An experiment was conducted to investigate the relative effectiveness of a combined behavior rehearsal anxiety relief conditioning paradigm with a more conventional behavioral rehearsal program in the treatment of deficient assertive behavior.

Autophagy is an evolutionarily conserved process across eukaryotes that is responsible for degradation of cargo such as aggregate-prone proteins, pathogens, damaged organelles, macromolecules etc. via its delivery to lysosomes. The process is known to involve the formation of a double-membraned structure, called autophagosome, that engulfs the cargo destined for degradation and delivers its contents by fusing with lysosomes. This process involves several proteins at its core which include two transmembrane proteins, ATG9 and VMP1. While ATG9 and VMP1 has been discovered for about a decade and half, the trafficking and function of these proteins remain relatively unclear. My work in this thesis identifies and characterises a novel trafficking route for ATG9 and VMP1 and shows that both these proteins traffic via the dynamin-independent ARF6-associated pathway. Moreover, I also show that these proteins physically interact with each other. In addition, the tools developed during these studies helped me identify a new role for the most common autophagy receptor protein, p62. I show that p62 can specifically associate with and sequester LC3-I in autophagy-impaired cells (ATG9 and ATG16 null cells) leading to formation of LC3-positive structures that can be misinterpreted as mature autophagosomes. Perturbations in the levels of p62 were seen to affect the formation of these LC3-positive structures in cells. This observation, therefore, questions the reliability of LC3-immunofluorescence assays in autophagy-impaired cells as method of assessing autophagy and points towards the homeostatic function played by p62 in autophagy-impaired cells.

The DNA Mismatch repair (MMR) pathway is responsible for the repair of base-base mismatches and insertion/deletion loops that arise during DNA replication. MMR deficiency is currently estimated to be present in 15-17% of colorectal cancer cases and 30% of endometrial cancers. MLH1 is one of the key proteins involved in the MMR pathway. MMR deficient tumours are often resistant to standard chemotherapies, therefore there is a critical need to identify new therapeutic strategies to treat MMR deficient disease. This study demonstrates that MLH1 deficient tumours are synthetically lethal with the mitochondrial-targeted agent Parthenolide which is known to induce reactive oxygen species (ROS) as one of its main mechanisms of action. Upon functional analysis, I show for the first time that loss of MLH1 is associated with deregulated mitochondrial function evidenced by a reduction in complex I expression and activity, reduced basal oxygen consumption rate and reduced spare respiratory capacity. This mitochondrial phenotype in the MLH1-deficient cell lines is accompanied by a reduction in mitochondrial biogenesis as evidenced by down regulation of pgc1β and decreased mitochondrial copy number. Furthermore, MLH1-deficient cancer cells have a decreased antioxidant defence capacity with reduced expression of the antioxidant genes NRF1, NRF2, Catalase, Glutathione peroxidase and SOD1 as well as increased ROS production when treated with Parthenolide. I further demonstrate that both MSH2- and MSH6-deficient cell lines also display deficiencies in complex I compared to their MMR-proficient counterparts. Taken together, the results of this study show a novel role for MLH1 in mitochondrial function and biogenesis. The MMR proteins MSH2 and MSH6 are also likely to have a role in the mitochondria. My results suggest that targeting the mitochondria may be a potential therapeutic strategy for the treatment of MMR and specifically MLH1 deficient disease.

It is currently believed that the etiology of inflammatory bowel disease involves an aberrant immune response towards the gastrointestinal microbial flora. In addition, an increase in intestinal paracellular permeability may also be a contributing factor of disease, as it precedes disease in several animal models. However, it remains unclear whether increased intestinal permeability is an epiphenomenon of disease or if it can lead to it. The goal of this thesis is to elucidate this cause-effect relationship. The IL-10-/- mouse is a model of IBD that spontaneously develops colitis after 12 weeks of age. We measured intestinal permeability in this mouse from 4-17 weeks of age and observed that there was a significant increase in small intestinal permeability early in life and before the onset of colitis. When small intestinal permeability was selectively decreased with AT-1001 (a ZOT antagonist peptide) colitis was significantly ameliorated. In contrast, when it was increased with AT-1002 (a ZOT agonist peptide) colitis worsened, indicating that modifications in the paracellular traffic of the small intestine had a significant effect on the severity of colonic disease. In order to study the possible mechanisms by which small intestinal permeability modulated disease in the colon, we measured the effect of increasing small intestinal permeability on the colonic microbial flora of IL-10-/- mice. After AT-1002 treatment from 4-12 weeks of age, there was an evident shift in colonic adherent flora. This effect was not a consequence of inflammation as there was a similar effect in wild type mice. We also studied the effect of increasing small intestinal permeability in the development of oral tolerance to dietary antigens. When wild-type mice were fed OVA under conditions of increased small intestinal permeability there was a significant increase in the proliferation of B cells in the spleen and an increase in OVA-specific humoral response, compared to animals fed OVA alone. Moreover, the production of IL-10 in response to oral OVA was prevented when OVA was given with AT-1002, both in the small intestine and the colon. The studies presented in the doctoral thesis suggest that small intestinal permeability has a critical role in the development of colitis in IL-10-/-mice, and that increasing paracellular traffic in the small intestine may lead to changes in colonic bacterial flora and the abrogation of tolerance to oral antigens, two features of inflammatory bowel disease in humans. / Experimental Medicine

No description available.

The beta+-decay half-lives of the neutron-deficient, odd-odd, N = Z nuclei, 74Rb, 78Y, 82NB and 86Tc were measured following the fragmentation of a primary 92Mo beam at an energy of 60 MeV per nucleon at the GANIL laboratory, France. They were measured by correlating beta+-decays with the implantation of unambiguously identified fragments. The deduced log10 ft1/2-values are consistent with 0+→ 0+, Fermi superallowed transitions, which together with the measured beta+-detection efficiencies suggest T = 1, 1pi- 0+ ground states for these odd-odd, N = Z nuclei. These data represent the heaviest N = Z systems for which Fermi superallowed decays have been established. The results suggest that these nuclei can be used to extend the mass range used to test the Conserved Vector Current hypothesis of the standard model. In addition, an experiment using the reaction 58Ni + 28Si at a beam energy of 200 MeV was performed at the Laboratorio Nationale di Legnaro, Italy, using the EUROBALL spectrometer together with a charged particle silicon detector ball and an array of liquid scintillator neutron detectors. The measured values of the energies of the evaporated, charged-particles and those of the gamma-ray multiplicity (deduced using the liquid scintillator detectors), are used to implement a novel technique for the channel selection of weakly populated evaporation channels.

Sorghum (Sorghum bicolor L. Moench) ranks fifth worldwide in production among cereals. It is a major staple food for millions in Africa and Asia, and a major livestock feed grain in developed countries. However, the sorghum grain is poor in lysine content, limiting its value as food and feed. In this study, I hypothesize that reduction of some of the major storage proteins that are inherently poor in lysine through in vitro manipulation will result in the enhanced expression of proteins with a better lysine profile and, thus, increased overall grain lysine content. Sorghum genotypes were screened for in vitro amenability and a sorghum genotype-tissue culture medium combination that yielded the highest somatic embryo callus formation and regeneration potential, was identified. This resulted in the establishment of a sorghum biolistic transformation method with a transformation efficiency of 3.36%, the highest reported to date. Using genetic engineering tools, the enhancement of the nutritional quality of grain sorghum was achieved by increasing the seed lysine content. An RNAi co-suppression strategy was employed and resulted in 45.23 and 77.55% increase in whole seed and endosperm lysine increase, respectively. The co-suppression RNAi constructs targeted the endosperm specific suppression of three lysine-poor storage proteins, namely ä-kaf-2, ã-kaf-1 and -2, and an enzyme that catalyzes seed lysine degradation, lysine keto-gluterate reductase (LKR). Seven independent transgenic events displayed successful transgene integration for both the selectable marker gene and the target constructs. However, the Southern blot hybridization analysis revealed two transgenic events that displayed transgene re-arrangement at the 5’promoter end, thus resulting in a lack of suppression of target proteins. Variations in target proteins co-suppression was observed with Western blot analysis and RT-PCR for both the target kafirins and LKR suppression, and no lysine improvement was observed where no kafirin suppression occurred. The transgenic co-suppression of the target kafirins resulted in the endosperm structural change from a hard, corneous endosperm to a soft, floury endosperm, consistent with ã-zein suppression in the Opaque-2 maize mutant. / Thesis (PhD)--University of Pretoria, 2010. / Plant Science / unrestricted

Post tensioning bridges is an efficient way of construction where segments of a bridge are strung together using small diameter high strength strands. The steel strands are passed through internal and/or external ducts within the concrete segments, jacked to high levels of stress, anchored using steel wedges, and then filled with a corrosion inhibiting filler. A cementitious material called grout has been widely used for over 50 years as this filler. The Federal Highway Administration design and construction manual provides recommendations in the properly filling techniques to ensure the tendons are full of grout. Despite the level of detail the manual provides, several Florida bridges that used this construction technique have seen severe corrosion related failures due to poor grout quality. Oil based products have been developed for the express purpose of reducing the corrosion rate in previously grouted tendons. Utilizing the interstitial spaces in the steel strands, low viscosity fluid is pumped through the strand which can also penetrate the surrounding grout and fill unintentional voids. However, by doing so, there is the concern that the bond strength could be compromised and increase the risk of failure. This study presents the findings of small scale grouted mono-strand tendons which determined the bond effects of defective grout, evaluated the injection capabilities of various penetrants, and determined whether or not the penetrant adversely affects bond after impregnation. No appreciable variation in bond was found.

A facile and efficient base-mediated protocol for debromination of vic-dibromides in perfluoroalkyl(aryl) compounds in aqueous medium has been demonstrated. With mild reaction conditions, the developed strategy has a good substrate scope and electron-deficient olefin products were obtained in good yields. A mechanistic explanation of the debromination is offered with three key experimental observations: (1) the reactions are accelerated by the more electron-rich nucleophiles, (2) the reactions are promoted by the more electron poor vic-dibromides in perfluoroalkyl compounds, and (3) the nucleophilic side reaction is preventable. It is evident that the electronic factors strongly dictate vic-dibromides elimination to the perfluorovinyl ethers, which are the precursors for various perfluorinated polymers. The different reaction conditions were tested in implicit solvent (water) conditions, which helped to confirm the E2-like mechanism.

Biomaterials are defined as non-viable materials whose functions strives to interact with biological systems, this makes biomaterials suitable for medical devices and applications. Biomaterials biological properties must be thoroughly investigated and evaluated in order to be approved for clinical usage. In vitro studies are used to characterize the materials biological properties, if promising results are achieved in vitro, in vivo studies may be performed to ensure that the material can interact with living animal models in the intended way. However, for a biomaterial such as calcium-deficient hydroxyapatite there is a gap between results observed in vitro and in vivo studies. Microfluidic systems have been highlighted as a possible evaluation model to achieve reliable results for in vitro studies. The aim of this thesis was to compare and evaluate a biomaterial- on-a-chip, i.e., a biomaterial integrated in a dynamic microfluidic system, with a traditional static in vitro system in the regards of drug release and protein adsorption. Two chemically identical calcium-deficient hydroxyapatites but with different topographies were integrated in the microfluidic system and manufactured as discs for static evaluation. Results from drug release and protein adsorption studies showed different behaviours for dynamic and static control, which is significant since it indicates that the outcome of the characterization correlates to the evaluation model used.