The development and op timisation of a Theiler’s murine encephalomyelitis virus antiviral assay
- Authors: Naidoo, Urisha Tirah
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424677 , vital:72174
- Description: Picornaviruses belong to the Picornaviridae family which are one of the largest and most diverse family of RNA viruses that cause a broad spectrum of infections in both humans and animals. These diseases range from severe infections such as poliomyelitis, meningitis, myocarditis to mild illnesses such as the common cold. Picornavirus outbreaks are a worldwide threat as they are continuously occurring. A recent outbreak of foot-and-mouth disease caused by a picornavirus occurred in South Africa, resulting in a temporary ban on the movement of cattle. Currently, the FDA has not approved any antiviral drugs against this virus, increasing the urgency for identifying effective antivirals. Picornaviruses have similar genomes and capsid organisation as such, those that are non-hazardous to humans can be used as a model system. A Theiler’s murine encephalomyelitis virus (TMEV) strain GDVII and Baby Hamster Kidney fibroblasts (BHK-21 cells) was used as a replication system to develop and optimise a medium-throughput antiviral screening assay. The TMEV GDVII replication system in BHK-21 cells was validated, and preliminary experiments were performed that were necessary for the development of the TMEV GDVII antiviral assay. This was achieved by conducting a CPE assay to visually monitor the onset and development of CPE induced by TMEV GDVII. Plaque assays accurately quantified the number of infectious virus particles required for calculating the MOI in downstream experiments. Lastly, indirect immunofluorescence and Western blot analysis detected the expression of viral proteins using previously generated antibodies against the TMEV GDVII VP1 capsid and 2C protein, thereby confirming infection in BHK-21 cells. The development of robust and reproducible assays is an essential component in antiviral drug discovery. Therefore, the confirmed replication system was then used as a foundation to develop a medium-throughput CPE-based TMEV GDVII antiviral assay whereby the parameters were optimised to produce one of high quality. Firstly, the quantitation of viral-induced CPE was examined and confirmed in a 96-well plate using resazurin as a cell viability indicator. Each parameter was tested at varying conditions, and the optimal was concluded as 2 % FBS in the assay media, a 15 000 cells/well seeding density, infecting the cells with TMEV GDVII at an MOI of 0.00625 and measuring resazurin at an endpoint of 72 hpi. Furthermore, the parameters were ii validated by calculating the Z’- factor, which consistently produced scores above 0.5, indicative of a reliable, robust, reproducible antiviral assay. Currently, there are no inhibitors against TMEV GDVII that have been reported or confirmed in cell lines, animal models or clinical trials. Therefore, once the optimal assay parameters were selected, it presented an opportunity to assess whether potential compounds, including itraconazole (ITZ) and dipyridamole (DIP), possessed antiviral activity that could firstly, be utilised as a control inhibitor when screening compounds against TMEV GDVII and secondly, contribute to research on this virus. Additionally, the previously produced anti-TMEV GDVII capsid antibody was shown to neutralise viral infection and was also included as a potential control. The sensitivity of the cells towards DMSO, a solution in which the compounds were solubilised, was first investigated. It was found that concentrations above 1 % are toxic to the cells; as such, the final DMSO concentrations were always kept below 1 % when screening compounds. Lastly, the generation of dose-response curves aided in the conclusion that the antibody was the most suitable control inhibitor as it displayed potent antiviral activity and no cytotoxicity towards the cells. In contrast, ITZ and DIP did not possess effective antiviral action and were toxic to cells at high concentrations. Finally, after all the components of the medium-throughput TMEV GDVII antiviral assay were identified, it was possible to screen 24 compounds from a coumarin and marine natural product library for cell cytotoxicity and antiviral activity. After generating dose-response curves, it was concluded that no compound effectively inhibited virus-induced CPE, and most were toxic to cells at relatively high concentrations. In conclusion, this is the first study that describes the development and optimisation of a robust medium-throughput CPE-based antiviral assay that has immense potential to screen other libraries of compounds for antiviral activity against TMEV GDVII. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2023
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Vernacular sound technologies: experimenting with reverb in isiXhosa choral recordings
- Authors: Ncanywa, Sibusiso
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/432147 , vital:72846
- Description: Access restricted. Expected release date 2025. , Thesis (MMus) -- Faculty of Humanities, Music and Musicology, 2023
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Visual representations of linear algebraic expressions: a case study in a Grade 9 after-school mathematics club
- Authors: Herbert, Sindisiwe
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424019 , vital:72116
- Description: Visualisation is commonly used as a tool in introducing algebra through visual or kinaesthetic sequences designed to prompt learners' development of a general rule for moving from a term's position to its output value. Fluency in both the concepts and the conventions of elementary algebra are essential to learners, as algebra forms the language in which many advanced mathematical ideas are encoded. Moreover, algebraic fluency is often associated with an ability to think abstractly about arithmetic processes. In many classrooms, however, research has shown that learners often focus on fluency in algebraic conventions rather than concepts, learning how to manipulate expressions without understanding the algorithms they are taught to follow. This trend can be linked to several causes, including teacher-centred mathematics classrooms in which learners are – whether implicitly or explicitly – encouraged to copy formulae and methods in order to ‘get it right in a test’ without necessarily grasping the underlying logical relationships. This case study, therefore, aimed to determine whether there was value in using visual, kinaesthetic models to broaden and deepen learners' use of algebra. To that end, in the context of an extra-curricular mathematics club that aimed to decentre the teachers and demand innovative ideas of the participants, six pairs of Grade 9 learners were tasked with creating visual representations of a linear algebraic expression using coloured building cubes. The responses to this task over the course of five assignments were many and varied and almost universally displayed a sustained internal logic that the learners were able to explain and develop. Most pairs began with a visual list of terms arranged in sets of towers, pyramids or, in one case, a spiral. At the end of the study, all but one of the pairs had settled on a Visual Expression, in which various colours were used to represent elements of the algebra such as the values of the coefficient, the variable and the constant term. The participants' representations grew in complexity over the course of the study and the conformity of the final responses showed that the club was a collaborative space in which learners shared ideas. However, the structure of the Visual Expressions and their own confessions of nerves about ‘getting it wrong’ in the interviews suggest that the participants were stuck in a mindset that led them to seek out and idealise the representation closest to the original algebra, even though that representation revealed little about the structural relationship underlying the expression. , Thesis (MEd) -- Faculty of Education, Secondary and Post-School Education, 2023
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Year-end oversight in Local Government: a case study of water and sanitation service delivery at Amathole District Municipality
- Authors: Seoke, Duncan
- Date: 2023-10-13
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/425206 , vital:72219
- Description: Poor service delivery is a challenge that has continued to plague the constitutional democracy of South Africa. Legislation has, in response, been passed to ensure efficient service delivery. Despite this legislation, municipal service delivery has been alarmingly poor, and reports on municipal performance and newspaper articles have corroborated this. On the causes of poor service delivery, the existing literature appears to have focused on financial mismanagement, corruption, and the incapacity of officials. Absent from existing literature are discussions on how municipal service delivery is affected by the lack of interrelatedness between municipal public resource management processes (such as revenue collection, expenditure management and oversight). This dissertation addresses this gap by examining the municipal public resource management processes from an oversight perspective. The study sought to analyse the effectiveness of year-end oversight at Amathole District Municipality (ADM) over water and sanitation services. This research aimed to study the year-end oversight over service delivery with reference to the Municipal Public Resource Management (MPRM) model. According to the model, effective service delivery includes inter alia effective oversight, as oversight forms part of the entire system of the municipal resource management cycle. Thus, for the MPRM model, oversight should be exercised on all these processes. The research was a desktop analysis that used both a qualitative and quantitative research methodology. The primary document analysed was the ADM Oversight Report (OR). This report is produced by the Municipal Public Accounts Committee (MPAC) and reflects the year-end oversight conducted by the MPAC on the municipality's activities. In the analysis of the OR, it was concluded that the ADM MPAC had failed to conduct effective year-end oversight of W&S services. The efficacy of the ADM MPAC's year-end oversight was measured against guidelines for effective year-end oversight from the National Treasury and the MPRM model. The findings suggested that there were inefficiencies in the manner that the ADM MPAC exercised its year-end oversight. , Thesis (MA) -- Faculty of Humanities, Sociology, 2023
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Biological longitudinal aspects of the Kabompo River, a significant tributary of the upper Zambezi sub-catchment, North-west Province, Zambia
- Authors: Janse van Rensburg, Lomarie Cathleen
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422292 , vital:71928
- Description: The landscape of the Upper Zambezi Catchment in the remote North-Western Province of Zambia retains many natural features, but current and future anthropogenic activities threaten its integrity. Lack of information relating to the basic ecological functioning of the region hampers conservation efforts. Flooding from the adjacent woody-savanna and open grasslands in the Upper-Zambezi Catchment in the wet season drives crucial processes in the aquatic ecosystems such as the Kabompo River. This thesis describes aquatic food webs and describes the nutrient cycling in this river. A general introduction was based on the seasonality, river morphology, and biodiversity of the Kabompo River. There is no aquatic food web information available on this river. To address this knowledge gap, sampling of the principal food web components– vegetation, invertebrates, and fish – were collected in the dry season from August to September 2019 across six sites in the Kabompo River. Stable isotope analyses provided proxies for the food web structure at each site, and were the basis of longitudinal comparisons. The analyses suggest that the Kabompo River food webs follow the trophic ordination and nutrient cycling characteristic of the revised-Riverine Productivity Model. Food webs and community assemblages remain structurally similar between sites and provide some evidence of bottom-up productivity-driven trophic dynamics. To predict the possible landscape-scale processing of nutrient changes present for the aquatic ecosystems of the Kabompo River, a longitudinal comparison between a herbivore, Labeo cylindricus, and omnivore, Synodontis spp., fish species and their resources (primary producers and invertebrates) was done and showed archetypal fish food web trophic separation (2 ‰ to3 ‰) between species which remains consistent for the headwater sites. The nutrient values change at the convergence between the Kabompo Bridge and the Mwinilunga branches and show a decrease in the trophic separation of the sites below. This change becomes apparent from the last site, Watopa, where the trophic separation re-establishes (2 ‰ to 3 ‰) between the species. The trend suggests primarily autochthonous production in the headwater reaches, changing to allochthonous and autochthonous nutrient incorporation after the convergence of the two main branches. Permanent wetlands surrounding the convergence zones may be conducive to more primary producer activity and increased nutrient turnover. Permanent wetlands are a common feature of the Upper Zambezi Catchment tributaries, with the most notable lentic system the Barotse Flood Plains in the south, which shares reach-adjacent characteristics with the Kabompo River. Trends from the 2015 dry season identified for the Barotse Flood Plains support the findings of this thesis, where the δ13C values recorded reach depletion of -45 ‰. Management in the Upper Zambezi Basin and the Kabompo River comprises biannual assessments of water quality and quantity by the Zambezi River Authority, but not of the ecosystem process. This thesis may provide information to address the ecological (food webs and nutrient cycling) dynamics of the Basin rivers. As the first baseline information on the river's biology and ecology, it may present a comparative basis for future assessments under conservation management strategies. , Thesis (MSc) -- Faculty of Science, Ichthyology and Fisheries Science, 2023
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Factors influencing the number of non-timber forest product species used at the individual, household and village levels in Vhembe, Limpopo Province, South Africa
- Authors: Mulaudzi, Takalani Rachel
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422481 , vital:71946
- Description: Non-Timber Forest Products (NTFPs) are wild biological resources obtained from natural landscapes, agricultural systems, and rural and urban regions for a variety of uses. NTFPs contribute significantly to livelihoods and living standards by offering a variety of socioeconomic benefits such as food, health, income generation, and safety nets. These multiple benefits are provided by a wide diversity of plant, animal and fungal species in different settings. However, the precise number of NTFP species used depends on the scale of assessment, namely use by individuals, by households and by villages. Consequently, it is difficult to compare numbers of species used between studies because of the differing scales. As a result, the objective of this study was to determine the number of NTFP species used at three different scales and how each is correlated with appropriate scale-dependent contextual variables. This objective was met by sampling individuals and whole households in five villages in the Vhembe District in Limpopo Province, South Africa. Twenty households were randomly selected in each village, providing a total sample of 100 households and 179 individuals in those households. In each household, every adult member was interviewed individually about the NTFPs that they used or knew. The major questions covered the NTFP species used, the respondents' socioeconomic level, the NTFP species used previously, and if the number of NTFP species are changing in the landscape. Data were reported per individual as well as per household and per village. The household level was derived by adding the responses of all the adults in each household but excluding duplicates. The same approach was used at the village scale, namely, the data across the 20 households were pooled, and duplicates were omitted. Patterns of NTFP use were examined by type of NTFP, and analysis using standard statistical tests, along with a Principal Components Analysis and a multivariate Detrended Correspondence Analysis of the NTFP species composition across villages. A total of 275 species of NTFPs were mentioned across the entire sample, including 81 species of firewood, 48 species of edible insects, 43 species of wild vegetables, 40 species of edible fruits, 38 species of wood utensils, 28 species of medicinal plants, 23 species of thatch grass, 19 species of poles, 19 species of wood for carvings, nine species of reeds, eight species of mushroom, and eight species of fodder. Of these species, some species had more than one use. Approximately 90% of the species used by the respondents were native species. Wild vegetables (e.g., Corchorus olitorius, Amaranthus capensis, Bidens pilosa), firewood (e.g., Colophospermum mopane, Parinari curatellifolia, Combretum hereroense), wild fruit (e.g., Landolphia kirkii, Vangueria infausta), and edible insects (e.g., Imbrasia belina, Macrotermes michaelseni, Encosternum delegorguei) were the most often used NTFPs by communities in the Vhembe area. Most of the households relied on NTFPs for subsistence. At the scale of an individual, the mean number of NTFP species used was 26.8±12.6 across the five villages. When considering a household, the mean number was 36.1±10.7 species. When aggregated across the 20 households per village, the mean number per village was 105.2±34.1 species. The species accumulation curve showed that 20 households per village were reasonably representative for four of the five villages. The results from the principal component analysis (PCA) and correlation analysis revealed that age, education, and employment status were significantly associated with the number of species used by individuals, whereas gender showed no clear correlation. At the household scale, household size was positively related with the number of species used, but income level and income source were not. At the village scale, attributes such as proximity to a town, settlement type, vegetation type, and distance to a nearby savanna all had some influence on the number of NTFP species listed. Most of the respondents felt that the number of species used had increased over the last 10–15 years, although some stated the opposite. Increases were attributed by traditional councils bans, the establishment of protected areas, and restoration. Activities eroding the number of NTFP species were identified as clear felling for farming, climate change, and over-harvesting. This study suggests that communities must be made more aware of the overharvesting of NTFP species in the savanna to avoid biodiversity loss. , Thesis (MSc) -- Faculty of Science, Environmental Science, 2023
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Mineral paragenesis of olmiite/poldevaartite and rhodochrosite/shigaite occurrences in the Kalahari Manganese Field, and their relation to the formation of high-grade manganese ore of Nchwaning II mine, Black Rock, South Africa
- Authors: Opperman, Alicia
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422448 , vital:71943
- Description: Access restricted. Access embargoed until 2025. , Thesis (MSc) -- Faculty of Science, Geology, 2023
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Pharmaco-chemical investigation of Erythrina caffra: extracts, isolated compounds and their biological activities
- Authors: Nogqala, Simnikiwe
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422459 , vital:71944
- Description: In this study, secondary metabolites isolated from Erythrina caffra, a medicinal plant indigenous to South Africa, were investigated. E. caffra is well-known for its healing properties and it is traditionally used for treating bacterial infections like tuberculosis (TB), abscesses, tooth aches and ear infections. Its extracts have also been used to treat cancer. Though many studies have been done on this plant, most of them tended to focus solely on the isolated compounds. In the present study however, extracts, fractions and isolated compounds from E. caffra were evaluated for their anticancer, anti-oxidant, anti-enzymatic, antibacterial and cytotoxicity. The methanol crude extract (B1) from the stem bark of E. caffra was used to extract alkaloidic fractions (B2 and B3) using ethyl acetate and n-butanol respectively, a third fraction (B4) was also extracted using ethyl acetate this fraction was called a neutral fraction. The neutral fraction (B4) was fractionated and through a sequence of column chromatography three active secondary metabolites were isolated. The isolated compounds included Lupeol (1), stigmasterol (2) and 5,7-Dihydroxy-4'-methoxy-3',5'-diprenylflavanone (3). These isolated compounds were characterized and identified using spectroscopic techniques including IR, NMR and high-resolution Mass Spectrometry. Using the cell line HCC-70, isolated from a primary ductal carcinoma, in vitro anticancer assays were carried out on the crude extract from the bark, fractions, isolated compounds and an unseparated mixture of two compounds. These samples were also evaluated for their anti-oxidant, anti-enzymatic, antibacterial and cytotoxicity activities. The crude extract inhibited the cell viability by over 30% and had no effect on the HeLa cells at concentrations of 20μM. Abyssinone V’ 4-methyl-ether (3) and the mixture of stigmasterol (2) and an unidentified compound exhibited potent anticancer activity against the HCC-70 cell line with IC50 of 18.05μM and 9.04μM respectively. Antibacterial assays were also carried out on the crude extracts, fractions and concoctions made from the fractions with the best activity combined with the ones that performed poorly. The concoctions were prepared as two separate series (S and N series). The crude extract inhibited more than 80% of the Staphylococcus aureus cells at a concentration of 20μM with only minimal damage to the HeLa cells. In the concoctions however, the N series managed to inhibit over 96% of the S. aureus while exhibiting no cytotoxicity towards HeLa cells. The extract and its fractions also showed good anti-oxidant activities. Molecular docking of these compounds was done on the Human estrogen receptor (PDB ID:3ERT) and Abyssinone V’ 4-methyl-ether (3) showed the best docking score of -6.6 Kcal/mol, for the simulation against Epidermal growth factor receptor (PDB ID: 1M17) Stigmasterol (2) showed the best docking score of -3.8 Kcal/mol. In silico docking on 3ERT and 1M17 were done to test the binding affinity of the isolated compounds to the proteins which are well known to be overexpressed in some types of cancer. Flavonoids isolated from Erythrina species have been reported to possess good antiplasmodial activity. However, due to the minute amounts isolated in the present study in-vitro assays could not be carried out. Nevertheless, in-silico assays were conducted on the most prominent protozoal parasite which causes malaria in the majority of African countries. In-silico simulations were done against Plasmodium falciparum protein (PDB ID: 7KJH), of the tested compounds Abyssinone V’ 4-methyl-ether (3) was found possess the best docking score of -4.4 Kcal/mol. The molecular docking of 7KJH was done to assess the inhibitory potential of the isolated compounds on protozoal parasites. Pharmacokinetic properties of the isolated compounds were also assessed in silico to assist in evaluating the drug likeness of these compounds. The compounds showed a percent human oral absorption of 100% except for Abyssinone V’ 4-methyl-ether (3), which showed 93.83%, this indicates a remarkable oral bioavailability. Stigamsterol (2) exhibited a Caco-2 cell permeability (QPPCaco) greater than 500 which indicates outstanding results for good intestinal absorption. The compounds also displayed a blood-brain partition co-efficient (QPlogBB) ranging from -1.433 to 0.128 suggesting they will have less potential to cross the blood-brain barrier, thus reducing any CNS related toxicity. Molecular networking of the crude extracts and the fractions was done through GNPS which allowed the identification of known compounds including one isolated in the present study, Abyssinone V’ 4-methyl-ether (3). Possible derivatives that have not been isolated from this plant before were also putatively identified. , Thesis (MSc) -- Faculty of Science, Chemistry, 2023
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Prosecution of forced marriage by international courts and tribunals in light of the principle of legality
- Authors: Okeyo, Julian Rebecca Atieno
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/420720 , vital:71772
- Description: Thesis embargoed. Probable release date in 2024. , Thesis (LLM) -- Faculty of Law, Law, 2023
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