This work is a preliminary attempt to study the effect between thermal shock and abrasive wear in WC-Co alloys. This was done by evaluating the thermal shock resistance of a WC-12wt%Co mining grade as a function of temperature, number of thermal shock cycles and making comparisons between the abrasive wear responses of samples subjected to thermal shock and samples not subjected to thermal shock. A furnace was designed for the thermal shock treatments. Abrasive wear tests were performed on a 2-body sliding wear apparatus using 80-grit SiC abrasive paper as a counter-face. Stereo and electron microscopy as well as microprobe techniques were used to analyse the effects of thermal shock. It is confirmed that thermal shock has a negative effect on the wear rate of WC-12wt%Co. The results showed an initial high mass loss rate during abrasive wear testing, which increased with increasing temperature and a decrease in wear rate with time until the wear rates converged for all samples. The surface analysis after thermal shock indicated voids on and below the surface, stained surfaces, a thin oxide layer and the possibility of WC decarburization which accelerated the wear response.

The human non-integrin laminin receptor (LamR) is a multifunctional protein which is localised to a number of sub-cellular locations. LamR is a component of the ribosome and has a number of intracellular functions; it also acts as an extracellular receptor for laminin, growth factors, pathogenic microorganisms, prion proteins, toxins and the anticarcinogen epigallocatechin gallate (ECGC). Although LamR is present in most cellular compartments, its overexpression in many types of cancer cells suggests a vital role for LamR in tumor-cell migration and invasion. There are two isoforms of laminin receptor: the monomeric 37-kDa laminin receptor precursor (37LRP) and the mature 67-kDa laminin receptor (67LR). Although the precise molecular nature of 67LR is unclear, accumulating evidence strongly suggest that 37LRP can undergo homo- and/or hetero-dimerization with Galectin-3 (Gal-3) to form mature 67LR. A recent study demonstrated that both homo- and heterodimer LamR forms are present on the cell surface, where they form distinct populations. Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae type b (Hib) are major causes of bacterial meningitis. The contribution of LamR in traversal of the blood brain barrier (BBB) by neurotropic viruses is well established and interaction with LamR was recently found to be critical for initiation of bacterial contact with the blood brain barrier (BBB). These bacteria bind LamR via the surface protein adhesins: meningococcal PilQ and PorA; pneumococcal CbpA; and H. influenzae OmpP2. Further investigations showed that the fourth and second extracellular loops of meningococcal PorA and OmpP2 of H. influenzae, respectively, are responsible for LamR binding. The work presented here consists of two complementary projects in which a number of approaches were taken to characterise the ligand-binding domains of LamR. The first project aimed to identify sites on LamR that are critical for binding the ligands of bacterial meningeal pathogens. The second project aimed to identify residues that contribute to the stability of LamR homodimers and the heterodimer with Gal-3. Several mutations were introduced into full-length human LamR, either by deletion mutations within the C-terminal domain (CTD) of LamR using inverse polymerase chain reaction (IPCR), or by single-amino acid substitution in the N-terminal domain (NTD) of LamR using site-directed mutagenesis. Protocols for large-scale expression of full-length and truncated LamR proteins in human cells were developed, as well as non-denaturing purification protocols. We hypothesised that bacteria-binding domains could be located on both the NTD and CTD of LamR. In vivo examination using ELISA assays, in which the interaction of LamR and whole bacteria or purified recombinant PorA or PilQ were tested identified several novel sites on LamR that contributed significantly to binding of the bacterial ligands. These sites include amino acids 206-229 and 263-282, located within the CTD, and Tyrosine 156 in the NTD, each of which contributed to the binding of meningococcal PorA, and more specifically it’s fourth extracellular loop. Furthermore, three sites on LamR comprising amino acids 206-229 and 263-282 within the CTD and Tyrosine 139 in the NTD were shown to contribute to binding pneumococcal CbpA, OmpP2 and Loop two of OmpP2 of H. influenzae. These results indicate that the three neuroinvasive bacteria share the same binding sites on LamR. Bimolecular fluorescence complementation (BifC) coupled with flow cytometry and confocal microscopy revealed the vital contribution of amino acid residues Arginine 155, Tyrosine 156 and Lysine 166 (all within the NTD of LamR) for the homodimerization and heterodimerization of LamR with Gal-3. The dimerization of two meningococcal host receptors, LamR and Gal-3, may help to extend spectrum of their bacterial adhesins, which may act cooperatively or synergistically at different stages of infection. Information about the residues in LamR that contribute to the stabilization of LamR dimers has implications for the role of LamR dimers in the pathogenesis of bacterial meningitis. Identification of bacteria-binding domains on LamR is of particular interest for development of vaccines or therapeutic interventions that provide protection against the three major meningitis-causing bacteria. The aim of the current work was first to localise the domains of LamR responsible for binding with PorA and PilQ of N. meningitidis; CbpA and OmpP2 of S. pneumonia, and OmpP2 of H. influenzae. Also, previous studies have shown conspicuous homodimerisation of 37LRP and heterodimerisation with Gal-3. Our second aim was to identify of amino acid residues involved in 37LRP self-association and heterodimer formation with Gal-3. In current work, several regions of LamR were hypothesised to constitute the binding site for the bacterial ligands; these predictions were based on previous studies on LamR binding domains and the crystal structure of laminin receptor. Also, to investigate both homo and heterodimerisation of LamR, the involvement of several putative amino acid residues within 37LRP in LamR dimerisation was was hypothesised.

HIV, syphilis, malaria, and anaemia are major causes of adverse pregnancy outcomes in sub-Saharan Africa (SSA). Despite global and national policies advocating for screening of these conditions, only HIV testing has achieved good coverage, precluding early detection and appropriate management in pregnancy. Rapid pointof-care tests (POCTs) provide an opportunity to integrate diagnosis and provide timely treatment of these conditions in rural antenatal care (ANC) settings. After an introductory chapter, a review of the literature on these four conditions in pregnancy is presented with a focus on SSA. The thesis then shifts attention to Kenya, a country that embodies many of the disease challenges and health system characteristics of the region. Kenyan ANC policy recommends testing for HIV, syphilis and anaemia and preventive strategies for malaria. The following chapters are comprised of three linked studies conducted in western Kenya, that use different methods to progressively investigate the implementation success of integrated point-of-care testing (POCT) for HIV, syphilis, malaria and anaemia at seven peripheral dispensaries. Baseline data confirmed that testing requirements for syphilis, malaria and anaemia are not currently met at dispensary level. We implemented an intervention where test kits were supplied and training plus supervision were provided to enable healthcare workers to conduct integrated POCT for pregnant women. Adoption and fidelity were measured quantitatively using exit interviews, antenatal registers and proficiency scores (Study 1: Integrating point-of-care testing (POCT) for HIV, syphilis, malaria and anaemia in antenatal care at dispensary level in western Kenya: an implementation study) while acceptability, appropriateness and feasibility were assessed qualitatively (Study 2: Exploring healthcare workers and pregnant women’s perspectives on appropriateness, acceptability and feasibility of integrating point-of care testing: A qualitative study). Our findings show that the innovation was highly adopted, meaning almost all pregnant women received the essential tests. This was supported by the qualitative findings where healthcare workers and pregnant women found the innovation acceptable and appropriate. However, fidelity to clinical management guidelines can still be improved. Our qualitative findings provide some explanation for these gaps. One common sentiment among interviews with healthcare workers was that workload was perceived to be a barrier to providing quality care. We explored this further with discrete-event simulation modelling (Study 3: Investigating the operational impact of integrating HIV, syphilis, malaria and anaemia point-of-care testing in antenatal care clinics in western Kenya: a discrete event simulation model) and found the healthcare workers were actually under-utilized. This suggests that nurses should, in theory, have sufficient time to deliver essential ANC services. While integrating POCT addresses one gap, additional interventions to support and supervise healthcare workers are needed to ensure appropriate and high quality of care. An integrated approach to health systems strengthening and more investment in implementation and translation research using multi-methods are needed.

The microstructures and performance of Ni-based alloys and Ni-WC (nickel-tungsten carbide) composite overlays deposited by plasma transferred arc welding have been studied. The Ni-alloy overlays had similar microstructures consisting of Ni dendrites, with interdendritic Ni-based eutectics, borides and carbides. Low hardness alloy overlays contained a smaller fraction of interdendritic phases relative to the high hardness alloys. The interdendritic regions make a significant contribution to the hardness since they are more than twice as hard as primary dendrites. The Ni-WC composites contained similar phases, however WC dissolution was observed leading to the formation of other carbides. Ni-alloys with low carbon and Cr content exhibited the lowest WC dissolution. The Ni-WC overlays produced using these dilute alloys generally performed better in ASTM G65 wear tests. This was likely due to the reduced dissolution which avoided formation of brittle secondary phases, maintaining a short mean free path between WC particles and allowing increased impact energy absorption. / Materials Engineering

In this dissertation, an electrical discharge coating uses an isolated sleeve to form a closed space between the end surface of electrode and the work to deposit a thin film onto a substrate. The discharge occurs at the location where the two surfaces are closest and the dielectric fluid ionizes at this location to create a path for the discharge. Hence, this closed space is heated to extremely high temperature, so that a small portion of the work surface is suddenly melted with the particles in the dielectric fluid and then coated to increase its coating speed and quality. The electrode material is made of brass, the work material SKD11, and the dielectric fluid is kerosene with the WC powder concentration of 50g/L. The pulse-on and pulse-off times are 25 and 500£gs, respectively. The effects of supply voltage, electrical discharge coating time, electrical discharge gap, and powder added cycle on the coating characteristics are investigated. According to the experimental results, the electrical discharge with isolated sleeve can achieve a complete coating layer onto the surface of work. The coating thickness increases with increasing electrical discharge coating time and gap as the supply voltage is larger than the threshold voltage of electrical discharge. At the supply voltage below 250V and the added powder cycle less than 10, the coating thickness increases with increasing supply voltage and cycle. The quality of coating layer is better at the low gap distance and the high supply voltage. The hardness of coating layer is about HV 1687 which is approximately 5 times of substrate hardness using micro-hardness test. The electrical discharge without isolated sleeve cannot achieve the above-mentioned advantages.

Infection with hepatitis C virus (HCV) is cleared spontaneously in only 20% of cases with the majority of individuals developing a chronic infection. This discrepancy in disease outcome is incompletely understood but current understanding of the immune response to HCV suggests that rapid induction of a broadly neutralising antibody (nAb) response leads to resolution of acute infection. The majority of nAb identified target the envelope glycoproteins, particularly E2, and most appear to inhibit binding of E2 to the cellular receptor CD81. Antibodies targeting other interactions, such as those with the receptor CLDN or the fusion determinant, are underrepresented in the repertoire of anti-HCV antibodies. However, the antibody discovery process may have been biased by the nature of the assays used. Therefore new assays are needed to enable the discovery and characterisation of antibodies in an unbiased manner. To facilitate this, a novel insect cell display library was developed for mapping antibody-binding epitopes. Cells expressing specific E2 mutants provided the necessary proof-of-principle that loss of antibody binding could be detected in this system before a library expressing randomly mutated E2 was developed. Sorting experiments demonstrated that single cells could be isolated and enriched based on loss of antibody binding. Secondly, a method for characterising the immunoglobulin (Ig) genes of HCV infected patients was developed; Ig genes were isolated from small numbers of B cells and the sequences analysed. Finally, a patient cohort was studied with a view to investigating the evolution of the antibody response during early infection. The unreliable nature of the samples prevented such analysis; however a DNA fingerprinting method of testing the origin and relatedness of serum samples was developed. This will improve the reliability of future studies. Together these methods provide a work model for the assessment of samples and the isolation and characterisation of antibodies.

No description available.

Student Number : 0109917P - PhD thesis - School of Process and Materials Engineering - Faculty of Engineering and the Built Environment / The aim of this project was to characterise new WC-10VC-Co powders, and to deposit WC-10VC-Co thermal spray coatings from these powders for characterisation and testing in adhesion, wear and corrosion tests. Throughout the project, the new powders and coatings were compared to commercial WC-Co powders of the same binder content and commercial WC-Co thermal spray coatings. All the powders i.e WC-10VC-Co and WC-Co powders, were produced by agglomeration (by spray drying) and sintering and characaterised by determining the sizes and size distributions of the powders' particles, the morphology, the flowability and the phase composition and grain size and size distribution of carbide grains. The vanadium carbide in the WC-10VC-Co powders occurred in the solution as the double carbide (V,W)C and the carbides present in the WC-10VC-Co powders were WC and (V,W)C. None of the starting VC was left in the powders. Coatings were deposited using high pressure high velocity oxy-fuel (HP/HVOF) spraying systems, and characterized by determining the microstructures, the phase compositions and the carbide grain sizes, as had been done for the powders. Three types of tests were done on the coatings: adhesion tests, (according to standard SNECMA 14 -008); dry abrasion, wet abrasion and slurry erosion tests; and corrosion tests, in synthetic mione water. Thermal spraying lead to some WC decarburization to W2C and eta phase, and to the formation of amorphous binder. The W2C grains from the WC decarburization formed in the amorphous binder matrix of coatings. All the coatinge were porous, but the new WC-10VC-Co coatings were more porous than the commercial Wc-Co coatings because the spray parameters had only been optimized for the WC-Co coatings. The carbide grains decreased in size by as much as 50% during decomposition. Evidence suggested that the WC grains in the coatings were subjected to different residual stresses that in the powders, probably due to the formation of the amorphous binder. Vanadium carbide in the Wc-10VC-Co coatings occurred as (V,W)C, just as in the powders, with as distribution that was reasonably homogeneous. The apparent hardness of the new Wc-10VC-Co coatings was slightly lower than that of WC-Co coatings of the same cobalt content, due to their higher porosity. The adhesion of the new Wc-10VC-Co coatings was as good as that of the Wc-Co coatings. The dry and wet abrasion resistance of the new Wc-10VC-Co coatings was better that for the Wc-Co coatings of equal Co wt%, on account of the Wc-10VC-Co coatings having a lower binder volume fraction, finer carbide grains, and (V,W)C grains. The (V,W)C grains are harder than WC grains and apparently slowed down the overall abrasion rate. In slurry erosion, the best performance of the Wc-10VC-Co coatings was as good as that of the commercial WC-Co coatings at equal cobalt mass content, due to the higher porosity of the Wc-10VC-Co coatings, apparent faster erosion of the harder but brittle (V,W)C grains, and, from what evidence appreared to suggest, generally slightly poorer erosion resistance of the fine WC grains under the test conditions used. Polishing the slurry erosion test specimens reduced mass losses in slurry erosion by factor of up to 10 compared to the unpolished specimens, and led to better erosion resistance of the WC-10VC-Co coating compared to the WC-12Co coating. The results of the tests done to investigate the corrosion properties of the coatings were conclusive. This is because the effects of cleaning procedures on mass loss after immersion corrosion were not explored, and it appeared, for some coatings, that the corrosion mechanisms in immersion corrosion could not be reproduced in electrochemical testing.

Les nanomatériaux sont utilisés dans de nombreux secteurs industriels, et plusieurs produits de consommation contenant des nanomatériaux sont d’ores et déjà commercialisés. Dans ce contexte d’exposition humaine croissante aux nanomatériaux, l’évaluation de leur potentiel génotoxique est d’une importance significative. Cependant, la pertinence des tests classiques de génotoxicité, développés pour des produits non nanoparticulaires, est fréquemment remise en question pour l’évaluation des nanomatériaux. Un témoin positif de référence sous forme nanoparticulaire pourrait donc constituer une avancée importante pour l’évaluation de la génotoxicité des nanomatériaux, permettant de s’assurer que les systèmes d’essais sont appropriés et/ou d’en valider de nouveaux.Dans un premier temps, nous avons étudié la possibilité d’utiliser des nanoparticules de carbure de tungstène – cobalt (WC-Co) commerciales, préalablement caractérisées sur le plan physico-chimique (distribution de taille et charge dans les milieux utilisés), comme témoin positif dans trois essais de génotoxicité in vitro. Le test de mutations géniques au locus thymidine kinase sur cellules de lymphome de souris, le test des micronoyaux étudiant les dommages chromosomiques et le test des comètes détectant les dommages primaires à l’ADN ont ainsi été réalisés, les deux derniers essais dans deux types cellulaires, la lignée de lymphome de souris L5178Y et des cultures primaires de lymphocytes humains. Nos résultats montrent que les nanoparticules de WC-Co pourraient être utilisées comme témoin positif dans ces essais de génotoxicité in vitro, selon le type cellulaire et le schéma de traitement.Nous avons ensuite étudié les mécanismes d’action impliqués dans la génotoxicité des nanoparticules de WC-Co. Le marquage des centromères dans les micronoyaux grâce à la technique d’hybridation in situ en fluorescence (FISH) montre l’implication d’évènements clastogènes et aneugènes. Ces résultats ont été confirmés par un essai d’aberrations chromosomiques sur lymphocytes humains bloqués en métaphase, avec l’observation de cassures de chromatides et de cellules polyploïdes. Par ailleurs, les mécanismes oxydants étant les plus décrits pour les nanomatériaux, nous avons étudié les lésions oxydatives à l’ADN en utilisant le test des comètes in vitro modifié avec l’enzyme de réparation de l’ADN formamidopyrimidine DNA glycosylase (FPG). Nous avons également détecté par résonance paramagnétique électronique une production de radicaux hydroxyles après mise en suspension des nanoparticules de WC-Co en présence et en absence de cellules. Dans le cadre d’études haut-débit des nanoparticules de WC-Co réalisées dans trois lignées cellulaires humaines correspondant aux principaux organes cibles pour les nanomatériaux (la lignée pulmonaire A549, la lignée hépatique Hep3B et la lignée rénale Caki-1), il a été confirmé que le stress oxydant joue un rôle important dans la toxicité des nanoparticules de WC-Co. En effet, la production d’espèces réactives de l’oxygène dans les cellules traitées avec les nanoparticules de WC-Co était corrélée avec l’observation d’une cytotoxicité et de génotoxicité, étudiée à l’aide du test de détection des foyers γH2AX.Finalement, nous avons appliqué les tests de génotoxicité les plus pertinents à l’étude de nanodiamants et de nanocapsules lipidiques, qui constituent des candidats prometteurs pour la vectorisation de principes actifs. Les tests des comètes et des micronoyaux in vitro ont ainsi été réalisés sur d’autres types cellulaires mimant des organes cibles : la lignée intestinale T84 et la lignée bronchique 16-HBE exposées à des nanodiamants de trois tailles différentes et des lymphocytes humains exposés à des nanocapsules lipidiques de 3 tailles et 3 charges différentes. / Nanomaterials are used in many industrial sectors, and many nanomaterial-containing consumer products are already available. In this context of increasing human exposure to nanomaterials, the evaluation of their genotoxicity is of significant importance. However, the relevance of routinely used genotoxicity assays, developed for non-nanoparticular products, is often questioned for the evaluation of nanomaterials. A nanoparticulate reference positive control would therefore constitute an important step to a better testing of nanomaterials genotoxicity, ensuring that test systems are actually appropriate and/or allowing the validation of new ones.Firstly, we studied the possibility of using commercially-available tungsten carbide-cobalt (WC-Co) nanoparticles, previously characterized for physicochemical properties (size distribution and charge in used media), as positive control in three in vitro genotoxicity assays. The mouse lymphoma thymidine kinase gene mutation assay, the micronucleus assay studying chromosomal aberrations and the comet assay detecting primary DNA damage were performed. The last two assays were realized in two cell types, the mouse lymphoma cell line L5178Y and primary cultures of human lymphocytes. Our results show that WC-Co nanoparticles could be used as positive control in these in vitro genotoxicity assays, according to cell type and treatment schedule.Secondly, we investigated the mechanisms of action involved in WC-Co nanoparticles genotoxicity. Detection of centromeres in micronuclei using fluorescence in situ hybridization (FISH) show the involvement of both clastogenic and aneugenic activities. This was correlated with the results of a chromosome aberration assay on human lymphocytes blocked in metaphase, showing chromatid breaks and polyploid cells. Moreover, as oxidative mechanisms are the most described for nanomaterials, we studied oxidative DNA damage using the modified in vitro comet assay with the DNA repair enzyme formamidopyrimidine DNA glycosylase (FPG). We also detected a production of hydroxyl radicals using electron paramagnetic resonance in suspensions of WC-Co nanoparticles with and without cells. While performing high-throughput assays on WC-Co nanoparticles in three human cell lines corresponding to the main target organs for nanomaterials (A549 lung cell line, Hep3B liver cell line and Caki-1 kidney cell line) it was confirmed that oxidative stress play a significant role in the toxicity of WC-Co nanoparticles. Indeed, the production of reactive oxygen species in cells exposed to WC-Co nanoparticles was correlated to the observation of cytotoxicity and genotoxicity, studied using the detection of γH2AX foci.Finally, we carried out the most relevant genotoxicity assays to study nanodiamonds and lipid nanocapsules, which constitute promising nanovectors for drug delivery. The in vitro comet and micronucleus assays were performed on other cell types mimicking target organs: the T84 intestinal epithelial cell line and the 16-HBE bronchial epithelial cell line exposed to nanodiamonds of three different sizes, and human lymphocytes exposed to lipid nanocapsules of three different sizes and three different charges.

Thesis ( MPH ) -- University of Limpopo (Medunsa Campus), 2013. / Background and introduction Disclosure of HIV sero-status is critical in the control of the spread of HIV and research. To better understand the factors influencing disclosure will enhance the development of prevention interventions and ultimately lead to better control of the spread of the disease. However literature shows that the rates of disclosure are generally low and vary substantially in different populations. Study purpose To determine the prevalence, reasons for disclosure, partner reaction to disclosure, and intentions of disclosure to sexual partners among HIV positive adults receiving antiretroviral treatment. Study design Cross sectional survey was conducted with 400 adult patients aged 18 years and above, who receive ART, and have known their HIV status for six more than six months. Structured close ended self-administered questionnaire was used to collect data. The study participants were recruited from a wellness clinic of an academic hospital in the City of Johannesburg, Gauteng province between October and November 2012. Descriptive and inferential statistics were performed using STATA 10 for analysis. Pearson X2 tests were used to determine variables associated with disclosure. Results A total of 400 HIV positive adults participated in the survey. There were slightly more female (n=229, 57%) than male (n=171, 43%), the mean age of participants was 39.9 years, (range 18-80 years). Almost half (n=176, 46%) had known of their HIV diagnosis for more than 5 years. High proportion (n=293, 73%) were sexually active three months prior to the survey, (n=250, 63%) knew their partner’s HIV status, more than a third (n=145, 36.3%) had more than one sexual partner, (n=263, 73.5%) reported condom use, (n=261, 75%) disclosed to their partners. Gender, discussing HIV testing with sexual partner, knowing partner’s HIV status, and living with partner were significantly associated with disclosure. iv The most common cited reasons for disclosure were that they needed to protect their partner from being infected with HIV, and needed support from their partner. Partner reactions to disclosure included support, shock, and denial of the test results, blame, abandonment, violence, anger, and divorce. The most cited reasons for nondisclosure were concerns that the partner might leave, partner might be afraid of catching HIV, partner might think they were unfaithful, partner might get angry, partner might hurt them physically and that partner might stop financial support. Conclusion The study concludes that the prevalence of disclosure to sexual partners among sexually active adults was high and that most respondents disclosed immediately after they were diagnosed with HIV. However, disclosure to multiple sexual partners was lower as compared to disclosure to the steady partner. Respondents disclosed to protect the partner from HIV infection and to receive support. Nondisclosure was mainly used to protect self from negative reactions from the partner. Recommendations Researchers and health care providers needs to take cognisant of the risk sexual behaviour an low condom use among HIV positive adults receiving ART. Secondary prevention efforts targeting risky sexual behaviour among HIV-positive persons need to receive greater attention.