<p><i>Lactococcus lactis,</i> prolidase (<i>Lla</i>prol) hydrolyzes Xaa-Pro dipeptides. Since Xaa-Pro is known as bitter peptides, <i>Lla</i>prol is potentially applicable to reduce bitterness of fermented foods. <i>Lla</i>prol shows allosteric behaviour and substrate inhibition, which are not reported in other prolidases. Computer models of <i>Lla</i>prol based on an X-ray structure of non-allosteric <i>Pyrococcus furiosus</i> prolidase showed that a loop structure (Loop^32-43) is located at the interface of the protomers of this homodimeric metallodipeptidase. This study investigated roles of four charged residues (Asp³⁶, His³⁸, Glu³⁹, and Arg⁴⁰) of Loop^32-43 in <i>Lla</i>prol using a combination of kinetic examinations of ten mutant enzymes and their molecular models. Deletion of the loop structure by Î36-40 mutant resulted in a loss of activity, indicating Loop^32-43 is crucial for the activity of <i>Lla</i>prol. D36S and H38S exhibited 96.2 % and 10.3 % activity of WT, whereas little activities (less than 1.0 % of WT activity) were observed for mutants E39S, D36S/E39S, R40S, R40E, R40K and H38S/R40S. These results implied that Glu³⁹ and/or Arg⁴⁰ play critical role(s) in maintaining the catalytic activity of <i>Lla</i>prol. These observations suggested that the loop structure is flexible and this attribute, relying on charge-charge interactions contributed by Arg⁴⁰, Glu³⁹ and Lys¹⁰⁸, is important in maintaining the activity of <i>Lla</i>prol. When the loop takes a conformation close to the active site (closed state), Asp³⁶ and His³⁸ at the tip of the loop can be involved in the catalytic reaction of <i>Lla</i>prol. The two active mutant prolidases (D36S and H38S) resulted in modifications of the unique characteristics; the allosteric behaviour was not observed for D36S, and H38S <i>Lla</i>prol showed no substrate inhibition. D36E/R293K, maintaining the negative charge of position 36 and positive charge of position 293, still possessed the allosteric behaviour, whereas the loss of the charges at these positions (D36S of this study and R293S of a previous study (Zhang et al., 2009 BBA-Proteins Proteom 1794, 968-975) eliminated the allosteric behaviour. These results indicated the charge-charge attraction between Asp³⁶ and Arg²⁹³ is important for the allostery of <i>Lla</i>prol. In the presence of either zinc or manganese divalent cations as the metal catalytic centre, D36S and H38S enzymes also showed different substrate preferences from WT <i>Lla</i>prol, implying the influence of Asp³⁶ and His³⁸ on the substrate binding. D36S and H38S also showed higher activities at pH 5.0 to 6.0, in which range WT <i>Lla</i>prol steeply decreased its activity, indicating Asp³⁶ and His³⁸ are involved in the active centre and influence the microenvironment of catalytic His²⁹⁶. The above observations are attributed to modifications of their local structure in the active centre since the temperature dependency and thermal denaturing temperature indicated little effects on the overall structure of the <i>Lla</i>prol mutants.</p> <p>From these results, we concluded that the unique behaviours of <i>Lla</i>prol are correlated to Loop^32-43 and Asp³⁶ and His³⁸ on it. When Loop^32-43 takes a closed conformation, Asp³⁶ interacts with Arg²⁹³ via charge-charge attraction to form an allosteric subsite. The saturation of the allosteric site with substrates further allowed the communications of His³⁸ with S₁ site residues to complete the active site. When the substrate concentration becomes higher than it is required to saturated productive S₁' site, His³⁸, Phe¹⁹⁰ and Arg²⁹³ would resemble the residue arrangement of S₁' site residues (His²⁹², Tyr³²⁹, and Arg³³⁷) and bind to the proline residue of substrates. This non-productive binding would prevent the conformational change of Loop^32-43, which further results in the substrate inhibition. For further confirmation of this mechanism, crystallographic studies will be conducted. In this thesis, we have indentified the conditions to produce crystals of <i>Lla</i>prol proteins.</p>

Site quality, or the ability of land to grow trees, is an important component for identifying the most appropriate locations for establishing plantations of fast growing tree species to meet societys demands for timber and other environmental benefits. The goal of this thesis project was to predict site quality for poplars using soil and site information in Saskatchewan, Alberta and Quebec and to examine the effect of site quality on competition control in hybrid poplar plantations in Saskatchewan.<p>The first study examined factors affecting trembling aspen productivity in the boreal shield ecozone of Quebec on till and fluvial parent materials using general map data and measured soil and site information. Relationships with productivity were stronger using measured soil and site variables for individual parent materials (R2>0.6) than using general map data only (R2<0.25). Including biological variables, such as overstory species composition, had a major impact on site quality with conifer dominance negatively impacting the growth of trembling aspen.<p>The second study examined the factors affecting trembling aspen productivity in the boreal transition ecoregion of Saskatchewan on three different soil parent materials: fluvial, lacustrine and till. Relationships with productivity were stronger using soil and site variables for individual parent materials (R2 0.48-0.58) than using agricultural capability classes or other soil properties for all plots combined (R2<0.2). For fluvial and lacustrine sites, increasing clay content and nutrient availability (e.g. pH and total N) were positively related to productivity while tree productivity was negatively related to poor drainage for till sites. <p>The third study examined the factors affecting site quality for a single hybrid poplar clone in industrial plantations in Alberta at both the local scale (between plantations) and the microsite scale (within plantations). At the local scale, foliar P and Cu concentrations, soil water availability and drainage, and Ca and Mn in the C horizon were related to hybrid poplar productivity. There were also curved relationships with productivity and soil texture in the B horizon and pH of the A horizon, indicating an optimal range for poplar growth. At the microsite scale, soil texture was the best predictor of productivity with different relationships at each site depending on where the sites were in relation to the optimal soil texture.<p>The final study examined the response of hybrid poplar plantations in Saskatchewan to interspecific competition control on a range of site productivities. Competition control greatly increased tree growth with the greatest benefit being on the best quality sites. Both water and nutrients were highly competed for between trees and weeds. In the weed-free plots, tree growth was positively related to the amount of silt and clay in the soil and foliar P concentrations. This series of studies has demonstrated that it should be possible to predict poplar productivity reasonably well using only soil and site information within limited areas across Canada. However, the important drivers of productivity varied between the regions studied and between site groupings, such as by parent material, within local areas. This information can now be used to help land managers make better decisions regarding the establishment and management of plantations of fast growing tree species, notably hybrid poplar plantations.

This work is a compilation of ideas intended as a framework for an alternative approach to engaging ‘site’ in the design process, an approach that maintains and explores the complexities and subtleties of a landscape, of a place. Through two parallel explorations - one that considers an expanded and inclusive interpretation of landscape as the frame through which we engage with, and design, our surroundings, and one that examines the specific nature of this engagement as exchange between the self and the milieu - such an approach has been developed. This approach is called RECONNAISSANCE. Through encouragement of explicit, conscious consideration of how we perceive and experience a landscape, how this contributes to an understanding of a particular place and how this relates to and informs the practice of landscape architecture (both the process and the outcome), RECONNAISSANCE contributes to a strengthening of our abilities and actions as landscape architects. / February 2008

Le développement d'applications s'appuyant sur le Web a connu ces dernières années une très forte croissance. Dans le secteur public, les sites Web sont devenus l'épine dorsale du système informatique du gouvernement car leur importance est vitale à cause des services essentiels qu'ils offrent aux citoyens et aux entreprises. Le besoin de mettre au point des principes et méthodes spécifiques au développement de ce type d'applications d'une manière générale, et à l'évaluation de leur qualité se fait de plus en plus ressentir dans l'industrie du génie logiciel. En effet, l'évaluation quantitative de la qualité des applications Web (critères et métriques) est très récente. Dans ce mémoire, nous présentons d'abord les deux principales méthodes traditionnelles d'évaluation de la qualité des sites Web du gouvernement du Canada, à savoir l'analyse de la conformité à des normes et directives gouvernementales du Canada et la méthode de calcul de l'indice de maturité des services publics en ligne puis par la suite les principales composantes d'une méthode nouvelle, WebQEM (Web Quality Evaluation Method), à savoir l'arbre de qualité basé sur la norme ISO 9126 et la structure mathématique du modèle WebQEM. Finalement, nous proposons une adaptation de la méthode WebQEM pour son application à l'environnement spécifique du ministère RHDSC, ce qui conduira à une évaluation du niveau de satisfactions de 10 facteurs de qualité du site Web de ce ministère. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Évaluation quantitative, Qualité, Arbre de qualité, WebQEM, ISO 9126.

Le secteur des aliments fonctionnels, nutraceutiques et produits de santé naturels est un secteur émergent très présent au Québec par le nombre d'entreprises qui s'y trouvent (Saives et Cloutier, 2003, 2006; Demol, 2001). Ces entreprises sont très hétérogènes et comprendre les modalités de leur développement au Québec à long terme est source de questionnement avec l'entrée en vigueur d'une nouvelle réglementation des produits de santé naturels en 2004. Aussi, dans une perspective territoriale, la présente recherche tente de comprendre pourquoi et comment les entreprises de ce secteur s'ancrent au Québec. Dans ce travail, l'ancrage correspond à une dynamique de co-création de ressources entre l'entreprise et le territoire où elle est localisée, par la co-construction de ressources et compétences dans des espaces partagés dits « Ba ». En effet, les entreprises possèdent des ressources qui peuvent être stratégiques, c'est-à-dire sources d'avantage concurrentiel durable (Wernerfelt, 1984; Barney, 1991). Parmi les différentes ressources de l'entreprise, les connaissances s'avèrent les plus stratégiques. En effet, « Aujourd'hui, les connaissances et la capacité à créer et utiliser les connaissances sont considérées comme la plus importance source d'avantage concurrentiel soutenu de la firme » (Nonaka et Toyama, 2003, p. 2). La gestion des connaissances devient donc par le fait même une compétence importante pour les entreprises. Emprunté au modèle de Nonaka et Takeushi (1997), le concept de Ba est utilisé dans la littérature récente en économie régionale (Gilly et Lung, 2004) et rapproché de la notion de proximité afin d'expliquer la création et l'exploitation de ressources plus ou moins ancrées. Simultanément, à partir d'une littérature classique sur le sujet (Nekka et Dokou, 2004), les facteurs du choix de la localisation des entreprises sont identifiés pour comprendre l'essence de leur ancrage. Pour répondre à la question de recherche dans le cadre théorique choisi, une étude multi-cas a été menée par le biais d'entrevues de type semi dirigé auprès de cinq entreprises du secteur au Québec. Les données textuelles recueillies ont par la suite été analysées par catégorisation à la fois manuelle et statistique. Une interprétation selon laquelle il existe différents modèles de choix de localisation des entreprises et que celles-ci évoluent dans deux sortes de Ba reliés à leur environnement est proposée. Le croisement entre les Ba et les modèles de choix de localisation a permis de faire ressortir des modèles d'ancrage territorial différents selon les entreprises et leurs stratégies respectives. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Nutraceutique, aliment fonctionnel, produit de santé naturel, territoire, facteurs de localisation.

Deuteron 1D and 2D NMR spectroscopy was used to study the dynamics of water molecules within the mesoporous material MCM-41. The deuteron spectra show three magnetization components for a sample hydrated to a 0.2 monolayer level. One component was assigned to the pore surface silanol group deuterons that exhibit a broad Gaussian line of 32.6 kHz FWHM and the other components were assigned to the water deuterons. At room temperature one water deuteron component has a powder pattern line shape (splitting of about 4.2 kHz and population of about 61%) and the other has a Lorentzian line shape (about 388 Hz FWHM and population of 39%). Magnetization exchange occurs between these components. An exchange model, based on multi-site exchange, was constructed and used to analyse the results for exchange. For the 0.2 monolayer sample the rate of magnetization exchange out of the hydration site where the water deuterons exhibit a Lorentzian line in the deuteron spectra is 1.3 ms. 2D measurements at 233 K and room temperature confirmed the magnetization exchange scenario for the two water deuteron sites. Combining the deuteron results with proton-silicon cross polarization magic angle spinning experiments together with heat treatment of the sample, definitive hydration site identification for MCM-41 was achieved. This study has shown that the water molecules bound to the hydrogen-bonded silanol groups produce the powder pattern while water molecules bound to the single silanol groups produce the Lorentzian line. This represents a necessary first step toward a meaningful modeling of NMR observables in terms of site-specific water molecule coordination and dynamics in MCM-41.

Progress in DNA sequencing of plant genomes has revealed that, in addition to microorganisms, a number of plants contain genes which share similarity to microbial 1-aminocyclopropane-1-carboxylate (ACC) deaminases. ACC deaminases break down ACC, the immediate precursor of ethylene in plants, into ammonia and α-ketobutyrate. We therefore sought to isolate putative ACC deaminase cDNAs from tomato plants with the objective of establishing whether the product of this gene is a functional ACC deaminase. It was demonstrated that the enzyme encoded by the putative ACC deaminase cDNA does not have the ability to break the cyclopropane ring of ACC, but rather that it utilizes D-cysteine as a substrate, and in fact encodes a D-cysteine desulfhydrase. Kinetic characterization of the enzyme has shown that it is similar to other previously characterized D-cysteine desulfhydrases. Using site-directed mutagenesis, it was shown that altering two amino acid residues within the predicted active site changed the enzyme from D-cysteine desulfhydrase to ACC deaminase. Concomitantly, it was shown that by altering two amino acids residues at the same position within the active site of ACC deaminase from Pseudomonas putida UW4 changed this enzyme into D-cysteine desulfhydrase.

This dissertation argues that a multi-site church that is not characterized by campus-wide gatherings does not have a sound biblical or theological basis for considering itself one church and is inconsistent with historic congregationalism. This argument is made in six steps. First, a framework for determining whether or not a church practice is biblical is put forward. Second, it is shown that the people of God as one assembly is a central theme in the Old Testament. Third, it is demonstrated that this theme of the people of God as one assembly is also a central theme of the New Testament. Fourth, it is argued that there is not precedent for multi-site church structure in the house churches of the New Testament. Fifth, it is shown that multi-site is not consistent with historic congregationalism as seen in Baptist ecclesiology. And sixth, it is argued that multi-site church structure is consistent with historic connectionalism as seen, for example, in Methodist ecclesiogy--a fact that only serves to heighten the problem with multi-site from a congregationalist point of view.

<p><i>Lactococcus lactis,</i> prolidase (<i>Lla</i>prol) hydrolyzes Xaa-Pro dipeptides. Since Xaa-Pro is known as bitter peptides, <i>Lla</i>prol is potentially applicable to reduce bitterness of fermented foods. <i>Lla</i>prol shows allosteric behaviour and substrate inhibition, which are not reported in other prolidases. Computer models of <i>Lla</i>prol based on an X-ray structure of non-allosteric <i>Pyrococcus furiosus</i> prolidase showed that a loop structure (Loop^32-43) is located at the interface of the protomers of this homodimeric metallodipeptidase. This study investigated roles of four charged residues (Asp³⁶, His³⁸, Glu³⁹, and Arg⁴⁰) of Loop^32-43 in <i>Lla</i>prol using a combination of kinetic examinations of ten mutant enzymes and their molecular models. Deletion of the loop structure by Î36-40 mutant resulted in a loss of activity, indicating Loop^32-43 is crucial for the activity of <i>Lla</i>prol. D36S and H38S exhibited 96.2 % and 10.3 % activity of WT, whereas little activities (less than 1.0 % of WT activity) were observed for mutants E39S, D36S/E39S, R40S, R40E, R40K and H38S/R40S. These results implied that Glu³⁹ and/or Arg⁴⁰ play critical role(s) in maintaining the catalytic activity of <i>Lla</i>prol. These observations suggested that the loop structure is flexible and this attribute, relying on charge-charge interactions contributed by Arg⁴⁰, Glu³⁹ and Lys¹⁰⁸, is important in maintaining the activity of <i>Lla</i>prol. When the loop takes a conformation close to the active site (closed state), Asp³⁶ and His³⁸ at the tip of the loop can be involved in the catalytic reaction of <i>Lla</i>prol. The two active mutant prolidases (D36S and H38S) resulted in modifications of the unique characteristics; the allosteric behaviour was not observed for D36S, and H38S <i>Lla</i>prol showed no substrate inhibition. D36E/R293K, maintaining the negative charge of position 36 and positive charge of position 293, still possessed the allosteric behaviour, whereas the loss of the charges at these positions (D36S of this study and R293S of a previous study (Zhang et al., 2009 BBA-Proteins Proteom 1794, 968-975) eliminated the allosteric behaviour. These results indicated the charge-charge attraction between Asp³⁶ and Arg²⁹³ is important for the allostery of <i>Lla</i>prol. In the presence of either zinc or manganese divalent cations as the metal catalytic centre, D36S and H38S enzymes also showed different substrate preferences from WT <i>Lla</i>prol, implying the influence of Asp³⁶ and His³⁸ on the substrate binding. D36S and H38S also showed higher activities at pH 5.0 to 6.0, in which range WT <i>Lla</i>prol steeply decreased its activity, indicating Asp³⁶ and His³⁸ are involved in the active centre and influence the microenvironment of catalytic His²⁹⁶. The above observations are attributed to modifications of their local structure in the active centre since the temperature dependency and thermal denaturing temperature indicated little effects on the overall structure of the <i>Lla</i>prol mutants.</p> <p>From these results, we concluded that the unique behaviours of <i>Lla</i>prol are correlated to Loop^32-43 and Asp³⁶ and His³⁸ on it. When Loop^32-43 takes a closed conformation, Asp³⁶ interacts with Arg²⁹³ via charge-charge attraction to form an allosteric subsite. The saturation of the allosteric site with substrates further allowed the communications of His³⁸ with S₁ site residues to complete the active site. When the substrate concentration becomes higher than it is required to saturated productive S₁' site, His³⁸, Phe¹⁹⁰ and Arg²⁹³ would resemble the residue arrangement of S₁' site residues (His²⁹², Tyr³²⁹, and Arg³³⁷) and bind to the proline residue of substrates. This non-productive binding would prevent the conformational change of Loop^32-43, which further results in the substrate inhibition. For further confirmation of this mechanism, crystallographic studies will be conducted. In this thesis, we have indentified the conditions to produce crystals of <i>Lla</i>prol proteins.</p>

Deuteron 1D and 2D NMR spectroscopy was used to study the dynamics of water molecules within the mesoporous material MCM-41. The deuteron spectra show three magnetization components for a sample hydrated to a 0.2 monolayer level. One component was assigned to the pore surface silanol group deuterons that exhibit a broad Gaussian line of 32.6 kHz FWHM and the other components were assigned to the water deuterons. At room temperature one water deuteron component has a powder pattern line shape (splitting of about 4.2 kHz and population of about 61%) and the other has a Lorentzian line shape (about 388 Hz FWHM and population of 39%). Magnetization exchange occurs between these components. An exchange model, based on multi-site exchange, was constructed and used to analyse the results for exchange. For the 0.2 monolayer sample the rate of magnetization exchange out of the hydration site where the water deuterons exhibit a Lorentzian line in the deuteron spectra is 1.3 ms. 2D measurements at 233 K and room temperature confirmed the magnetization exchange scenario for the two water deuteron sites. Combining the deuteron results with proton-silicon cross polarization magic angle spinning experiments together with heat treatment of the sample, definitive hydration site identification for MCM-41 was achieved. This study has shown that the water molecules bound to the hydrogen-bonded silanol groups produce the powder pattern while water molecules bound to the single silanol groups produce the Lorentzian line. This represents a necessary first step toward a meaningful modeling of NMR observables in terms of site-specific water molecule coordination and dynamics in MCM-41.