Made available in DSpace on 2016-03-04T13:55:12Z (GMT). No. of bitstreams: 2 7.pdf: 2671790 bytes, checksum: 3ea789720d5aa831716c6bfe2ca4f9b6 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2014 / Fundação Oswaldo Cruz. Instituto de Tecnologia em Fármacos/Farmanguinhos. Rio de Janeiro, RJ, Brasil. / O Instituto de Tecnologia em Fármacos (Farmanguinhos), unidade técnico-científica da Fundação Oswaldo Cruz, representa o maior laboratório farmacêutico oficial vinculado ao Ministério da Saúde brasileiro responsável pela produção de mais de um bilhão de medicamentos por ano. As legislações sanitárias vigentes definem que devem ser utilizadas instalações segregadas e dedicadas para a produção de medicamentos, como penicilinas, minimizando os riscos de contaminação cruzada, que consequentemente possam gerar danos à saúde. Por isso, Farmanguinhos possui, atualmente, uma área dedicada à produção de amoxicilina, assim como o sistema gerador de água purificada, que atende à referida fabricação, também é dedicado à mesma. A água é a matéria-prima de mais elevado volume empregada na produção farmacêutica, exercendo profundo impacto na qualidade do produto e na segurança do paciente. O controle microbiológico da qualidade da água utilizada em processos farmacêuticos torna-se fundamental, já que micro-organismos podem sobreviver e proliferar em sistemas de água, tornando-se fontes de contaminação microbiana e pirogênica. A identificação destes micro-organismos e as informações obtidas sobre os mesmos podem ser extremamente úteis na identificação da fonte de contaminação microbiana de um produto ou processo, alem de direcionar ações corretivas, se necessário. O objetivo deste trabalho foi avaliar a diversidade microbiana presente em sistema gerador de água purificada, dedicado à produção de penicilínicos, de Farmanguinhos. Para isto, as amostras de água coletadas a partir do ponto de entrada, ponto de água potável, e do ponto de saída do sistema, ponto de água purificada, foram analisadas microbiologicamente quanto à contagem de micro-organismos mesófilos e quanto à presença de patógenos: Escherichia coli; coliformes totais e Pseudomonas aeruginosa. As amostras foram analisadas entre janeiro e maio de 2013. Todas as colônias obtidas foram isoladas e submetidas à identificação, através de equipamento MALDI-TOF. Não foram observados resultados acima da especificação para ambos os pontos, no período analisado. Do ponto de água potável foram isoladas apenas três espécies bacterianas: Bacillus cereus, Delftia acidovorans e Acinetobacter sp. Uma diversidade microbiana maior foi isolada a partir do ponto de água purificada, como: Burkholderia sp, Delftia acidovorans, Stenotrophomonas maltophilia, Bacillus cereus, Pseudomonas oryzihabitans e Pseudomonas putida. A espécie Delftia acidovorans, associada a fenômenos de biocorrosão foi o micro-organismo mais encontrado, sendo identificado em cerca de 1/3 do total de isolados. O sistema de água de Farmanguinhos é sanitizado mensalmente com sanitizante à base de glutaraldeído. Pode-se concluir que a sanitização está sendo eficiente para manter a contagem total de micro-organismos mesófilos dentro dos limites especificados. No entanto, algumas ações podem ser tomadas para evitar a presença de biofilmes, o que aprimorará ainda mais a qualidade da água purificada gerada. / The Instituto de Tecnologia em Fármacos (Farmaguinhos), technical- scientific unit of Fundação Oswaldo Cruz, is the largest official pharmaceutical laboratory under the Brazilian Ministry of Health responsible for producing more than one billion drugs per year. The current sanitary laws define that segregated facilities must be dedicated and used for the production of drugs such as penicillin, thus the risk of serious damage to health, due to crosscontamination, can be minimized. Because of this, Farmanguinhos has a dedicated production of amoxicillin, as well as a purified water generator system that caters to such manufacturing is also dedicated to this area. Water is the raw material of the highest volume used in pharmaceutical production so that it has a profound impact on the product quality and patient safety. Therefore, the microbiological quality control of water used in pharmaceutical processes becomes crucial, since microorganisms can survive and proliferate in water systems, becoming sources of microbial and pyrogenic contamination. The identification of these microorganisms and the information obtained about them can be extremely useful in identifying the source of microbial contamination of a product or process, in addition to direct corrective actions, if necessary. The objective of this study was to evaluate the microbial diversity present in the purified water generator system, dedicated to the area of manufacturing penicillin-based drugs of Farmanguinhos. To aim this objective, water samples collected from the entry point of the system, point of drinking water, and the point of departure, point of purified water, were analyzed microbiologically for the count of mesophilic and for the presence of pathogens: Escherichia coli; coliforms and Pseudomonas aeruginosa, respectively. The samples were analyzed between January and May of 2013. All colonies obtained were isolated and subjected to identification by MALDI - TOF equipment. No results above specification for both points were observed in the analyzed period. From the point of drinking water only three bacterial species were isolated: Bacillus cereus, Acinetobacter sp and Delftia acidorovans. A higher microbial diversity was observed from the purified water source, such as Burkholderia sp, Delftia acidovorans, Stenotrophomonas maltophilia, Bacillus cereus, Pseudomonas putida and Pseudomonas oryzihabitans. The species Delftia acidovorans associated with biocorrosion phenomena was the microorganism most frequently found and was identified in about 1/3 of the total isolates. The water system of Farmaguinhos water is sanitized monthly with sanitizing agent comprising glutaraldehyde. It can be concluded that sanitization is being effective to maintain the total count of mesophilic within the specified limits. However, some actions can be taken to avoid the presence of biofilms, which will improve even more the quality of purified water generated.

A água empregada na produção de doses inseminantes (DIs) de suínos pode ser um dos veículos de introdução de bactérias contaminantes, podendo levar à perda de qualidade das mesmas. O objetivo do presente estudo foi avaliar o número de mesófilos aeróbios totais (MAT) na água utilizada em centrais de inseminação de suínos e sua influência nas contagens bacterianas das DIs produzidas. Foram conduzidos três ciclos de amostragem em seis centrais de inseminação (A-F), intencionalmente incluídas no estudo de acordo com os critérios: estar localizada no sul do Brasil; produzir >1.000 DIs mensais; e concordar em participar do estudo. A cada ciclo, eram colhidas amostras de água antes da purificação, água após a passagem pelo sistema de purificação, água após armazenamento, diluente, sêmen in natura de três machos distintos de fertilidade comprovada; e das respectivas doses inseminantes preparadas com sêmen e diluente amostrados. A água colhida antes do sistema de purificação foi avaliada quanto ao número mais provável (NMP) de coliformes totais e Escherichia coli, em 100 mL. pela técnica dos tubos múltiplos. Todas as amostras colhidas foram avaliadas quanto ao número de mesófilos aeróbios totais (MAT), por meio da semeadura em profundidade em Ágar para Contagem (PCA). Todas as 18 amostras de água colhidas antes da purificação apresentaram <1,1 NMP.100 mL-1 de Escherichia coli. A média de MAT nas amostras de água variou entre 0,1 log UFC.mL-1 na água após-purificação da central E e 3,5 log.UFC.mL-1 na água antes da purificação da central F. Não houve diferença significativa (P>0,05) entre as contagens médias de MAT obtidas em diferentes tipos de amostra de água e entre as centrais de inseminação. A média de MAT nas amostras de diluente variou de 0,16 log UFC. mL-1 à 2,78 log UFC.mL-1. As centrais D e F apresentaram diluentes significativamente (P<0,05) mais contaminados. No sêmen, a média de MAT variou de 1,75 log. UFC.mL-1 à 3,79 log UFC.mL-1 e nas DIs entre 0,73 log.UFC.mL-1 e 2,88 log UFC.mL-1. A central de inseminação F apresentou média de MAT significativamente (P<0,05) maior que as demais centrais. Considerando os padrões existentes, conclui-se que a água captada, purificada e armazenada apresentou boa qualidade microbiológica em todas as centrais de inseminação e não influenciou a média de mesófilos aeróbios totais das DIs preparadas. O incremento de bactérias, quando observado, foi provavelmente resultante da contaminação de origem ambiental durante o preparo do diluente e das doses inseminantes. / The water employed in the production of insemination doses (IDs) of swine may be one of the vehicles for introduction of contaminant bacteria, and can lead to their quality loss. The objective of this study was to assess the number of total aerobic mesophilic (TAM) in water used in swine artificial insemination centers (SAIC) and its influence in the total bacterial counting of the IDs produced. Three cycles of sampling were conducted in six SAIC (A-F) intentionally included in the study according to the following criteria: to be located in the south of Brazil, to produce > 1.000 (IDs) monthly, and to agree in participating of the study. At each cycle, samples were collected from water before the purification, water after passage through the purification system, water after storage, extenders, fresh semen collected from three different boars and the respective (ID) prepared with the semen and extender sampled. The water collected before the purification system was evaluated for total coliforms and Escherichia coli most probable number (NMP) in 100 mL using the multiple tubes technic. All samples collected were evaluated for the number of total aerobic mesophilic (TAM) by the pour plate technic. All the 18 samples of water collected before the purification presented < 1.1 NMP. 100 mL-1 of Escherichia coli. The average of TAM in the water samples varied between 0.1 log.CFU. ml-1 in the water after purification of the central E and 3.5 log. CFU.mL-1 in the water before purification of the central F. There was no significant difference (P>0.05) among the average counting of TAM obtained in different water samples types and among SAICs. The average TAM in samples of diluents varied from 0.16 log.CFU.mL-1 to 2.78 log.CFU.mL-1; the SAICs D and F presented diluents significantly (P< 0.05) more contaminated. Among the semen samples, the TAM average varied from 1.75 log.CFU.mL-1 to 3.79 log CFU.mL-1 and in the DIs between 0.73 log.CFU.mL-1 and 2.88 log.CFU.mL-1. The SAIC F presented average of TAM significantly (P<0.05) higher than the others SAICs. Considering the available standards, it was concluded that water collected, purified and stored presented a good microbiological quality in all insemination centers and did not influence the number of TAM in the prepared IDs. The increase on bacterial population may have resulted from contamination of environmental origin during the preparation of the extender and insemination doses.

A água empregada na produção de doses inseminantes (DIs) de suínos pode ser um dos veículos de introdução de bactérias contaminantes, podendo levar à perda de qualidade das mesmas. O objetivo do presente estudo foi avaliar o número de mesófilos aeróbios totais (MAT) na água utilizada em centrais de inseminação de suínos e sua influência nas contagens bacterianas das DIs produzidas. Foram conduzidos três ciclos de amostragem em seis centrais de inseminação (A-F), intencionalmente incluídas no estudo de acordo com os critérios: estar localizada no sul do Brasil; produzir >1.000 DIs mensais; e concordar em participar do estudo. A cada ciclo, eram colhidas amostras de água antes da purificação, água após a passagem pelo sistema de purificação, água após armazenamento, diluente, sêmen in natura de três machos distintos de fertilidade comprovada; e das respectivas doses inseminantes preparadas com sêmen e diluente amostrados. A água colhida antes do sistema de purificação foi avaliada quanto ao número mais provável (NMP) de coliformes totais e Escherichia coli, em 100 mL. pela técnica dos tubos múltiplos. Todas as amostras colhidas foram avaliadas quanto ao número de mesófilos aeróbios totais (MAT), por meio da semeadura em profundidade em Ágar para Contagem (PCA). Todas as 18 amostras de água colhidas antes da purificação apresentaram <1,1 NMP.100 mL-1 de Escherichia coli. A média de MAT nas amostras de água variou entre 0,1 log UFC.mL-1 na água após-purificação da central E e 3,5 log.UFC.mL-1 na água antes da purificação da central F. Não houve diferença significativa (P>0,05) entre as contagens médias de MAT obtidas em diferentes tipos de amostra de água e entre as centrais de inseminação. A média de MAT nas amostras de diluente variou de 0,16 log UFC. mL-1 à 2,78 log UFC.mL-1. As centrais D e F apresentaram diluentes significativamente (P<0,05) mais contaminados. No sêmen, a média de MAT variou de 1,75 log. UFC.mL-1 à 3,79 log UFC.mL-1 e nas DIs entre 0,73 log.UFC.mL-1 e 2,88 log UFC.mL-1. A central de inseminação F apresentou média de MAT significativamente (P<0,05) maior que as demais centrais. Considerando os padrões existentes, conclui-se que a água captada, purificada e armazenada apresentou boa qualidade microbiológica em todas as centrais de inseminação e não influenciou a média de mesófilos aeróbios totais das DIs preparadas. O incremento de bactérias, quando observado, foi provavelmente resultante da contaminação de origem ambiental durante o preparo do diluente e das doses inseminantes. / The water employed in the production of insemination doses (IDs) of swine may be one of the vehicles for introduction of contaminant bacteria, and can lead to their quality loss. The objective of this study was to assess the number of total aerobic mesophilic (TAM) in water used in swine artificial insemination centers (SAIC) and its influence in the total bacterial counting of the IDs produced. Three cycles of sampling were conducted in six SAIC (A-F) intentionally included in the study according to the following criteria: to be located in the south of Brazil, to produce > 1.000 (IDs) monthly, and to agree in participating of the study. At each cycle, samples were collected from water before the purification, water after passage through the purification system, water after storage, extenders, fresh semen collected from three different boars and the respective (ID) prepared with the semen and extender sampled. The water collected before the purification system was evaluated for total coliforms and Escherichia coli most probable number (NMP) in 100 mL using the multiple tubes technic. All samples collected were evaluated for the number of total aerobic mesophilic (TAM) by the pour plate technic. All the 18 samples of water collected before the purification presented < 1.1 NMP. 100 mL-1 of Escherichia coli. The average of TAM in the water samples varied between 0.1 log.CFU. ml-1 in the water after purification of the central E and 3.5 log. CFU.mL-1 in the water before purification of the central F. There was no significant difference (P>0.05) among the average counting of TAM obtained in different water samples types and among SAICs. The average TAM in samples of diluents varied from 0.16 log.CFU.mL-1 to 2.78 log.CFU.mL-1; the SAICs D and F presented diluents significantly (P< 0.05) more contaminated. Among the semen samples, the TAM average varied from 1.75 log.CFU.mL-1 to 3.79 log CFU.mL-1 and in the DIs between 0.73 log.CFU.mL-1 and 2.88 log.CFU.mL-1. The SAIC F presented average of TAM significantly (P<0.05) higher than the others SAICs. Considering the available standards, it was concluded that water collected, purified and stored presented a good microbiological quality in all insemination centers and did not influence the number of TAM in the prepared IDs. The increase on bacterial population may have resulted from contamination of environmental origin during the preparation of the extender and insemination doses.

A água empregada na produção de doses inseminantes (DIs) de suínos pode ser um dos veículos de introdução de bactérias contaminantes, podendo levar à perda de qualidade das mesmas. O objetivo do presente estudo foi avaliar o número de mesófilos aeróbios totais (MAT) na água utilizada em centrais de inseminação de suínos e sua influência nas contagens bacterianas das DIs produzidas. Foram conduzidos três ciclos de amostragem em seis centrais de inseminação (A-F), intencionalmente incluídas no estudo de acordo com os critérios: estar localizada no sul do Brasil; produzir >1.000 DIs mensais; e concordar em participar do estudo. A cada ciclo, eram colhidas amostras de água antes da purificação, água após a passagem pelo sistema de purificação, água após armazenamento, diluente, sêmen in natura de três machos distintos de fertilidade comprovada; e das respectivas doses inseminantes preparadas com sêmen e diluente amostrados. A água colhida antes do sistema de purificação foi avaliada quanto ao número mais provável (NMP) de coliformes totais e Escherichia coli, em 100 mL. pela técnica dos tubos múltiplos. Todas as amostras colhidas foram avaliadas quanto ao número de mesófilos aeróbios totais (MAT), por meio da semeadura em profundidade em Ágar para Contagem (PCA). Todas as 18 amostras de água colhidas antes da purificação apresentaram <1,1 NMP.100 mL-1 de Escherichia coli. A média de MAT nas amostras de água variou entre 0,1 log UFC.mL-1 na água após-purificação da central E e 3,5 log.UFC.mL-1 na água antes da purificação da central F. Não houve diferença significativa (P>0,05) entre as contagens médias de MAT obtidas em diferentes tipos de amostra de água e entre as centrais de inseminação. A média de MAT nas amostras de diluente variou de 0,16 log UFC. mL-1 à 2,78 log UFC.mL-1. As centrais D e F apresentaram diluentes significativamente (P<0,05) mais contaminados. No sêmen, a média de MAT variou de 1,75 log. UFC.mL-1 à 3,79 log UFC.mL-1 e nas DIs entre 0,73 log.UFC.mL-1 e 2,88 log UFC.mL-1. A central de inseminação F apresentou média de MAT significativamente (P<0,05) maior que as demais centrais. Considerando os padrões existentes, conclui-se que a água captada, purificada e armazenada apresentou boa qualidade microbiológica em todas as centrais de inseminação e não influenciou a média de mesófilos aeróbios totais das DIs preparadas. O incremento de bactérias, quando observado, foi provavelmente resultante da contaminação de origem ambiental durante o preparo do diluente e das doses inseminantes. / The water employed in the production of insemination doses (IDs) of swine may be one of the vehicles for introduction of contaminant bacteria, and can lead to their quality loss. The objective of this study was to assess the number of total aerobic mesophilic (TAM) in water used in swine artificial insemination centers (SAIC) and its influence in the total bacterial counting of the IDs produced. Three cycles of sampling were conducted in six SAIC (A-F) intentionally included in the study according to the following criteria: to be located in the south of Brazil, to produce > 1.000 (IDs) monthly, and to agree in participating of the study. At each cycle, samples were collected from water before the purification, water after passage through the purification system, water after storage, extenders, fresh semen collected from three different boars and the respective (ID) prepared with the semen and extender sampled. The water collected before the purification system was evaluated for total coliforms and Escherichia coli most probable number (NMP) in 100 mL using the multiple tubes technic. All samples collected were evaluated for the number of total aerobic mesophilic (TAM) by the pour plate technic. All the 18 samples of water collected before the purification presented < 1.1 NMP. 100 mL-1 of Escherichia coli. The average of TAM in the water samples varied between 0.1 log.CFU. ml-1 in the water after purification of the central E and 3.5 log. CFU.mL-1 in the water before purification of the central F. There was no significant difference (P>0.05) among the average counting of TAM obtained in different water samples types and among SAICs. The average TAM in samples of diluents varied from 0.16 log.CFU.mL-1 to 2.78 log.CFU.mL-1; the SAICs D and F presented diluents significantly (P< 0.05) more contaminated. Among the semen samples, the TAM average varied from 1.75 log.CFU.mL-1 to 3.79 log CFU.mL-1 and in the DIs between 0.73 log.CFU.mL-1 and 2.88 log.CFU.mL-1. The SAIC F presented average of TAM significantly (P<0.05) higher than the others SAICs. Considering the available standards, it was concluded that water collected, purified and stored presented a good microbiological quality in all insemination centers and did not influence the number of TAM in the prepared IDs. The increase on bacterial population may have resulted from contamination of environmental origin during the preparation of the extender and insemination doses.

This master’s thesis project, conducted in Industrial Design Engineering at KTH Royal Institute of Technology in Stockholm and in collaboration with Trangia, addresses the increasing significance of home preparedness due to global conflicts and natural disasters. The relevance of home preparedness is growing both in Swedish society and for Trangia, prompting the company to participate in this expanding market. Therefore, the mission is to develop customized solutions for cooking, heating up a room, and water purification to assist urban users in preparing for emergencies. The project utilized the Double Diamond methodology which offers a clear structure that systematically leads through the complex phases of product development; Discover, Define, Develop and Deliver. The initial phase of the project, Discover, included comprehensive information gathering, user studies, and branding exploration to expand Trangia’s product line. The project prioritized to identify the most suitable and healthy fuel sources for the user and delved into the technical principles enabling cooking, heating, and water purification. In the second phase, Define, the target group was narrowed down to urban users within three behavioral groups that exhibited distinct needs. Here, a design space was created that addressed the practical and emotional needs, ultimately shaping the functional requirements. The third phase, Develop, was initiated by the design space. Here, the concept development activities started, which subsequently led to the creation of Trangia KRIS in the Deliver-phase. A home preparedness kit comprising a crisis kitchen, Krisköket, a crisis heater, Krisvärmaren and a service proposal, including a disaster page educating potential buyers on the risks of being unprepared. Along with a coffee table book including an info card collection offering user-friendly instructions and emergency preparedness tips. The project emphasizes the importance of home preparedness, focusing on essential actions such as access to food, clean water, and warmth during crises like power outages or conflicts, rather than relying solely on products for survival. / Detta examensarbete inom Teknisk Design vid Kungliga Tekniska Högskolan i Stockholm och i samarbete med Trangia, tar upp den ökande betydelsen av hemberedskap på grund av globala konflikter och naturkatastrofer. Relevansen av hemberedskap växer både i det svenska samhället och för Trangia, vilket har fått företaget att vilja delta i denna växande marknad. Uppdraget är att utveckla en lösning som möjliggör förberedelser för matlagning, uppvärmning av ett rum och vattenrening under en kris för urbana användare. Projektet utgick från design processen Double Diamond som erbjuder en tydlig struktur som systematiskt leder genom de komplexa faserna av produktutveckling; Discover, Define, Develop och Deliver. Den initiala fasen av projektet, Discover, inkluderade omfattande informationsinsamling, användarstudier och varumärkesutforskning för att utöka Trangias produktlinje. Projektet prioriterade att identifiera de mest lämpliga och hälsosamma bränslekällorna för användaren och fördjupade sig i de tekniska principerna som möjliggör matlagning, uppvärmning och vattenrening. I den andra fasen, Define, begränsades målgruppen till stadsanvändare inom tre beteendegrupper som uppvisade distinkta behov. Här skapades ett design space som adresserade bland annat de praktiska och psykologiska behoven, vilket slutligen formade en kravspecifikation. Den tredje fasen, Develop, initierades av design spacet. Här började konceptutvecklingsaktiviteterna, vilket senare ledde till skapandet av Trangia KRIS i Deliver-fasen. Ett hemberedskapskit bestående av ett kriskök, Krisköket, en krisvärmare, Krisvärmaren och en tjänst. Tjänsten består av en katastrofsida med syfte att informera potentiella köpare om riskerna med att vara oförberedd. Tillsammans med en coffee table book innehållande en samling informationskort som erbjuder användarvänliga instruktioner och tips för hemberedskap. Projektet betonar vikten av hemberedskap, med fokus på väsentliga åtgärder såsom tillgång till mat, rent vatten och värme under kriser som strömavbrott eller krig, snarare än att enbart förlita sig på produkter för överlevnad.

L’un des objectifs du programme de recherche Opticook est d’équiper les fours avec des capteurs spectroscopiques qui permettront de piloter de façon non intrusive la cuisson de viande et de poisson. Le développement de ces capteurs est basé sur l’hypothèse que les protéines de ces aliments sont de bons marqueurs du degré de cuisson. Dans ce contexte, dans un premier temps, plusieurs outils ont été mis en œuvre pour caractériser l’effet de différents barèmes de cuisson sur les propriétés macroscopiques (texture, couleur) d’échantillons de bœuf, de poulet et de dos de cabillaud. Puis, pour étudier les transformations que subissent ces échantillons à l’échelle moléculaire, nous avons utilisé la calorimétrie et différentes spectroscopies. La spectroscopie dans le domaine du visible nous a permis d’observer la dénaturation des hémoprotéines grâce à l’évolution de l’état (oxy à met ). Ces protéines sont responsables du changement de couleur observé entre 50 et 75 °C. Les spectroscopies dans le moyen infrarouge et de fluorescence ont été utilisées pour suivre les changements de structures associées à la dénaturation des protéines fibrillaires. La myosine, purifiée à partir du blanc de poulet, a été étudiée en fonction de la température par fluorescence. Les résultats nous ont montré que des évolutions de la structure protéique se produisent dès 50 °C. À l’échelle microscopique, nous avons réalisé des expériences sur des échantillons de bœuf par RMN bas champ (séquences d’impulsion et gradient de champ), et par tomographie de neutrons pour rendre compte de la contraction des fibres et de la libération du jus selon le degré de cuisson. L’analyse des images 3D enregistrées par tomographie de neutrons et le traitement des données de RMN montrent des changements importants de la morphologie et de la densité de fibres des échantillons après la cuisson à haute température. Pour associer les deux approches (moléculaire et microscopique) des mesures couplées de spectroscopie et d’imagerie de neutrons ont été menées dans le four Opticook pendant la cuisson de tranches de bœuf. Ce couplage nous a permis de suivre l’évolution de la morphologie due à la contraction des fibres, la migration du jus, ainsi que le changement de couleur associé à la dénaturation des hémoprotéines. Nous en concluons qu’il est en effet possible de suivre le degré de cuisson de viande et de poisson à l’aide de différentes spectroscopies (la fluorescence et le visible) grâce aux signatures spectroscopiques de plusieurs marqueurs internes à ces matrices : la myosine, le collagène et les hémoprotéines. / One of the purposes of the Opticook project is to install spectroscopic sensors in ovens, so equipping them with non intrusive tools allowing following cooking process of meat and fish. The hypothesis on which sensors were developed was: are muscular proteins proper probes to discriminate among several cooking degrees? Thus, at the beginning of the project several tools were used to characterise effect of several cooking degrees on macroscopic properties (texture and colour) of beef, chicken and cod fillet samples. Following, calorimetry and spectroscopic techniques were used to study transformations at molecular scale. In particular, visible spectroscopy followed denaturation of haemproteins observing their evolution from oxy to met state. Colour change was observed between 50 and 75 °C. Middle infrared and fluorescence spectroscopies were used to observe evolution of fibrillar proteins which are associated with meat texture changes. One of myofibrils, myosin, was purified from chicken breast and was studied depending on temperature by fluorescence spectroscopy. Observations showed evolution of protein structure occurred around 50 °C. At microscopic scale, low field NMR (spin echo and gradient field configurations) and neutron tomography were used to observe morphology changes and juice release on beef samples according to cooking degree. 3D image analysis and NMR data showed morphology and fibre density changes at high temperature.We associated the two approaches (molecular and microscopic) in the last part of the research. So coupled measures were realised by combining neutron imaging and spectroscopy techniques during cooking process on beef slices by Opticook oven. Coupled measures allowed following morphology evolution because of fibre contractions, juice migration and colour modification as well associated to haemprotein denaturation. In conclusion, it is possible to follow cooking process of meat and fish by following several spectroscopic signatures (fluorescence and visible ones) of matrix own probes: myosin, collage and haemproteins.

Les polyphénols, micronutriments abondants dans notre alimentation, sont bien connus pour leurs propriétés antioxydantes et anti-inflammatoires et leur consommation a été associée à un moindre risque de maladies chroniques (maladies cardiovasculaires, cancers…). Ces composés suscitent un intérêt croissant dans le domaine de la santé digestive. En effet, les polyphénols sont faiblement absorbés au niveau de l’intestin grêle et sont présents en quantité relativement importante dans le côlon. Les polyphénols et leurs métabolites pourraient donc exercer des effets bénéfiques à ce niveau. Des études expérimentales récentes suggèrent que les polyphénols pourraient aider à prévenir ou retarder la progression de l’inflammation colique. Le raisin est l’un des fruits les plus riches en polyphénols. Divers co-produits du raisin ont montré des effets bénéfiques vis-à-vis de différentes situations d'inflammation, effets souvent attribués à leur richesse en composés polyphénoliques. Les objectifs de cette thèse étaient tout d’abord d’évaluer l’impact du régime de base sur le développement de l’inflammation colique induite par le sulfate de dextran sodique (DSS) chez le rat (1) puis d’évaluer les effets d’une supplémentation nutritionnelle en extraits de marc de raisin (EMR) riches en polyphénols dans la prévention de l’inflammation colique induite par le DSS (2). Pour cela, des rats Wistar ont été nourris ad libitum durant 21 jours avec un régime standard ou un régime semi-synthétique (1) ou les animaux ont été nourris avec un régime semi-synthétique supplémenté ou non avec un EMR (2). Dans tous les cas, l’inflammation colique a été induite par administration de DSS à 4% dans l’eau de boisson pendant les 7 derniers jours de l’expérimentation. Les effets ont été évalués au niveau colique par analyse histologique, détermination de différents marqueurs de l'inflammation et de la défense antioxydante et évaluation de l'expression de différents gènes impliqués dans l’inflammation et le stress oxydant.La consommation préventive de trois EMRs, issus de deux cépages différents (Alicante et Pinot) exerce des effets bénéfiques sur le développement de l’inflammation colique induite par le DSS. Globalement, la consommation à titre préventif d’EMRs prévient la perte de poids, retarde l’apparition des symptômes de la maladie et limite le développement des atteintes tissulaires induites par le DSS. Les EMRs atténuent l’inflammation comme en témoigne la diminution de l’activité de la MPO et le maintien des taux coliques en cytokines pro-inflammatoires au niveau des animaux témoins. De plus, ces EMRs modulent les activités des enzymes antioxydantes. Les EMRs limitent également la surexpression de plusieurs gènes impliqués dans la réponse inflammatoires et le stress oxydant (COX-2, iNOS, ICAM-1, MMP-9…). De plus, nous avons montré que les EMRs provenant du cépage Alicante exercent des effets protecteurs plus marqués que l’EMR Pinot.Ainsi, ces résultats soulignent d’une part l’influence du régime alimentaire de base dans l’induction de l’inflammation colique. D’autre part, la consommation à titre préventif d’EMRs exerce des effets protecteurs sur le développement de l'inflammation colique induite par le DSS via la modulation de marqueurs de l’inflammation et de la défense antioxydante. Ces résultats pourraient permettre d’envisager de nouvelles stratégies nutritionnelles dans la prévention des maladies inflammatoires de l’intestin. / Polyphenols are abundant micronutrients in plant foods, where they have been associated with lower risk of chronic diseases from cancer to cardiovascular disease. The beneficial health effects of polyphenols have been attributed to their antioxidant and anti-inflammatory properties. Several studies point to a promising role of these micronutrients in inflammatory bowel disease (IBD), as given that a large fraction of dietary polyphenols reach the colon, polyphenols and their metabolites could exert beneficial effects locally at this level. Recent experimental data suggest that polyphenols could help prevent or delay the progression of intestinal inflammation. Grape is one of the richest sources of polyphenols, and various grape products have demonstrated beneficial effects in various chronic diseases. These effects are often attributed to grape polyphenolic compounds.This thesis aimed (1) to evaluate the impact of purified and non-purified basal diets on the development of dextran sodium sulfate (DSS)-induced colitis in the rat, then (2) to evaluate the effects of preventive consumption of polyphenol-rich red grape pomace extracts (GPEs) on DSS-induced colitis. Rats were fed for 21 days with (1) a non-purified or purified diet or (2) GPE-enriched purified diet, and colitis was induced by DSS administration in drinking water (4% w/v) during the last 7 days of the experimental design. The effects on colitis development were assessed by histological analysis, determination of various markers of inflammation and oxidative stress, and evaluation of the expression of various genes implicated in this disorder.DSS induced a more marked colitis in rats receiving the purified diet than the non-purified diet. Indeed, rats receiving the purified diet, the inflammation was characterized by decreased appetite and weight gain, early onset of disease (diarrhea and rectal bleeding), and colon shortening. In rats fed the purified diet, DSS also induced marked histological colonic damage and increased MPO activity. In these animals, DSS also modulated oxidative status by affecting antioxidant enzyme activities. Furthermore, DSS-induced inflammation was associated with increased pro-inflammatory cytokine levels and decreased anti-inflammatory cytokine levels in colonic tissue, together with upregulation of the expression of various genes (iNOS, ICAM-1 and MMP-9).Preventive consumption of three GPEs from two different grape varieties (Alicante and Pinot) showed beneficial effects on DSS-induced colitis development. Overall, preventive dietary GPE supplementation delayed the onset of colitis symptoms and limited the extent of weight loss and colonic mucosa damage. GPE administration reduced inflammation (as evidenced by a decrease in MPO activity) and kept colonic pro-inflammatory cytokines at near-control levels. In addition, the three GPEs modulated antioxidant enzyme activities and limited the overexpression of several genes involved in DSS-induced inflammatory response and oxidative stress (COX-2, iNOS, ICAM-1, MMP-9). Moreover, Alicante-variety GPEs exerted stronger protective effects than Pinot-variety GPEs.These results highlighted (1) the influence of basal diet on induction of colonic inflammation and (2) that preventive consumption of GPEs exerted protective effects against colitis development by modulating inflammatory processes and oxidative stress markers. These findings open up perspectives for new preventive nutritional strategies in colonic inflammation management.

Multipotent mesenchymal stem cells (MSCs), found in many adult tissues, may be useful for regenerative medicine applications. Their identification and purification have been difficult due to their low frequency and lack of unambiguous markers. Using a magnetic micro-beads negative selection technique to remove contaminating hematopoietic cells from mouse bone marrow stromal cells (BMSCs), our lab recently isolated a highly purified osteoprogenitor (HipOp) population that was also enriched for other mesenchymal precursors, including MSCs (Itoh and Aubin, 2009). To further enhance enrichment, we positively selected BMSCs and HipOps for CD73, a putative MSC marker, which resulted in no significant additional enrichment for osteoprogenitors when the population was tested in vitro. However, we also found that HipOps were enriched in vascular endothelial cells, and that removing these cells by further negative selection with CD31/PECAM resulted in a CD31(-) HipOp population with higher osteogenic capacity than HipOps in vitro and in vivo.

Multipotent mesenchymal stem cells (MSCs), found in many adult tissues, may be useful for regenerative medicine applications. Their identification and purification have been difficult due to their low frequency and lack of unambiguous markers. Using a magnetic micro-beads negative selection technique to remove contaminating hematopoietic cells from mouse bone marrow stromal cells (BMSCs), our lab recently isolated a highly purified osteoprogenitor (HipOp) population that was also enriched for other mesenchymal precursors, including MSCs (Itoh and Aubin, 2009). To further enhance enrichment, we positively selected BMSCs and HipOps for CD73, a putative MSC marker, which resulted in no significant additional enrichment for osteoprogenitors when the population was tested in vitro. However, we also found that HipOps were enriched in vascular endothelial cells, and that removing these cells by further negative selection with CD31/PECAM resulted in a CD31(-) HipOp population with higher osteogenic capacity than HipOps in vitro and in vivo.

Thesis (DPhil (History))--University of Stellenbosch, 2010. / ENGLSIH ABSTRACT: This study is a political biography of D.F. Malan (1874–1959), the first of the apartheid-era Prime Ministers, and covers the years 1874 to 1954, when Malan retired from politics. It endeavours to provide a warts-and-all account of D.F. Malan which challenges prevalent myths and stereotypes surrounding his public persona and his political orientation. While the overwhelming focus is on Malan’s political career, special attention is paid to his personal life in order to paint a multi-faceted picture of his character. The biography is written in the form of a seamless narrative and employs a literary style of writing. It is based on archival research which utilised Malan’s private collection, as well as the private collections of his Nationalist contemporaries. Malan takes the centre stage at all times, as the biography focuses on his perceptions and experiences. Malan’s views regarding Afrikaner nationalism, which was his foremost political priority, are described, and are related to his views of British imperialism as well as other ideologies such as communism and totalitarianism. This study demonstrates that there is a notable link between Malan’s perceptions of race relations and his concerns about the poor white problem. It reveals that Malan’s racial policy was, to some extent, fluid, as were his views on South Africa’s constitutional position. Debates about South Africa’s links to Britain and the nature of the envisioned republic preoccupied Afrikaner nationalists throughout the first half of the twentieth century – and served as an outlet for regional and generational tensions within the movement. Malan’s clashes with nationalists such as Tielman Roos, J.B.M. Hertzog and J.G. Strijdom are highlighted as an indication of the internecine power struggles within the National Party (NP). By emphasising these complexities, this study seeks to contribute to a nuanced understanding of the South African past. / AFRIKAANSE OPSOMMING: Hierdie studie is politieke biografie van D.F. Malan (1874–1959), die eerste van die apartheid-era Eerste Ministers, en dek die jare 1874 tot 1954, toe Malan uit die politiek getree het. Dit poog om onversuikerde beeld van Malan te skets wat heersende mites en stereotipes aangaande sy openbare beeld en sy benadering tot die politiek uitdaag. Die fokus is hoofsaaklik op Malan se politieke loopbaan, maar besondere aandag word aan sy private lewe geskenk om sodoende veelsydige portret van sy karakter te skilder. Die biografie is in die vorm van naatlose narratief geskryf en maak van literêre skryfstyl gebruik. Dit is gebaseer op argivale navorsing, waartydens daar van D.F. Malan se privaat versameling gebruik gemaak is, sowel as die privaat versamelings van sy tydgenote. Malan is ten alle tye die sentrale figuur en die biografie fokus op sy persepsies en ervarings. Malan se denke oor Afrikaner nasionalisme, wat sy vernaamste prioriteit was, word beskryf en in verband gebring met sy opinie van Britse imperialisme, sowel as ander ideologieë soos kommunisme en totalitarisme. Die studie wys op die verband tussen Malan se denke oor rasseverhoudinge en sy besorgdheid oor die armblanke vraagstuk. Dit dui daarop dat Malan se rassebeleid tot sekere mate vloeibaar was. Dit was ook die geval met sy benadering tot Suid-Afrika se konstitusionele posisie. Afrikaner nasionaliste het tydens die eerste helfte van die twintigste eeu baie aandag geskenk aan debatte oor Suid-Afrika se verhouding tot Brittanje en die aard van die voorgenome republiek. Dit was tot mate weerligafleier vir reeds bestaande spanning tussen die onderskeie streke en generasies. Malan se botsings met nasionaliste soos Tielman Roos, J.B.M. Hertzog en J.G. Strijdom word belig as aanduiding van die diepgewortelde magstryd binne die Nasionale Party (NP). Deur op hierdie kompleksiteite klem te lê, poog die studie om bydrae te lewer tot meer genuanseerde begrip van die Suid-Afrikaanse verlede.