (5.8) was discovered in collaboration with colleagues at the University of British Columbia (UBC) and how this discovery inspired us to design a synthetic route to the dibrominated bisindole, isobromotopsentin (5.20) in an attempt to increase the bioactivity displayed by 5.8. We devised a fast and high yielding synthetic route using microwave assited organic synthesis. We first tested this synthesis using simple aryl glyoxals (5.27-5.32) as precursors to synthesize biphenylimidazoles (5.21-5.26), which later allowed us to synthesize the ascidian natural product 5.111. This method was sucessfully extended to the synthesis of deoxytopsentin (5.33) from an N-Boc protected indole methyl ketone (5.89). We subsequently were able to effectively remove the carbamate protection via thermal decomposition by heating the protected bisindole imidazole (5.90) in a microwave reactor for 5 min under argon. The synthesis of 5.20 resulted in an inseparable mixture of monoprotected and totally deprotected topsentin products, and due to time constraints we were not able to optimise this synthesis. Nonetheless our synthesis of the marine natural product 5.33 which was faster and higher yielding than previously reported routes could be extended to the synthesis of other topsentin bisindoles (5.138-5.140). Work towards this goal continues in our laboratory.
- Full Text:
- Date Issued: 2012
Towards development of a malaria diagnostic: Generation, screening and validation of novel aptamers recognising Plasmodium falciparum lactate dehydrogenase
- Authors: Frith, Kelly-Anne
- Date: 2020
- Subjects: Plasmodium falciparum , Malaria -- Chemotherapy , Oligonucleotides , Lactate dehydrogenase , Biochemical markers , Systematic evolution of ligands through exponential enrichment (SELEX)
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/142247 , vital:38062
- Description: Malaria, caused by infection with the Plasmodium parasite, is one of the leading causes of death in under-developed countries. Early detection is crucial for the effective treatment of malaria, particularly in cases where infection is due to Plasmodium falciparum. There is, therefore, an enduring need for portable, sensitive, reliable, accurate, durable, self-validating and cost-effective techniques for the rapid detection of malaria. Moreover, there is a demand to distinguish between various infectious species causing malaria. Research in the area of malarial biomarkers has identified a unique, species-specific, epitope of P. falciparum lactate dehydrogenase (PfLDH), enhancing prospects for the development of diagnostics capable of identifying the species causing malarial infection. In recent years, improvements have been made towards the development of rapid diagnostic tests for detecting malarial biomarkers. Owing to their low cost, ease of labeling, and high thermal stability (relative to antibodies), the development and synthesis of aptamers that target the malarial lactate dehydrogenase represents one of the key innovations in the field of rapid diagnostics for malaria. This study explored the generation of aptamers that specifically target P. falciparum. Two sets of aptamers with diagnostically-supportive functions were generated independently, through parallel SELEX of recombinantly-expressed, full-length Plasmodium falciparum lactate dehydrogenase (rPfLDH), and an oligopeptide comprising the P. falciparum-specific epitope on lactate dehydrogenase (LDHp). The latter offers a promising solution for generating aptamers capable of binding with high specificity to P. falciparum. In this work, an rLDH class of aptamers was generated when SELEX was performed using the full-length rPfLDH protein as the target and the LDHp class of aptamers was generated when SELEX was performed using the oligopeptide LDHp as a target. Aptamers were successfully generated through the process of SELEX (systematic evolution of ligands through exponential enrichment) following the study and application of several optimisation steps, particularly during the amplification stage of SELEX. Optimisation steps included the study of improvements in PCR conditions; role of surfactants (Triton-X), modifying the PCR clean-up protocol; and agarose gel excision. Structurally-relevant moieties with particular consensus sequences (GGTAG and GGCG) were found in aptamers both reported here and previously published, confirming their importance in recognition of the target. Novel moieties particular to this work (ATTAT and poly-A stretches) were identified. Clades of consensus sequences were identified in both the rLDH and LDHp groups of aptamers, where sequences in the rLDH clade did not show preferential binding to rPfLDH while those in the LDHp clade (particularly LDHp 3 and 18) were able to recognise and bind only LDHp. Of the 19 sequences returned from the parallel SELEX procedures for rPfLDH (11 sequences) and LDHp (8 sequences), six rPfLDH and all eight LDHp sequences underwent preliminary screening and those with low responses eliminated. Of the eight LDHp-targeting aptamer sequences, five were preliminarily shown to bind to LDHp, whereas only two rPfLDH-targeting sequences were shown to bind to the target (rLDH 4 and 7). To this small selection of rPfLDH oligonucleotide sequences, two more (rLDH 1 and 15) were chosen for further study based on their sequences, secondary and predicted tertiary conformations. Sequences chosen for further study were therefore: rLDH 1, 4, 7 and 15 in the rLDH class, and LDHp 1, 3, 11, 14 and 18 in the LDHp class. Binding properties of the aptamers towards their targets were investigated using enzyme-linked oligonucleotide assays (ELONA), fluorophore-linked oligonucleotide assays (FLONA), electromobility shift assays (EMSA), surface plasmon resonance (SPR), and GelRed dissociation assays, while applications towards aptasensors were explored using electrochemical impedance spectroscopy (EIS) and fluorescent microscopy. Some inconsistencies were seen for specific aptamer to target binding interactions using specific techniques; however, generally, binding to the targets was observed across the techniques assessed. These varied responses demonstrate the need to screen and validate aptamers using a variety of techniques and platforms not necessarily specific for the proposed application. From the aptamer binding screening studies using ELONA, the most promising aptamers generated were identified as LDHp 11, rLDH 4, rLDH 7 and rLDH 15. Aptamer rLDH 4, which was generated against rPfLDH, exhibited preferential and specific binding to the lactate dehydrogenase from P. falciparum, over the recombinantly-expressed lactate dehydrogenase from Plasmodium vivax (rPvLDH), albeit with lowered responses compared to LDHp 11 in ELONA and EMSA studies. However, in kinetic ELONA studies rLDH 4 showed binding to both rPfLDH and rPvLDH. Aptamer rLDH 7 showed high affinity for rPfLDH and rPvLDH in kinetic studies using ELONA. However, screening studies with ELONA indicates that aptamer rLDH 7 may not be suitable for diagnostic tests in serum samples given its non-specific binding to human serum albumin (HSA). Aptamer rLDH 15 exhibited species specificity for rPfLDH in screening studies using ELONA but showed affinity towards rPvLDH (albeit lower relative to its affinity for rPfLDH) in kinetic studies using ELONA. LDHp 11, generated against the PfLDH peptide, showed a clear preference for rPfLDH when compared to rPvLDH and other control proteins, in both sets of ELONA studies conducted, as well as EMSA, thus possessing a strong ability to identify the presence of Plasmodium falciparum owing to its generation against the species-specific epitope. While LDHp 1 demonstrated binding to plasmodial LDH in a flow-through system (SPR), so reiterating ELONA responses, it did not perform well in the remaining methodologies. Aptamers rLDH 1 and 15 and LDHp 3, 14 and 18 exhibited a mixed set of results throughout the target protein screening analyses and were, thus, not considered for selective binding in P. falciparum parasite bodies. In studies aimed at exploring biosensor assemblies utilising the developed aptamers, both rLDH 4 and LDHp 11, along with rLDH 7, LDHp 1 and pL1, demonstrated in situ binding to the native PfLDH in fluorescent microscopy. LDHp 11 exhibited FITC-based fluorescence equivalent to the anti-rPfLDHp IgY antibody in confocal fluorescent microscopy indicating superior binding to the native PfLDH compared to the remaining aptamers. An examination of electrochemical impedance as a platform for a biosensor assembly did not, in these studies, exhibit the required sensitivity using physiologically relevant concentrations of analyte expected for pLDH following infection with Plasmodium spp. Malstat/LDH activity was explored for application in a colorimetric aptasensor. A decrease in both rPfLDH and rPvLDH activity was observed following incubation with the tested aptamers, but rLDH 1, rLDH 7 and LDHp 14 did not exhibit similar decreases in rPvLDH activity. Aptamers rLDH 1, 4 and 7 and LDHp 11 and 14 were, therefore, not selected as candidates for LDH capture in LDH activity-based diagnostic devices for P. falciparum. The decreases in pLDH activity in the presence of aptamers could hold promise as direct or antagonistic malaria therapeutic agents. Preliminary studies on the application of aptamers as malaria therapeutic agents, while of interest, should be viewed with due caution given the challenges of aptamers reaching the intracellular native plasmodial LDH hosted within the red blood cells. In conclusion, this work has shown the ability of the LDHp 11 aptamer, generated in these studies, to selectively bind rPfLDH over rPvLDH, and to bind to the native PfLDH in fluorescent microscopy, indicating that this aptamer holds promise as a biorecognition element in malaria biosensors and other diagnostic devices for the detection, and differentiation, of P. falciparum and P. vivax. The use of a species-specific epitope of P. falciparum as a target in aptamer generation paves the way for similar such studies aimed at generating aptamers with species selectivity for other Plasmodium species.
- Full Text:
- Date Issued: 2020
- Authors: Frith, Kelly-Anne
- Date: 2020
- Subjects: Plasmodium falciparum , Malaria -- Chemotherapy , Oligonucleotides , Lactate dehydrogenase , Biochemical markers , Systematic evolution of ligands through exponential enrichment (SELEX)
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/142247 , vital:38062
- Description: Malaria, caused by infection with the Plasmodium parasite, is one of the leading causes of death in under-developed countries. Early detection is crucial for the effective treatment of malaria, particularly in cases where infection is due to Plasmodium falciparum. There is, therefore, an enduring need for portable, sensitive, reliable, accurate, durable, self-validating and cost-effective techniques for the rapid detection of malaria. Moreover, there is a demand to distinguish between various infectious species causing malaria. Research in the area of malarial biomarkers has identified a unique, species-specific, epitope of P. falciparum lactate dehydrogenase (PfLDH), enhancing prospects for the development of diagnostics capable of identifying the species causing malarial infection. In recent years, improvements have been made towards the development of rapid diagnostic tests for detecting malarial biomarkers. Owing to their low cost, ease of labeling, and high thermal stability (relative to antibodies), the development and synthesis of aptamers that target the malarial lactate dehydrogenase represents one of the key innovations in the field of rapid diagnostics for malaria. This study explored the generation of aptamers that specifically target P. falciparum. Two sets of aptamers with diagnostically-supportive functions were generated independently, through parallel SELEX of recombinantly-expressed, full-length Plasmodium falciparum lactate dehydrogenase (rPfLDH), and an oligopeptide comprising the P. falciparum-specific epitope on lactate dehydrogenase (LDHp). The latter offers a promising solution for generating aptamers capable of binding with high specificity to P. falciparum. In this work, an rLDH class of aptamers was generated when SELEX was performed using the full-length rPfLDH protein as the target and the LDHp class of aptamers was generated when SELEX was performed using the oligopeptide LDHp as a target. Aptamers were successfully generated through the process of SELEX (systematic evolution of ligands through exponential enrichment) following the study and application of several optimisation steps, particularly during the amplification stage of SELEX. Optimisation steps included the study of improvements in PCR conditions; role of surfactants (Triton-X), modifying the PCR clean-up protocol; and agarose gel excision. Structurally-relevant moieties with particular consensus sequences (GGTAG and GGCG) were found in aptamers both reported here and previously published, confirming their importance in recognition of the target. Novel moieties particular to this work (ATTAT and poly-A stretches) were identified. Clades of consensus sequences were identified in both the rLDH and LDHp groups of aptamers, where sequences in the rLDH clade did not show preferential binding to rPfLDH while those in the LDHp clade (particularly LDHp 3 and 18) were able to recognise and bind only LDHp. Of the 19 sequences returned from the parallel SELEX procedures for rPfLDH (11 sequences) and LDHp (8 sequences), six rPfLDH and all eight LDHp sequences underwent preliminary screening and those with low responses eliminated. Of the eight LDHp-targeting aptamer sequences, five were preliminarily shown to bind to LDHp, whereas only two rPfLDH-targeting sequences were shown to bind to the target (rLDH 4 and 7). To this small selection of rPfLDH oligonucleotide sequences, two more (rLDH 1 and 15) were chosen for further study based on their sequences, secondary and predicted tertiary conformations. Sequences chosen for further study were therefore: rLDH 1, 4, 7 and 15 in the rLDH class, and LDHp 1, 3, 11, 14 and 18 in the LDHp class. Binding properties of the aptamers towards their targets were investigated using enzyme-linked oligonucleotide assays (ELONA), fluorophore-linked oligonucleotide assays (FLONA), electromobility shift assays (EMSA), surface plasmon resonance (SPR), and GelRed dissociation assays, while applications towards aptasensors were explored using electrochemical impedance spectroscopy (EIS) and fluorescent microscopy. Some inconsistencies were seen for specific aptamer to target binding interactions using specific techniques; however, generally, binding to the targets was observed across the techniques assessed. These varied responses demonstrate the need to screen and validate aptamers using a variety of techniques and platforms not necessarily specific for the proposed application. From the aptamer binding screening studies using ELONA, the most promising aptamers generated were identified as LDHp 11, rLDH 4, rLDH 7 and rLDH 15. Aptamer rLDH 4, which was generated against rPfLDH, exhibited preferential and specific binding to the lactate dehydrogenase from P. falciparum, over the recombinantly-expressed lactate dehydrogenase from Plasmodium vivax (rPvLDH), albeit with lowered responses compared to LDHp 11 in ELONA and EMSA studies. However, in kinetic ELONA studies rLDH 4 showed binding to both rPfLDH and rPvLDH. Aptamer rLDH 7 showed high affinity for rPfLDH and rPvLDH in kinetic studies using ELONA. However, screening studies with ELONA indicates that aptamer rLDH 7 may not be suitable for diagnostic tests in serum samples given its non-specific binding to human serum albumin (HSA). Aptamer rLDH 15 exhibited species specificity for rPfLDH in screening studies using ELONA but showed affinity towards rPvLDH (albeit lower relative to its affinity for rPfLDH) in kinetic studies using ELONA. LDHp 11, generated against the PfLDH peptide, showed a clear preference for rPfLDH when compared to rPvLDH and other control proteins, in both sets of ELONA studies conducted, as well as EMSA, thus possessing a strong ability to identify the presence of Plasmodium falciparum owing to its generation against the species-specific epitope. While LDHp 1 demonstrated binding to plasmodial LDH in a flow-through system (SPR), so reiterating ELONA responses, it did not perform well in the remaining methodologies. Aptamers rLDH 1 and 15 and LDHp 3, 14 and 18 exhibited a mixed set of results throughout the target protein screening analyses and were, thus, not considered for selective binding in P. falciparum parasite bodies. In studies aimed at exploring biosensor assemblies utilising the developed aptamers, both rLDH 4 and LDHp 11, along with rLDH 7, LDHp 1 and pL1, demonstrated in situ binding to the native PfLDH in fluorescent microscopy. LDHp 11 exhibited FITC-based fluorescence equivalent to the anti-rPfLDHp IgY antibody in confocal fluorescent microscopy indicating superior binding to the native PfLDH compared to the remaining aptamers. An examination of electrochemical impedance as a platform for a biosensor assembly did not, in these studies, exhibit the required sensitivity using physiologically relevant concentrations of analyte expected for pLDH following infection with Plasmodium spp. Malstat/LDH activity was explored for application in a colorimetric aptasensor. A decrease in both rPfLDH and rPvLDH activity was observed following incubation with the tested aptamers, but rLDH 1, rLDH 7 and LDHp 14 did not exhibit similar decreases in rPvLDH activity. Aptamers rLDH 1, 4 and 7 and LDHp 11 and 14 were, therefore, not selected as candidates for LDH capture in LDH activity-based diagnostic devices for P. falciparum. The decreases in pLDH activity in the presence of aptamers could hold promise as direct or antagonistic malaria therapeutic agents. Preliminary studies on the application of aptamers as malaria therapeutic agents, while of interest, should be viewed with due caution given the challenges of aptamers reaching the intracellular native plasmodial LDH hosted within the red blood cells. In conclusion, this work has shown the ability of the LDHp 11 aptamer, generated in these studies, to selectively bind rPfLDH over rPvLDH, and to bind to the native PfLDH in fluorescent microscopy, indicating that this aptamer holds promise as a biorecognition element in malaria biosensors and other diagnostic devices for the detection, and differentiation, of P. falciparum and P. vivax. The use of a species-specific epitope of P. falciparum as a target in aptamer generation paves the way for similar such studies aimed at generating aptamers with species selectivity for other Plasmodium species.
- Full Text:
- Date Issued: 2020