There are an estimated 2 billion individuals infected with Mtb, and 37.7 million people living with HIV (PLWH) worldwide. HIV/Mtb co-infection increases the risk of developing active tuberculosis by over 20-fold, and 210,000 of 1.5 million deaths from TB were among co-infected PLWH in 2020. Therefore, development of effective TB vaccination, particularly within the vulnerable PLWH population, is an urgent global issue. With limited in vivo models to study co-infection, humanized NRG (huNRG) mice and humanized DRAG-A2 mice (a next-generation of huNRG mice expressing HLA class I and II transgenes with improved human immune reconstitution, huDRAG-A2) are promising tools for HIV and TB reserach as they develop robust human immune cell populations and recapitulate many aspects of HIV or TB clinical disease. HIV/Mtb co-infection was investigated using huNRG and hu-DRAG-A2 mice in separate experiments where intravaginal (with DMPA pre-treatment) or intraperitoneal HIV-1 infection was administered, respectively, and intranasal infection of Mtb was administered 3.5 weeks later. Both huNRG and huDRAG-A2 mice recapitulated hallmark features of HIV/Mtb co-infection such as severe granuloma pathology, hCD4+ T cell depletion in lung and spleen tissue, and human like lung pathology such as Mtb-infected foamy macrophages in the granuloma. Co-infected huDRAG-A2 mice also displayed significantly higher bacterial burden in the lungs, increased extrapulmonary dissemination into spleen and liver, and significantly lower hCD4+ T cells in the peripheral blood post-Mtb infection when compared to the Mtb-only infected group. To investigate TB vaccine immunogenicity, huNRG and huDRAG-A2 mice were immunized with a novel trivalent vaccine, AdCh68MV. Upon intranasal immunization, both models showed trends of developing higher Mtb antigen-specific hCD4+ T cell responses in the lung and spleen. Overall, this project sets the initial stages of a pre-clinical HIV/Mtb co-infection model in huNRG and huDRAG-A2 mice appropriate for immune investigations, therapeutic and vaccination development. / Thesis / Master of Science in Medical Sciences (MSMS) / There are over 2 billion individuals infected with TB and 37.7 million people living with HIV (PLWH) worldwide. When someone is co-infected with both diseases, the risk of death is greatly increased. Research in co-infection and developing effective TB vaccination for PLWH are urgent global issues. Animal studies are currently limited because studying HIV requires human immune cells. Our lab has established humanized mice (hu-mice) that develop many different human immune cells and are useful for HIV/Mtb co-infection research. When hu-mice were co-infected, they showed more dying lung tissue, immune cell loss, and bacteria in the lungs. Hu-mice were also used to study human immune responses to a novel TB vaccine delivered to the lungs. Trends of higher immune responses towards TB were observed in the lung and spleen of immunized hu-mice. Overall, this project shows the utility of hu-mice as pre-clinical models of HIV/Mtb co-infection and Mtb vaccine studies.

Doctor of Philosophy / Department of Chemistry / Christer B. Aakeröy / Co-crystallization allows the manipulation of physical properties of a given compound without affecting its chemical behavior. The ability to predict hydrogen bonding interactions, provides means to the rational design of supramolecular architectures. It also makes it possible to select with a degree of accuracy, a few co-formers that have a high probability of forming co-crystals with a compound of interest, instead of blindly screening against a large number of candidates. To study the effects of changing electronic environment on the ability to form co-crystals, five symmetric dioximes of different hydrogen bond donating ability were synthesized with different functional groups on the carbon α to the oxime moiety. It was shown that the supramolecular yield increase with the positive MEP value on the donor site. In order to further explore this relationship between calculated MEP values and supramolecular selectivity three asymmetric ditopic donors containing phenol carboxylic acid and aldoxime groups were screened against a series of asymmetric ditopic acceptors. Nine crystal structures show that the supramolecular outcome can be predicted according to Etter’s rules by ranking donors and acceptors according to calculated MEP values. To explore the possibility of using the same approach with other hydrogen bond donors, three asymmetric ditopic donor ligands containing cyanooxime groups were synthesized and screened against a series of asymmetric ditopic acceptors. Nine out of ten times the supramolecular outcome could be predicted by MEP calculations 1-deazapurine exists in two tautomeric forms (1H and 3H) in aqueous solution, which have very different hydrogen bonding environments. The 3H tautomer forms a self-complementary dimer involving a donor and an acceptor site leaving a second acceptor site vacant. In order to stabilize this tautomer the molecule was screened against a of series hydrogen and halogen bond donors. Four out of five structures obtained showed 3H tautomer. The 1H tautomer is the geometric complement of urea. Therefore the molecule was screened against a series of N,N-diphenylureas and all five structures showed the 1H tautomer.

The purpose of the study was to describe and analyze the living condition for individuals with co-morbidity in different spheres of life based on a survey conducted in Blekinge county council. The aim has also been to examine the relation between individuals with co-morbidity and the care and support that is available under current legislation. The study examined how two hypotheses about comorbidity is related to different spheres of life as well as health care and support. Both hypotheses was confirmed by the results of the study. Individuals with co-morbidity has often a troublesome living situations and they do not receive adequate care and support. The study's perhaps most unexpected result is the result that emerged in addition to the hypotheses. The young group of people with co-morbidity is high and the group has a great need for extended care and support activities in all areas of life. Individuals with co-morbidity needs to be a continued research focus, especially the young persons.

The remittances transferred from Ecuadorian emigrants to Ecuador represent the second largest source of foreign device for the country. It is clear that such large amount of money makes an important impact on the economy. This study aims at evaluating the sustainability of the remittance generated development in the receiving households and also to find evidence if the organizations, authorities and other actors who work in relation to migration, remittances and development are able to increase this level of sustainability by implementing projects. Both quantitative and qualitative methods are used in order to approach these issues. Questionnaires are carried out with remittance receivers and the results are compared to the results from previous studies in order to identify the usage of remittances. Now, as before it shows that the large majority of the remittances are used in order to cover the household’s daily expenses. The rate of investments and savings is on the other hand low, a situation that creates a fragile development that is heavily dependent on the continued inflows of remittances. This is not a new discovery and several actors including governmental authorities, NGO’s as well as migrant’s associations and international institutions have therefore contributed with development strategies in order to intent increase the sustainability. In order to identify these actors and learn about their objectives and development ideas are semi-structured interviews carried out. One main concern among these actors is to increase the level of investments in productive activities such as micro businesses. Such investments are for example incentivized by providing training for micro entrepreneurs, issuing seed money, promoting local financial structures and facilitating access to micro credits.

La fibrose kystique (FK) est une maladie génétique autosomale récessive conduisant à une défaillance pulmonaire mortelle. Celle-ci est causée par l’expression dysfonctionnelle de l’allèle CFTR codant pour une protéine transmembranaire impliquée dans le transport Cl- des cellules de l’épithélium pulmonaire vers la lumière des voies respiratoires. Cette mutation induit la production d’un mucus visqueux qui entrave les voies respiratoires et favorise l’accumulation de bactéries pathogènes. L’éradication de cette présence bactérienne est maintenant l’enjeu primordial chez les patients FK afin de procurer un bien-être et de prolonger l’espérance de vie de ceux-ci. Le cheval de bataille de cette lutte au mieux-être du patient FK s’appuie en partie sur l’efficacité des antibiotiques. Cependant, nous nous heurtons à la capacité d’adaptation des bactéries face à l’antibiothérapie, ce qui nous conduit à développer sans cesse de nouvelles molécules thérapeutiques. Ainsi, l’essence de ce projet de recherche a été de caractériser l’efficacité de la tomatidine (TO); l’aglycone de la tomatine (un glycoalcaloïde), produit par les plants de tomate et de son analogue : la molécule FC04-100. Par le passé, notre laboratoire a établi que la TO possède une action antibactérienne contre Staphylococcus aureus prototype via l’inhibition de l’expression des facteurs de virulences associés au système de régulation ARG ainsi qu’en agissant comme potentialisateur d’action des aminoglycosides (AMI). De plus, la TO est également un inhibiteur de la réplication intracellulaire de S. aureus small-colony variant (SCV) infectant des cellules épithéliales pulmonaires différenciées (Calu-3). Le premier volet de mon projet a été consacré à l’évaluation du potentiel d’action de la TO contre S. aureus et Pseudomonas aeruginosa; deux bactéries fréquemment co-isolées des poumons des patients. Ainsi, lors de cette étude, j’ai démontré pour la première fois que la TO peut être employée seule contre S. aureus en tant qu’agent bactéricide lorsque celle-ci est utilisée en présence de P. aeruginosa. Ce phénomène dépend de la production par P. aeruginosa, du 2-heptyl-4-quinolone-N-oxide (HQNO) de l’endopeptidase LasA. De plus, j’ai évalué la possibilité d’utiliser un AMI et TO lorsque S. aureus et P. aeruginosa sont en co-culture afin de réduire la population bactérienne de ces deux pathogènes. Les résultats obtenus ont permis de démontrer qu’une combinaison, de tobramycine (TOB) et TO, permet d’inhiber significativement la croissance bactérienne d’un S. aureus résistant à la méthiciline (MRSA) résistant à la TOB et P. aeruginosa, co-cultivés en condition planctonique. Le deuxième volet de ma maîtrise visait à mesurer l’efficacité antibactérienne de FC04-100 en collaboration avec ma collègue Isabelle Guay. Pour ce projet, j’ai démontré l’efficacité de la combinaison FC04-100 et la gentamicine (GEN) contre un biofilm de S. aureus. Pour ce faire, j’ai utilisé une méthode de culture en microplaque 96 puits, permettant ainsi de former plusieurs bioflims et de tester plusieurs concentrations d’antibiotiques. Les résultats suite à l’exposition des biofilms à la combinaison TO-GEN ont démontré qu’il était possible de réduire significativement la viabilité bactérienne de S. aureus en biofilm. De plus, j’ai comparé l’efficacité de FC04-100 à TO dans un essai d’infection de cellules Calu-3 différenciées et infectées par une souche clinique de S. aureus SCV. Les résultats révèlent que ces deux composés diminuent significativement la viabilité bactérienne, et ce, en proportion similaire par rapport aux cellules infectées non traitées. L’ensemble des résultats obtenus dans ces deux projets a permis de démontrer clairement le potentiel antimicrobien de la TO et de son analogue FC04-100. Cette découverte apporte donc un nouveau squelette de molécule qui pourrait s’avérer utilisable comme antibiotique.

With the current trends in integrated management of childhood diseases in developing countries, it is important to resolve the controversies of coinfections between helminths and malaria, and properly evaluate the epidemiology of diarrhegenic parasites with molecular study, which sometimes cause overlapping infections. Again, the major challenge facing the global success of mass deworming initiative aimed at controlling helminths is lack of sustainability due to limited donor funds. We therefore decided to evaluate the effectiveness of a school based deworming program using only the school teachers without spending money on training and logistics. Demographic information, height and weight were measured and stool samples were collected from pupils in a semi-rural area of Nigeria during the initial visit by the study team. Malaria cases were recorded over a 3 month malaria transmission period prior to stool sampling. Four hundred and seventy six (33%) of the study population was infected with one Soil transmitted helminth (STH) or the other, especially with Askaris lumbricoides (26.0%) and Hookworm (8.4%). We found a negative association between malaria and STH in this community. Helminth infection rate of 18.3% was observed in children with malaria compared to 34.4% in controls. We also found a high carriage rate of Giardia (37.2%), low Cryptosporidium (1%) and no E. histolytica infection contrary to previous studies that were based on traditional diagnostic techniques. There was 7.9% reduction in the number children with low weight-for-age in the helminth infected children at 6 months after mass deworming, the number of uninfected children with low weight-for-age also reduced by 3.2%. There was also a reduction in the number of children with more than 25% absenteeism among both helminth infected (13.9%) as well as uninfected (7.2%). The association between malaria and STH in our study calls for the need for integrated approach to health problem in Africa instead of the common vertical campaigns. Results from our molecular study also shows the need to strengthen collaborations between researchers from developed and developing countries to be able to map out the true epidemiology of these parasites and hopefully produce novel, inexpensive diagnostics that circumvent the need for advance technological infrastructure

Crafting and stimulating service innovation is considered a main research priority and remains a challenge for service providers. One suggested component of stimulating service innovation is customer creativity. Customers who adapt, modify and transform services or products to better suit themselves are increasingly being recognized as a source of competitive value and innovation. It has been proposed that understanding and supporting the customer’s value creating practices is the key to creating and sustaining value over time in health care. Health services directly address a customer’s well-being and have a significant impact on his or her quality of life. In these types of services, the service outcome is highly dependent on the activities of the individual customer. Health care services often require customers to participate extensively, over long periods of time, with limited support and control. Health services also stretch far beyond the particular service setting into the customer’s daily life. While research, policy, and legislation have all emphasized the active role of health care customers, such customers have traditionally had few opportunities to design their health care services. Nevertheless, health care customers solve health-related problems and engage in self-care and medical decision-making on a day-to-day basis, although this creativity is often unknown to the service provider. To understand how health care customers can enable service innovation, this thesis seeks to conceptualize and investigate the concept of customer creativity in health care. The thesis focuses on customer creativity, not only as an outcome, but also as a dynamic and contextualized process that can be enhanced. The thesis combines insights from health care research with service and innovation research to provide build a framework for health care customer creativity. Building on five papers, the research develops an understanding for health care customer creativity. The individual papers are based on systematic literature reviews as well as empirical data in the form of customers’ ideas for service innovation collected through diaries. The results of the thesis suggest that despite the negative nature of the service, health care customers are creative. Given the opportunity, health care customers can provide creative ideas and solutions on a multitude of aspects, both within and outside the health care setting. This provides the potential to view the health care experience through the customers’ eyes and take part in their creativity in spheres where the service providers have not traditionally had any access. This thesis contributes to the literature by providing a framework for health care customer creativity that recognizes the concept as a complex interplay of factors operating at the individual, contextual, and situational levels. The proposed framework specifies the health care specific factors upon which customer creativity depends, with the intention of positing potential research directions and developing an enriched theory of health care customer creativity.

Résumé : La présente thèse porte sur le développement d’un modèle d’intervention au profit des agents de développement économique méso dans un contexte de co-innovation et au niveau de l’entrepreneuriat technologique. Grâce à une expérience de plus de quinze ans comme agent de développement économique, une problématique managériale a émergé tranquillement. Les agents de développement économique n’avaient pas à leur disposition les outils d’intervention qui leur permettent de faire face aux nouvelles réalités du développement économique par l’innovation. Une première question peut se poser: qu’est-ce qui a changé dans l’innovation ? Tout d’abord, les savoirs sont de plus en plus complexes et diffus. En effet, les sciences et les technologies, surtout si elles ont atteint un certain degré de maturité, se développent de plus en plus en fonction d’usages qu’elles pourraient remplir (ces derniers étant fonction des besoins exprimés par les usagers). Par ailleurs, tout en se complexifiant à cause de leurs ramifications, les technologies sont de plus en plus accessibles aux usagers, ce qui leur permet de ne plus être seulement consommateurs de technologie, mais également producteurs et codéveloppeurs de produits et services. C’est en ce sens que l’on voit exploser le développement des applications mobiles. En définitive, on passe d’une innovation qui est linéaire, développée en vase clos ou en collaboration entre entreprises et centres de recherche triés sur le volet, qui est par la suite valorisée puis éventuellement commercialisée, à une innovation de nature systémique à la fois portée par des entreprises, des gouvernements et des usagers, ce qui est convenu d’appeler désormais la co-innovation. Par contre, les moyens dont disposent les agents de développement économique ne sont plus adaptés à cette nouvelle réalité. En effet, les outils dont bénéficient ces derniers sont basés sur les principes suivants : 1) la spécialisation sectorielle selon le modèle de Porter (1993), avec la stratégie des grappes où les agents de développement économique faisaient de la concertation et de l’animation du milieu ou 2) la diversité sectorielle et individuelle (Florida, 2005; Jacobs, 1969), où l’on s’est concentré sur la formation entrepreneuriale et sur les visites aux entreprises pour les aider dans leur croissance. Ces stratégies ont été payantes en termes de concertation et de développement hyperlocal, mais sont à la recherche d’un nouveau souffle, notamment en matière d’intergrappes et de cohérence supralocale. Au tournant des années 1990, on voit apparaître la notion de systèmes régionaux d’innovation, qui sont une première tentative de penser le développement économique sous forme systémique. En revanche, les aspects normatifs ont vite disparu des recherches universitaires, ce qui n’a pas permis aux agents de développement économique de s’approprier ces notions, la plupart les trouvant trop compliquées pour y voir de réelles applications en matière d’intervention en développement économique. Enfin, depuis les années 2000, la notion d’innovation ouverte apparaît et suscite beaucoup d’intérêt chez les agents de développement économique, mais ceux-ci manquent encore de modèle afin de pouvoir l’utiliser de façon plus structurée, au-delà de la simple stratégie de gestion de la propriété intellectuelle, comme véritable outil de co-innovation. Ainsi, la question de recherche suivante a été dégagée des éléments issus de la profession et d’une revue de littérature exhaustive: quel nouveau modèle de développement économique (DÉ) basé sur la co-innovation et au niveau de l’entrepreneuriat technologique peut-on développer et quelles seront alors les nouvelles méthodes d’intervention pour les agents de DÉ ? Pour répondre à cette question, la théorie enracinée est apparue comme une méthode de recherche appropriée, car elle permettait la théorisation, essentielle dans le développement d’un modèle. Vingt et une entrevues ont été menées entre août et décembre 2012 entre le Québec, les États-Unis et la France. Les résultats de cette recherche sont les suivants : 1) le niveau d’intervention adéquat en matière de développement économique par l’innovation est le niveau régional, soit les agents de développement économique de type méso (entre le macro et le micro), 2) le processus de développement économique par la co-innovation est un processus intermédié en deux temps. En effet, les agents de DÉ méso travaillent à identifier des problématiques et des besoins communs à différents écosystèmes, et ce, en comptant sur leur réseau. Grâce à ce mécanisme, les agents de DÉ créent des communautés (surtout technologiques) autour desquelles des opportunités intrapreneuriales apparaissent. Celles-ci pourront se transformer en projets structurants qui, à leur tour, pourront déboucher sur des opportunités entrepreneuriales. 3) L’agent de développement économique est à la fois un intrapreneur (qui défend ses projets dans sa propre structure organisationnelle) et un exopreneur (qui défend ses projets dans différentes communautés). Les résultats de cette recherche ont d’ores et déjà servi à la mise sur pied de nouvelles formes d’intervention au niveau méso avec le développement du programme de financement de la Ville de Montréal PR@M-Est, qui est basé directement sur les principes de co-innovation. Par ailleurs, les recherches suscitent de l’intérêt dans plusieurs agglomérations en France et auprès des professionnels en développement économique au Québec et en France. Enfin, des recherches subséquentes ont été identifiées afin de venir compléter le corpus de savoir en matière de développement économique par la co-innovation, notamment en ce qui concerne le partage de rente économique (monétisable ou non) entre les initiateurs de démarches co-innovantes et leurs contributeurs. / Abstract : For a few years, economic developers have integrated innovation to their practice as the engine for economic development. It is now well understood that innovation and economic development are intertwined with the seminal work of Schumpeter more than a century ago. Many researches have been done since, but the very dynamic nature of innovation, and thus the economic developers’ actions, needs constant reassessment. Innovation is changing from a linear perspective where firms, research centers and universities try to develop and commercialize new products and services, based on scientific researches, the so-called Public-Private Partnership, to a systemic form where firms, research centers and universities are still involved in the innovation process, but where more and more Public-Private-People Partnerships can occur. Why is innovation becoming a systemic process? One of the explanations lays in the greater complexity of knowledge and a greater distribution of it. In the meantime, more people can access technologies, as it is seen with the blooming of mobile applications. Since then, the way of innovation process occurs is dramatically changing, and more contributors can join the process, including as usual private companies or government, but also users. What is new with the users is that they are not only involved for commercialization purpose, but now also as co-developers and co-creators. By acknowledging those shifts, economic developers should also change the way they are acting to foster economic development and innovation. So, it is time to develop new forms of intervention in a context of co-innovation. They have to step back to see that their actions are done at the macro level (even if it has been practiced at the regional level) and generally based on industrial concentration with MAR and his industrial districts, on one side and specialization, on the other side with Porter and his clusters’ strategy. The idea is to nourish innovation by geographical, cultural, institutional and social proximity. In doing so, it helps tacit knowledge to flow from one company to another and to bring to the market new products and services more rapidly. Economic developers works with geographic proximity and implements clusters strategies all over the world with different results. So the mechanistic aspect of the strategy is not that simple. Actually, clusters are now shifting to a systemic perspective and try to open-up collaboration between clusters and enhance cross sectorial projects. It also has been demonstrated that innovation is more about people than organizations. Since Jacobs, and later with Florida, the spotlight has been put on the individual as the engine of creativity and innovation. Jacobs thought that knowledge is embedded in the individual expertise, and Florida worked on the creative class which represent about 30% of the population, and who is made of people who promote creativity and changes in some place, generally cities. With Jacobs and Florida, the quality of place is critical because it can help to attract talents. At that micro level also, works on the entrepreneur-opportunity dyad is important. With the neo-classic theory, entrepreneurs can use information asymmetry to exploit opportunities and bring their creativity to the market. Entrepreneurs are very special in essence, even though it has not been establish that they have some special traits, it seems that they have in common a way to act creatively and transform this creativity into opportunities and economic development. For economic developers, it means to develop supportive actions towards entrepreneurs: training, entrepreneurs club, special funding for example. But, the difficulties lay in: how can economic developers develop a more structured model for their interventions to entrepreneurs? Moreover, some do believe that these micro levels approaches can be self-organized and that there is no need for public policy towards entrepreneurs. At the meso level, economic developers can improve, particularly regarding the Regional Innovation System (RIS), the way they can foster business serendipity across networks. Since Chesbrough has defined the concept of open innovation, a more systemic approach to innovation seems to be accepted by economic developers. But, once again, there is lot of work to do to better understand how economic developers can put into practice those new approaches. Thus, neither the macro level approach, which is too mechanistic, nor the meso approaches level, which is to complex, nor the micro level, which doesn’t allow public policy, seem to offer a proper answer to economic developers with the systemic approach of innovation. This dissertation tries to answer the following question: What model can be developed for economic developers in context of co- innovation and technological entrepreneurship? Grounded theory as presented by Corbin and Strauss (2008) was the methodology in this survey. As it is permitted in grounded theory, the literacy has been completed by works on complexity theory, network theory and structural holes and co-innovation. Twenty-one interviews have been conducted in Montréal, the Province of Québec province, France and The United States between August and December 2012 with economic developers, scientific and technological parks managers, co-creation spaces organizers and entrepreneurs. The main conclusions of this research in an academic perspective are: 1. The correct level to support co-innovation is the meso level: this level allows macro and micro level actions to be coherent in a more interdependent perspective. The meso level is a medium term perspective at the regional level. The economic developers mostly use projects and communities of practice to do their actions regarding co-innovation; 2. The process for the economic development through co-innovation is intermediated and two-fold: a) Economic developers identify common needs in their ecosystems through their networks. In doing so, they create communities in which intrapreneurial opportunities occur. Some of these intrapreneurial opportunities lead to structuring projects. b) These projects open-up entrapreneurial opportunities that help regional economic development; 3. Finally, Economic developers are both intrapreneurs (leading project in their own organisations and exopreneurs (leading projects in communities). But as we want to develop something operational for economic developers, we also created an algorithm, which will help them to structure co-innovation actions (how to identify needs, communities, partners, etc.), but also a guide to use the right co-innovative tool at the right moment in their process of co-innovation. Some results of this research has been put into practice with the new program as PR@M-Est at the City of Montréal, this program is based on business serendipity and on co-innovation principles. This research shows some interest in France also. Of course this research has some limits, particularly with the size and the diversity of the sample, but we are confident it could open the way to further research in the field.

Le cancer de l’ovaire est le plus mortel des cancers gynécologiques. CA125 est le marqueur de progression utilisé pour effectuer le suivi des patientes atteintes de ce dernier. La protéine CA125 est surexprimée au niveau des tissus ovariens tumoraux, mais elle est non détectable avec les moyens d'aujourd'hui au niveau des tissus ovariens normaux. CA125 est une protéine dont les fonctions demeurent inconnues sinon hypothétiques. Dans notre laboratoire, la recherche est axée sur la détermination des fonctions de l’antigène tumoral CA125. Pour cela un projet de maîtrise précédent au miens a permis la mise au point d’un outil nommé anticorps monovalent modifié (ScFv) dans une lignée de cellules tumorales de l’ovaire. Cet outil permet la liaison de la protéine cible, dans notre cas CA125, et empêche sa localisation physiologique à la membrane, éliminant ainsi sa fonction dans la cellule ciblée. Grâce à cet outil, quelques rôles potentiels ont pu être proposés. CA125 aurait des implications dans la prolifération, l’adhésion cellule-cellule, la résistance à l’apoptose et la migration des cellules tumorales de l’ovaire. Pour étudier cette protéine d’un point de vue moléculaire, mon projet principal consistait à effectuer un criblage double-hybride nucléaire chez la levure pour découvrir des interactions protéiques à une échelle cellulaire. En parallèle, l’utilisation de la méthode d’immunoprécipitation avec des cellules tumorales de l’ovaire en tant que projet secondaire avait pour but de vérifier des voies de signalisation spécifiques. Grâce aux résultats précédents obtenus par des expériences antérieures dans le laboratoire ainsi qu'à ceux obtenus pour d’autres mucines, certaines voies de signalisation démontraient un potentiel intéressant pour l’implication de la protéine CA125. Par des expériences d'immunoprécipitation, nous avons découvert que des complexes protéiques contenant la protéine E-cadhérine et la protéine β-caténine contiennent aussi CA125. Ces protéines sont impliquées dans la prolifération et l’adhésion cellulaire. Certaines fonctions hypothétiques de CA125 sont reliées à ces processus cellulaires, c’est-à-dire l’adhésion cellule-cellule et la prolifération cellulaire. Ces interactions protéiques permettront de mieux comprendre l’implication de CA125 dans ces processus cellulaires. Mon projet principal était la mise au point d’un double-hybride nucléaire chez la levure. Pour y parvenir, nous avons dû modifier les protocoles proposés normalement pour ce type de double-hybride. Nous avions choisi d’effectuer notre criblage avec le domaine cytoplasmique de CA125. Par contre, étant donné que ce domaine est très court seulement 31 a.a., le domaine transmembranaire de CA125 a été ajouté au premier domaine mentionné pour favoriser une conformation protéique adéquate. Le domaine cytoplasmique de CA125 a été choisi pour tenter de mieux comprendre d’un point de vue moléculaire les voies de signalisation dans lesquelles cet antigène tumoral est impliqué. Lors du premier essai pilote de double-hybride effectué, nous avons repêché une protéine qui interagit probablement de manière non spécifique avec CA125 selon des statistiques recueillies lors de criblages pour d’autres protéines transmembranaires. Pour remédier à ce problème majeur, nous avons recherché la source de ce problème. Nous avons alors réalisé que les levures utilisées n’étaient pas les Saccharomyces cerevisiae PJ69-4A que nous avions choisies pour effectuer notre double-hybride. Par la suite, les levures ont été maintenues sur des milieux sélectifs pour les PJ69-4A, c’est-à-dire en absence de lysine (Lys -). Pour parvenir à obtenir des colonies positives lors de la transformation de l’ADN de la librairie avec notre construction, il nous a fallu amplifier les transformants en milieu liquide toute la nuit en sélectionnant pour les vecteurs et la levure. Une fois les transformants amplifiés, nous avons obtenu des candidats potentiels qui ont franchi les différentes étapes de sélection du double-hybride. D’autres techniques ont dû être adaptées à nos besoins pour parvenir à mettre au point notre double-hybride nucléaire chez la levure pour notre protéine transmembranaire. Grâce à ces efforts, certains candidats ont été séquencés et la première protéine découverte par ce criblage est RNF5 (ring finger protein 5). Cette dernière est impliquée dans la régulation de la motilité cellulaire. CA125 semble être aussi impliqué dans ce processus. Il est alors possible de relier CA125 à RNF5. Ce premier résultat nous suggère que nous avons réussi à mettre au point un double-hybride nucléaire chez la levure pour l’antigène tumoral CA125.

Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Gasification provides a proven alternative to the dependence on petroleum for the production of high value products such as liquid fuels and chemicals. Syngas, the main product from gasification can be converted to fuels and chemicals via a number of possible synthesis processes. Coal and natural gas are currently the main feedstock used for syngas production. In South Africa (SA), Sasol operates the largest commercial coal-to-liquids conversion process in the world, based on updraft fixed bed gasification of low grade coal to syngas. Co-utilizing alternative and more sustainable feedstock (such as biomass and wastes) with coal in existing coal-based plants offers a realistic approach to reducing the costs and risks associated with setting up dedicated biomass conversion plants. An experimental and modelling investigation was performed to assess the impacts of co-gasifying two of the most commonly available agricultural wastes in SA (sugarcane bagasse and corn residue) with typical low grade SA coals, on the main products of updraft fixed bed gasification, i.e. liquid condensates and syngas. Condensates are produced in the pyrolysis section of the updraft gasifier, whereas syngas is a result of residual char conversion. An experimental set-up that simulates the pyrolysis section of the gasifier was employed to investigate the yield and composition of devolatilized products at industrially relevant conditions of 26 bars and 400-600°C. The results show that about 15 wt% of coal and 70 wt% of biomass are devolatilized during the pyrolysis process. The biomass derived condensates were determined to comprise of significantly higher quantities of oxygenates such as organic acids, phenols, ketones, and alcohols, whereas coal derived hydrocarbon condensates were dominated by polycyclic aromatic hydrocarbons, creosotes and phenols. Results of investigation into the influence of coal-biomass feedstock mix ratio on yields of products from pyrolysis show limited evidence of non-additive or synergistic behaviour on the overall distribution of solid, liquid and gas yields. On the other hand, in terms of the distribution of specific liquid phase hydrocarbons, there was significant evidence in favour of non-additive pyrolysis behaviour, as indicated by the non-additive yield distribution of specific chemicals. Synergistic trends could also be observed in the thermogravimetric (TGA) study of pyrolysis under kinetically controlled non-isothermal conditions. Model free and model fitting kinetic analysis of the TGA data revealed activation energies ranging between 94-212 kJ mol-1 for the biomass fuels and 147-377 kJ mol-1 for coal. Synergistic interactions may be linked to the increased presence of hydrogen in biomass fuels which partially saturates free radicals formed during earlier stages of devolatilization, thereby preventing secondary recombination reactions that would have produced chars, allowing for the increased formation of volatile species instead. Analysis of char obtained from the co-pyrolysis experiments revealed that the fixed carbon and volatile content of the blended chars is is proportional to the percentage of biomass and coal in the mixture. CO2 reactivity experiments on the chars showed that the addition of biomass to coal did not impose any kinetic limitation on the gasification of blended chars. The blended chars decomposed at approximately the same rate as when coal was gasified alone, even at higher biomass concentrations in the original feedstock blend. Based on these observations, a semi-empirical equilibrium based simulation of syngas production for co-gasification of coalbiomass blends at various mix ratios was developed using ASPEN Plus. The model showed that H2/CO ratio was relatively unaffected by biomass addition to the coal fuel mix, whereas syngas heating value and thermal efficiency were negatively affected. Subsequent evaluation of the production cost of syngas at biomass inputs ranging between 0-20 wt% of coal reflected the significant additional cost of pretreating biomass (3.3% of total capital investment). This resulted in co-gasification derived syngas production costs of ZAR146/tonne (ZAR12.6/GJ) at 80:20 coalbiomass feedstock ratio, compared to a baseline (coal only) cost of ZAR130/tonne (ZAR10.7/GJ). Sensitivity analysis that varied biomass costs from ZAR0 ZAR470 revealed that syngas production costs from co-gasification remained significantly higher than baseline costs, even at low to zero prices of the biomass feedstock. This remained the case even after taking account of a carbon tax of up to ZAR117/tCO2. However, for range of carbon tax values suggested by the SA treasury (ZAR70 tCO2 to ZAR200 tCO2), the avoided carbon tax due to co-feeding biomass can offset between 40-96% of the specific retrofitting cost at 80:20 coal-biomass feedstock mass ratio. In summary, this dissertation has showed that in addition to the widely recognized problems of ash fouling and sintering, co-feeding of biomass in existing coal based updraft gasification plants poses some challenges in terms of impacts on condensates and syngas quality, and production costs. Further research is required to investigate the potential in ameliorating some of these impacts by developing new high value product streams (such as acetic acid) from the significant fraction of condensates derived from biomass. / AFRIKAANSE OPSOMMING: Vergassing bied 'n beproefde alternatief vir die afhanklikheid van petroleum vir die produksie van hoë waarde produkte soos vloeibare brandstof en chemikalieë. Sintese gas, die belangrikste produk van vergassing, kan omgeskakel word na brandstof en chemikalieë deur 'n aantal moontlike sintese prosesse. Steenkool en aardgas is tans die belangrikste grondstowwe wat gebruik word vir sintese gas produksie. In Suid-Afrika (SA) bedryf Sasol die grootste kommersiële steenkool-totvloeistof omskakelingsproses in die wêreld, gebaseer op stygstroom vastebed vergassing van laegraadse steenkool na sintese gas. Die gebruik van alternatiewe en meer volhoubare grondstowwe (soos biomassa en afval) saam met steenkool in die bestaande steenkool-gebaseerde aanlegte bied 'n realistiese benadering tot die vermindering van die koste en risiko's wat verband hou met die oprigting van toegewyde biomassa omskakelingsaanlegte. 'n Eksperimentele en modelleringsondersoek is uitgevoer om die impak van gesamentlike vergassing van twee van die mees algemeen beskikbare landbouafvalprodukte in Suid-Afrika (suikerriet bagasse en mieliereste) met tipiese laegraadse SA steenkool op die vernaamste produkte van stygstroom vastebed vergassing, dws vloeistof kondensate en sintese gas, te evalueer. Kondensate word geproduseer in die piroliese gedeelte van die stygstroomvergasser, terwyl sintese gas 'n resultaat is van die omskakeling van oorblywende houtskool. 'n Eksperimentele opstelling wat die piroliese gedeelte van die vergasser simuleer is gebruik om die opbrengs en die samestelling van produkte waarvan die vlugtige komponente verwyder is by industrie relevante toestande van 26 bar en 400-600°C te ondersoek. Die resultate toon dat ongeveer 15% (massabasis) van die steenkool en 70% (massabasis) van die biomassa verlore gaan aan vlugtige komponente tydens die piroliese proses. Daar is vasgestel dat die kondensate afkomstig van biomassa uit aansienlik hoër hoeveelhede suurstofryke verbindings soos organiese sure, fenole, ketone, en alkohole bestaan, terwyl koolwaterstofkondensate afkomstig uit steenkool oorwegend bectaan uit polisikliese aromatise verbindings, kreosote en fenole. Die resultate van die ondersoek na die invloed van die verhouding van steenkool tot biomassa grondstof op piroliese opbrengste toon beperkte bewyse van nie-toevoegende of sinergistiese gedrag op die algehele verspreiding van soliede, vloeistof en gas opbrengste. Aan die ander kant, in terme van die verspreiding van spesifieke vloeibare fase koolwaterstowwe, was daar beduidende bewyse ten gunste van 'n sinergistiese piroliese gedrag. Sinergistiese tendense is ook waargeneem in die termogravimetriese (TGA) studie van piroliese onder kineties beheerde nieisotermiese toestande. Modelvrye en modelpassende kinetiese analise van die TGA data het aan die lig gebring dat aktiveringsenergieë wissel tussen 94-212 kJ mol-1 vir biomassa brandstof en 147-377 kJ mol-1 vir steenkool. Ontleding van die houtskool verkry uit die gesamentlike piroliese eksperimente het aan die lig gebring dat die onmiddellike kenmerke van die gemengde houtskool die geweegde gemiddelde van die individuele waardes vir steenkool en biomassa benader. CO2 reaktiwiteitseksperimente op die houtskool het getoon dat die byvoeging van biomassa by steenkool nie enige kinetiese beperking op die vergassing van gemengde houtskool plaas nie. Die gemengde houtskool ontbind teen ongeveer dieselfde tempo as wanneer steenkool alleen vergas is, selfs teen hoër biomassa konsentrasies in die oorspronklike grondstofmengsel. Op grond van hierdie waarnemings is 'n semi-empiriese ewewig-gebaseerde simulasie van sintese gas produksie vir gesamentlike vergassing van steenkool-biomassa-mengsels vir verskeie mengverhoudings ontwikkel met behulp van Aspen Plus. Die model het getoon dat die H2/CO verhouding relatief min geraak is deur biomassa by die steenkool brandstofmengsel te voeg, terwyl sintese gas se verhittingswaarde en termiese doeltreffendheid negatief geraak is. Daaropvolgende evaluering van die produksiekoste van sintese gas vir biomassa insette wat wissel tussen 0-20% (massabasis) van die hoeveelheid steenkool het die aansienlike addisionele koste van die vooraf behandeling van biomassa (3.3% van die totale kapitale belegging) gereflekteer. Dit het gelei tot 'n produksiekoste van ZAR146/ton (ZAR12.6/GJ) vir sintese gas afkomstig uit gesamentlike-vergassing van 'n 80:20 steebkool-biomassa grondstof mengesl, in vergelyking met 'n basislyn (steenkool) koste van ZAR130/ton (ZAR10.7/GJ). Sensitiwiteitsanalise wat biomassa koste van ZAR0 - ZAR470 gevarieër het, het aan die lig gebring dat sintese gas produksiekoste van gesamentlike vergassing aansienlik hoër bly as die basislyn koste, selfs teen 'n lae of nul prys van biomassa grondstof. Dit bly die geval selfs nadat koolstof belasting van tot ZAR117/tCO2 in ag geneem is. In opsomming het hierdie verhandeling getoon dat, bykomend tot die wyd-erkende probleme van as besoedeling en sintering, die gesamentlike gebruik van biomassa in bestaande steenkool stygstroom vergassingsaanlegte groot uitdagings inhou in terme van die impak op die kwaliteit van kondensate en sintese gas, asook produksiekoste. Verdere navorsing is nodig om die potensiaal te ondersoek vir die verbetering van sommige van hierdie impakte deur die ontwikkeling van nuwe hoë waarde produkstrome (soos asynsuur) uit die beduidende breukdeel van kondensate wat verkry word uit biomassa.