Bibliography: leaves 82-111. This thesis analyses the molecular composition of the surface components of the Endoparasitoid "Venturia canescens" using serological methods and specific sugar-binding lectins as diagnostic tools. The data reveals that the protective layer consists of at least two parts: a mucin-like glycoprotein and additional components from the wasp calyx fluid and the host hemolymph. The study suggests that the wasp larval cuticle is protected in a similar fashion to the egg chorion, except that the calyx-specific VLPs are probably replaced on the larval cuticle by host hemolymph proteins. The findings suggest that the mechanism of passive immune evasion could emerge during the evolution of the wasp-host interactions. The implication is that structurally conserved components may have similar functions in the parasitic and non parasitic species and could constitute a useful pre-adaptations for an endoparasitoid lifestyle.

Includes bibliographical references. The aim of this work is the further characterisation of the citrus exocortis viroid (CEV) replication complex in a tomato host system. The study employed three approaches. i. To raise polyclonal antiserum against a putative catalytic component of the CEV replication complex, the second largest subunit of tomato RNA polymerase II--ii. To develop a cell-free system for CEV RNA synthesis--iii. To use the polyclonal antiserum to the second largest subunit of RNA polymerase II, and a commercially available monoclonal antibody to the largest subunit, for antibody inhibition and immunoprecipitation experiments using the cell-free system.

Bibliography: leaves 348-366. Elucidates the nature and extent of genetic and environmental contributions to variation in permanent tooth crown size. Sibling correlations are compared to find evidence of sex-linked genes contributing to crown size. This hypothesis was tested by comparing mean tooth size in female-male opposite-sex twins with same-sex twins, and singletons.

Mycobacterium avium subsp. hominissuis (MAH) is a pervasive environmental bacterium that can cause opportunistic infections in humans. Among the most robust and hardy members of the Mycobacterium genus, M. avium can persist and thrive in a range of challenging environments, including many which place it in direct contact with humans. Surface-exposed proteins are central to the bacterial processes involved in both environmental persistence and pathogenesis. These proteins also play a critical role in how the immune system of the host recognizes and responds to pathogens. Mycobacteria have evolved a specialized mechanism for protein export, a Type VII Secretion System (T7SS), in order to transport their proteins through their thick and impermeable cell envelope. This system is responsible for the export of several classes of proteins, many of which play an integral role in virulence. A central focus of this dissertation is the characterization of a conserved element of the T7SSs in pathogenic mycobacteria, a PPE family protein, whose deletion attenuates virulence in M. avium. Specifically, we examined the localization of this PPE protein (MAV_2928) within the bacterium, screened potential protein-protein interactions with other conserved elements in the adjacent T7SS loci and analyzed the transcriptional regulation of the gene in response to environmental changes. Seeking to more thoroughly characterize the surface-exposed proteome of M. avium, particularly in the context of early infection, we then developed a method, based on selective biotinylation and affinity purification, to profile the of surface-exposed proteome of the bacterium. We employed this method to analyze the surface-exposed proteomes of M. avium 109 that had been exposed to macrophages to those of M. avium 109 that had been cultured in media. This comparison detected several proteins whose presence at the bacterial surface appeared to be dependent on particular growth conditions. Lastly, in order to establish a more efficient method to isolate biotinylated surface proteins from complex mixtures, we developed a testing paradigm to identify modifications to the original method that might improve our coverage of identified proteins. Through this process, we developed a more robust methodology that yielded improved coverage and depth. We then utilized this technology to profile the surface-exposed proteome of another clinical isolate of M. avium subsp. hominissuis, M. avium 104. Beyond improving our understanding of the basic biology of M. avium, this new data provides independent evidence that PPE family proteins are indeed exported to the surface of M. avium, where they remain associated with the bacterial cell envelope. In total, this analysis represents the most comprehensive profile of the surface-exposed proteins of M. avium generated to date. / Graduation date: 2012

Plantago ovata Forssk. (Plantaginaceae) is a winter annual species which, in North America, inhabits desert and Mediterranean habitats of the southwest United States, northwest Mexico and the Channel Islands of California and Mexico. In the eastern hemisphere P. ovata inhabits desert regions ranging from the Canary Islands, across northern Africa to western India. The wide disjunction between P. ovata in the western and eastern hemispheres poses an interesting question as to the origin and biogeography of the species. Previous authors have hypothesized that P. ovata was introduced to North America over the Bering land bridge, from Asia, during the Miocene, or introduced anthropogenically from Europe during the 18th century by Spanish settlers. In this study we examined sequence data from the chloroplast trnL-trnF, trnS-trnG and psbA-trnH regions, the nuclear ribosomal internal transcribed spacer (ITS) and a region 5' of the TCP region of a CYCLOIDEA gene. Using a molecular clock based on an ITS calibration within the Plantago genus, and a clock for plant chloroplast, we date a non-anthropogenic introduction event, from the Old World to North America, approximately 200,000-650,000 years ago. This is consistent with a Pleistocene origin, and does not support a Miocene origin of the disjunction. Based on a morphological survey of 552 specimens, from throughout the world range of P. ovata, we suggest the recognition of four subspecific taxa. Molecular phylogenetic analysis of chloroplast DNA and nuclear ribosomal DNA ITS sequences support this taxonomic treatment. Furthermore, phylogenetic sequences of the CYCLOIDEA gene support the morphological data. Both suggest the origin of North American P. ovata as a result of hybridization between Old World P. ovata varieties. This event provides further evidence that hybridization may serve as a predictor of invasiveness in plants. / Graduation date: 2005

The major goals of this work were to: develop a new methodology for the preparation of site-isolated catalytic sites on a silica surface, prepare the first truly single-site supported metallocene/CGC polymerization catalyst, and develop structure-reactivity relationships for these new systems. To synthesize these novel catalysts, the approach taken was to develop a protocol which allows for the synthesis of an aminosilica material with isolated, uniform amine sites. This patterned aminosilica was then used as a scaffold to support a constrained geometry catalyst. These functionalizations occurred at essentially a quantitative level, in stark contrast to previous literature reports. The patterned catalysts were evaluated in the polymerization of ethylene and compared to densely loaded literature materials. Overall, it was found the patterned materials were 5-10 times more active than traditional immobilized CGC catalysts. The patterned catalysts were also found to be effective catalysts for the copolymerization of norbornenes (including functionalized norbornenes) and ethylene, the first reported use of a tethered CGC for the production of ethylene-norbornene copolymers. The control materials were inactive in these polymerizations, providing further evidence that the patterning protocol allows for the synthesis of unique highly active, isolated catalytic sites. Various structural components of the immobilized CGC developed in this work were tested for their impact on catalyst synthesis and reactivity in ethylene polymerizations. The results showed the patterned materials in general behaved according to the trends seen in homogeneous CGC polymerizations. These results, while congruent with similar homogeneous CGC studies, are in direct conflict with previous work on supported CGCs reported in the literature. This discrepancy is likely the result of the difference between the isolated, possibly single-site patterned catalysts developed in the course of this work and the multi-sited catalysts prepared by traditional supporting protocols. This also further illustrates the difficulty in developing structure-reactivity relationships when ill-defined solid catalysts are used.

published_or_final_version / Anatomy / Master / Master of Philosophy

published_or_final_version / Medicine / Master / Master of Research in Medicine

published_or_final_version / Medicine / Doctoral / Doctor of Philosophy

published_or_final_version / Biochemistry / Master / Master of Philosophy