published_or_final_version / Paediatrics and Adolescent Medicine / Doctoral / Doctor of Philosophy

Gram-negative bacteria decorate their outermost surface structure, lipopolysaccharide, with elaborate chemical moieties, which effectively disguises them from immune surveillance and protects them from the onslaught of host defenses. Many of these changes occur on the lipid A component of lipopolysaccharide, which is crucial for host recognition of Gram-negative infection. Despite its highly inflammatory nature, LPS is a molecule with remarkable therapeutic potential. Lipid A is a glycolipid that serves as the hydrophobic anchor of LPS and constitutes a potent ligand of the TLR4/MD2 receptor of the innate immune system. A less toxic mixture of mono-phosphorylated lipid A species (MPL) recently became the first new FDA-approved adjuvant in over 70 years. Whereas wild-type E. coli LPS provokes strong inflammatory MyD88-mediated TLR4 signaling, MPL preferentially induces less inflammatory TRIF-mediated responses. Here, we developed a system for combinatorial structural diversification of E. coli lipid A yielding a spectrum of bioactive variants that display distinct TLR4 agonist activities and cytokine induction. Mice immunized with engineered lipid A/antigen emulsions exhibited robust IgG titers indicating the efficacy of these molecules as adjuvants. Other types of modification to the lipid A domain, such as altering the length of the fatty acyl chains that anchor LPS to the cell membrane, were found to affect bacterial fitness but not drastically influence detection by the TLR4/MD2 receptor. Overall, this combinatorial approach demonstrates how engineering lipid A can be exploited to generate a spectrum of immunostimulatory molecules for vaccine and therapeutics development as well as for a deeper understanding of bacterial membrane biogenesis. / text

The impact of adjuvants on atomization and patternation of spray mixtures was evaluated. The data showed that certain adjuvants, in particular drift control agents, could potentially detrimentally affect the distribution of herbicide dose across the sprayed swath. The present research sets out to evaluate the impact of this distribution and to seek ways of improving the way researchers and users characterize and possibly mitigate these effects with a view to minimizing the potential detriments and maximizing the efficiency of herbicide active ingredient (AI) utilization. Different formulations of glyphosate with and without a novel polymeric drift control agent (AgRho DR 2000) applied to contrasting broad-leaved and grass weeds were used to evaluate several effects of polymer use. Variables included nozzle type (XR TeeJet extended range flat spray tips, TT Turbo TeeJet wide angle flat spray tips, and TurboDrop air induction nozzles) sampling position (principally under the nozzle centers and under the overlap between two adjacent nozzles), boom height (30, 45, and 60 cm above the target), spray delivery (the volume of spray arriving at the target), spray retention ( the volume of spray actually retained by the target foliage), and herbicide efficacy ( the response of the target weeds to the herbicide dose applied). The data showed that when the polymer was included in the spray mixture, the nozzle used, boom height, presence of the adjuvant, sampling position and certain interactions between these variables were all significant. Spray retention was affected by plant type and retention of coarse sprays was improved by the inclusion of DR 2000. Very coarse sprays reduced glyphosate efficacy on both grasses and broad-leaved weeds although that effect was reduced by use of DR. Addition of drift control agents always ii resulted in increased variability in spray distribution with concomitant increases in both retention and efficacy variability. Variability was shown to decrease with decreasing boom height. There was little correlation between spray delivery and herbicide efficacy. Deposit structure was shown to be a highly important factor in understanding herbicide dose transfer. A novel methodology utilizing digital imaging technology and diversity statistics was developed and evaluated to improve the way we measure and characterize deposit structures. Separation of qualitatively different treatments with similar volumetric distributions was possible. This methodology will be of use to both biologists and fOnTIulation chemists for prediction or explanation of biological results relating to deposit structure. Use of Scanning electron microscopy, and epi-fluorescence microscopy was used to characterize deposit morphology. Differences in deposit morphology were observed and documented leading to a possible explanation for the enhanced glyphosate activity in the presence of DR 2000 iii

Thesis (Ph. D.)--University of Hong Kong, 2007. / Also available in print.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 4 uppsatser.

Thesis (Ph. D.)--University of Hong Kong, 2007. / Also available online.

La tuberculose humaine, causée par Mycobacterium tuberculosis, est responsable de plus d'un million de décès par an et on estime qu'un tiers de la population mondiale est infecté de façon latente. Lors de leur arrivée dans les poumons, les mycobactéries sont détectées par les cellules du système immunitaire inné grâce à de nombreux récepteurs, tels que les lectines de type C. Le but de mes travaux était de caractériser les bases moléculaires et les conséquences fonctionnelles des interactions entre les mycobactéries et trois lectines de type C : Mincle, Dectine-1 et Dectine-2. Nous avons ainsi défini les relations structure/fonction de la reconnaissance des ligands de nature glycolipidique par Mincle et Dectine-2 et synthétisé des ligands de structure minimale de Mincle qui pourraient avoir de potentielles applications comme adjuvants de vaccination. Enfin, nous avons identifié Dectine-1 comme une possible voie d'entrée silencieuse des mycobactéries dans les cellules phagocytaires. / Mycobacterium tuberculosis is the causative agent of human tuberculosis leading to more than 1 million deaths every year and infecting latently one third of the population worldwide. Once in the lungs, mycobacteria are detected by innate immune cells through several pattern recognition receptors such as C-type lectins. The aim of my thesis was to characterize the molecular bases and the functional consequences mycobacteria recognition by three C-type lectins: Mincle, Dectin-1 and Dectin-2. We have characterized the structure/function relationships of the glycolipid ligands recognition by Mincle and Dectin-2 and synthesized ligands of Mincle with a minimal structure that could be used as vaccine adjuvants inducing Th1 and Th17 responses. We also identified Dectin-1 as a possible silent entry gate for mycobacteria into phagocytic cells allowing binding to immune cells without triggering intracellular signaling.

Depuis plusieurs années, la perte causée par le rebond en béton projeté fait l'objet de préoccupations environnementales. L'optimisation continue de l'équipement et de la mise en place permet de rendre la technique de projection plus efficiente. Le laboratoire de béton projeté de l'Université Laval enchaine ainsi les projets de recherche afin d'avoir une meilleure compréhension et un contrôle accru du phénomène. Les connaissances acquises dirigent cette recherche vers une caractérisation plus complète de la projection par voie sèche et de son substrat récepteur à l'état frais. Un montage à échelle réduite incluant les avancées récentes de l'équipement a été développé afin d'offrir des avantages technologiques et environnementaux non négligeables. Cette implantation permet de s'affranchir de certains problèmes associés à la méthode traditionnelle à l'échelle 1 :1 et d'évaluer plusieurs mélanges à dosages d'adjuvants variés, et ce, rapidement et sans la production et l'utilisation excessive de matériau. Le béton mis en place de façon gravitaire est généralement caractérisé à l'état frais à l'aide de paramètres rhéologiques. Cependant, les appareils traditionnels de mesures, tel que les rhéomètres, ne peuvent être utilisés en béton projeté par voie sèche à cause la consistance du matériau mis en place. Une méthodologie a alors été proposée afin de pouvoir étudier et de vérifier l'applicabilité des paramètres rhéologiques. La complexité des phénomènes présents lors de la mise en place du béton projeté par voie sèche a cependant montré la difficulté de capturer les effets visés. Cela a permis d'introduire le concept d'historique rhéologique. Le concept d'historique rhéologique présenté permet d'unir plusieurs paramètres théoriques et diverses constatations expérimentales ainsi que de proposer une approche différente de la description du matériau à l'état frais et de sa mise en place. Il identifie deux zones distinctes sur la surface réceptrice par une fréquence énergétique critique. Ces zones sont définies selon l'état du substrat : fluide ou élastoplastique. Ces deux états sont incontournables et se comportent différemment, ce qui nécessite de les caractériser séparément. L'évaluation des propriétés de la couche élastoplastique a ensuite été abordée avec une caractérisation énergétique du matériau. Le concept d'historique rhéologique mène à une évaluation et compréhension plus détaillée du matériau de même qu'à l'optimisation de sa composition et de sa mise en place, qui se matérialisera vraisemblablement à travers une démarche la perspective de modélisation et de prédiction du rebond. / For several years, the loss of material caused by rebound in shotcrete has been the subject of environmental concerns. Continuous optimization of equipment and placement makes the spaying technique more efficient. Université Laval's shotcrete laboratory has been carrying out a number of research projects in order to gain a better understanding and better control over the phenomenon. The knowledge acquired is orienting this research towards a more complete characterization of dry-mix shotcrete and its fresh substrate. A small-scale assembly, including recent technological advances on equipment, has been developed to provide significant technological and environmental benefits. Therefore, this implantation makes it possible to overcome certain problems of the traditional method and evaluate several mix-design with multiple dosages of admixtures quickly and without production and excessive use of material. Cast-in-place concrete is generally characterized in its fresh state using rheological parameters. However, traditional testing devices, such as rheometers, cannot be used with dry-mix shotcrete because of the consistency of the material spayed. Thus, a methodology was proposed in order to study and verify the applicability of rheological parameters. The complexity of the phenomenon present during placement of dry shotcrete, however, has proven this methodology inadequate to successfully capture the sought effects. This permitted to introduce the rheological history concept. The rheological history concept unites several theoretical parameters and experimental observations and offers a different approach for the description of the fresh state material and its placement. It identifies two areas separated by a critical energetic frequency. These areas are defined according to the condition of the substrate: fluid or elastoplastic. These two states are unavoidable and behave differently, which requires characterizing them separately. The evaluation of the elastoplastic layer's properties was approached with an energetic characterization of the material. The rheological history concept leads to a more detailed evaluation and understanding of the material, and the optimization of its composition and placement while bringing closer the perspective of rebound modeling and prediction.

published_or_final_version / abstract / Zoology / Master / Master of Philosophy

published_or_final_version / Ecology and Biodiversity / Doctoral / Doctor of Philosophy