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Crude gliadins were extracted from Canada Western Red Spring (CWRS) wheat flour with 70% (v/v) aqueous ethanol solutions and then lyophilized. Lyophilized crude gliadins were dissolved in 70% (v/v) aqueous ethanol (EtOH) or 4 mM acetic acid (HAc) and the density and ultrasound properties were measured at 20 °C. Good linear relationships of density, ultrasound velocity and ultrasound attenuation with solution concentrations were found. Solvent and sonication effects on the crude gliadins were discussed in terms of the values of the partial specific volume and the partial specific adiabatic compressibility coefficient for crude gliadins. The ethanol soluble crude gliadins had a larger partial specific volume and larger partial specific adiabatic compressibility coefficient than those for acidic soluble crude gliadins. These large values for the physical properties of ethanol soluble crude gliadins were thought to be evidence for the existence of complexes formed by some proteins (ethanol soluble LMW-glutenins and gliadins) and lipids in ethanol solutions and it was also found that the protein-lipid complexes were not destroyed by sonication treatment. Besides, there was no evidence showing that gliadins change with different wheat flours and cause different volume and compressibility properties of crude gliadins.

This thesis is an experimental investigation on the binding of self-assembling peptides to oligodeoxynucleotides (ODNs) and the characterization of the resulting peptide-ODN complexes/aggregates, the first key step in the development of a peptide-based gene delivery system. Effects of pH, charge distribution along the peptide backbone, and oligonucleotide sequences on the peptide-ODN binding were investigated by a series of physicochemical methods. UV-Vis absorption and fluorescence anisotropy experiments demonstrate that aggregates are formed after mixing the peptide and ODN in aqueous solution. The aggregates in solution can be centrifuged out. Based on this property, the fraction of ODNs incorporated in the peptide-ODN aggregates can be obtained by comparing the UV-Vis absorption of the solution before and after centrifugation. Binding isotherms are generated by a binding density function analysis of the UV absorbance results. The binding parameters are extracted from the analysis of the binding isotherms based on the McGhee and von Hippel model. Equilibrium binding parameter studies show that the binding of two self-assembling peptides, EAK16-II and EAK 16-IV, to model single and double-stranded ODNs at pH 4 is stronger than at pH 7, and that no binding occurs at pH 11. These results demonstrate that electrostatic interactions play an important role in the EAK-ODN binding because EAKs are more positively charged at low pH. EAKs bind more strongly to dG16 than to the other ODN sequences dC16 and dGC16. This demonstrates that the hydrogen bond might be involved because they promote the binding of the lysine residues of the peptide to dG16 to a greater extent than to dC16. The charge distribution along the peptides is found to have an effect on the binding. EAK16-IV, whose positively charged residues are clustered at one end of the peptide, binds to the ODNs more strongly than EAK16-II, whose positively charged residues are distributed throughout the peptide chain, at the same pH. The binding process of EAKs to the ODNs was investigated by fluorescence anisotropy and static light scattering experiments. The results show that individual EAK and ODN molecules complex first, followed by the aggregation of these complexes into large aggregates. The nature of the resulting peptide-ODN complexes/aggregates is examined by UV-Vis absorption, fluorescence anisotropy, and PAGE experiments. The results demonstrate that free EAK, free ODNs, and small EAK-ODN complexes, which can not be centrifuged out, exist in the supernatant, and that large aggregates are collected in the pellets after centrifugation of the solution. The size of the resulting EAK-ODN complexes/aggregates measured by AFM and DLS is around a few hundreds of nanometers at low EAK concentrations. The accessibility of the ODNs to the quencher in the solution is reduced by 40 % and 60 % after binding to EAK16-II and EAK16-IV, respectively, as determined by fluorescence quenching experiments on EAK-ODN mixture solutions. An ODN protection from Exonuclease 1 degradation is provided by the EAK16-II or EAK16-IV matrix when they are mixed with the ODNs at pH 4. However, the ODNs are protected to a much lower degree when the EAK-ODN aggregates are prepared at pH 7. The EAK-ODN aggregates prepared at pH 7 are found to dissociate more easily than those prepared at pH 4 when they are incubated with exonuclease I solution at pH 9.5. These results suggest that the ODN protection afforded by the EAK-ODN aggregates is correlated with their structural stability after being incubated with the nuclease solution. The stability of the EAK-ODN aggregates after dilution is determined by UV-Vis absorption. No detectable dissociation of the aggregates is observed over 20 hrs after a 5- and 10-fold dilution of the solution in the same buffer used for their preparation. The EAK-ODN aggregates remain stable after the solutions are centrifuged, and re-dissolved in fresh buffer solutions. The ability of an EAK matix to protect ODNs from nuclease degradation together with its biocompatibility and low-toxicity suggests that EAK self-assembling peptides could be used as carriers for gene delivery.

Host factors can have important consequences for HIV risk and disease progression. Two separate projects relevant to Aboriginal populations in Manitoba were undertaken. The Solvent Use Project investigated solvent use in Winnipeg through an interdisciplinary multi-phase approach that integrated community based research and basic science. From interviews with solvent users and key informants we learned that solvent users experience many health and social disparities. We demonstrated that there is support within the community to work with solvent users and study solvent use further. The HLA-B*35 Project aimed to identify the diversity of HLA-B*35 allele subtypes, in HIV+ patients that presented to care between 2007 and 2010 in Manitoba. We observed 11 distinct HLA-B*35 allele subtypes. Case studies reflected the overrepresentation of Aboriginal people infected with HIV in Manitoba, and the pressing issues of either late presentation to care or rapid disease progression within patients who are HLA B*35.

Crude gliadins were extracted from Canada Western Red Spring (CWRS) wheat flour with 70% (v/v) aqueous ethanol solutions and then lyophilized. Lyophilized crude gliadins were dissolved in 70% (v/v) aqueous ethanol (EtOH) or 4 mM acetic acid (HAc) and the density and ultrasound properties were measured at 20 °C. Good linear relationships of density, ultrasound velocity and ultrasound attenuation with solution concentrations were found. Solvent and sonication effects on the crude gliadins were discussed in terms of the values of the partial specific volume and the partial specific adiabatic compressibility coefficient for crude gliadins. The ethanol soluble crude gliadins had a larger partial specific volume and larger partial specific adiabatic compressibility coefficient than those for acidic soluble crude gliadins. These large values for the physical properties of ethanol soluble crude gliadins were thought to be evidence for the existence of complexes formed by some proteins (ethanol soluble LMW-glutenins and gliadins) and lipids in ethanol solutions and it was also found that the protein-lipid complexes were not destroyed by sonication treatment. Besides, there was no evidence showing that gliadins change with different wheat flours and cause different volume and compressibility properties of crude gliadins.