- ItemThe Relationship between Phenylpropanoid Metabolism and Drought in Phaseolus vulgaris(University of Guelph, ) Peña Barrena, Luis Eduardo; Bozzo, GaleDrought events are expected to become more frequent and more severe with climate change. To date, the extent of metabolic changes in Phaseolus vulgaris L. that are associated with drought are largely unexplored. This thesis investigated whether alterations in flavonol, isoflavone and phenolic acid metabolites are linked to the drought response in two white bean recombinant inbred lines BT6 and BT44 when cultivated under severe drought relative to well-watered conditions. In general, the fresh weight and dry weight biomass of both recombinant inbred lines was reduced under severe drought, but some root/shoot dry weight ratios were increased. Drought stress induced the accumulation of isoflavone-derived phytoalexins in the roots of the white bean plants. Some phenolic acids and a flavonol glycoside accumulated at higher concentrations in the leaves of drought-stressed plants relative to those grown under well-watered conditions. This research identified key phenylpropanoid indicators of severe drought in white bean plants.
- ItemVANL-100: Shaking up the Game of Glycogen Synthase Kinase-3beta Inhibition in SH-SY5Y Human Neuroblastoma Cells(University of Guelph, ) Pfeifer, Julia; Kalisch, Bettina; Saleh, TarekThis thesis introduces VANL-100, a conjugate of alpha-lipoic acid and naringenin, as a novel inhibitor of glycogen synthase kinase-3beta (GSK3B). Immunoblotting analysis in SH-SY5Y cells evidenced this through increased GSK3B phosphorylation at serine-9. Interestingly, neither of the parent compounds of VANL-100, individually or combined without a chemical bond, achieved inhibition of GSK3B, underscoring the significance of conjugate therapy. VANL-100 was then benchmarked against CHIR99021, a gold-standard GSK3B inhibitor. Immunofluorescence illustrated that both compounds seemed to notably promote nuclear migration of beta-catenin, a key GSK3B target. Cell viability assays highlighted VANL-100 and its ability to boost neuroblastoma cell viability, whereas CHIR99021 diminished it. Critically, VANL-100 outperformed CHIR99021 in counteracting cell loss due to amyloid beta toxicity. These findings spotlight the significant potential of VANL-100 for GSK3B inhibition and advocate for its continued exploration across diverse disease models.
- ItemThe Relationship Between State Funded Resource Development Infrastructure and Indigenous Jurisdiction: An Analysis of Two Development Projects in Nunavut, Canada(University of Guelph, ) Peres, Megan; Stanley, Anna; Moola, FaisalCanada is a resource-rich nation within a context of unfinished settler colonialism. The settler state is consistently working to manage the reality of Indigenous jurisdiction while attempting to assert sovereignty over the land. Indigenous jurisdiction threatens the settler state’s ability to extract value from the land. This research aims to understand the present-day relationship between Indigenous jurisdiction and resource development infrastructure (RDI). Specifically, it aims to understand how two current infrastructure projects in Nunavut engage Inuit jurisdiction. RDI is intended to expand access to resources for extraction, especially in remote areas. The findings suggest that resource development infrastructure developed with Inuit organizations as proponents is a method through which the settler state manages the threat posed by Inuit jurisdiction to expand access to Inuit lands. This research provides a foundation for further Indigenous-led, empirical, and theoretical research on ongoing settler colonialism and publicly funded resource development infrastructure in Canada that could help illuminate pathways toward decolonization.
- ItemCoccidiosis and Necrotic Enteritis (NE): Testing Molecular Tools for Quantifying Clostridium and Eimeria Simultaneously in Experimentally Challenged Broiler Chickens(University of Guelph, ) Heibein, Will; Barta, JohnVarious species of Eimeria infect the intestinal tract of poultry, with Eimeria maxima commonly associated with the development of necrotic enteritis (NE). Restrictions to prophylactic use of Category 2 antibiotics in Canadian poultry make outbreaks of NE caused by Clostridium perfringens more likely. Infections with Eimeria species, combined with poor diet and sanitation, are key drivers of C. perfringens infection and NE. This study quantified C. perfringens and E. maxima DNA from within the intestinal tract of experimentally infected broilers using a newly developed, duplexed, probe-based qPCR assay. C. perfringens showed little change in quantification but expressed lesion development in birds infected with E. maxima, particularly from 5 days-post-infection. Inoculation of day-of-age chicks with C. perfringens permitted colonization and persistence throughout the intestinal tract for several weeks. Birds inoculated with C. perfringens prior to E. maxima, and vice versa, were more likely to exhibit lesions consistent with NE.
- ItemExtending the Longevity of AAV-mediated Lung Gene Therapy(University of Guelph, ) Thomas, Sylvia; Wootton, SarahAAV-mediated gene therapy has progressed rapidly in the past three decades, permitting the use of this vector for the treatment of several diseases. Although the first administration of an AAV vector into a human patient occurred in 1995 to correct cystic fibrosis in the lungs, this organ remains a challenging treatment site and elusive target for gene therapy. Limitations are attributed to the requirement for lung tropic vectors, the defensive barriers of the lungs, immune system considerations, the obstacles in the journey to reach the target cell and be granted entry, and cell turnover. In monogenic disorders such as surfactant protein B deficiency, additional hurdles appear in achieving long term expression due to the developing lung. This disease affects neonates, causing respiratory distress leading to death within a year of birth. Novel vectors such as capsid engineered AAV6.2FF may be able to deliver the gene of interest, but the gene addition delivery of AAV will be maintained in an episomal fashion and eventually wane in expression. Therefore, persistent expression will need to be achieved through either readministration or permanent gene correction. The following chapters feed into each other in progressing this vector forward and extending its persistence in the lungs. In this thesis, we assess the vector performance of AAV6.2FF, analyzing its biodistribution and stability, along with investigating mechanisms to extend expression in the lungs. Here, we administered 1e11 vector genome (vg) of AAV6.2FF with an alkaline phosphatase reporter gene by five routes of administration into C57BL/6 mice and viewed robust and localized expression in the lungs through intranasal instillation. Separately, we examined the thermal stability of this vector and determined that an excipient formulation change to include the non-ionic surfactant chemical PF-68 improved stability under different exposure conditions. In addition, we investigated mechanisms to extend the persistence of AAV6.2FF gene expression in lungs by mitigating the immune response with a stealth design change to the genome that permitted readministration along with demonstrating that permanent gene correction with AAV6.2FF is possible, providing optimistic conclusions that both readministration and gene editing can extend survival in surfactant protein B deficiency.