Previously we have identified the glycosphingolipid globotriaosylceramide (Gb3/Pk) as an inhibitor and resistance factor against HIV-1 infection in vitro. Here we show that a novel, soluble, completely synthetic Gb3 analogue, FSLGb3, inhibits infection of X4 strains of HIV-1 in the Jurkat T-cell line and both R5 and X4 strains in PBMCs. FSLGb3 absorbs into cellular plasma membranes and membrane adsorbed FSLGb3 was able to inhibit subsequent HIV-1 infection. We have also developed a mouse model to test in vivo the efficacy of soluble Gb3 analogs in the prevention of mucosal viral infection. Soluble Gb3 was incorporated into gel or alone and applied directly to the vaginal and rectal mucosal tissue of mice. We have not yet shown a statistically significant reduction in infection, although a trend towards inhibition is evident. Our studies show synthetic Gb3 to be an inhibitor of HIV-1 infection and further exploration of therapeutic strategies are warranted.

Previously we have identified the glycosphingolipid globotriaosylceramide (Gb3/Pk) as an inhibitor and resistance factor against HIV-1 infection in vitro. Here we show that a novel, soluble, completely synthetic Gb3 analogue, FSLGb3, inhibits infection of X4 strains of HIV-1 in the Jurkat T-cell line and both R5 and X4 strains in PBMCs. FSLGb3 absorbs into cellular plasma membranes and membrane adsorbed FSLGb3 was able to inhibit subsequent HIV-1 infection. We have also developed a mouse model to test in vivo the efficacy of soluble Gb3 analogs in the prevention of mucosal viral infection. Soluble Gb3 was incorporated into gel or alone and applied directly to the vaginal and rectal mucosal tissue of mice. We have not yet shown a statistically significant reduction in infection, although a trend towards inhibition is evident. Our studies show synthetic Gb3 to be an inhibitor of HIV-1 infection and further exploration of therapeutic strategies are warranted.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Thu Annelise Nguyen / Preclinical animal models of mammary carcinoma formation are vital for the advancement of cancer research, specifically in drug development. Two different types of animal models were utilized to determine the efficacy of combinational treatment of common antineoplastic drugs and the new class of primaquines that act as gap junction enhancers (PQs) at attenuating mammary tumor growth. The classic xenograft mouse model was used to show that PQs could increase the efficacy of cisplatin and paclitaxel. Combinational treatment induced an upregulation of connexin and caspase expression in the isolated tumor. Next the transgenic PyVT mouse model was characterized by multiple factors, including hormone receptor status, molecular markers for survival and proliferation, tissue histopathology, and secondary metastases during multiple stages of tumor development. This model showed limited therapeutic response to the antineoplastic drugs tested. PQ1 effectively attenuated tumor growth at all stages of tumorigenesis in the PyVT model, while PQ7 was determined to be an effective chemopreventive compound rather than chemotherapeutic. The PQs altered the expression profiles of connexins during tumorigenesis. Together the results indicate that PQs have an anticancer effect that is more efficient at attenuating tumor growth than the common antineoplastic compounds. Lastly the PyVT mouse model was used to determine the efficacy of antineoplastic compounds on male mammary carcinoma development. Interestingly, the antineoplastic compound that attenuated female mammary carcinoma growth did not produce a therapeutic response in the males and vice versa, suggesting a need for further studies into the male response to therapy.

Thrombosis is a vascular pathological process which frequently affects the veins of the lower limbs, and embolisation of thrombotic material is common. Thromboembolic disease is a common cause of death in Western societies, predominantly affecting the elderly. Numerous risk factors and co-morbidities have been identified. Safe and effective means of prophylaxis are available but are underutilised. Anticoagulant drugs are very effective in preventing thromboembolic disease, but their effects on the evolution of the thrombotic process remain poorly documented. These drugs are difficult to use, and treatment errors and treatment failures are common. The factors which determine the embolisation of deep vein thrombosis are poorly understood. When such embolisation occurs it is usually asymptomatic. Symptomatic pulmonary embolism presents in 3 general ways-pleuritic pain, shortness of breath, or collapse. Tachypnea is the commonest sign. Tests which are available to assist in the diagnosis of thromboembolic disease include the ventilation perfusion lung scan, ultrasound of the lower limbs, pulmonary angiography and echocardiography. The commonest investigation requested in this context is the lung scan. Although the results are often inconclusive, this is frequently the only specific investigation which is performed. The accurate interpretation of lung scans requires consideration of the pretest probability of pulmonary embolism. The D-dimer assay is another test which may be useful. The finding of a normal D-dimer level substantially reduces the probability of thromboembolic disease and may render a lung scan unnecessary. / Subscription resource available via Digital Dissertations only.

Thrombosis is a vascular pathological process which frequently affects the veins of the lower limbs, and embolisation of thrombotic material is common. Thromboembolic disease is a common cause of death in Western societies, predominantly affecting the elderly. Numerous risk factors and co-morbidities have been identified. Safe and effective means of prophylaxis are available but are underutilised. Anticoagulant drugs are very effective in preventing thromboembolic disease, but their effects on the evolution of the thrombotic process remain poorly documented. These drugs are difficult to use, and treatment errors and treatment failures are common. The factors which determine the embolisation of deep vein thrombosis are poorly understood. When such embolisation occurs it is usually asymptomatic. Symptomatic pulmonary embolism presents in 3 general ways-pleuritic pain, shortness of breath, or collapse. Tachypnea is the commonest sign. Tests which are available to assist in the diagnosis of thromboembolic disease include the ventilation perfusion lung scan, ultrasound of the lower limbs, pulmonary angiography and echocardiography. The commonest investigation requested in this context is the lung scan. Although the results are often inconclusive, this is frequently the only specific investigation which is performed. The accurate interpretation of lung scans requires consideration of the pretest probability of pulmonary embolism. The D-dimer assay is another test which may be useful. The finding of a normal D-dimer level substantially reduces the probability of thromboembolic disease and may render a lung scan unnecessary. / Subscription resource available via Digital Dissertations only.

Thrombosis is a vascular pathological process which frequently affects the veins of the lower limbs, and embolisation of thrombotic material is common. Thromboembolic disease is a common cause of death in Western societies, predominantly affecting the elderly. Numerous risk factors and co-morbidities have been identified. Safe and effective means of prophylaxis are available but are underutilised. Anticoagulant drugs are very effective in preventing thromboembolic disease, but their effects on the evolution of the thrombotic process remain poorly documented. These drugs are difficult to use, and treatment errors and treatment failures are common. The factors which determine the embolisation of deep vein thrombosis are poorly understood. When such embolisation occurs it is usually asymptomatic. Symptomatic pulmonary embolism presents in 3 general ways-pleuritic pain, shortness of breath, or collapse. Tachypnea is the commonest sign. Tests which are available to assist in the diagnosis of thromboembolic disease include the ventilation perfusion lung scan, ultrasound of the lower limbs, pulmonary angiography and echocardiography. The commonest investigation requested in this context is the lung scan. Although the results are often inconclusive, this is frequently the only specific investigation which is performed. The accurate interpretation of lung scans requires consideration of the pretest probability of pulmonary embolism. The D-dimer assay is another test which may be useful. The finding of a normal D-dimer level substantially reduces the probability of thromboembolic disease and may render a lung scan unnecessary. / Subscription resource available via Digital Dissertations only.

Thrombosis is a vascular pathological process which frequently affects the veins of the lower limbs, and embolisation of thrombotic material is common. Thromboembolic disease is a common cause of death in Western societies, predominantly affecting the elderly. Numerous risk factors and co-morbidities have been identified. Safe and effective means of prophylaxis are available but are underutilised. Anticoagulant drugs are very effective in preventing thromboembolic disease, but their effects on the evolution of the thrombotic process remain poorly documented. These drugs are difficult to use, and treatment errors and treatment failures are common. The factors which determine the embolisation of deep vein thrombosis are poorly understood. When such embolisation occurs it is usually asymptomatic. Symptomatic pulmonary embolism presents in 3 general ways-pleuritic pain, shortness of breath, or collapse. Tachypnea is the commonest sign. Tests which are available to assist in the diagnosis of thromboembolic disease include the ventilation perfusion lung scan, ultrasound of the lower limbs, pulmonary angiography and echocardiography. The commonest investigation requested in this context is the lung scan. Although the results are often inconclusive, this is frequently the only specific investigation which is performed. The accurate interpretation of lung scans requires consideration of the pretest probability of pulmonary embolism. The D-dimer assay is another test which may be useful. The finding of a normal D-dimer level substantially reduces the probability of thromboembolic disease and may render a lung scan unnecessary. / Subscription resource available via Digital Dissertations only.

Thrombosis is a vascular pathological process which frequently affects the veins of the lower limbs, and embolisation of thrombotic material is common. Thromboembolic disease is a common cause of death in Western societies, predominantly affecting the elderly. Numerous risk factors and co-morbidities have been identified. Safe and effective means of prophylaxis are available but are underutilised. Anticoagulant drugs are very effective in preventing thromboembolic disease, but their effects on the evolution of the thrombotic process remain poorly documented. These drugs are difficult to use, and treatment errors and treatment failures are common. The factors which determine the embolisation of deep vein thrombosis are poorly understood. When such embolisation occurs it is usually asymptomatic. Symptomatic pulmonary embolism presents in 3 general ways-pleuritic pain, shortness of breath, or collapse. Tachypnea is the commonest sign. Tests which are available to assist in the diagnosis of thromboembolic disease include the ventilation perfusion lung scan, ultrasound of the lower limbs, pulmonary angiography and echocardiography. The commonest investigation requested in this context is the lung scan. Although the results are often inconclusive, this is frequently the only specific investigation which is performed. The accurate interpretation of lung scans requires consideration of the pretest probability of pulmonary embolism. The D-dimer assay is another test which may be useful. The finding of a normal D-dimer level substantially reduces the probability of thromboembolic disease and may render a lung scan unnecessary. / Subscription resource available via Digital Dissertations only.

Doctor of Philosophy / Department of Agronomy / Tesfaye Tesso / Fusarium stalk rot and charcoal rot caused by Fusarium thapsinum and Macrophomina phaseolina respectively are devastating global diseases in sorghum that lead to severe quality and yield loss each year. In this study, three sets of interrelated experiments were conducted that will potentially lead to the development of resistance based control option to these diseases. The first experiment was aimed at identifying sources of resistance to infection by M. phaseolina and F. thapsinum in a diverse panel of 300 sorghum genotypes. The genotypes were evaluated in three environments following artificial inoculation. Out of a total of 300 genotypes evaluated, 95 genotypes were found to have resistance to M. phaseolina and 77 to F. thapsinum of which 53 genotypes were resistant to both pathogens. In the second experiment, a set of 79,132 single nucleotide polymorphisms (SNPs) markers were used in an association study to identify genomic regions underlying stalk rot resistance using a multi-locus mixed model association mapping approach. We identified 14 loci associated with stalk rot and a set of candidate genes that appear to be involved in connected functions controlling plant defense response to stalk rot resistance. The associated SNPs accounted for 19-30% of phenotypic variation observed within and across environments. An analysis of associated allele frequencies within the major sorghum subpopulations revealed enrichment for resistant alleles in the durra and caudatum subpopulations compared with other subpopulations. The findings suggest a complicated molecular mechanism of resistance to stalk rots. The objective of the third experiment was to determine the functional relationship between stay-green trait, leaf dhurrin and soluble sugar levels and resistance to stalk rot diseases. Fourteen genotypic groups derived from a Tx642 × Tx7000 RIL population carrying combinations of stay-green quantitative trait loci were evaluated under three environments in four replications. The stg QTL had variable effects on stalk rot disease. Genotypes carrying stg1, stg3, stg1,3 and stg1,2,3,4 expressed good levels of resistance to M. phaseolina but the combination of stg1 and stg3 was required to express the same level of resistance to F. thapsinum. Other stg QTL blocks such as stg2 and stg4 did not have any impact on stalk rot resistance caused by both pathogens. There were no significant correlations between leaf dhurrin, soluble sugar concentration, and resistance to any of the pathogens.

Master of Science / Department of Human Nutrition / Denis M. Medeiros / Copper deficiency has been linked with many health problems, among them cardiac hypertrophy. Because of its potential for causing oxidative damage, copper within the cell must be bound to chaperone proteins. In this thesis, we examined the role of dietary copper deficiency in the regulation of select copper chaperone proteins in cardiac tissue of rats. Sixteen weanling male Long-Evans rats were randomized into treatment groups, one group receiving a copper deficient diet (< 1 mg Cu/kg diet) and one group receiving a diet containing adequate copper (6 mg Cu/kg diet) for 5 weeks. Rats were sacrificed and a small blood sample was removed to determine hematocrit. Also, heart and liver tissues were removed for subsequent analysis. Rats fed the copper deficient diet had lower body weights but greater heart weights and heart:body weight. Hematocrit levels and liver copper concentrations were markedly decreased in copper deficient rats. These variables indicated that the copper deficient diet did in fact induce a copper deficiency in these animals. Non-myofibrillar proteins from the hearts were removed and separated by SDS-PAGE. Western Blotting was used to determine the concentrations of CTR1, CCS, Cox17, SCO1, Cox1 and Cox4. No changes were observed in the concentrations of CTR1 and Cox17. CCS and SCO1 were up-regulated as a result of copper deficiency, while Cox1 and Cox4 were both down-regulated. However, use of another antibody against Cox subunits suggested that only the nuclear encoded subunits including subunit IV were decreased, but not subunits I and II. These data provide new insight into the cardiac hypertrophy observed in copper deficiency, which suggests that select chaperone proteins may be up-regulated by a dietary copper deficiency.