Solid-phase extraction based sample preparation for the determination of drug and organic pollutant residue
- Authors: Pule, Bellah Oreeditse
- Date: 2011 , 2011-02-08
- Subjects: Food contamination , Drugs -- Analysis , Pharmaceutical chemistry , Extraction (Chemistry) , Sorbents
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4406 , http://hdl.handle.net/10962/d1006711 , Food contamination , Drugs -- Analysis , Pharmaceutical chemistry , Extraction (Chemistry) , Sorbents
- Description: This thesis presents solid phase extraction (SPE) methodologies based on mixed-mode polymeric sorbents; a mixed mode strong anion exchanger (Agilent SampliQ SAX) and a mixed mode strong cation exchanger (Agilent SampliQ SCX). Furthermore, dispersive-SPE based on a quick, easy, cheap, effective, rugged and safe (QuEChERS) method was assessed for applicability in the determination of drug residues. The mixed-mode polymeric sorbents were evaluated for the simultaneous fractionation of drugs that exhibit diverse polarities with acidic, basic and neutral functionalities in biological matrices (plasma and urine). The polymeric skeleton of these sorbents entails an exchanger group and therefore provides two retention mechanisms, strong cation or anion exchange retention mechanisms with hydrophobic interactions. It was demonstrated that with a sequential elution protocol for sample clean-up analytes were fractionated into acidic, basic and neutral classes. The SAX was employed for analysis of ketoprofen, naproxen (acidic drugs), nortriptyline (basic) and secobarbital (neutral) from urine sample. The SCX was used for fractionating phenobarbital, p-toluamide (acidic), amphetamine, m-toluidine (basic) and acetaminophen (neutral drug) from plasma sample. QuEChERS method was employed for quantitative determination of 16 polycyclic aromatic hydrocarbons (PAHs) from fish fillets and soil; 9 sulfonamides (SAs) from chicken muscles and acrylamide (AA) in cooking oil. The analyte recoveries ranged from 79.6 - 109% with RSDs ranging from 0.06 - 1.9% at three different fortification levels. Good linearity (r2 > 0.9990) was attained for most analytes. The limits of detection and quantification ranged from 0.03 - 0.84 μg/ml and 0.81 - 1.89 μg/ml respectively for analytes in biological samples. LODs and LOQs for analytes in food and environmental samples ranged from 0.02 to 0.39 and 0.25 to 1.30 ng/g respectively.
- Full Text:
- Date Issued: 2011
- Authors: Pule, Bellah Oreeditse
- Date: 2011 , 2011-02-08
- Subjects: Food contamination , Drugs -- Analysis , Pharmaceutical chemistry , Extraction (Chemistry) , Sorbents
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4406 , http://hdl.handle.net/10962/d1006711 , Food contamination , Drugs -- Analysis , Pharmaceutical chemistry , Extraction (Chemistry) , Sorbents
- Description: This thesis presents solid phase extraction (SPE) methodologies based on mixed-mode polymeric sorbents; a mixed mode strong anion exchanger (Agilent SampliQ SAX) and a mixed mode strong cation exchanger (Agilent SampliQ SCX). Furthermore, dispersive-SPE based on a quick, easy, cheap, effective, rugged and safe (QuEChERS) method was assessed for applicability in the determination of drug residues. The mixed-mode polymeric sorbents were evaluated for the simultaneous fractionation of drugs that exhibit diverse polarities with acidic, basic and neutral functionalities in biological matrices (plasma and urine). The polymeric skeleton of these sorbents entails an exchanger group and therefore provides two retention mechanisms, strong cation or anion exchange retention mechanisms with hydrophobic interactions. It was demonstrated that with a sequential elution protocol for sample clean-up analytes were fractionated into acidic, basic and neutral classes. The SAX was employed for analysis of ketoprofen, naproxen (acidic drugs), nortriptyline (basic) and secobarbital (neutral) from urine sample. The SCX was used for fractionating phenobarbital, p-toluamide (acidic), amphetamine, m-toluidine (basic) and acetaminophen (neutral drug) from plasma sample. QuEChERS method was employed for quantitative determination of 16 polycyclic aromatic hydrocarbons (PAHs) from fish fillets and soil; 9 sulfonamides (SAs) from chicken muscles and acrylamide (AA) in cooking oil. The analyte recoveries ranged from 79.6 - 109% with RSDs ranging from 0.06 - 1.9% at three different fortification levels. Good linearity (r2 > 0.9990) was attained for most analytes. The limits of detection and quantification ranged from 0.03 - 0.84 μg/ml and 0.81 - 1.89 μg/ml respectively for analytes in biological samples. LODs and LOQs for analytes in food and environmental samples ranged from 0.02 to 0.39 and 0.25 to 1.30 ng/g respectively.
- Full Text:
- Date Issued: 2011
An investigation of a framework to evaluate computer supported collaborative work
- Beauvais, Erik Alexander Maoui
- Authors: Beauvais, Erik Alexander Maoui
- Date: 1999
- Subjects: Computer networks , Teams in the workplace -- Data processing , Technology -- Data processing , Information technology -- Economic aspects
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:1135 , http://hdl.handle.net/10962/d1002763 , Computer networks , Teams in the workplace -- Data processing , Technology -- Data processing , Information technology -- Economic aspects
- Description: Rapidly changing technology constantly modifies the way in which tasks are conceived and executed. Furthermore, leading organisations also encourage the use of new technology to support and establish innovative ways of doing business. For example, technology has helped to drive the globalization and tighter integration of world markets. This has facilitated, and even necessitated, business organisations to compete for work across national borders. For geographically dispersed organisations, collaboration has become critical. But this has presented time and space work complexities. Communication and collaboration technologies, for example, are needed to support such emerging co-operative business practices. The short development cycle of technology, communication and collaborative technologies included, leaves little time for testing and less for understanding the impact of new technology. To compound the problem, no established formulae exist for matching technology with work: over-investment is a waste of resources, while under-investment could lead to work failure. This research proposes a framework to assist organisations in selecting the appropriate level of technology with the work required. In this study, the methodology developed by the Evaluation Working Group, the DARPA Intelligent Collaboration and Visualization program is extended, and a matrix framework is constructed which compares the success of generic work tasks against a range of technology resources. The framework is tested using a low resource configuration, with selected work task types generally found in software development. (The Joint Application Development (JAD) methodology for software development is used as the contextual basis of the experiment.) All activities are evaluated according to selected measure components of success. A collection of research methods known as ethnography is used to examine and test the framework. Methods employed include questionnaires, interviews, interaction analysis and ethnomethodology. The results indicate that even at a low resource level, given selected criteria, collaborative technology successfully supports certain collaborative work activities. Findings also indicate that softer people issues require much more attention in order for technology to support natural collaborative work. Finally, user defined parameter testing has indicated that the framework functions as expected and designed.
- Full Text:
- Date Issued: 1999
- Authors: Beauvais, Erik Alexander Maoui
- Date: 1999
- Subjects: Computer networks , Teams in the workplace -- Data processing , Technology -- Data processing , Information technology -- Economic aspects
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:1135 , http://hdl.handle.net/10962/d1002763 , Computer networks , Teams in the workplace -- Data processing , Technology -- Data processing , Information technology -- Economic aspects
- Description: Rapidly changing technology constantly modifies the way in which tasks are conceived and executed. Furthermore, leading organisations also encourage the use of new technology to support and establish innovative ways of doing business. For example, technology has helped to drive the globalization and tighter integration of world markets. This has facilitated, and even necessitated, business organisations to compete for work across national borders. For geographically dispersed organisations, collaboration has become critical. But this has presented time and space work complexities. Communication and collaboration technologies, for example, are needed to support such emerging co-operative business practices. The short development cycle of technology, communication and collaborative technologies included, leaves little time for testing and less for understanding the impact of new technology. To compound the problem, no established formulae exist for matching technology with work: over-investment is a waste of resources, while under-investment could lead to work failure. This research proposes a framework to assist organisations in selecting the appropriate level of technology with the work required. In this study, the methodology developed by the Evaluation Working Group, the DARPA Intelligent Collaboration and Visualization program is extended, and a matrix framework is constructed which compares the success of generic work tasks against a range of technology resources. The framework is tested using a low resource configuration, with selected work task types generally found in software development. (The Joint Application Development (JAD) methodology for software development is used as the contextual basis of the experiment.) All activities are evaluated according to selected measure components of success. A collection of research methods known as ethnography is used to examine and test the framework. Methods employed include questionnaires, interviews, interaction analysis and ethnomethodology. The results indicate that even at a low resource level, given selected criteria, collaborative technology successfully supports certain collaborative work activities. Findings also indicate that softer people issues require much more attention in order for technology to support natural collaborative work. Finally, user defined parameter testing has indicated that the framework functions as expected and designed.
- Full Text:
- Date Issued: 1999
The effect of diet type and feeding rate on growth, morphological development and behaviour of larval and juvenile goldfish Carassius auratus (L.)
- Authors: Paulet, Timothy Guy
- Date: 2004
- Subjects: Goldfish -- Growth , Goldfish -- Nutrition , Goldfish -- Development , Goldfish -- Larvae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5271 , http://hdl.handle.net/10962/d1005115 , Goldfish -- Growth , Goldfish -- Nutrition , Goldfish -- Development , Goldfish -- Larvae
- Description: Intensive grow-out of goldfish, Carassius auratus (L.), larvae and juveniles in closed recirculating systems requires the control of environmental conditions and feeding. This study investigated the effect of different diets, environmental condition and feeding level on growth, development and survival of goldfish larvae and juveniles. Prey preference for Artemia nauplii or decapsulated Artemia cysts as well as agonistic behaviour was recorded. The micrometer device used to measure mouth-gape was shown to produce accurate measurements which could be used to estimate the maximal particle size that can be ingested by goldfish larvae of a particular age. Goldfish completed metamorphosis earlier with an increased feeding level of Artemia cysts and by making the cysts more accessible to the fish using up-welling water movement. Morphometric plasticity in goldfish larvae was exhibited within two weeks of growth and it may be possible to induce changes in morphology by manipulating diet and rearing environment. Fish that fed on moving prey items had a significantly larger mouth-gape than those that browsed cysts from the tank bottom or dry food items. The development of mouth-gape was not affected by the feeding level of cysts. Condition factor increased with an increase in the number of cysts fed per fish per day. Goldfish larvae and juveniles grew faster and had a higher survival when fed on decapsulated Artemia cysts than on instar I Artemia nauplii or a mixed live/dry diet of Artemia nauplii and dry food. Feeding at least 155 cysts per fish per day, in tanks with upwelling water movement, gave the best growth and survival and the smallest size variation. In addition, cysts remained available to the fish for longer periods, and were easier to prepare and feed. Goldfish larvae preferred decapsulated Artemia cysts to nauplii and rejected fewer prey items as they grew older. The frequency of agonistic behaviour increased as fish grew but no cannibalism was recorded for cyst-fed fish. This study showed that decapsulated Artemia cysts are a good alternative to Artemia nauplii as a diet for larval goldfish. Good growth and high survival was achieved for cyst-fed goldfish larvae and juveniles at 23 ± 1.5°C and at an initial stocking density of 12 fish per litre. This research also contributes to an understanding of feeding behaviour and attempts to minimise under- or over-feeding of Artemia cysts in order to reduce grow-out costs due to the high value of the feed type.
- Full Text:
- Date Issued: 2004
- Authors: Paulet, Timothy Guy
- Date: 2004
- Subjects: Goldfish -- Growth , Goldfish -- Nutrition , Goldfish -- Development , Goldfish -- Larvae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5271 , http://hdl.handle.net/10962/d1005115 , Goldfish -- Growth , Goldfish -- Nutrition , Goldfish -- Development , Goldfish -- Larvae
- Description: Intensive grow-out of goldfish, Carassius auratus (L.), larvae and juveniles in closed recirculating systems requires the control of environmental conditions and feeding. This study investigated the effect of different diets, environmental condition and feeding level on growth, development and survival of goldfish larvae and juveniles. Prey preference for Artemia nauplii or decapsulated Artemia cysts as well as agonistic behaviour was recorded. The micrometer device used to measure mouth-gape was shown to produce accurate measurements which could be used to estimate the maximal particle size that can be ingested by goldfish larvae of a particular age. Goldfish completed metamorphosis earlier with an increased feeding level of Artemia cysts and by making the cysts more accessible to the fish using up-welling water movement. Morphometric plasticity in goldfish larvae was exhibited within two weeks of growth and it may be possible to induce changes in morphology by manipulating diet and rearing environment. Fish that fed on moving prey items had a significantly larger mouth-gape than those that browsed cysts from the tank bottom or dry food items. The development of mouth-gape was not affected by the feeding level of cysts. Condition factor increased with an increase in the number of cysts fed per fish per day. Goldfish larvae and juveniles grew faster and had a higher survival when fed on decapsulated Artemia cysts than on instar I Artemia nauplii or a mixed live/dry diet of Artemia nauplii and dry food. Feeding at least 155 cysts per fish per day, in tanks with upwelling water movement, gave the best growth and survival and the smallest size variation. In addition, cysts remained available to the fish for longer periods, and were easier to prepare and feed. Goldfish larvae preferred decapsulated Artemia cysts to nauplii and rejected fewer prey items as they grew older. The frequency of agonistic behaviour increased as fish grew but no cannibalism was recorded for cyst-fed fish. This study showed that decapsulated Artemia cysts are a good alternative to Artemia nauplii as a diet for larval goldfish. Good growth and high survival was achieved for cyst-fed goldfish larvae and juveniles at 23 ± 1.5°C and at an initial stocking density of 12 fish per litre. This research also contributes to an understanding of feeding behaviour and attempts to minimise under- or over-feeding of Artemia cysts in order to reduce grow-out costs due to the high value of the feed type.
- Full Text:
- Date Issued: 2004
The feeding biology and potential impact of introduced giraffe (Giraffa camelopardalis) in the Eastern Cape Province, South Africa
- Authors: Parker, Daniel Matthew
- Date: 2004
- Subjects: Giraffe -- South Africa -- Eastern Cape , Animal-plant relationships -- South Africa -- Eastern Cape , Giraffe -- Food -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5694 , http://hdl.handle.net/10962/d1005380
- Description: Giraffe (Giraffa camelopardalis) are extralimital (non-native) to the Eastern Cape Province, South Africa yet they have and continue to be introduced to the region. Financial gain has arguably been the driving force behind these introductions as foreign tourists associate giraffe with Africa and the African wildlife experience. This raises a number of ethical, ecological and philosophical questions especially when it is considered that the impact of these browsers on the indigenous vegetation has remained largely unquantified. In this study I assessed the diet and potential impact of three populations of giraffe in the Eastern Cape Province between January 2002 and October 2003. The diet was assessed by both direct observations and faecal analysis. There was no significant difference (P > 0.05) between the results of the two methods of analysis, although direct observations appeared to be a superior method for assessing the diet of giraffe. The diet of giraffe in the Eastern Cape Province was similar to that within their native range with a deciduous species from the genus Acacia (Acacia karroo) being the most important component of the diet. However, giraffe in the Eastern Cape Province consumed more evergreen plant species than those within their native range. The relative lack of deciduous species in the Eastern Cape Province provides a likely explanation for such a result. Seasonal variation in the consumption of the most important species in the diet was evident and this was attributed to the deciduous nature of A. karroo and the seasonal growth of new shoots which were more palatable. The vegetation of the areas most commonly utilised by giraffe at each site was sampled using the point-centred-quarter method and the results related to the frequency of each species in the diet to calculate preference indices. Giraffe preference was strongest for A. karroo and this was attributed to the highly favourable chemical composition of the species. The browse utilisation of giraffe at each site was determined using the twig-length method and intake rates for the three most important species in the diet calculated using a pre-existing regression equation. Male giraffe fed at a higher rate than females. This was probably due to males adopting a “time-minimising” strategy to their feeding in order to allow more time for reproductive pursuits. Giraffe browse utilisation was highest where giraffe density was highest. However, several species were more heavily browsed than others even when giraffe density was low, suggesting that giraffe are capable of negatively affecting the indigenous flora of the province. I conclude that giraffe numbers should be reduced relative to property size in the Eastern Cape Province and that research into the impact of not only giraffe but the combined effects of giraffe and other extralimital herbivores on the indigenous flora and fauna be continued.
- Full Text:
- Date Issued: 2004
- Authors: Parker, Daniel Matthew
- Date: 2004
- Subjects: Giraffe -- South Africa -- Eastern Cape , Animal-plant relationships -- South Africa -- Eastern Cape , Giraffe -- Food -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5694 , http://hdl.handle.net/10962/d1005380
- Description: Giraffe (Giraffa camelopardalis) are extralimital (non-native) to the Eastern Cape Province, South Africa yet they have and continue to be introduced to the region. Financial gain has arguably been the driving force behind these introductions as foreign tourists associate giraffe with Africa and the African wildlife experience. This raises a number of ethical, ecological and philosophical questions especially when it is considered that the impact of these browsers on the indigenous vegetation has remained largely unquantified. In this study I assessed the diet and potential impact of three populations of giraffe in the Eastern Cape Province between January 2002 and October 2003. The diet was assessed by both direct observations and faecal analysis. There was no significant difference (P > 0.05) between the results of the two methods of analysis, although direct observations appeared to be a superior method for assessing the diet of giraffe. The diet of giraffe in the Eastern Cape Province was similar to that within their native range with a deciduous species from the genus Acacia (Acacia karroo) being the most important component of the diet. However, giraffe in the Eastern Cape Province consumed more evergreen plant species than those within their native range. The relative lack of deciduous species in the Eastern Cape Province provides a likely explanation for such a result. Seasonal variation in the consumption of the most important species in the diet was evident and this was attributed to the deciduous nature of A. karroo and the seasonal growth of new shoots which were more palatable. The vegetation of the areas most commonly utilised by giraffe at each site was sampled using the point-centred-quarter method and the results related to the frequency of each species in the diet to calculate preference indices. Giraffe preference was strongest for A. karroo and this was attributed to the highly favourable chemical composition of the species. The browse utilisation of giraffe at each site was determined using the twig-length method and intake rates for the three most important species in the diet calculated using a pre-existing regression equation. Male giraffe fed at a higher rate than females. This was probably due to males adopting a “time-minimising” strategy to their feeding in order to allow more time for reproductive pursuits. Giraffe browse utilisation was highest where giraffe density was highest. However, several species were more heavily browsed than others even when giraffe density was low, suggesting that giraffe are capable of negatively affecting the indigenous flora of the province. I conclude that giraffe numbers should be reduced relative to property size in the Eastern Cape Province and that research into the impact of not only giraffe but the combined effects of giraffe and other extralimital herbivores on the indigenous flora and fauna be continued.
- Full Text:
- Date Issued: 2004
Localisation of Theiler's Murine Encephalomyelitis virus non-structural proteins 2B, 2C, 2BC and 3A in BHK-21 cells, and the effect of amino acid substitutions in 2C on localisation and virus replication
- Authors: Murray, Lindsay
- Date: 2007
- Subjects: Encephalomyelitis -- Genetic aspects , Amino acid sequence , Picornaviruses , Viruses -- Reproduction
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4090 , http://hdl.handle.net/10962/d1007722 , Encephalomyelitis -- Genetic aspects , Amino acid sequence , Picornaviruses , Viruses -- Reproduction
- Description: The picornavirus family includes significant human and animal viruses such as poliovirus (PV), human rhinovirus (HRV) and foot-and-mouth-disease virus (FMDV). Current disease treatment and control strategies are limited by an incomplete understanding of the interactions between the non-structural, replicative picornavirus proteins and host cell components. To investigate these interactions, Theiler's murine encephalomyelitis virus (TMEV) 2B, 2C, 2BC and 3A proteins were transiently expressed in BHK-21 cells and detected by indirect immunostaining and laser-scanning or epifluorescence microscopy. The signal of the 2B protein overlapped with that of the ER marker protein, ERp60, as well as that of the peripheral Golgi marker protein, β-COP. The 2C protein overlapped with ERp60 in a faint reticular stain, and localised to large punctate structures that partially overlapped with β-COP at higher levels of expression. The 2BC protein located to large perinuclear structures that overlapped exclusively with β-COP. The TMEV 3A protein signal overlapped with both ERp60 and β-COP stains, in addition in cells expressing the 3A protein the ER appeared swollen and bulbous while the Golgi was dispersed in some cells. 2C and 2BC proteins with C-terminal deletions localised in the same manner as the wild type proteins indicating that the localisation signals that determine subcellular localisation of the proteins are within the N-terminal 60 amino acids of the 2C protein. The significance of the high degree of conservation of the N-terminal domain of the 2C protein throughout the Picornaviridae was investigated through the introduction of amino acid substitution mutations at highly conserved residues in the N-terminal domain of 2C into the viral cDNA. Upon transfection of the viral RNA into BHK-21 cells, it was observed that substitution of amino acid residues 8, 18 and 29 abolished the ability of TMEV to induce cytopathic effect (CPE), while substitution of residues 4, 14 and 23 only attenuated the ability of TMEV to induce CPE. To determine whether amino acid substitution mutations would affect the localisation of the 2C protein, 2C proteins with substitution mutations at amino acids 4, 8, 14, 18, 23 and 29 were transiently expressed in BHK-21 cells and detected by indirect imrnunostaining and examination by laser-scanning confocal and epifluorescence microscopy. The 2C mutant 4, 8 and 29 proteins showed slightly altered localisation patterns compared to the wild type protein with a significant portion of the proteins localising in a perinuclear stain suggesting possible localisation to the nuclear envelop. The 2C mutant 14 and 18 proteins localised to a diffuse pattern in BHK-21 cells while the 2C mutant 23 protein located to small punctate structures that partially overlapped with the ERp60 stain but were completely separate from the β-COP stain. Finally, a hydrophilic, antigenic region of the 2C protein was expressed in frame with an N-terminal GST tag and was successfully purified on a pilot-scale and detected by Western analysis. This 2C178 peptide will be used to generate antibodies against the 2C and 2BC proteins for use in future studies. This study has furthered our knowledge of the localisation of the picornavirus 2B, 2C, 2BC and 3A proteins in host cells and identified a possible link between this localisation and an ability of TMEV to replicate in BHK-21 cells.
- Full Text:
- Date Issued: 2007
- Authors: Murray, Lindsay
- Date: 2007
- Subjects: Encephalomyelitis -- Genetic aspects , Amino acid sequence , Picornaviruses , Viruses -- Reproduction
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4090 , http://hdl.handle.net/10962/d1007722 , Encephalomyelitis -- Genetic aspects , Amino acid sequence , Picornaviruses , Viruses -- Reproduction
- Description: The picornavirus family includes significant human and animal viruses such as poliovirus (PV), human rhinovirus (HRV) and foot-and-mouth-disease virus (FMDV). Current disease treatment and control strategies are limited by an incomplete understanding of the interactions between the non-structural, replicative picornavirus proteins and host cell components. To investigate these interactions, Theiler's murine encephalomyelitis virus (TMEV) 2B, 2C, 2BC and 3A proteins were transiently expressed in BHK-21 cells and detected by indirect immunostaining and laser-scanning or epifluorescence microscopy. The signal of the 2B protein overlapped with that of the ER marker protein, ERp60, as well as that of the peripheral Golgi marker protein, β-COP. The 2C protein overlapped with ERp60 in a faint reticular stain, and localised to large punctate structures that partially overlapped with β-COP at higher levels of expression. The 2BC protein located to large perinuclear structures that overlapped exclusively with β-COP. The TMEV 3A protein signal overlapped with both ERp60 and β-COP stains, in addition in cells expressing the 3A protein the ER appeared swollen and bulbous while the Golgi was dispersed in some cells. 2C and 2BC proteins with C-terminal deletions localised in the same manner as the wild type proteins indicating that the localisation signals that determine subcellular localisation of the proteins are within the N-terminal 60 amino acids of the 2C protein. The significance of the high degree of conservation of the N-terminal domain of the 2C protein throughout the Picornaviridae was investigated through the introduction of amino acid substitution mutations at highly conserved residues in the N-terminal domain of 2C into the viral cDNA. Upon transfection of the viral RNA into BHK-21 cells, it was observed that substitution of amino acid residues 8, 18 and 29 abolished the ability of TMEV to induce cytopathic effect (CPE), while substitution of residues 4, 14 and 23 only attenuated the ability of TMEV to induce CPE. To determine whether amino acid substitution mutations would affect the localisation of the 2C protein, 2C proteins with substitution mutations at amino acids 4, 8, 14, 18, 23 and 29 were transiently expressed in BHK-21 cells and detected by indirect imrnunostaining and examination by laser-scanning confocal and epifluorescence microscopy. The 2C mutant 4, 8 and 29 proteins showed slightly altered localisation patterns compared to the wild type protein with a significant portion of the proteins localising in a perinuclear stain suggesting possible localisation to the nuclear envelop. The 2C mutant 14 and 18 proteins localised to a diffuse pattern in BHK-21 cells while the 2C mutant 23 protein located to small punctate structures that partially overlapped with the ERp60 stain but were completely separate from the β-COP stain. Finally, a hydrophilic, antigenic region of the 2C protein was expressed in frame with an N-terminal GST tag and was successfully purified on a pilot-scale and detected by Western analysis. This 2C178 peptide will be used to generate antibodies against the 2C and 2BC proteins for use in future studies. This study has furthered our knowledge of the localisation of the picornavirus 2B, 2C, 2BC and 3A proteins in host cells and identified a possible link between this localisation and an ability of TMEV to replicate in BHK-21 cells.
- Full Text:
- Date Issued: 2007
The role of computational thinking in introductory computer science
- Authors: Gouws, Lindsey Ann
- Date: 2014
- Subjects: Computer science , Computational complexity , Problem solving -- Study and teaching
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4690 , http://hdl.handle.net/10962/d1011152 , Computer science , Computational complexity , Problem solving -- Study and teaching
- Description: Computational thinking (CT) is gaining recognition as an important skill for students, both in computer science and other disciplines. Although there has been much focus on this field in recent years, it is rarely taught as a formal course, and there is little consensus on what exactly CT entails and how to teach and evaluate it. This research addresses the lack of resources for integrating CT into the introductory computer science curriculum. The question that we aim to answer is whether CT can be evaluated in a meaningful way. A CT framework that outlines the skills and techniques comprising CT and describes the nature of student engagement was developed; this is used as the basis for this research. An assessment (CT test) was then created to gauge the ability of incoming students, and a CT-specfic computer game was developed based on the analysis of an existing game. A set of problem solving strategies and practice activities were then recommended based on criteria defined in the framework. The results revealed that the CT abilities of first year university students are relatively poor, but that the students' scores for the CT test could be used as a predictor for their future success in computer science courses. The framework developed for this research proved successful when applied to the test, computer game evaluation, and classification of strategies and activities. Through this research, we established that CT is a skill that first year computer science students are lacking, and that using CT exercises alongside traditional programming instruction can improve students' learning experiences.
- Full Text:
- Date Issued: 2014
- Authors: Gouws, Lindsey Ann
- Date: 2014
- Subjects: Computer science , Computational complexity , Problem solving -- Study and teaching
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4690 , http://hdl.handle.net/10962/d1011152 , Computer science , Computational complexity , Problem solving -- Study and teaching
- Description: Computational thinking (CT) is gaining recognition as an important skill for students, both in computer science and other disciplines. Although there has been much focus on this field in recent years, it is rarely taught as a formal course, and there is little consensus on what exactly CT entails and how to teach and evaluate it. This research addresses the lack of resources for integrating CT into the introductory computer science curriculum. The question that we aim to answer is whether CT can be evaluated in a meaningful way. A CT framework that outlines the skills and techniques comprising CT and describes the nature of student engagement was developed; this is used as the basis for this research. An assessment (CT test) was then created to gauge the ability of incoming students, and a CT-specfic computer game was developed based on the analysis of an existing game. A set of problem solving strategies and practice activities were then recommended based on criteria defined in the framework. The results revealed that the CT abilities of first year university students are relatively poor, but that the students' scores for the CT test could be used as a predictor for their future success in computer science courses. The framework developed for this research proved successful when applied to the test, computer game evaluation, and classification of strategies and activities. Through this research, we established that CT is a skill that first year computer science students are lacking, and that using CT exercises alongside traditional programming instruction can improve students' learning experiences.
- Full Text:
- Date Issued: 2014
A synthetic and spectrometric study of the initial phases in urea-formaldehyde resin formation
- Authors: Nocanda, Xolani Wittleton
- Date: 1998
- Subjects: Phenolic resins , Gums and resins, Synthetic
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4383 , http://hdl.handle.net/10962/d1005048 , Phenolic resins , Gums and resins, Synthetic
- Description: In a series ofpreIiminary studies of urea-formaldehyde reactions, the influence of the U:F molar ratio, the temperature and the catalyst were investigated. Subsequently, in a more rigorous evaluation of the influence of four variables on urea-formaldehyde resin formation, viz., U:F molar ratio, pH, temperature and reaction time, a set of twenty reactions were performed using a statistical approach. The results indicate that high resin viscosity is best achieved by using a high U:F molar ratio (1:2) and conducting the reaction at 90°C. Several basic components produced in urea-formaldehyde reactions have been synthesised, characterised by NMR spectroscopy, silylated using bis(trimethylsilyl)trifluoroacetamide and analysed by low resolution mass spectrometry. The use oftriazones as cross-linking agents has been investigated, and a series of 5-substituted triazones have been prepared and their mass fragmentation patterns explored using a combination of low and high resolution mass spectrometry. In order to facilitate interpretation of ¹³C NMR data obtained for U-F resins, internal rotation in various urea derivatives was investigated using variable temperature ¹H and ¹³C NMR spectroscopic study. Signal splittings observed below 247 K have been rationalised in terms of various rotameric structures.
- Full Text:
- Date Issued: 1998
- Authors: Nocanda, Xolani Wittleton
- Date: 1998
- Subjects: Phenolic resins , Gums and resins, Synthetic
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4383 , http://hdl.handle.net/10962/d1005048 , Phenolic resins , Gums and resins, Synthetic
- Description: In a series ofpreIiminary studies of urea-formaldehyde reactions, the influence of the U:F molar ratio, the temperature and the catalyst were investigated. Subsequently, in a more rigorous evaluation of the influence of four variables on urea-formaldehyde resin formation, viz., U:F molar ratio, pH, temperature and reaction time, a set of twenty reactions were performed using a statistical approach. The results indicate that high resin viscosity is best achieved by using a high U:F molar ratio (1:2) and conducting the reaction at 90°C. Several basic components produced in urea-formaldehyde reactions have been synthesised, characterised by NMR spectroscopy, silylated using bis(trimethylsilyl)trifluoroacetamide and analysed by low resolution mass spectrometry. The use oftriazones as cross-linking agents has been investigated, and a series of 5-substituted triazones have been prepared and their mass fragmentation patterns explored using a combination of low and high resolution mass spectrometry. In order to facilitate interpretation of ¹³C NMR data obtained for U-F resins, internal rotation in various urea derivatives was investigated using variable temperature ¹H and ¹³C NMR spectroscopic study. Signal splittings observed below 247 K have been rationalised in terms of various rotameric structures.
- Full Text:
- Date Issued: 1998
A psychobiographical study of Antwone Fisher
- Authors: Wannenburg, Nicola
- Date: 2014
- Subjects: Fisher, Antwone Quenton , Psychology -- Biographical methods , Psychoanalysis , Abused children -- Biography
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3219 , http://hdl.handle.net/10962/d1012939
- Description: Psychobiographical researchers methodically formulate life histories and interpret them by means of psychological theories. The research typically focuses on exemplary and completed lives. The cases that are studied are usually of individuals who are of particular interest to society as a result of excelling in their particular fields, be they to society’s benefit or detriment. Antwone Fisher was chosen for this study using purposive sampling as he meets the psychobiographical requirement of being an extraordinary individual. Fisher grew up in the Foster Care System and faced many abusive experiences as child. Despite difficult beginnings, Fisher developed into a stable and creative adult who excels as a poet, screenwriter and film director. The primary aim of this study is to describe and interpret the life of Antwone Fisher through Erikson’s theory of psychosocial development. The data was processed and analysed using Miles and Hubermans’ general view of qualitative analyses as well as Alexander’s method of asking the data questions. This study contributes to the development of psychobiographical research in South Africa as well as to personality and developmental theory.
- Full Text:
- Date Issued: 2014
- Authors: Wannenburg, Nicola
- Date: 2014
- Subjects: Fisher, Antwone Quenton , Psychology -- Biographical methods , Psychoanalysis , Abused children -- Biography
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3219 , http://hdl.handle.net/10962/d1012939
- Description: Psychobiographical researchers methodically formulate life histories and interpret them by means of psychological theories. The research typically focuses on exemplary and completed lives. The cases that are studied are usually of individuals who are of particular interest to society as a result of excelling in their particular fields, be they to society’s benefit or detriment. Antwone Fisher was chosen for this study using purposive sampling as he meets the psychobiographical requirement of being an extraordinary individual. Fisher grew up in the Foster Care System and faced many abusive experiences as child. Despite difficult beginnings, Fisher developed into a stable and creative adult who excels as a poet, screenwriter and film director. The primary aim of this study is to describe and interpret the life of Antwone Fisher through Erikson’s theory of psychosocial development. The data was processed and analysed using Miles and Hubermans’ general view of qualitative analyses as well as Alexander’s method of asking the data questions. This study contributes to the development of psychobiographical research in South Africa as well as to personality and developmental theory.
- Full Text:
- Date Issued: 2014
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