Aujourd’hui les matériaux plastiques sont indissociables de notre quotidien, tandis que leur fabrication dépend des ressources pétrolières qui ne sont pas durables sur le long terme. Dans l’idée de produire des nouveaux intermédiaires pour matériaux à partir de matière première renouvelable, les dérivés furaniques sont des candidats de choix puisqu’ils sont facilement synthétisables à partir des sucres constitutifs de la biomasse végétale. Le poly(éthylène-2,5-furandicarboxylate) ou PEF est l’un des plastiques qui peut être fabriqué à partir du diacide 2,5-furandicaboxylique (FDCA). Il peut être produit selon des procédés similaires au poly(éthylène téréphtalate) (PET), son homologue petro-sourcé, mais a récemment fait l’objet d’une attention grandissante, notamment pour sa perméabilité aux gaz plus basse et son module mécanique et sa température de transition vitreuse plus élevées. Ces caractéristiques rendent le PEF particulièrement intéressant pour la fabrication de matériaux d’emballage, en particulier dans l’alimentaire. Dans la première partie de ce travail, nous étudions la cristallisation du PEF, aspect particulièrement important pour la production et la manipulation des granulés de plastique, ainsi que pour la transparence et les propriétés thermiques requises par les applications finales, et ce, en fonction de la masse moléculaire et du type de catalyseur utilisé. L’application de certains modèles mathématiques a permis de décrire à la fois la cinétique de cristallisation du PEF en mode isotherme et non isotherme du PEF. Cette cristallisation est généralement plus lente que celle du PET. La cristallisation du PEF à partir de l’état vitreux a montré des résultats atypiques qui ont été modélisés avec succès à l’aide de modèles non-conventionnels et de la méthode isoconversionnelle. Les différences ont été expliquées par la nucléation qui peut se produire à température relativement basse. Cette nucléation à basse température a notamment été exploitée judicieusement pour accélérer la cristallisation froide. La deuxième partie de ce travail concerne le comportement thermomécanique du PEF, important notamment pour les procédés de mise en forme et les applications finales. La température de transition vitreuse du PEF, pourtant plus élevée que celle du PET, n’a pas montré d’augmentation particulière avec la cristallinité ce qui a été attribuée à une réduction de la mobilité de la chaîne compensée par une augmentation du volume libre. Le faible taux d’enchevêtrement des chaînes de PEF pourrait s'expliquer par la réduction des dimensions de la chaîne non perturbée, ce qui est peut être directement lié à la diminution de la taille du monomère FDCA en comparaison avec celle de l’acide p-téréphtalique. Aucune restriction d’origine conformationnelle n'a été trouvée ce qui implique, que la réduction de la mobilité doit être principalement intermoléculaire. Pour des PEF de masses molaires diverses ou élaborés à partir de différents catalyseurs, une dépendance plus importante de la viscosité avec la température et la vitesse de déformation est constatée. Ce comportement qui peut être décrit mathématiquement peut également être expliqué par une diminution de la densité d’enchevêtrement des chaînes. Les propriétés mécaniques des phases amorphes et leurs dépendances avec la vitesse de déformation indiquent un faible réseau d'enchevêtrement du PEF par rapport au PET, bien que la réduction de la mobilité intrinsèque de la chaine de PEF puisse également jouer un rôle. L'orientation biaxiale du PEF dans l’état caoutchoutique a été étudiée en comparaison avec celle du PET, ce qui s’avère pertinent pour la production de films et de bouteilles. Les résultats ont montré que des taux d’étirage plus élevés sont nécessaires dans le cas du PEF pour atteindre le même durcissement structural. / Plastics have become an integral part of our lives, while the petrochemical feedstocks used to make them are not sustainable on the long term. In pursuit of production processes starting from renewable feedstocks, furanics were found to form quite readily from abundant plant-based carbohydrates and to bring new functionality as intermediates. Poly(ethylene 2,5-furandicarboxylate) or PEF is one of the plastics that can be made through 2,5-furandicaboxylic acid (FDCA) as an intermediate. It can be produced analogously to the ubiquitous material Poly(ethylene terephthalate) (PET) but has only recently been gaining more attention including the finding that it has greatly reduced gas permeability and a higher modulus and glass transition temperature, rendering it interesting as a packaging material. In the first part of this work we study the crystallization behavior of PEF, relevant for production and handling of pellets as well as transparency and thermal properties in end-use applications, as a function of molecular weight and the type of catalyst used. Mathematical models were found that describe both isothermal crystallization kinetics and non-isothermal kinetics for PEF, which is generally slower than PET. PEF crystallization from the glass found to be atypical and was modeled using unconventional models and the isoconversional approach. The origin of this behavior was found to be nucleation at low temperatures, which can be influenced to accelerate its crystallization. The second part of this work relates to the thermomechanical behavior of PEF, relevant for its processing and application in particular. The higher glass transition temperature was found to not increase as much by crystallinity as PET, and could be attributed to a reduced chain mobility compensated by increased free volume. The loose entanglement of PEF could be explained by reduced unperturbed chain dimensions following quite directly from the reduced bond length of FDCA. No significant conformational restictions were found, thus any mobility reduction should be intermolecular. A higher temperature and strain rate dependence of the melt viscosity was found for PEF across various molecular weights and catalyst types, which was described mathematically and can also be explained by a more loosely entangled network. The amorphous mechanical properties and higher strain rate dependence at room temperature also point to a low entanglement network although mobility reduction may also play a role. Biaxial orientation of PEF in the rubbery state, relevant for producing films and bottles, showed that higher draw ratios are needed than for PET until molecular orientation is maximized and strain hardening begins. However, it was found that upon using higher stretch ratios, oriented PEF can exhibit increased strength and Tg compared to oriented PET and further reduced gas permeability.

Calpains are intracellular enzymes that merge cysteine protease and calcium sensing activities together in one molecule. They respond to Ca2+ signals and modify the activity of their targets by selective proteolysis. Calpains are involved in normal cellular process like cell migration and apoptosis. The over-activation of calpain due to disturbances in Ca2+ homeostasis or inactivation due to mutations, contribute to diseases like ischemic injury and muscular dystrophy. The classical calpains 1 and 2 are heterodimeric enzymes containing a large (80 kDa) subunit and a small subunit (28 kDa). Dimerization occurs through the 5th EF-hand of penta-EF-hand (PEF) domains present in both large and small subunits. In this study, I have used structural genomics approaches to explore the PEF and C2-like regulatory domains of some of the other 12 human calpain isoforms. I have shown that recombinant PEF domain of skeletal muscle-specific calpain 3 exists as a stable homodimer when produced alone. Modelling studies suggest that there would be no barriers for dimerization of the full-length enzyme through the PEF domains which would place the protease cores at opposite ends of the dimer. Co-expression studies using small subunit were performed with PEF domains of calpains 1, 3, 8, 9, 11, 12 and 13. A differential tagging system was devised to differentiate heterodimers from homodimers. The PEF domains of calpains 1, 3, 9 and 13 co-expressed with the small subunit, while the others failed to express. The PEF domains of calpains 1 and 9 formed heterodimers. Conversely, the PEF domain of calpain 3 formed a homodimer and that of calpain 13 predominantly formed a homodimer with a small amount of heterodimer. Homodimerization of calpains implies they are less-likely to be inhibited by the endogenous calpain inhibitor, calpastatin. C2-like regulatory domains of calpains 5-13 were also studied. The structure of the distal C2-like domain of calpain 7 was solved. It is markedly different from canonical C2 domains and may not bind Ca2+. / Thesis (Ph.D, Biochemistry) -- Queen's University, 2009-02-11 12:30:29.18

High voltage treatment technology has been developed in this thesis and had initially shown promise in its effectiveness in reducing microorganisms found in water supplies. Initial testing found that the high voltage could destroy over 99.9% of the bacteria S. marcescens (a 3-log reduction). Cited literature on the effects of high voltage pulsed electric fields (PEFs) on various microorganisms have shown that high destruction rates of up to 9-log can be achieved. Thus by increasing the electric field strength or exposure time, or by improving the design of the electrode flow chamber, better results should be achieved using high voltage on water. However, contrary to this, upon further design improvements the 99.9% destruction threshold was rarely increased. The initial slow flow device of one litre-per-minute (1 LPM) was scaled up to flows of 10 LPM and 33 LPM. However, these faster flow devices were even less effective in the destruction of bacteria, destroying only 99% of S. marcescens (2-log reduction). No physical or technical design parameters could account for this low performance. One possible reason for these low results was in the preparation of the bacteria themselves. It was discovered that the growth stage of bacteria prepared for experiments had a large effect on the results. Bacteria harvested in the early growth stage could be nearly all destroyed by the high voltage (greater then 4-log reduction), whereas those harvested in the late stationary stage were much more resistant (less than 0.5-log reduction). Bacteria naturally occurring in water supplies will mostly be in a non-metabolising state. This implies that they will be more resistant to high voltage exposure than bacteria grown in a laboratory under standard testing procedures. Thus standard testing procedures for this device do not give accurate results. Further research into the mechanism behind the bacterial resistance is required to improve the performance of high voltage devices. A combination of different technologies may also prove effective in overcoming the resistance mechanism. These improvements are required before high voltage treatment can be properly developed and commercially exploited.

L’interaction avec les cellules hôtes est une étape primordiale du cycle infectieux des Salmonella. Elle implique entre autres des fimbriae permettant l’attachement des Salmonella aux cellules et des invasines permettant leur internalisation. Généralement, l’expression de ces facteurs est réprimée et ils ne sont exprimés qu’in vivo en des sites bien spécifiques. Notre objectif a été d’étudier les mécanismes de répression de l’expression des fimbriae Pef et de l’invasine Rck. Nos résultats montrent que les nucléoprotéines H-NS, Hha et YdgT régulent négativement l’expression de ces deux facteurs de virulence. Le mécanisme de répression de la transcription de l’opéron pef a été caractérisé et montre un rôle prépondérant d’H-NS. De plus, la régulation de cet opéron par Hha et YdgT semble être dépendante ou indépendante de H-NS en fonction des conditions de culture de la bactérie. La répression de l’opéron pefI-srgC, portant l’ORF rck, semble, quant à elle, plus complexe. A l’heure actuelle, seule la région cible de la répression par Hha et YdgT a pu être identifiée. Ces résultats renforcent l’hypothèse de l’existence de régulations par Hha-YdgT indépendantes de H-NS. / The interaction with host cells is an essential step of Salmonella infection cycle. Among other virulence factors, fimbriae allow attachment of Salmonella to eukaryotic cells and invasins enable cell internalisation. Generally, the expression of these factors is suppressed and they are only expressed in vivo at very specific locations. Our objective was to study the mechanisms of Pef fimbriae and Rck invasin repression. Our results show that the nucleoproteins H-NS, Hha and YdgT negatively regulate the expression of these two virulence factors. The mechanism of pef operon transcription repression was characterized and shows a predominant role of H-NS. Moreover, the regulation of this operon by Hha and YdgT appears to bedependent or independent of H-NS depending on bacterial culture conditions. Repression of the pefI-srgC operon, carrying rck ORF, shows a higher degree of complexity. Currently, only the Hha and YdgT targeted region for the repression has been identified. These results reinforce the hypothesis that Hha-YdgT can act independently of H-NS.

Esta pesquisa analisa os recentes processos de caráter geopolítico que vêm ocorrendo na Amazônia brasileira e confere especial ao papel desempenhado pelas Forças Armadas nas áreas fronteiriças desta região. A argumentação que desenvolvemos volta seu foco para a atuação dos Pelotões Especiais de Fronteira, no contexto sócioeconômico e político de Roraima, no período que vai desde sua implantação do Território Federal, acontecida nos anos de 1940, até os dias atuais. Para que possamos traçar a contento esse panorama, investigamos mais detidamente as políticas de governo estabelecidas após os anos 60, quando as unidades militares assumiram papel central na transformação deste território, definido como estratégico para os eventos que estavam ocorrendo no País e no subcontinente sul americano. Assim, tivemos que dar ênfase a alguns fatos e eventos importantes para podermos demonstrar o papel específico que as Forças Armadas, notadamente, o Exército e a Aeronáutica, desempenharam no cotidiano de Roraima, através de suas ações, de seu ethos, e da atuação da administração direta dos oficiais da Aeronáutica, durante o regime militar. Os resultados que obtivemos nos permitiram constatar que a organização do espaço social de Roraima continua a ser pensada e concebida a milhas de distância deste espaço. Prova disso é que as Forças Armadas ainda desempenham importante papel no Estado, apesar do seu distanciamento corporativo e do antagonismo em conciliar a hierarquia militar com a gestão participativa, base do discurso hegemônico, desde os anos 90. A investigação nos permitiu concluir que as Forças Armadas ainda desfrutam de status e desempenham um papel ainda muito importante na Amazônia. Pode-se a isso acrescentar que, no momento, elas protagonizam um novo e importante papel, que se consubstancia por meio dos Pelotões Especiais de Fronteira/PEF, bem como de outros mecanismos, recentes implantados em áreas importantes à soberania do País, agora também tornadas relevantes pelos seus aspectos ambientais, étnicos e culturais. / This research analyses recent processes of political feature that are happening in the Brazilian Amazon, with emphasis to the role of the Armed Forces in border areas of this region. Our argumentation focused on the performance of the Border Special Platoons, in the socio-economic and political context of Roraima, since the foundation of the Federal Territory, in the 1940s. We concentrated, however, on policies established after the 1960s, when military unities start to play central role in the transformation of this territory, defined as strategic for events that were happening in the country and in the South American subcontinent. We retrieved some facts and events in order to demonstrate the specific role that the Armed Forces, especially, the Army and the Air Force have played in Roraima everyday life, through their activities, their ethos, and the direct administration of officials belonging to the Air Force, during the military regime. We found out that the organization of the Roraima social space continues to be conceived from miles away of this space, so much so that the Armed Forces still play important role in the Roraima State, despite its corporative distance, and the antagonism in conciliating the military hierarchy, with the participative administration, since the 1990s. We concluded that the Armed Forces, with a widened role in the Amazon, represent a new moment through Border Special Platoons/BSP and others recent mechanisms, in important areas to the sovereignty of the country, now also relevant for their environmental, ethnic and cultural aspects.

Esta pesquisa analisa os recentes processos de caráter geopolítico que vêm ocorrendo na Amazônia brasileira e confere especial ao papel desempenhado pelas Forças Armadas nas áreas fronteiriças desta região. A argumentação que desenvolvemos volta seu foco para a atuação dos Pelotões Especiais de Fronteira, no contexto sócioeconômico e político de Roraima, no período que vai desde sua implantação do Território Federal, acontecida nos anos de 1940, até os dias atuais. Para que possamos traçar a contento esse panorama, investigamos mais detidamente as políticas de governo estabelecidas após os anos 60, quando as unidades militares assumiram papel central na transformação deste território, definido como estratégico para os eventos que estavam ocorrendo no País e no subcontinente sul americano. Assim, tivemos que dar ênfase a alguns fatos e eventos importantes para podermos demonstrar o papel específico que as Forças Armadas, notadamente, o Exército e a Aeronáutica, desempenharam no cotidiano de Roraima, através de suas ações, de seu ethos, e da atuação da administração direta dos oficiais da Aeronáutica, durante o regime militar. Os resultados que obtivemos nos permitiram constatar que a organização do espaço social de Roraima continua a ser pensada e concebida a milhas de distância deste espaço. Prova disso é que as Forças Armadas ainda desempenham importante papel no Estado, apesar do seu distanciamento corporativo e do antagonismo em conciliar a hierarquia militar com a gestão participativa, base do discurso hegemônico, desde os anos 90. A investigação nos permitiu concluir que as Forças Armadas ainda desfrutam de status e desempenham um papel ainda muito importante na Amazônia. Pode-se a isso acrescentar que, no momento, elas protagonizam um novo e importante papel, que se consubstancia por meio dos Pelotões Especiais de Fronteira/PEF, bem como de outros mecanismos, recentes implantados em áreas importantes à soberania do País, agora também tornadas relevantes pelos seus aspectos ambientais, étnicos e culturais. / This research analyses recent processes of political feature that are happening in the Brazilian Amazon, with emphasis to the role of the Armed Forces in border areas of this region. Our argumentation focused on the performance of the Border Special Platoons, in the socio-economic and political context of Roraima, since the foundation of the Federal Territory, in the 1940s. We concentrated, however, on policies established after the 1960s, when military unities start to play central role in the transformation of this territory, defined as strategic for events that were happening in the country and in the South American subcontinent. We retrieved some facts and events in order to demonstrate the specific role that the Armed Forces, especially, the Army and the Air Force have played in Roraima everyday life, through their activities, their ethos, and the direct administration of officials belonging to the Air Force, during the military regime. We found out that the organization of the Roraima social space continues to be conceived from miles away of this space, so much so that the Armed Forces still play important role in the Roraima State, despite its corporative distance, and the antagonism in conciliating the military hierarchy, with the participative administration, since the 1990s. We concluded that the Armed Forces, with a widened role in the Amazon, represent a new moment through Border Special Platoons/BSP and others recent mechanisms, in important areas to the sovereignty of the country, now also relevant for their environmental, ethnic and cultural aspects.

In this study conducted at Food Science Australia (FSA) and Berlin University of Technology (BUT), the effects of pulsed electric field (PEF) treatment, a novel non-thermal processing technology on bovine milk microflora and native enzymes and on the rheological and textural properties of rennet-induced milk gels was investigated. The PEF treatments were conducted at field intensities of 25-37 kV cm-1 (up to 50 kV cm-1)and temperature range of 30°C to 75ºC. Native milk enzymes selected for the study included alkaline phosphatase, lipase, xanthine oxidase and plasminand microbiological study included determining Total Plate Count (TPC) and Pseudomonas and Enterobacteriaceae counts in skim milk. At 30ºC PEF treatment at maximum field intensity inactivated AlP by 42% while at 60oC inactivation was higher (67%). Under these treatment conditions less than1 log reduction in TPC and Pseudomonas count and 2.1 logs reduction in the Enterobacteriaceae count was achieved at 30oC while at 60ºC TPC dropped by 2.4 logs and Pseudomonas and Enterobacteriaceae counts were reduced by 5.9 and 2.1 logs, respectively to below the detection limit of 1 CFU mL-1. Combining PEF treatment with heat increased the inactivation level of all enzymes which showed an increasing trend with increasing field intensity and temperature. Treatment time (4.8, 9.6, 19.2, 28.8 and 38.4 µs) was controlled by either changing the pulse frequencies (100-400 Hz) or product flow rate (30-240 mL min-1) at a constant field intensity of 31 kV cm-1 and it was found that changing the flow rate was a more effective way of enzyme inactivation than changing the frequency due to longer exposure time of enzymes to heat and field intensity. The size of casein micelles and fat globules was not affected by PEF treatment while severe heating of milk at 97oC for 10 min decreased both micelle and fat globule sizes marginally. The coagulation time of rennet-induced gels made from PEF-treated (35 to 50 kV cm-1) milks (whole and skim) increased as the treatment intensity increased, but remained shorter than gels made from pasteurised milk. The PEF treatment of milk at various field intensities and temperatures adversely affected the G′, G′′ and firmness of gels, but the effects were less pronounced than in gels made from pasteurised milks. This study concludes that for successful application in milk processing the PEF treatment needs to be combined with mild heat treatment. This approach could achieve safer milk with less damage to milk functionality. However, the quest for a suitable quality assurance indicator enzyme will need more extensive studies.

The PEF treatments were conducted at field intensities of 25-37 kV cm-1 (up to 50 kV cm-1)and temperature range of 30°C to 75ºC. Native milk enzymes selected for the study included alkaline phosphatase, lipase, xanthine oxidase and plasminand microbiological study included determining Total Plate Count (TPC) and Pseudomonas and Enterobacteriaceae counts in skim milk. At 30ºC PEF treatment at maximum field intensity inactivated AlP by 42% while at 60oC inactivation was higher (67%). Under these treatment conditions less than1 log reduction in TPC and Pseudomonas count and 2.1 logs reduction in the Enterobacteriaceae count was achieved at 30oC while at 60ºC TPC dropped by 2.4 logs and Pseudomonas and Enterobacteriaceae counts were reduced by 5.9 and 2.1 logs, respectively to below the detection limit of 1 CFU mL-1. Combining PEF treatment with heat increased the inactivation level of all enzymes which showed an increasing trend with increasing field intensity and temperature. Treatment time (4.8, 9.6, 19.2, 28.8 and 38.4 µs) was controlled by either changing the pulse frequencies (100-400 Hz) or product flow rate (30-240 mL min-1) at a constant field intensity of 31 kV cm-1 and it was found that changing the flow rate was a more effective way of enzyme inactivation than changing the frequency due to longer exposure time of enzymes to heat and field intensity. The size of casein micelles and fat globules was not affected by PEF treatment while severe heating of milk at 97oC for 10 min decreased both micelle and fat globule sizes marginally. The coagulation time of rennet-induced gels made from PEF-treated (35 to 50 kV cm-1) milks (whole and skim) increased as the treatment intensity increased, but remained shorter than gels made from pasteurised milk. The PEF treatment of milk at various field intensities and temperatures adversely affected the G′, G′′ and firmness of gels, but the effects were less pronounced than in gels made from pasteurised milks. This study concludes that for successful application in milk processing the PEF treatment needs to be combined with mild heat treatment. This approach could achieve safer milk with less damage to milk functionality. However, the quest for a suitable quality assurance indicator enzyme will need more extensive studies.

Pulsed electric field (PEF) processing has been demonstrated to be an effective non-thermal pasteurization method for food-treatment applications. With this method, high voltage, short-duration pulses are applied to a chamber through which liquid food is passed. If the voltage applied and the corresponding electric field develops a potential higher than a critical trans-membrane potential, the pores expand, and the membrane of the living cell is ruptured. Due to the lower amount of energy consumed during a PEF process, the temperature of the liquid is kept much lower than as opposed to conventional pasteurization. The PEF method thus kills bacteria and other microorganisms while preserving the nutrition and taste of the liquid foods. Although the parameter responsible for inactivation is the voltage applied, for any given voltage, the conductivity of the liquid defines a current through the liquid that causes the temperature to rise. Therefore, preventing excessive heating of the liquid requires the application of an efficient waveform. According to the literature, the most efficient waveform is a square wave since the entire energy applied would be used for the inactivation process. Although some power supplies are capable of generating such a waveform, the generation of an efficient waveform that satisfies all the requirements for producing a viable product for PEF applications is still a challenging problem. In this research, a cascadable pulse generator, based on a Marx generator design, was designed and implemented in order to generate a pulsed waveform for the treatment of liquid food. IGBT switches were used to charge capacitors in parallel and to discharge them in series as a means of generating a high voltage at the output. The design was implemented and tested for two stages, generating up to 6 kV and 1.6 kA square pulses with a controllable pulse width from 1 µs to 10 µs. Up to 3 switches were connected in parallel to enhance the current capability of the system. Also investigated are ways to improve the transient time by enhancing the IGBT driver circuit. The effect of design parameters such as pulse width, voltage, and current on the temperature rise in the liquid was also studied. A variety of liquid foods with different conductivities were tested in order to confirm the functionality of the system.

Pulsed electric field (PEF) processing has been demonstrated to be an effective non-thermal pasteurization method for food-treatment applications. With this method, high voltage, short-duration pulses are applied to a chamber through which liquid food is passed. If the voltage applied and the corresponding electric field develops a potential higher than a critical trans-membrane potential, the pores expand, and the membrane of the living cell is ruptured. Due to the lower amount of energy consumed during a PEF process, the temperature of the liquid is kept much lower than as opposed to conventional pasteurization. The PEF method thus kills bacteria and other microorganisms while preserving the nutrition and taste of the liquid foods. Although the parameter responsible for inactivation is the voltage applied, for any given voltage, the conductivity of the liquid defines a current through the liquid that causes the temperature to rise. Therefore, preventing excessive heating of the liquid requires the application of an efficient waveform. According to the literature, the most efficient waveform is a square wave since the entire energy applied would be used for the inactivation process. Although some power supplies are capable of generating such a waveform, the generation of an efficient waveform that satisfies all the requirements for producing a viable product for PEF applications is still a challenging problem. In this research, a cascadable pulse generator, based on a Marx generator design, was designed and implemented in order to generate a pulsed waveform for the treatment of liquid food. IGBT switches were used to charge capacitors in parallel and to discharge them in series as a means of generating a high voltage at the output. The design was implemented and tested for two stages, generating up to 6 kV and 1.6 kA square pulses with a controllable pulse width from 1 µs to 10 µs. Up to 3 switches were connected in parallel to enhance the current capability of the system. Also investigated are ways to improve the transient time by enhancing the IGBT driver circuit. The effect of design parameters such as pulse width, voltage, and current on the temperature rise in the liquid was also studied. A variety of liquid foods with different conductivities were tested in order to confirm the functionality of the system.