La BSP1, principale protéine du plasma séminal bovin, interagit avec les membranes des spermatozoïdes et joue un rôle crucial dans les événements qui conduisent à la fécondité des spermatozoïdes, lors du phénomène de la capacitation. Le but de cette recherche est d’investiguer la nature de ces interactions. Ce travail vise à démontrer l’influence des lipides qui composent les membranes sur l’action de la protéine BSP1. À l’aide de la fluorescence intrinsèque de la protéine, l’affinité de la protéine a été caractérisée pour quatre systèmes lipidiques. Les résultats montrent que la composition lipidique affecte significativement l'affinité de la protéine pour les membranes. Nous avons observé l'ordre suivant : 1-palmitoyl-2-oléoyl-sn-glycéro-3-phosphocholine (POPC) > POPC/1-palmitoyl-2-oléoyl-sn-glycéro-3-phosphoéthanolamine (POPE) ≈ POPC/1-palmitoyl-2-oléoyl-sn-glycéro-3-phospho-L-sérine (POPS) > POPC/cholestérol. La protéine interagit préférentiellement avec POPC. La présence de POPE, POPS, ou cholestérol dans la membrane diminue systématiquement l’affinité. Il est connu que la présence de POPE ou cholestérol augmente l’empilement des lipides dans les membranes. Cet effet de condensation des chaînes pourrait être défavorable à l’insertion de la partie hydrophobe de la protéine dans les membranes et réduire ainsi l'affinité. La diminution de l’affinité de la protéine induite par la présence de POPS, un lipide chargé négativement, pourrait être associée aux interactions électrostatiques répulsives car la protéine porte une charge globale négative. La littérature mentionne que la BSP1 extrait sélectivement les phospholipides de type choline et le cholestérol lors de son association avec les membranes de spermatozoïdes. Un efflux lipidique est aussi observé avec des membranes modèles. Nous avons désiré caractériser la « solubilisation » des membranes par la BSP1, par diffusion dynamique de la lumière. Comme étape préliminaire, nous avons étudié comment le détergent Triton X-100 solubilise les membranes en utilisant cette technique. Les mesures démontrent que la composition lipidique des membranes (POPC, POPC/POPE, POPC/1-palmitoyl-2-oléoyl-sn-glycéro-3- [phospho-rac-(1-glycérol)] (POPG)) n’affecte pas le mécanisme général de solubilisation/reconstitution des membranes modèles. Il a été montré qu'il existe trois régions lors des processus de solubilisation pour les différents systèmes lipidiques : i) le détergent se distribue dans les membranes, ii) une coexistence de membranes saturées en détergents et de micelles mixtes de phospholipides/Triton X-100 et iii) exclusivement des micelles mixtes de phospholipides/Triton X-100. Nos résultats montrent que la forme conique de POPE augmente la résistance des membranes à la solubilisation. La présence de POPG, apportant une charge négative à l’interface des membranes, n’induit aucun changement aux processus de solubilisation/reconstitution des membranes par Triton X-100. La diffusion dynamique de la lumière a également permis d’observer si la protéine BSP1 induit des modifications morphologiques des membranes suite à son interaction avec les membranes de POPC. Nos observations n'ont montré aucune variation significative de la taille des particules lors du titrage des vésicules de POPC par la protéine, sur une gamme de rapport molaire de POPC/BSP1 variant de 20 à 0.6. Avec des compositions aussi différentes, on suppose une transition des vésicules saturées en protéine à des complexes de protéine avec un peu de lipides. Cependant, il semble impossible avec la diffusion dynamique de la lumière de différencier ces particules. / BSP1, the main protein in bovine seminal plasma, interacts with sperm membranes and plays a crucial role in events that lead to sperm fertility, during the capacitation. The purpose of this research is to investigate the nature of these interactions. This work aims to demonstrate the influence of the lipids that compose membranes on the action of the BSP1 protein. Using the intrinsic fluorescence of the protein, the affinity of the protein was characterized for four lipid systems. The results show that the lipid composition significantly affects the affinity of the protein for membranes. We observed the following order: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) > POPC/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) ≈ POPC/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS) > POPC/cholesterol. The protein interacts preferentially with POPC. The presence of POPE, POPS, or cholesterol in membranes decreases systematically the affinity. It is established that the presence of POPE or cholesterol increases the packing of lipids in membranes. This condensation effect could be detrimental to the insertion of the hydrophobic part of the protein into the membranes and reduces, as a consequence, the affinity. The decrease in protein affinity induced by the presence of POPS, a negatively charged lipid, could be associated with repulsive electrostatic interactions as the protein global charge is negative. The literature mentions that BSP1 selectively extracts choline phospholipids and cholesterol when combined with sperm membranes. A lipid efflux is also observed with model membranes. We characterized membrane "solubilisation" by BSP1, using dynamic light scattering. As a preliminary step, we studied how Triton X-100 detergent solubilizes membranes using this technique. The measurements showed that the lipid composition of the membranes does not affect the general solubilization/reconstitution mechanism of the model membranes (POPC, POPC/POPE, POPC/1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1-rac-glycerol) (POPG)). It is known that three different regions exist during the solubilization process for the different lipid systems: i) the detergent is distributed in the membranes, ii) a coexistence of membranes saturated with detergents and mixed phospholipid/Triton X-100 micelles and iii) exclusively mixed phospholipid/Triton X-100 micelles. Our results show that the conical shape of POPE increases the resistance of the membranes to solubilization. The presence of POPG, bringing a negative charge at the membrane interface, does not induce any change in solubilization/reconstitution processes. Dynamic light scattering also made it possible to observe if the BSP1 protein induces morphological changes in the membranes following its interaction with POPC membranes. Our observations showed no significant variation in particle size during the titration of POPC vesicles by the protein, over a molar ratio range of POPC/BSP1 from 20 to 0.6. Considering such different compositions, a transition from vesicles saturated with protein to protein complexes with some lipids is assumed. However, it appeared impossible with dynamic light scattering to differentiate these particles.

No description available.

This work deals with the current topic of carrier systems. Since the biggest problem is the passage of hydrophobic drugs through the bloodstream, or through universal body barriers (eg blood-brain), it is necessary to chemically modify these carriers in order to be able to administer hydrophobic substances effectively. Based on a literature search, several systems are designed and subsequently studied, in which there is a presumption of possible use for carrier systems and at the same time they have biosurfactants incorporated in them due to their ability to solubilize hydrophobic molecules. The theoretical part of this work will describe the individual biosurfactants, the process of their production, their physicochemical properties, and the possibility of their use in carrier systems. At the same time, the individual carrier systems, the procedure of their preparation, the possibilities of their use are described, and their advantages and disadvantages are also compared. In the practical part, the screening of both individual substances and their mutual interactions, as well as methods used to study the emerging structures is then performed. The study of molecular interactions is primarily performed using the technique of dynamic light scattering. The next part describes the optimization of hydrogel formation with incorporated biosurfactants in their structure and then the formed gels are subjected to rheological and solubilization tests. The study of the internal structure of these gels is performed using a scanning electron microscope.

This diploma thesis deals with the study of physical hydrogels based on positively-charged polyelectrolyte. The study is to investigate the interaction of selected positively-charged polyelectrolytes with oppositely charged surfactants in water and in physiologicla saline solution (0.15 M NaCl). The influence of the process for preparing hydrogels. Hydrogels were prepared by dry or wet. Were tested solubilization abilities hydrogels prepared using the hydrophobic dye oil red O were also measured rheological properties of the hydrogels prepared using frequency oscillatory tests and flow tests, depending on the concentration of the polymer or surfactant. For the rheological experiments suggest that chitosan is capable of forming rigid hydrogels with better mechanical properties than the dextran in the presence of both surfactants. Selected samples were also carried out experiments using fluorescent sold prodan, nile red and -naphthol as fluorescent probes. And in selected experiments was determined by solids content in percentage depending on the concentration used polyelectrolytes or surfactants.

Plus de 30% des enzymes présentent un ion métallique dans leur site actif qui est la clé pour les réactions de catalyse. Une sous-famille d’enzymes comporte un site actif où un seul ion métallique (Zn2+ ou Cun+) est coordiné à un cœur polyhistidine. L’élaboration de composés modèles est important pour la compréhension des mécanismes fondamentaux impliqués dans le cycle biocatalytique de ces enzymes mononucléaires. Dans ce contexte, nous avons développé des ligands basés sur des calix[6]arènes présentant un chapeau polyazoté lié de manière covalente au macrocycle. Ces ligands offrent un site de coordination pour le métal et une cavité hydrophobe bien définie. Cet assemblage permet et contrôle la liaison d’un ligand exogène dans la cavité. Afin de moduler les propriétés des complexes métalliques calixaréniques basés sur une unité coordinante tren (tris(2-aminoéthyl)amine), nous avons synthétisé avec succès et étudié plusieurs nouveaux récepteurs calix[6]aréniques. La synthèse de récepteurs bisétagés, le calix[6]amido-tren et le calix[6]amido-tacn, a permis de modifier la seconde sphère de coordination par incorporation de groupements amide entre la cavité et le site de coordination. Cette stratégie s’est avérée efficace. En effet, la coordination d’anions au centre métallique a été mise en évidence et en particulier celle simultanée de deux Cl¯ pour le Cu-calix[6]amido-tren. Cette coordination d’un ou plusieurs invités anioniques était impossible dans le cas du calix[6]tren parent qui présente une seconde sphère de coordination électroniquement dense et un chapeau trop petit pour y accueillir plusieurs invités. La présence de groupements carbonyle entre la chapeau tren et le calixarène modifie également le comportement des complexes : leur coordination au centre métallique entre en compétition avec celle d’invités exogènes. L’oxydation d’amines encombrées sous O2 en hydroxylamines et oximes a été réalisée par la réduction du complexe cuivrique du calix[6]amido-tren par électrochimie. Ce type d’amines n’est pas oxydable par l’adduit superoxo du complexe parent, dû à la taille de la cavité qui empêche la coordination d’invités encombrés. De plus, la formation d’un adduit superoxo a été mise en évidence qualitativement au départ du complexe cuivreux isolé de ce même composé. Ces résultats permettent d’envisager des études de réactivité mimant le site actif des enzymes PHM et DβM par exemple. Enfin, une comparaison des propriétés hôte-invité et de la réactivité des complexes du calix[6]amido-tren avec celles des complexes du calix[6]amido-tacn a été réalisée afin d’évaluer l’impact de la nature du chapeau polyazoté sur les propriétés hôte-invité et la réactivité de ces complexes. Une méthodologie de déméthylation assistée de manière supramoléculaire a été développée pour différents calix[6]azacryptands chapeautés. Celle-ci a ensuite été appliquée avec succès à la déméthylation du calix[6]tren pour obtenir le calix[6]trentrisPhOH. Ce récepteur a montré un comportement très différent de celui du récepteur parent. En effet, la complexation d’anions et d’ammoniums biologiquement pertinents a été possible par ce récepteur protonné. L’interaction entre le centre métallique et les phénates en présence de base a conduit à la réalisation d’un switch moléculaire multi-étapes entre différents états de protonation du récepteur et du complexe zincique. Son complexe cuivrique a montré la présence d’un radical phénoxyle après oxydation du phénate. Cette espèce CuII-radical phénoxyle mime la forme oxydée du site actif de la galactose oxydase, catalysant l’oxydation d’alcools en aldéhydes. Ces résultats ouvrent donc la voie à de nouvelles perspectives pour les études de réactivité de ce composé. Un analogue du calix[6]tren, porteur d’un bras avec une fonction réactive sur le chapeau, a été synthétisé et complexé au cuivre avant d’être immobilisé sur surface via la formation de monocouches. (...) / More than 30% of all enzymes present a metal ion in their active site. Interesting sub-families present a mononuclear active site where a single metal ion (Zn2+ or Cun+) is coordinated to a polyhistidine core. The elaboration of model compounds is important for understanding the fundamental mechanisms involved in their bio-catalytic cycles. In this context, we have developed calix[6]arene based ligands presenting an aza cap covalently linked to the calixarene moiety. These supramolecular assemblies present several advantages: they offer a coordination site for a metal ion through the nitrogen donor atoms of the cap, the small rim of the calixarene moiety defines the second coordination sphere, and the hydrophobic cavity presents an access channel for exogenous ligands. In these systems the coordinating nitrogen atoms are separated from the oxygen rich small rim by two carbon atoms, which allows the controlled coordination of one single guest inside the cavity. Wanting to modulate the properties of the calix[6]arene tren-based (tris(2-aminoethyl)amine) ligand, we successfully synthetized several new receptors and studied their reactivity. The synthesis of “two-story” receptors, the calix[6]amido-tren and calix[6]amido-tacn, allowed the modification of the first and second coordination sphere of a coordinated metal ion by introducing an additional spacer between the tripodal aza-cap and the calixarene macrocycle. The complexes of CuI, CuII and ZnII of these new ligands were synthesized and their reactivity was studied. The presence of three potentially coordinating carbonyl groups changed the host-guest reactivity of these metal complexes as they are in competition with exogenous ligands. A comparative study of the host-guest properties and of the reactivity of the metal complexes of both ligands, calix[6]amido-tren and calix[6]amido-tacn was realized. The cavity of these “two-story” ligands is larger than the one of the parent calix[6]tren and more flexible, allowing not only the coordination of an anion, but the simultaneous coordination of two Cl¯ ligands. The oxidation of sterically hindered amines into hydroxylamines and oximes in the presence of O2 was electrochemically realized by the reduction of the cupric center forming a CuII-superoxo adduct as the reactive species. Amine oxidation has been shown for the parent complex, however, the scope of this reaction was limited due to the smaller cavity. The formation of a CuII-superoxo adduct of the calix[6]amido-tren complex was qualitatively evidenced at low temperature upon addition of O2 to the isolated cuprous complex. This type of adduct has attracted much attention in recent years since it is considered to be a reactive intermediate in the catalytic cycle of copper monooxygenases, such as PHM and DβM. A supramolecular assisted demethylation methodology was developed for different capped calix[6]azacryptands. This methodology was then successfully applied to the demethylation of calix[6]tren in order to obtain calix[6]trentrisPhOH. This receptor showed a very different behavior in comparison to the parent one. Indeed, the complexation of anions and biologically relevant ammonium ions was evidenced by the monoprotonated receptor. The metal complexes of CuII and ZnII were synthesized. The interaction between the metal center and the phenate groups in presence of base has been evidenced and has conducted to a multi-step molecular switch. The cupric complex showed the presence of a phenoxyl radical resembling the oxidized form of galactose oxidase, catalyzing the oxidation of alcohols into aldehydes. These results open the way to new perspectives in reactivity studies. An analog of calix[6]tren, bearing a functionalized reactive arm on the tren cap, was synthesized and the corresponding CuII complex was immobilized on a gold surface through a monolayer formation. (...)

Der Mangel an Nährstoffen im Boden, hauptsächlich an Phosphor (P) und Stickstoff (N), verbunden mit einem hohen Salzgehalt und der generellen Verarmung landwirtschaftlicher Böden , sind ein ernstes Problem für die landwirtschaftliche Produktion weltweit. Daher besteht ein dringender Bedarf an Forschung und Entwicklung geeigneter landwirtschaftlicher Praktiken, um ungünstige Bodenbedingungen zu verringern und wenn möglich die Fruchtbarkeit von Kulturland wiederherzustellen. Die Verwendung von Rhizobakterien, die das Pflanzenwachstum (PGPR) fördern, kann sich bei der Entwicklung von Strategien zur Erleichterung des Pflanzenwachstums unter normalen Wachstumsbedingungen sowie unter abiotischen Stress als nützlich erweisen. Diese Bakterien bieten ihren pflanzlichen Wirten Vorteile, indem sie die Aufnahme von Bodenmineralien fördern und Pflanzen vor schädlichen Umwelteinflüssen schützen. Die vorliegende Arbeit bewertet die Rolle von in Kamerun natürlich vorkommenden PGPR an Mais und untersucht deren Potenzial als Bioimpfstoffe zur Steigerung des Pflanzenwachstums in Kamerun. Wir prüfen die Hypothese, dass einheimische Bakteriengemeinschaften aus Kamerun einen hohen Anteil an Bakterien aufweisen, deren Eigenschaften Kulturpflanzen helfen, mit ungünstigen Bedingungen umzugehen. In der vorliegenden Arbeit wurden dazu Bakteriengemeinschaften der Rhizosphäre von in Kamerun angebautem Mais isoliert und untersucht. Zum ersten Mal erfolgte eine umfassende phylogenetische Zuordnung aller kultivierbaren Bakterien, auf Grundlage ihrer potenziellen Fähigkeiten zur Förderung des Pflanzenwachstums. / Nutrient deficiencies in soil, mainly in phosphorus (P) and nitrogen (N), coupled to salinity and the impoverishment of agricultural soils, are a severe problem for agricultural production worldwide. Therefore, there is an urgent need for research and development of more suitable agricultural practices in order to reduce unfavorable conditions, and if possible, to restore the fertility of cultivated lands. The use of rhizobacteria, which promote plant growth (PGPR), can prove useful in developing strategies to facilitate plant growth under normal as well as under abiotic stress conditions. These bacteria offer benefits to plant hosts by promoting the uptake of soil minerals and protecting plants from environmental stresses. The thesis evaluates the role of native PGPR associated with maize as potential bio-inoculants for plants growth in Cameroon. We hypothesized that native bacterial communities from Cameroon include a high potential of bacteria helping the plant cope with unfavorable conditions. Here, we provide for the first time a comprehensive phylogenetic affiliation of cultivable bacterial communities associated with maize rhizosphere grown in Cameroon in relationship to their potential plant growth-promoting abilities.

In der Alzheimer-Forschung (engl.: Alzheimer Disease, AD) wird immenser Aufwand zur Entwicklung von den Krankheitsverlauf verändernden Medikamenten betrieben. Studien zeigten, dass die abnormale Aggregation des Tau-Proteins offenbar zum Zusammenbruch der Zellkommunikation führt. Niedermolekulare Substanzen, die die Tau-Protein-Aggregation inhibieren und sogar bereits gebildete Aggregate wieder auflösen können wurden entwickelt, sind jedoch oft aufgrund von schlechten Wasserlöslichkeiten nur unter Zusatz von Dimethylsulfoxid (DMSO) in Biotests einsetzbar. Durch das Design maßgeschneiderter Peptid-Polyethylenglykol (PEG)-Konjugate war die spezifische Bindung und anschließende Freisetzung ausgewählter potentieller anti-AD-Wirkstoffe in DMSO-freien Biotests möglich. Für den Entwurf der Wirkstoff-Transporter wurden Peptidbibliotheken mit Raman- und Fluoreszenz-Mikroskopie-basierten Methoden hinsichtlich der Anreicherung der Wirkstoffe an Peptiden mit hoher Wirkstoff-Bindekapazität getestet. Mithilfe von Matrix-unterstützter Laser-Desorption/Ionisation (MALDI)-Massenspektrometrie (MS/MS)-Fragmentierung konnten die Peptidsequenzen der positiven Treffer identifiziert werden. Die zugehörigen Konjugate wurden synthetisiert, mit den Wirkstoffen beladen und die entstehenden sehr gut wasserlöslichen Wirkstoff-Konjugat-Komplexe analysiert. Für biomedizinische Anwendungen sind kompakte und definierte Systeme von Vorteil. Zur Strukturaufklärung der Wirkstoff-Konjugat-Komplexe konnten zahlreiche Untersuchungen erfolgreich durchgeführt werden. Viele Komplexe wurden zudem in DMSO-freien Biotests der Tau-Protein-Aggregation eingesetzt. Die Bioverfügbarkeit der schwerlöslichen anti-AD-Wirkstoffe konnte durch die Solubilisierung mit maßgeschneiderten Peptid-PEG-Konjugaten enorm verbessert werden. Die auf Raman-aktive Substanzen erweiterte Screeningprozedur kann wahrscheinlich auf eine Großzahl von Wirkstoffen mit ungünstigen pharmakologischen Eigenschaften angewendet werden. / Considerable efforts are devoted in Alzheimer Disease (AD) research to develop disease modifying drugs. Various studies have demonstrated that abnormal aggregation of Tau protein probably interrupts communication between cells. Tau protein aggregation can be inhibited and even preformed aggregates can be redissolved by small-molecule compounds. Unfortunately, these molecules often can only be applied in limited biotests using dimethyl sulfoxide (DMSO) as co-solvent due to their poor water solubility and bioavailability. The solubilization of selected potential anti-AD drugs by tailored peptide-poly(ethylene glycol) (PEG) conjugates enabled the specific binding und subsequent release of these drugs in DMSO-free biotests. For the design of the drug conjugate carriers, large peptide libraries have been screened using Raman or fluorescence microscopy-based methods to follow drug enrichment on certain peptide library beads which exhibit high drug affinity. Identification of peptide sequences of positive hits was performed by Matrix-assisted Laser Desorption/Ionization (MALDI)-mass spectrometry (MS/MS) fragmentation. The corresponding conjugates were synthesized; loaded with the potential drugs and the resulting highly water-soluble drug transporter complexes were analyzed. Compact and defined complexes are desirable with regard to biomedical applications. Various studies on drug-peptide interactions, specifity of drug binding and influence of the different parts of the conjugates for drug capacities were performed successfully. Generated drug transporter complexes were finally tested in DMSO free bioassays. Depending on drug and peptide structures, the complexes could reach effects comparable to the drugs solubilized by DMSO. The bioavailability of poor water-soluble anti-AD compounds was largely improved. Presumably, the new developed Raman-screening procedure can be expanded to a great extent of compounds suffering from unfavorable pharmacological characteristics.

Heteropolymeric O antigen (O-Ag)-capped lipopolysaccharide is the principal constituent of the Gram-negative bacterial cell surface. It is assembled via the integral inner membrane (IM) Wzx/Wzy-dependent pathway. In Pseudomonas aeruginosa, Wzx translocates lipid-linked anionic O-Ag subunits from the cytoplasmic to the periplasmic leaflets of the IM, where Wzy polymerizes the subunits to lengths regulated by Wzz1/2. The Wzx and Wzy IM topologies were mapped using random C-terminal-truncation fusions to PhoALacZα, which displays PhoA/LacZ activity dependent upon its subcellular localization. Twelve transmembrane segments (TMS) containing charged residues were identified for Wzx. Fourteen TMS, two sizeable cytoplasmic loops (CL), and two large periplasmic loops (PL3 and PL5 of comparable size) were characterized for Wzy. Despite Wzy PL3–PL5 sequence homology, these loops were distinguished by respective cationic and anionic charge properties. Site-directed mutagenesis identified functionally-essential Arg residues in both loops. These results led to the proposition of a “catch-and-release” mechanism for Wzy function. The abovementioned Arg residues and intra-Wzy PL3–PL5 sequence homology were conserved among phylogenetically diverse Wzy homologues, indicating widespread potential for the proposed mechanism. Unexpectedly, Wzy CL6 mutations disrupted Wzz1-mediated regulation of shorter O-Ag chains, providing the first evidence for direct Wzy–Wzz interaction. Mutagenesis studies identified functionally-important charged and aromatic TMS residues localized to either the interior vestibule or TMS bundles in a 3D homology model constructed for Wzx. Substrate-binding or energy-coupling roles were proposed for these residues, respectively. The Wzx interior was found to be cationic, consistent with translocation of anionic O-Ag subunits. To test these hypotheses, Wzx was overexpressed, purified, and reconstituted in proteoliposomes loaded with I−. Common transport coupling ions were introduced to “open” the protein and allow detection of I− flux via reconstituted Wzx. Extraliposomal changes in H+ induced I− flux, while Na+ addition had no effect, suggesting H+-dependent Wzx gating. Putative energy-coupling residue mutants demonstrated defective H+-dependent halide flux. Wzx also mediated H+ uptake as detected through fluorescence shifts from proteoliposomes loaded with pH-sensitive dye. Consequently, Wzx was proposed to function via H+-coupled antiport. In summary, this research has contributed structural and functional knowledge leading to novel mechanistic understandings for O-Ag biosynthesis in bacteria. / Bookmarks within the document have been provided for ease of access to a particular section in the body of the thesis. Each entry in the Table of Contents, List of Tables, and List of Figures has been "linked" to its respective position and as such can be clicked for direct access to the entry. Similarly, each in-text Figure or Table reference has been "linked" to its respective figure/table for direct access to the entry. / 1.) Canadian Institutes of Health Research (CIHR) Frederick Banting and Charles Best Canada Graduate Scholarship doctoral award, 2.) CIHR Michael Smith Foreign Study Award, 3.) Cystic Fibrosis Canada (CFC) doctoral studentship, 4.) University of Guelph Dean's Tri-Council Scholarship, 5.) Ontario Graduate Scholarship in Science and Technology, 6.) Operating grants to Dr. Joseph S. Lam from CIHR (MOP-14687) and CFC