>Magister Scientiae - MSc / The therapeutic goal of cancer treatment is to trigger selective cell death in cancer cells. To eliminate cancerous cells effectively, the anti–cancer drugs must be targeted to the affected cells. However, anti–cancer drugs are often distributed non–specifically giving rise to systemic toxicities and other adverse effects. Cancer specific peptides are useful cancer targeting agents that can be used for the targeted delivery of anti-cancer drugs. Several cancer targeting peptides and some of their corresponding protein targets have been identified. Previous work investigated the specific binding of five of these peptides (p.C, p.H, p6.1, Frop-1 and p.L) conjugated to fluorescent nanoparticles (quantum dots) to a panel of human cell lines, which included four cancerous cell lines (Caco-2, HeLa, HT29 and HepG2) and one non-cancerous cell line (KMST-6). Flow cytometry showed that the p.L peptide preferentially bind to HT29 cells; suggesting that the expression levels of the target for the p.L peptide are higher in these cells. The objective of this study was to make use of target specific functionalised quantum dots (QDs) to deliver Second mitochondria-derived activator of caspases/ Direct AIP binding protein with low PI (Smac/DIABLO) to HT29 cells with the aim of enhancing the effects of pro-apoptotic drugs. Smac/DIABLO is a pro-apoptotic peptide that is able to interact with inhibitor of apoptosis proteins (IAPs), thereby inducing pro-apoptotic signalling. Methodology: CdSe/ZnS core-shell QDs were synthesised using the one-pot synthesis method. These QDs were characterised using photoluminescence (PL) spectroscopy, high resolution transmission electron microscopy (HR-TEM) and energy dispersive x-ray spectroscopy (EDS). The CdSe/ZnS core-shell QDs were solubilised with L-cysteine (Cys- QDs). The Cys-QDs were bi-conjugated to the p.L peptide and Smac peptide using 1-ethyl-3- (30-dimethylamino) carbodiimide (EDC) chemistry. Cultured HT29 cells were exposed to the 10 | P a g e QD peptide bi-conjugates and fluorescence microscopy was employed to assess targeting and internalisation. The cytotoxicity of the QD peptide bi-conjugates in combinatorial treatment with ceramide was evaluated using the WST-1 Cell Proliferation assay. A commercially available QD with similar chemistry was used to carry out a comparative study to relate the efficiency of the in-house synthesized QD.

Fluoride ion is an accompanying impurity in a wide variety of chemical gypsum throughout the world. In this study, the Ion Selective Electrode (ISE) method, the Ion Chromatography (IC) method and the standard Willard and Winter method of fluoride analysis were adapted and compared for use in the quantification of fluoride in South African chemical gypsum. During the use of ISE, the pH of sample solutions was found to be a critical parameter for the results to be meaningful. An operating pH of approximately 5 was suitable for consistency of results. It was important to ensure the existence of the ionised form of fluorine in solution, because the detection was based on the sensitivity of the membrane electrode to this species. In the case of the Willard and Winter method, the traditional visual indicator titration was replaced by a more sensitive spectrophotometric detection, because of low fluoride levels in the chemical gypsum. The parameter sensitive reaction rate approach was adapted, and the reaction allowed to go to completion to enable measurement with a bench top spectrophotometer. The IC method required a good separator since fluoride ions usually eluted too early for detection on common ion exchange columns. The data handling of the chromatographic software was thoroughly examined and consistent integration of the chromatograms maintained. Sample preparation of the chemical gypsum involved particle size reduction through grinding. No trend between fluoride impurity and the particle size of the sample was observed. <p. The quantity of fluoride in Kynoch and Omnia phosphogypsum were found to be 0.10% and 0.04% respectively. The Tioxide chemical gypsum, titangypsum, was found to contain in the region of 0.02% fluoride. Generally, the level of fluorine (F) has to be reduced to about 0.2% before phosphogypsum can be used as substitute for natural gypsum in the cement industry. The aim of this study is to critically evaluate three analytical methods, namely, the Willard and Winter standard method of fluoride analysis, the ISE method, and IC as applied in the quantification of fluoride in chemical gypsum. Secondly, the efficiency of treatment of the chemical gypsum with water, lime and sulphuric acid was investigated. The ISE method was found to be faster and relatively simpler versus both Willard and Winter and the IC methods. The IC method was quite superior for indicating general complexity of the sample and it was faster than the Willard and Winter method. The standard Willard and Winter method was generally found to be long and tedious. The three methods validated one another as percentages of fluoride in the chemical gypsum samples were the same. The study indicated that a sulphuric acid pretreatment of the chemical gypsum was the most effective (90%) in the removal of fluoride impurity compared to the lime treatment and water washing (9%). However, the water washing would be inexpensive for practical treatment of chemical gypsum at plant level. / Thesis (PhD(Chemistry))--University of Pretoria, 2006. / Chemistry / unrestricted

L’adénovirus (AdV) est un virus non enveloppé à ADN double brin qui entre dans la cellule par endocytose. Dans l’endosome un désassemblage partiel de la capside permet la libération d’une protéine interne de la capside, la protéine VI (PVI). Cette protéine code une hélice amphipathique qui va permettre la rupture de l’endosome. Des travaux antérieurs du laboratoire ont montré que le transport des particules virales vers le noyau nécessite la présence du motif conservé PPxY dans la PVI qui permet le recrutement d’ubiquitines ligases de la famille des Nedd4 (telles que Nedd4.1 et Nedd4.2). Il a précédemment été montré que la rupture des membranes induite lors d’infections bactériennes activait l’autophagie afin d’éliminer le pathogène intracellulaire via une dégradation lysosomale. Nos résultats démontrent que l’AdV induit également l’autophagie lors de son entrée dans la cellule. L’utilisation de différents AdV mutants nous a permis de démontrer que la rupture de l’endosome était responsable de l’induction de l’autophagie. De plus nos résultats montrent que le virus sauvage est capable d’éviter sa dégradation en contrôlant l’autophagie grâce au recrutement de la ligase Nedd4.2 via le motif PPxY de la PVI. Au contraire, un virus mutant dépourvu du motif PPxY et donc incapable de recruter la Nedd4.2 est séquestré dans les vésicules autophagiques puis dégradé par la fusion de ces vésicules avec les lysosomes. Ainsi le motif PPxY constitue un déterminant moléculaire permettant au virus de contourner les défenses cellulaires antivirales. / Adenoviruses (AdV) are linear ds-DNA containing, non-enveloped viruses that enter cells by receptor-mediated endocytosis. Once in the endosome it occurs a partial disassembly of the capsid allowing the releases of the membrane lytic capsid protein VI, which encodes an N-terminal amphipathic helix responsible for the endosome rupture. Our previous work showed that transport to the nucleus requires a conserved PPxY motif in PVI, which recruits ubiquitin ligases of the Nedd4 family (e.g. Nedd4.1 and 4.2). Previous work has shown that membrane damage induced by invasive bacteria elicits selective cellular autophagy to eliminate the pathogen via lysosomal degradation. In our current work we show that Adv also induce autophagy upon entry. Using a set of mutant AdV’s attenuated at each step of the membrane penetration process we show that indeed the membrane damage induced by the virus is causative for autophagy induction. Moreover the data show that wildtype AdV limit the level of autophagy induction and evade autophagic degradation by using a Nedd4.2 dependent process. In contrast, mutant viruses mutated for its PPxY and that fail to recruit Nedd4.2 are subject to autophagic degradation. Our data suggest that the presence of the PPxY motif in the virus subverts the autophagic process and thus identify the PPxY motif as an integral part of the virus to undermine cellular antiviral mechanism.

Since the 1960s and in reaction to its increasing popularity within Latino populations in the U.S., mariachi has become a common component of curriculum-based music courses in a growing number of public schools. Even though one of the principal purposes for the existence of mariachi programs is to improve students’ academic performance, music scholars have yet to address how mariachi education encourages higher academic achievement. On the other hand, anthropological educators argue that children underperform in school due to cultural difference or cultural mismatch; therefore, it is necessary to incorporate the cultural heritage of students into the curricula for academic success. The purpose of this study is to gain an increased understanding of the impact of mariachi programs on high school students' experiences and academic achievement and look for the relationship between achievement and cultural identity. This study examines two mariachi programs in the Tucson Unified School District as microcosms within the larger mariachi community in Tucson. In this interdisciplinary study, I use both ethnomusicology and anthropology of education as frameworks. I argue that students enrolled in the two mariachi programs are re-creating their cultural identities in response to the sociocultural context of the music education in the United States. This process of identity formation allows them to move more smoothly from one setting to another in their family, peer, and school worlds. The interrelationship between students and their peers, parents, and teachers facilitates their academic achievement.

Insomnia is a common health complaint that often becomes a persistent problem. The theoretical frameworks for understanding and treating insomnia have mostly been behavioural, yet the importance of cognitive processes has received greater attention over the years. The overall aim of this dissertation was to expand the knowledge on the processes from the Cognitive Model of Insomnia by investigating them in novel contexts. Study I examined the outcomes from cognitive therapy for insomnia on adolescents. Study II explored the relationship between cognitive processes and the association with remission and persistence of insomnia in the general population. Lastly, Study III investigated if cognitive processes mediated between cognitive behavioural therapy for insomnia (CBT-I) and outcomes of insomnia and depressive severity in a sample of people with insomnia comorbid with depressive problems. The findings show that cognitive therapy for insomnia affected sleep for adolescents, thus this is a promising treatment option for this age group. Further, it was found that cognitive processes distinguished between adults with normal sleep and persistent insomnia. For people with insomnia, elevated sleep-related worry at baseline increased the risk of reporting persistent insomnia later on, whereas a lowering of selective attention and monitoring, and safety behaviours over time increased the likelihood of remission from insomnia. This has clinical implications for insomnia assessment and treatment, as well as theoretical implications, and warrants further research. CBT-I was associated with greater reductions in dysfunctional beliefs and sleep-related safety behaviours compared to control treatment. Dysfunctional beliefs mediated between CBT-I and insomnia severity and depressive severity respectively. This supports the importance of negative thought content in both insomnia and depression.

Catalytic ethylene oligomerization is a well understood industrially viable process. The large majority of scientific literature and patents concerning this process has been developed with the use of chromium catalysts. Commercial systems producing selective tri/tetramerization, non-selective oligomerization and polymerization are all based on this metal with the exception of a few systems based on other transition metals (Zr, Ti, Ni etc.). This versatility raises interesting questions about chromium’s unique behaviour. Essentially, selective or non-selective oligomerization and polymerization processes could be regarded as belonging to the same category of C-C bond forming reactions, though different mechanisms are involved. The first part of this thesis explores a variety of chromium complexes for ethylene oligomerization purposes. In order to gather further information about the unique behaviour of chromium, we have explored a variety of nitrogen and phosphorus containing ligands. We started with a simple bi-dentate anionic amidophosphine (NP) ligand and assessed the role of the ligand’s negative charge and number of donor atoms in determining the type of catalytic behaviour in relation to the metal oxidation state. This ligand proved capable of generating a series of chromium dimeric, tetrameric or polymeric and even heterobimetallic chromium-aluminate complexes in different valence states. This allowed us to isolate a “single component” self activating Cr(II) complex as well as a rare example of mixed valence Cr(I)/Cr(II) species. Additionally, each of these species acted as switchable catalyst depending on the type of co-catalyst

Fluoxetine, a selective serotonin re-uptake inhibitor (SSRI) and the active ingredient in Prozac®, is found in the environment and disrupts feeding and metabolism in exposed fish. The objective of this research was to investigate the mechanisms involved in the feeding and metabolism disruption in the model goldfish (Carassius auratus). Two short-term waterborne fluoxetine exposures (7- and 14-days) were performed using two environmentally relevant doses of fluoxetine (0.5 and 1 μg/L) and metabolic effects at the level of the brain, liver, serum and bile in goldfish were investigated. Abundances of mRNA transcripts coding for six feeding neuropeptides were examined to determine which may be involved in the initial neural changes associated with decreased appetite in goldfish. The 7-day fluoxetine exposure at 1 μg/L caused corticotropin-releasing factor (CRF) mRNA levels to increase by 2-fold in female hypothalamus and telencephalon, indicating that CRF may be one of the first of the feeding neuropeptides to be altered. Six hepatic miRNAs were also evaluated in the goldfish liver that were previously associated with fluoxetine exposure in zebrafish (Danio rerio). Following the 7-day exposure at 1 μg/L, miR-22b, miR-140, miR-210, miR-301a and miR-457b levels increased in the female goldfish liver by 4-6 fold. The 14-day fluoxetine exposure at 1 μg/L caused 2-fold increases in miR-210, miR-301a, miR-457b and let-7d in male goldfish liver. These miRNAs were associated with the down-regulation of anabolic metabolic pathways in zebrafish, indicating a conservation of miRNA and fluoxetine effect between fish species. Serum and bile metabolite profiles of fluoxetine exposed goldfish were evaluated using ultra performance liquid chromatography coupled to quadrupole time of flight mass spectrometry. Following the 14-day exposure at 1 μg/L, the bile metabolite profiles of male goldfish were significantly different from controls as detected by cluster analysis and fluoxetine was tentatively identified in the serum. No other discriminant metabolites were identified as of yet. The data presented suggest that fluoxetine causes metabolic disruption in goldfish at multiple organ levels. Because of the widespread detection of fluoxetine and other emerging SSRIs in the aquatic environment, future research is required to firmly establish this pharmaceutical class as a metabolic and endocrine disrupting chemical.

It is well established that acquiring large amounts of range data with vision sensors can quickly lead to important data management challenges where processing capabilities become saturated and pre-empt full usage of the information available for autonomous systems to make educated decisions. While sub-sampling offers a naïve solution for reducing dataset dimension after acquisition, it does not capitalize on the knowledge available in already acquired data to selectively and dynamically drive the acquisition process over the most significant regions in a scene, the latter being generally characterized by variations in depth and surface shape in the context of 3D imaging. This thesis discusses the development of two formal improvement measures, the first based upon surface meshes and Ordinary Kriging that focuses on improving scene accuracy, and the second based upon probabilistic occupancy grids that focuses on improving scene coverage. Furthermore, three selection processes to automatically choose which locations within the field of view of a range sensor to acquire next are proposed based upon the two formal improvement measures. The first two selection processes each use only one of the proposed improvement measures. The third selection process combines both improvement measures in order to counterbalance the parameters of the accuracy of knowledge about the scene and the coverage of the scene. The proposed algorithms mainly target applications using random access range sensors, defined as sensors that can acquire depth measurements at a specified location within their field of view. Additionally, the algorithms are applicable to the case of estimating the improvement and point selection from within a single point of view, with the purpose of guiding the random access sensor to locations it can acquire. However, the framework is developed to be independent of the range sensing technology used, and is validated with range data of several scenes acquired from many different sensors employing various sensing technologies and configurations. Furthermore, the experimental results of the proposed selection processes are compared against those produced by a random sampling process, as well as a neural gas selective sensing algorithm.

L’objectif de ce travail est de proposer un mécanisme réactionnel pour l’oxydation aérobie d’hydrocarbures lourds, alcanes et alcènes, catalysée par les nanoparticules d’or. La co-oxydation du stilbène et du méthylcyclohexane est utilisée comme réaction modèle afin de comprendre les nombreux mécanismes mis en jeu. Le criblage initial d’une large gamme de catalyseurs d’or nous permet de mettre en évidence des effets de support dans cette réaction et d’établir un cahier des charges bien défini pour l’élaboration d’un catalyseur de référence. Une méthode chimique simple est mise au point pour préparer un tel catalyseur. Ce catalyseur, optimisé pour les réactions en milieux organiques apolaires, est ensuite utilisé pour réaliser des études macrocinétiques en variant de nombreux paramètres expérimentaux : température, concentration des réactifs, quantité de catalyseur. Un intermédiaire réactionnel clef, l’hydroperoxyde de méthylcyclohexane, est identifié. Après dosage, l’étude de l’évolution de sa concentration au cours du temps dans les différentes conditions de réaction permet de valider le mécanisme réactionnel existant et de mieux comprendre l’importance de certaines étapes élémentaires / The aim of this work is to propose a mechanism for the gold-catalyzed aerobic oxidation of bulky hydrocarbons, alkanes and alkenes. The co-oxidation of stilbene and methylcyclohexane is used as a model reaction to study different mechanisms which can take place simultaneously. After an initial screening of different gold catalysts in this reaction, essential characteristics of a reference catalyst for organic reactions in apolar media are identified. Based on these requirements, a straight-forward, chemical bottom-up method is designed to prepare this reference catalyst. This catalyst is used for macro-kinetic studies of the co-oxidation by modifying experimental parameters, such as temperature, alkane/alkene ratio, and reactants initial concentration. One key reaction intermediate, 1-methylcyclohexyl hydroperoxide, is identified. After titration, following the evolution of the concentration of this intermediate with time under the various reaction conditions considered validates the existing mechanism and highlight the importance of some elementary steps of the co-oxidation

Animal personality can be defined as a set of physiological and behavioral characteristics that differ between individuals, but are consistent over time and across situations. The evolution of individual differences in behavior that are consistent over time and situations is still not clear. Our understanding of why animals have personality can be improved by investigating the underlying physiological mechanisms of animal behavior. Serotonin is a key monoamine that serves as a physiological modulator of animal behavior. Selective serotonin reuptake inhibitors are a group of chemicals that increase levels of serotonin in the brain. Fluoxetine is one such chemical and is used to treat depression in humans. In the field cricket (Gryllus bimaculatus), increased levels of serotonin have been linked to higher activity and boldness, which are both personality traits. In the current study, the effects of induced serotonin on activity, exploration, boldness and aggression was investigated. My results show that injecting fluoxetine causes substantial changes in behavioral traits used to describe personality in field crickets. This result is opposite to previous studies, as serotonin induced individuals were less active, less explorative, and won less fights, compared to control individuals. This could be due to serotonin existing naturally within the circulatory system of the field cricket, whereas fluoxetine is a manufactured chemical intended for human receptors, or that fluoxetine has a similar effect in modulating personality in field crickets as in humans. Since fluoxetine acts similarly in field crickets as in humans, an increased understanding of the effects of induced serotonin on different behaviors in field crickets could be beneficial for treating psychological illnesses.