Studies towards the development of novel antimalarial agents
- Authors: Adeyemi, Christiana Modupe
- Date: 2015
- Subjects: Antimalarials , Malaria , Drug resistance , Drug development , Enzyme inhibitors , Plasmodium
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54645 , vital:26596
- Description: Considerable efforts have been made in the modification of existing antimalarial drugs, and the support of incentive programmes have led to a drastic decrease in malaria cases reported by WHO during the past 6 years. However, the development of drug resistance threatens the eradication of this deadly disease and has prompted research on the synthesis of novel antimalarial drugs. Our research has involved the design and synthesis of novel benzylated phosphonate esters as potential 1-deoxy-D-xylose-5-phosphate reductoisomerase (DXR) inhibitors. A series of amidoalkylphosphonate esters were obtained by reacting various 3-subsituted anilines and heterocyclic amines with chloroalkanoyl chlorides and reacting the resulting chloroalkanamides with triethyl phosphite using Michaelis-Arbuzov methodology. Benzylation of the phosphonate esters afforded a series of novel N-benzylated derivatives in good yields and these compounds were fully characterised by NMR and HRMS methods. Several approaches to the introduction of a benzyl group at the C-2 position of the phosphonate ester derivatives have been explored, leading unexpectedly to the isolation of unprecedented tetrahydrofuranyl derivatives. Studies towards the preparation of potential bi-functional PfDXR / HIV-1 RT inhibitors have also been initiated. Preliminary in silico docking studies of selected non-benzylated and benzylated phosphonated derivatives into the Pf-DXR active-site has provided useful insight into the binding potential of these ligands. Bioassays have revealed a very low toxicity for all the synthesised phosphonated compounds and a number of these ligands also exhibit a promising inhibitory activity against the Plasmodium parasite.
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Synthesis and biological evaluation of truncated sarganaphthoquinoic acid derivatives as Hsp90 inhibitors
- Authors: Chiwakata, Maynard T
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64708 , vital:28594
- Description: Hsp90 inhibition has been at the centre of attention in current research due to the possibility of “cracking down” on the entire process leading to the development of malignant cancers. Small underlying principles common in all types of cancers have been determined that govern the transformation of normal human cells into cancerous cells, with all relying on the ATPase activity of Hsp90 protein. Hsp90 protein is therefore an attractive drug target that if successfully inhibited can result in the remission of cancer tumours by one form of treatment. To date, no Hsp90 inhibitor has been sanctioned for cancer treatment as most are still in clinical development. Our research was therefore inspired by reports that indicated the potential of quinones / naphthoquinones to act as Hsp90 inhibitors. Preliminary results of a few selected marine natural product quinone systems i.e. sargaquinoic acid (SQA) (2.47) and lapachol (3.6) showed moderate cytotoxicity and weak interactions with the Hsp90 molecular chaperone, and evidence suggested C-terminal binding of these molecules. No correlation has been determined yet between cytotoxicity and Hsp90 inhibition, hence we aimed to develop natural product inspired molecules that exhibit both cytotoxic and Hsp90 inhibition properties. Due to limited amounts of the natural product that can be acquired from natural sources, synthetic analogues were opted for. Isolation of a few selected quinones was conducted to have material that could be used in biological assays. For structural modifications, a series of truncated naphthoquinone systems were prepared adopting the sarganaphthoquinoic acid (3.5) scaffold. The naphthoquinones were prepared via Diels-Alder reactions of relevant benzoquinones with myrcene, followed by aromatization reactions using MnO2. Various alkyl and aryl amines were then coupled to the C-2/3 position of the naphthoquinone using Michael’s addition reactions. Tricyclic naphthoquinones were also synthesized from reactions with hypotaurine and citral. Design of the analogues incorporated functionalities from known Hsp90 inhibitors e.g. geldanamycin (2.28) and its analogues. Preliminary results obtained showed that coupling of naphthoquinones with aryl-amines resulted in the most cytotoxic compounds (4.14-4.19) with IC50 values as low as 0.3 μM against Hs578T breast cancer carcinoma (triple negative). Most of the alkyl amines (4.20-4.25) had IC50 values greater than 50 μM except for 4.20 and 4.21 that showed IC50 values of 7.6 μM and 2.6 μM respectively. Tricyclic naphthoquinones (4.28-4.29) showed moderate cytotoxic activity of approximately 10 μM. Hsp90 inhibition was assessed by client protein degradation assays, of which SQA (2.47), showed the best Hsp90 inhibition properties, followed by compound 4.20. The most cytotoxic arylamino-naphthoquinone (4.16) and tricyclic naphthoquinones (4.28-4.29) showed only moderate inhibition. None of the compounds led to Hsp70 induction, suggesting possible binding to the C-terminus of Hsp90. Interactions at the binding site were assessed by molecular docking studies and saturation transfer difference (STD) NMR. Docking studies were conducted on the N-terminus of Hsp90 and better binding was observed for arylamino naphthoquinones (4.14-4.19) than for other series of compounds. Unfortunately, the co-crystal structure for the C-terminus of Hsp90 is unavailable, hence docking study comparisons on both domains could not be conducted. However, STD NMR offered a platform to assess binding interactions between the naphthoquinones and the N- or C-terminal domains of Hsp90. However no interactions were observed at both the N- and C- termini of Hsp90 due to either weak binding of ligands to the protein or poor water solubility of the ligands. From these preliminary results, naphthoquinones bind to Hsp90 protein but conclusive remarks to which terminal domain they bind to could not be made. The best candidate from amongst the series of naphthoquinones prepared that showed moderate cytotoxicity and promising Hsp90 inhibition was compound 4.20. We therefore succeeded in developing a new series of naphthoquinones that possess moderate cytotoxicity and show Hsp90 inhibition.
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The development of biological tools to aid in the genetic investigation of the black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros mitochondrial genomes
- Authors: Parsons, Michelle
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/56059 , vital:26769
- Description: The black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros are found in South Africa. A decline in the populations of these species has resulted due to human activities such as habitat fragmentation and poaching. This has contributed to the loss of genetic diversity amongst the black and white rhinoceros. Conservation and anti-poaching efforts are needed to help maintain genetic diversity. These efforts could be improved through the development of non-invasive techniques to examine DNA from threatened animals. The aim of this research was to develop a molecular technique which would allow for the identification of the black and white rhinoceros and to develop a molecular technique which would allow for intraspecies genetic variation to be examined. DNA extractions were performed on matched faecal and tissue samples that were collected from two regions in South Africa. Polymerase chain reaction (PCR) primer sets were designed to investigate several regions of the rhinoceros mitochondrial genome. PCR optimisation was completed for the target regions. Sequencing was conducted on all final PCR products. The cytochrome c oxidase subunit 1 (COIi) gene allowed for the rhinoceros family to be identified. This region was digested with the HindIII restriction enzyme, which allowed for the specific identification of either the black or white rhinoceros. A subsequent region of the cytochrome c oxidase subunit 1 (COIii) as well as the D-loop, hypervariable regions (HV1 and HV2), cytochrome b (cytb) and 16s rRNA regions were investigated. These regions displayed potential for establishing geographic origin for black rhinoceros samples, whereas the D-loop and HV2 show potential for the white rhinoceros. The white rhinoceros displayed sequence variation in the HV2 and cytb region, while variation was observed in the COIi and HV1 for the black rhinoceros. All investigated target regions allowed for the rhinoceros family to be identified. The COI (COIi and COIii), HV2 and cytb regions allowed for the subspecies of rhinoceros to be identified, however the D-loop was not able to identify the white rhinoceros species. The 16s rRNA and HV1 regions allowed for the correct subspecies of rhinoceros to be identified, however as the primers were only compatible for the black rhinoceros therefore a subsequent investigation is required for the white rhinoceros. The establishment of this novel PCR based technique to identify white and black rhinoceros will allow for efficient species identification in wildlife forensic cases. A biological method was established to study intraspecies variation for the white and black rhinoceros; however the investigated target regions did not yield sufficient genetic variation. The core techniques developed in this study will be valuable for future studies that wish to investigate genetic variation in mammal species.
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The diets of co-occuring anurans in a small South African river: assessments using stomach contents, stable isotope ratios and fatty acid profiles
- Authors: Sikutshwa, Likho
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54405 , vital:26562
- Description: During their life cycle amphibians change their habitat and feeding, and are important consumers in both terrestrial and aquatic habitats. The trophic ecology of anurans has been studied intensively using gut content analysis, but very little work has been done using biochemical techniques such as stable isotope and fatty acid analyses. These biochemical techniques produce data that allow for additional inferences about the trophic ecology of anurans, as they reveal what the organisms assimilated over time rather than what was recently ingested. The investigation of the feeding ecology and trophic interactions of anurans using gut content analyses together with stable isotope and fatty acid analyses will help us to better understand their ecological roles. The objective of this thesis was to assess the feeding ecology of four anurans (Amietia angolensis, Amietophyrnus rangeri, Strongylopus grayii and Xenopus laevis) in a temperate river using a combination of gut content, stable isotope and fatty acid analyses. Frogs, tadpoles and potential food sources were sampled in two sites (upstream and downstream) in the Kowie River, Eastern Cape Province, South Africa. Gut content analyses identified 147 prey items belonging to 12 prey orders in the stomachs of the sampled specimens. In both A. angolensis and S. grayii the most important prey category was Coleoptera, followed by Hemiptera, Diptera and Hymenoptera at both sites. Aquatic prey items (aquatic hemipterans, chirinomids and blackflies) were the most important food sources for X. laevis (Alimentary Index (IAi) = 6.4; 5.2; 4.2). In Am. rangeri, Hymenoptera was the most important prey category (IAi = 8.3). The trophic niche overlap between A. angolensis and S. grayii was biologically significant (> 0.6), and in the remaining species there was no significant trophic niche overlap. Amietia angolensis showed a larger trophic niche (Levin’s measure; B = 7.7 and Bst = 0.84 downstream, and upstream B = 7.6 (Bst = 0.82) compared to the other species. The gut content analyses showed that frogs feed on a variety of prey items that constitutes food sources from both aquatic and terrestrial habitats. Stable isotopes indicated that aquatic derived sources contributed significantly more towards the diets of X. laevis, A. angolensis tadpoles and S. grayii tadpoles compared with the other anurans, whereas aquatic and terrestrial derived food sources contributed equally to the diets of A. angolensis and S. grayii. Increased trophic positions in A. angolensis and S. grayii occurred throughout their development. The four different species had similar fatty acid profiles in the upstream region, and fairly similar δ13C values, suggesting that they probably consumed similar food. Fatty acid profiles of anurans in the downstream region showed distinct separations among the species. Tadpoles had high levels of diatom-associated fatty acids (20:5ω3; A. angolensis tadpole – 8.4 %, S. grayii tadpole – 9.4 % upstream and downstream; 9.1 and 6.1 % total fatty acids (TFA), respectively). All four species had substantial contributions from bacterial fatty acids, and large proportions of saturated fatty acids (30.6 - 50.0 %) including those with 14 and 18 carbons, indicating that bacterial and detritus food sources played an important role in their diets. The fatty acid profiles revealed high proportions of polyunsaturated fatty acids (PUFAs) and essential fatty acids (EFAs) in all species, indicating a good quality of food and that the quality of food consumed was similar among species. The results demonstrated the usefulness of a combination of traditional techniques (gut content techniques) and biochemical techniques (stable isotopes and fatty acid analysis) for assessing consumption and assimilation. The amphibian assemblages examined derived much of their energy from terrestrial and aquatic sources. This information will allow more precise and comprehensive assessments of trophic interactions in freshwater habitats, along with aiding in future amphibian conservation and management efforts.
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The economics of institutions, institutional governance and efficiency: the case of water distribution in Lower Sundays River Valley
- Authors: Madigele, Patricia Kefilwe
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MCom
- Identifier: http://hdl.handle.net/10962/54777 , vital:26611
- Description: The institutional dynamics, policies and legislation that were prevalent during the apartheid era have left imprints that are difficult to ignore as they still dictate the interaction between different elements in the water sector to date. During the apartheid era, the formulation of policies was informed by racial segregation, resulting in a socio-economic pattern that dictated the distribution and access of resources for the people of different races in the country. Post-apartheid, the National Water Act has established the basis for management of water resources on a catchment basis (for equity, efficiency and sustainability), and the Water Services Act aims to ensure everybody has access to basic water supply and sanitation services. Regardless of the improvements in water supply to the rural sector made by the South African government, many of the current patterns of water use are still characterised by inequality, inefficiency, and inadequacy. The poor remain marginalised, and emerging farmers and poor rural communities have limited access to water resources while water continues to be used inefficiently by some farmers in the agricultural sector with few incentives to improve its water use efficiency. Despite the existence of the thriving citrus industry in the area, around 60% of people in the Sundays River Valley Municipality (SRVM) live below the poverty datum line. The inequality between the municipal populace and the commercial citrus industry is noticeable and the inequitable water redistribution is prevalent in the Lower Sundays River Valley (LSRV). The problem of disrupted water supply is prevalent in the catchment. However, there is also currently no physical shortage of water in the catchment. Therefore, the currently experienced problems with water supply in the LSRV are consequence of a lack of effective institutions and infrastructure, not of physical water scarcity. It is argued in this paper that there is a notable lack of understanding about the design of institutions for water management in developing countries. The vast majority of research on water management and access is premised on neoclassical economics ideas related to water markets and pricing among others. The neoclassical economics approach, however, does not adequately define the role of institutions in shaping the direction of water access and supply. This study uses new institutional economics (NIE) arguments to define the institutional arrangements and dynamics defining the water sector in South Africa, using the Lower Sunday River Water Users Association (LSR-WUA) as the case study. It aims at analysing the institutional governance and performance of the using equity, efficiency and effectiveness as key indicators. The various research methods employed in this study include; interpretive and post-positivist paradigms, quantitative and qualitative research, the case study research method and in-depth key informant interviews. It is concluded that that the current and future decisions made by the LSR-WUA are not entirely independent of those made in the past under Sundays River Irrigation Board (SRIB). The thesis argues that such factors as old effective networks, vested interests of commercial farmers, sunk costs towards the building of canals, among other factors, may have influenced the dependence of the LSR-WUA on the SRIB’s set path. It is further concluded that the absence of contractual agreement between the LRS-WUA which acts as the bulk water supplier, and the SRVM which acts as both the water services authority (WSA) and the water service provider (WSP) creates an institutional arrangement deficiency. Such an institutional arrangement vacuum can lead to a failure of the water institutions in the catchment to provide water resources effectively. The study further argues that because the post-apartheid National Water Policy of South Africa is largely influenced by neoclassical economics foundations, the desired results in the water sector, such as equitable distribution of water resources, have not yet been fulfilled completely.
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The effect of extracellular Hsp90β and TGF-β1 on colon cancer biology
- Authors: Perks, Tamarin
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/55896 , vital:26753
- Description: The TGF-β signaling pathway is known to be one of the most commonly mutated pathways in human cancers, while Hsp90 is a bone fide drug target that is involved in regulating the conformation and activity of many oncoproteins. The role of intracellular Hsp90 in cancer has thus far been established and there is a growing link between extracellular Hsp90 and cancer metastasis, as well as the role of TGF-β in metastasis. This study aimed to analyse the interaction between Hsp90 (both intracellular and extracellular) and the TGF-β machinery in cancer cells, as well as to determine the effect of these proteins on cellular responses on the biology of cancer cells. This was achieved by studying the expression of Hsp90; TGF-βRII and TGF-β1 in cancer cell lines of various origins using flow cytometry, ELISA, and western blot analysis. The genetically paired SW480 and SW620 colon cancer cell lines, derived from a primary tumour and lymph node metastasis, respectively, were selected for further study due to differences in expression levels and activation of the TGF-β1 pathway. SW480 cells expressed double the level of TGF-βRII compared to SW620 cells, while SW620 expressed two times more extracellular TGF-β1 than SW480 cells. A direct interaction between TGF-β1 and Hsp90β was determined in vitro, and confirmed in vivo in SW620 cells. Growth, adhesion and migration were analysed in SW480 and SW620 cells. SW480 cells adhered significantly faster than SW620 cells, while SW620 cells had a greater rate of migration. Inhibiting the TGF-β pathway, specifically TGF-βRI, using SB 431542, as well as inhibiting Hsp90 with novobiocin, caused an increase in migration in SW480 cells. Only the addition of TGF-β1 in combination with Hsp90 as well as SB 431542 caused an increase in migration in SW620 cells. The canonical TGF-β1/TGF-βRI/TGF-βRII pathway may be constitutively active in SW620 cells and the inhibition of TGF-βRI may suggest an alternate pathway or receptor in both SW480 and SW620 cells.
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The isolation, characterisation and chemotaxonomic significance of secondary metabolites from selected South African Laurencia spp. Rhodophyta
- Authors: Fakee, Jameel
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64696 , vital:28593
- Description: Bioprospection of marine organisms as a potential source for lead drugs is becoming increasingly popular. The secondary metabolome of these organisms consists of structurally diverse molecules possessing unprecedented carbon skeletons, the biosynthesis of which occurs via complex metabolomic pathways driven by specialist enzymes. This structural novelty is highly influential on the favourable bioactivity these compounds display. A prominent example of such a compound is trabectedin marketed as Yondelis®. Registered for the treatment of soft tissue sarcomas, this marine drug was developed from extracts of the tunicate Ecteinascidia turbinata. South Africa is renowned for possessing a highly diverse marine biota including several endemic species of marine red algae belonging to the Laurencia sensu stricto genus, which falls within the Laurencia complex. Despite having a good reputation for fascinating secondary metabolites, the taxonomy of Laurencia natural products is proving challenging for reasons including the presence of cryptic species, as well as individual species displaying morphological variability. The aim of this study was thus to isolate secondary metabolites from various South African Laurencia spp. and subsequently assess their chemotaxonomic significance by analysis of a parallel plastid rbcL phylogeny study of Laurencia spp. This study reports the first phycochemical investigation into Laurencia natalensis Kylin, Laurencia cf. corymbosa J.Agardh, Laurencia complanata (Suhr) Kützing, Laurencia sodwaniensis Francis, Bolton, Mattio and Anderson submitted, Laurencia multiclavata Francis, Bolton, Mattio and Anderson submitted, and a South African specimen of Laurenciella marilzae Gil-Rodríguez, Sentíes, Díaz-Larrea, Cassano and M.T. Fujii (basionym: Laurencia marilzae) originally described from Spain. Additionally, the chemical profiles of previously explored species Laurencia flexuosa Kützing and Laurencia glomerata Kützing were re-investigated. The organic extracts of the above species afforded 31 compounds belonging to a wide array of structural classes including halo-chamigranes, linear C15 acetogenins, indole alkaloids, cuparanes and cyclic bromo-ethers. A new tri-cyclic keto-cuparane (4.4) was isolated from L.cf. corymbosa alongside the new cuparanes 4.1 and 4.7. Algoane (5.9), a unique marker compound isolated from L. natalensis, was previously only reported from a sea-hare. Such marker compounds which are exclusive to an individual algal species increase the ease of their subsequent identification. The feasibility of chemotaxonomy as an additional tool to classify Laurencia spp. Was established as broad predictions of a specimen’s phylogeny, based on representatives of its secondary metabolome, proved viable. The study specimens were shown to possess similar chemical profiles to their sister species e.g. L. complanata, L. sodwaniensis and L. multiclavata produced similar metabolites to their sister species as inferred by an rbcL phylogeny tree. Finally, a 1H NMR profiling study on the crude organic extracts of various Laurencia spp. generated distinctive, reproducible spectra, exposing the value of NMR spectroscopy as a rudimentary species discernment tool.
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The role of leadership style and organisational structure in organisational effectiveness: a case study
- Authors: Messaris, Annette
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MCom
- Identifier: http://hdl.handle.net/10962/54452 , vital:26566
- Description: This research explores the role leadership style and organisational structure play in organisational effectiveness. Organisational effectiveness is one of the more complex terms to define although essential to understand in order for an organisation to grow and develop. A small to medium wine sales and distribution organisation, its leadership style and organisational structure were chosen as a focus of this case study. Organisational effectiveness in this study is understood by the terms growth and development using the Greiner Theory of Evolution and Revolution (1983). The literature explores the role of leadership style in organisational effectiveness by exploring various theories and focusing on The Full Range Leadership Model by Bass and Avolio (1994). In order to analyse the role of organisational structure in organisational effectiveness, the different forms of organisational structure and the role they have on the organisation’s ability to grow and develop are explored. The primary purpose of this study is to investigate the role leadership style and organisational structure play in organisational effectiveness. A qualitative content analysis paradigm was used with a Case Study method. The data was gathered using structured interviews conducted on all 39 permanent staff members, day to day observation and the Multifactor Leadership Questionnaire (MLQ). The MLQ is used as a supporting tool to verify the leadership findings in the interviews. The data was analysed using NVivo10 and STATISTICA to gain insight into the leadership style and organisational structure of the organisation. The findings reflected that certain themes were repeatedly mentioned in the interviews and the definition of organisational effectiveness: structured leadership, more active leadership, lack of communication, staff motivation, staff training and development and organisational culture. As regards organisational structure, the following themes arose: The need for regular meetings, increased team work and more structured job descriptions. Through the analysis of all the components, leadership style and organisational structure were identified as having a significant role in organisational effectiveness which will be further analysed in this study.
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Thermal physiology and behavioural ecology of the white shark, carcharodon carcharias
- Authors: Gennari, Enrico
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64002 , vital:28522
- Description: Expected release date-May 2019
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Towards a Mobile Bioethanol Unit for point of source conversion of sugar sources to bioethanol: design and feasibility study for South Africa
- Authors: Cech, Alexandra Louise
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59141 , vital:27439
- Description: Restricted access-thesis embargoed for 5 years
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“Bold Distinguished You”: a critical discourse analysis of the representation of masculinity in Destiny Man, 2012 - 2013
- Authors: Hutcheson, Lauren
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/56032 , vital:26767
- Description: Radical social and economic changes have transformed the employment opportunities available to black men in South Africa, and consumer magazines like Destiny Man have been quick to respond. Historical shifts in the configuration of an ideal black masculinity have been proposed over time in magazine representations of black men. Critiques of the representations of black men in consumer magazines pre- and post- independence interrogate the politics of the ways in which black masculinity has been, and is, configured around classed identities. In this study, I present the findings of a critical discourse analysis (CDA) on the construction and representation of masculinity within a contemporary South African’s men’s lifestyle magazine, Destiny Man. This study presents an analysis of the subject positions that this magazine constructs for its male readers. Destiny Man proposes particular kinds of discursive subjects in its sets of representations, both in word and image. The images and copy validate a neo-liberal elite masculinity achieved through self-surveillance and a variety of technologies of self, with the ultimate focus on material success. I argue that these representations have implications for the development of masculinities in a context marked by extreme inequality.
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