Le but général de ce travail correspond à l’amélioration de la biodisponibilité de principes actifs connus pour leur faible biodisponibilité (testostérone) ou pour leur caractère lipidique (acides gras oméga 3). Des systèmes nanoparticulaires à base de lipides et des systèmes nanocristaux ont été développés notamment dans le cas de la testostérone, afin d’obtenir une biodisponibilité supérieure au système oral actuellement commercialisé, Andriol Testocaps®. L’autre partie de ce travail consistait en la conception d’une formule d’acides gras omega -3 dans des nanoparticules lipidiques, susceptibles malgré l’utilisation d’une huile de poisson bon marché comme source d’acides gras omega-3, de n’avoir que peu d’effets sur le goût et l’odorat tout en s’avérant stable. Le développement de systèmes oraux à base de testostérone a été possible autant sur la base de la technologie des nanoparticules lipidiques (Nanostructured Lipid Carriers, NLC), que sur celle des nanocristaux. Dans les deux cas, les systèmes développés ont permis de répondre aux exigences en matière d’incorporation (NLC) et de stabilité (NLC et suspensions Nano). Les NLC ont permis d’incorporer jusqu’à 30% d’undecanoate de testostérone en phase lipidique. La formulation a également été possible avec différents lipides, susceptibles, d’augmenter l’absorption lymphatique et de ce fait également la biodisponibilité de l’hormone. Les nanocristaux ont pu être produits à partir de la testostérone (T) ainsi que d’undecanoate de testostérone (TU), avec des tailles moyennes respectives d’environ 470 nm (TU) et 860 nm (T). Cette taille particulaire doit permettre une absorption lymphatique accrue. Les biodisponibilités des systèmes développés à base de NLC, se sont avérées, chez le rat Wistar, toujours plus élevées que la formulation commerciale, quand ils ont été administrés sans lipides additionnels ce qui permet de supposer, que l’influence simultanée de l’absorption de nourriture sur la biodisponibilité devrait être moins prononcée qu’elle ne l’est pour l’Andriol Testocaps®. En partant de ces résultats, à savoir l’augmentation de la biodisponibilité orale avec les nanoparticules lipidiques, le développement d’un système de nanoparticules avec les acides gras oméga-3 d’huile de poisson bon marché était un pas logique. La biodisponibilité orale des acides gras oméga-3 est largement supérieure à celle du TU (environ 70 %). L’utilisation de la technologie NLC a ainsi permis la réduction de l’odeur et du goût du produit. La formulation avait un pourcentage remarquablement élevé de 70 % en phase lipidique tout en restant pâteuse et redispersible. Ceci permet d’envisager son utilisation dans des boissons et dans la nutrition ce qui facilitera l’assistance des patients (et donc la biodisponibilité) avec les acides gras omega-3 essentiels. / The overall goal of this work consisted in ameliorating the bioavailability of drugs known for their poor hydrosolubility (testosterone) or for their lipidic character (omega-3 fatty acids). This was achieved using lipid nanoparticle systems and nanocrystals In case of testosterone the work consisted of the development of an oral dosage form with superior properties compared to the currently commercially available oral system (Andriol Testocaps®). The other part of this work was the design of a lipid nanoparticle-based omega- 3 fatty acid formulation, which, despite the use of cheap fish oil as source of omega-3 fatty acids, has low smell and taste properties while nevertheless being stable. The development of the oral testosterone drug delivery system was accomplished on the basis of lipid nanoparticles technology and also using drug nanocrystal technology. In both cases, systems could be developed that met the requirements with regards to drug loading (NLC) and stability (NLC and drug nanocrystals). Up to 30 % of testosterone undecanoate could be incorporated into the lipid phase of the NLC. Furthermore, the production of particles with different lipids, which are supposed to promote lymphatic absorption and hence the bioavailability of the hormone. Drug nanocrystals of testosterone (T) and testosterone undecanoate (TU) were prepared with a mean size of about 470 nm (TU) and 860 nm (T). Also with this system, an enhanced lymphatic absorption was expected. The bioavailabilites of the developed NLC based drug delivery systems were all higher than the bioavailability of the product on the market when no additional lipid was supplied. This gives reason to believe, that the influence of co-administered food on the bioavailability of the systems is less pronounced than with Andriol Testocaps®. Based on the findings that lipid nanoparticles can improve oral bioavailability, the development of an omega-3 fatty acids nanoparticulate system (NLC) out of cheap fish oil was a logic step. The oral bioavailability of the omega-3 fatty acids is much higher than the one of TU (about 70 %). Through the use of NLC technology, the taste and smell is even more reduced. It was rather unexpected that we achieved to have a formulation that consisted of 70 % lipid phase (and 30 % water) but still was paste-like and easy to redisperse. This makes the use of the paste as an additive in food and beverages possible to better supply the patient with essential omega-3 fatty acids.

No description available.

MA Cum Laude / Thesis presented in partial fulfilment of the requirements for the degree of Master of Arts (International Studies) at Stellenbosch University / ENGLISH ABSTRACT: The objective of this thesis is to explain the differing levels of rural activism in Brazil and South Africa. As both countries are plagued with similar land and poverty disparities, the varying intensity and national organisation of rural movements is striking. In Brazil a strong and nationally organised rural movement, the Movimento dos Trabalhadores Rurais Sem- Terra (MST), established itself as the leading rural movement; whereas South Africa’s National Land Committee (NLC) remained weak and ultimately collapsed. Today, South Africa is characterised by a complete lack of a national representation of rural interests and shows only timid attempts at the local level. In order to address the issue systematically and comprehensively, the thesis first provides a historical outline of both countries, thereby discerning similarities and differences in social, economic and political development. Subsequently, and based upon these findings, a systematic comparison of the NLC and MST is conducted. Utilising contemporary social movement theory, a synthesised theoretical framework of political opportunities, resource mobilisation and framing processes is proposed to methodically compare movement dynamics. Applying this synthesised framework the protest cycles of the NLC and the MST are compared, namely the emerging phase, the stabilisation and decline/resurgence phase. The study points to a complex network of reasons for varying rural activism. In South Africa an overall demobilising constellation of important movement dynamics led to the collapse of the NLC and the weakening of the rural grassroots. Political opportunities changed from overly exclusive to overly inclusive in South Africa whereby the NLC’s resource mobilisation became narrowly institutionalised; containing most oppositional forces at the national and local level. In Brazil, in contrast, political opportunities remained ambivalent throughout MST existence; thereby providing enough loopholes to achieve partial success and yet maintaining the critical distance and constraints which necessitates and legitimates grassroots mobilisation. In Brazil, land distribution has been singled out early as the prime source for deprivation and consequently served as a vantage point for framing processes which stimulated a coherent idea of landlessness and the legitimation of land occupations. The exclusive/inclusive dichotomy of the South African society with its strong racial overtones led to framing processes which interpret land reform as an exclusive state affair; thereby discouraging land occupations and merging land with the broad context of social injustice in South Africa. The thesis concludes that the historically constructed and contemporarily continued racial dichotomy of South Africa’s society has ultimately hampered rural movement dynamics in South Africa. / AFRIKAANSE OPSOMMING: Die doel van die tesis is om die verskille in aktivisme dinamiek van grondhervormingsbewegings in Suid-Afrika en Brasilië te verduidelik. Die verskillende in terme van nasionale organisasie en intensiteit is merkwaardig gegewe dat beide state gekenmerk word deur soortgelyke grond en armoede ongelykhede. In Brasilïe is ’n sterk en nasionaal georganiseerde beweging, die Movimento dos Trabalhadores Rurais-Sem Terra (MST) gevestig as die leidende grondhervormingsbeweging, terwyl Suid-Afrika se Nasionale Grond Komitee (National Land Committee, NLC) swak gebly het en eindelik as ’n beweging verval het. Suid-Afrika word vandag gekenmerk deur die afwesigheid van ’n nasionale artikulasie van die belange van grondloses met gebrekkige pogings om hul belange op plaaslike vlak te verteenwoordig. Ten einde die kwessie sistematies en omvattend aan te spreek, verskaf die tesis eerstens ’n historiese konteks van die politieke ekonomie van grond in beide state ten einde verskille en soortgelykhede uit te wys. Hierna word die MST en die NLC sistematies vergelyk. Deur gebruik te maak van kontemporêre sosiale bewegingsteorie word ‘n gesintetiseerde teoretiese raamwerk – wat fokus op Politieke Geleenthede, Hulpbron Mobilisering en Orienteringsprosesse – voorgestel om metodologies die dinamiek van die bewegings te ontleed. Deur die gesintetiseerde raamwerk toe te pas, word die protes siklusse van die NLC en die MST vergelyk, naamlik die ontstaan fase, die stabiliseringsfase en die verval/herlewingsfase. Die studie ontrafel ‘n kompleks netwerk van redes vir gedifferensieerde grondaktivisme. In Suid-Afrika het ‘n reeks demoboliserende faktore gelei tot die verval van die NLC en die verswakking van plattelandse organiasies op voetsoolvlak. Politieke geleenthede het verander van eksplisiet eksklusief na eksplisiet inklusiewe prosesse waardeur die NLC se basis vir hulpbron mobilisering baie nou geinstitusionaliseerd geword het en waardeur meeste aktiviste op nasionale en plaaslike vlak gekoopteer is. In Brasilïe in teenstelling het politieke geleenthede tydens die MST se bestaan ambivalent gebly en as gevolg daarvan voldoende ruimte gebied om ‘n kritieke afstand teenoor die staat in te neem. In Brasilïe is grondhervorming reeds lank gelede geidentifiseer as die oorsaak vir ontneming en het gevolglik gedien as die basis vir mobilisering rondom grondbesit en die legitimering van onwettige grond okkupasie. Die eksklusief/inklusief dichotomie van die Suid-Afrikaanse samelewing met gepaardgaande ras-kompleksiteit het gelei tot prosesse waardeur grondhervorming as ‘n ekslusiewe staats kwessie gesien is wat daardeur onwettige grond besettings verminder het en die debat rondom grondhervorming vetroebel het as net nog ‘n geval van sosiale ongeregtigheid. Daar word tot die gevolgtrekking gekom dat die historiese konstruksie en voortgesette rasse konteks waarbinne grondhervoming in Suid-Afrika plaasvind, die moontlikheid vir ‘n soortgelyke aktivistiese grondhervormingsbeweging soos in Brasilïe kniehalter.

Negative Linear Compressibility (NLC), where a material expands in a given direction when subjected to hydrostatic compression, is a rare elastic property that has received much attention recently, but has yet to be used in practical applications. What are the mechanisms responsible for this property in crystals and man-made structures? Are all mechanisms somehow related to the wine-rack model? Can we find an even simpler and more fundamental elucidation of NLC? Following this mechanistic approach, can we then identify “engineering” materials with NLC? To answer these questions, I have used a combination of analytical modelling based on beam theory and finite element analysis, to investigate several structures. At first, I examine in great detail the standard wine-rack in 2D and equivalents in 3D and identify the aspect ratio (close to two) at which NLC is maximum. By adding spacers I demonstrate that a cross is not a necessary condition, and that simpler angle changes in chains are sufficient to generate NLC. Looking for materials with intersecting straight chains, “zig-zag” chains or quasi-helical structures, I find that carbon fibre mats, some extruded polymers and some woods exhibit NLC. Finally, I show that elliptical voids in 2D sheets can also generate NLC in a way related to the wine-rack. This thesis improves the understanding of the mechanism(s) responsible for NLC by proving that a wine-rack is not necessary. Perhaps more importantly it suggests that the property can be exploited in several relatively common materials.

No description available.

No description available.

A película adquirida (PA) é um filme formado pela adsorção seletiva de proteínas, glicoproteínas e lipídeos à superfície dentária. A presença de proteínas na PA forma uma interface protetora sobre a superfície do dente, participando em todos os eventos interfaciais que ocorrem na cavidade bucal, tais como des- e remineralização, lubrificação das superfícies dos dentes, e aderência bacteriana. Com o advento da proteômica, tem havido um aumento considerável no conhecimento acerca do perfil proteico de PAs adquiridas formadas sobre o esmalte dentário, em diferentes situações, mas nenhum trabalho até o momento descreveu o perfil proteômico de PAs formadas sobre a dentina. Este estudo foi pioneiro em comparar o perfil proteico de PAs formadas in situ sobre o esmalte e a dentina, nos tempos de 10 minutos e 2 horas, utilizando análise proteômica quantitativa livre de marcadores. Os experimentos foram realizados por três dias consecutivos. Em cada dia, os 9 voluntários receberam profilaxia dentária e em seguida utilizaram um aparelho vestibular com 6 blocos de esmalte e 6 de dentina humanos por 10 minutos ou 2 horas. Após esses períodos, a PA formada era coletada com auxílio de um papel filtro de eletrodos embebido em ácido cítrico 3%. Para as análises foi realizado um pool com os papéis dos 9 voluntários de todos os dias, para cada substrato e tempo de formação. Após a extração e digestão das proteínas, a separação dos peptídeos foi realizada por nano-HPLC (nano-Cromatografia Líquida de Alta Performace), interligada a um espectrômetro de massa (nLC-ESI-MS/MS). Os dados MS/MS obtidos foram processados e pesquisados em bancos de dados de proteínas humanas (UniProt e TrEMBL), utilizando o algoritmo SEQUEST no software Proteome Discoverer 1.3. Para a PA formada sobre o esmalte, foram identificadas 160 e 64 proteínas, nos tempos de formação de 10 minutos e 2 horas, respectivamente. Os respectivos números de proteínas identificadas para a dentina foram 86 e 52, respectivamente. Nos tempos de 10 minutos e 2 horas, respectivamente, 25 e 11 proteínas foram comuns a ambos os substratos e foram submetidas à quantificação livre de marcadores (SIEVE), revelando que a maioria das proteínas com diferença de expressão entre os dois substratos teve sua expressão aumentada na dentina. Foram identificadas ainda, no tempo de 10 minutos de formação da PA, 135 e 61 proteínas exclusivas ao esmalte ou à dentina, respectivamente. O número correspondente de proteínas exclusivas para o tempo de 2 horas foi de 53 e 41 proteínas, para o esmalte e dentina, respectivamente. Dentre as proteínas exclusivas da dentina, foram identificadas várias proteínas relacionadas ao complexo cálcio/calmodulina, assim como proteínas associadas à tumorigênese e à fosforilação/desfosforilação de proteínas. Em adição, muitas das proteínas identificadas no presente estudo, tanto para o esmalte quanto para a dentina, ainda não foram caracterizadas e, portanto, não têm função conhecida na PA. Sua caracterização e estudos funcionais futuros poderão trazer novos horizontes no entendimento da importância da PA para a proteção da estrutura dentária, bem como do papel da PA como sítio de biomarcadores para doenças bucais e sistêmicas. / The acquired pellicle (AP) is a film that results from selective adsorption of proteins, glicoproteins and lipids on the tooth surface. The presence of proteins in the AP forms a protective interface on the tooth surface that participates in all the surface events occurring in the oral cavity, such as de- and remineralization, lubrification of the tooth surfaces and bacterial adherence. With the advent of Proteomics, considerable increase in the knowledge of the protein profile of the AP formed on tooth enamel, under different circunstances, has been observed. However, so far the proteomic profile of the AP formed on dentin has not been described. This is the first study to compare the proteomic profile of APs formed in situ for 10 minutes and 2 hours, on enamel and dentin, using quantitative label-free proteomics. The experiments were conducted for 3 consecutive days. Each day, 9 volunteers were submitted to dental prophylaxis and in sequence wore a vestibular device containing 6 human enamel and 6 human dentin blocks for 10 minutes or 2 hours. After these periods, the PA formed was collected with an electrode filter paper soaked in 3% citric acid. The papers from the 9 volunteers, for each substrate and time of pellicle formation were pooled and used for analysis. After protein extraction and digestion, peptides were separated by nano-HPLC (High-performance liquid chromatography) coupled to a mass spectrometer (nLC-ESI- MS/MS). The obtained MS/MS spectra were searched against human protein databases (UniProt and TrEMBL) using SEQUEST algorithm in Proteome Discoverer 1.3 software. For the AP formed on enamel, 160 and 64 proteins were identified for the times of pellicle formation of 10 minutes and 2 hours, respectively. The respective numbers of identified proteins for dentin were 86 and 52, respectively. For the times of 10 minutes and 2 hours, respectively, 25 and 11 proteins were common to both substrates. They were submitted to label-free quantification, which revealed that most of the proteins with differential expression were overexpressed in the dentin. For APs formed for 10 minutes, 135 and 61 proteins were identified exclusively for enamel or dentin, respectively. The corresponding number for the 2-hour APs was 53 and 41 proteins, respectively. Among the proteins identified exclusively in dentin, many proteins related with calcium/calmodulin complex, as well as proteins associated with tumorigenesis and protein phosphorylation/dephosphorylation were found. In addition, many of the identified proteins, both for enamel and dentin, remain uncharacterized and, therefore have no described function in the AP. In the future, their characterization and functional studies might open new avenues for the understanding of the importance of the AP for the protection of the dental structure, as well as for the use of the AP as a site for biomarkers of oral and systemic diseases.

A película adquirida (PA) é um filme formado pela adsorção seletiva de proteínas, glicoproteínas e lipídeos à superfície dentária. A presença de proteínas na PA forma uma interface protetora sobre a superfície do dente, participando em todos os eventos interfaciais que ocorrem na cavidade bucal, tais como des- e remineralização, lubrificação das superfícies dos dentes, e aderência bacteriana. Com o advento da proteômica, tem havido um aumento considerável no conhecimento acerca do perfil proteico de PAs adquiridas formadas sobre o esmalte dentário, em diferentes situações, mas nenhum trabalho até o momento descreveu o perfil proteômico de PAs formadas sobre a dentina. Este estudo foi pioneiro em comparar o perfil proteico de PAs formadas in situ sobre o esmalte e a dentina, nos tempos de 10 minutos e 2 horas, utilizando análise proteômica quantitativa livre de marcadores. Os experimentos foram realizados por três dias consecutivos. Em cada dia, os 9 voluntários receberam profilaxia dentária e em seguida utilizaram um aparelho vestibular com 6 blocos de esmalte e 6 de dentina humanos por 10 minutos ou 2 horas. Após esses períodos, a PA formada era coletada com auxílio de um papel filtro de eletrodos embebido em ácido cítrico 3%. Para as análises foi realizado um pool com os papéis dos 9 voluntários de todos os dias, para cada substrato e tempo de formação. Após a extração e digestão das proteínas, a separação dos peptídeos foi realizada por nano-HPLC (nano-Cromatografia Líquida de Alta Performace), interligada a um espectrômetro de massa (nLC-ESI-MS/MS). Os dados MS/MS obtidos foram processados e pesquisados em bancos de dados de proteínas humanas (UniProt e TrEMBL), utilizando o algoritmo SEQUEST no software Proteome Discoverer 1.3. Para a PA formada sobre o esmalte, foram identificadas 160 e 64 proteínas, nos tempos de formação de 10 minutos e 2 horas, respectivamente. Os respectivos números de proteínas identificadas para a dentina foram 86 e 52, respectivamente. Nos tempos de 10 minutos e 2 horas, respectivamente, 25 e 11 proteínas foram comuns a ambos os substratos e foram submetidas à quantificação livre de marcadores (SIEVE), revelando que a maioria das proteínas com diferença de expressão entre os dois substratos teve sua expressão aumentada na dentina. Foram identificadas ainda, no tempo de 10 minutos de formação da PA, 135 e 61 proteínas exclusivas ao esmalte ou à dentina, respectivamente. O número correspondente de proteínas exclusivas para o tempo de 2 horas foi de 53 e 41 proteínas, para o esmalte e dentina, respectivamente. Dentre as proteínas exclusivas da dentina, foram identificadas várias proteínas relacionadas ao complexo cálcio/calmodulina, assim como proteínas associadas à tumorigênese e à fosforilação/desfosforilação de proteínas. Em adição, muitas das proteínas identificadas no presente estudo, tanto para o esmalte quanto para a dentina, ainda não foram caracterizadas e, portanto, não têm função conhecida na PA. Sua caracterização e estudos funcionais futuros poderão trazer novos horizontes no entendimento da importância da PA para a proteção da estrutura dentária, bem como do papel da PA como sítio de biomarcadores para doenças bucais e sistêmicas. / The acquired pellicle (AP) is a film that results from selective adsorption of proteins, glicoproteins and lipids on the tooth surface. The presence of proteins in the AP forms a protective interface on the tooth surface that participates in all the surface events occurring in the oral cavity, such as de- and remineralization, lubrification of the tooth surfaces and bacterial adherence. With the advent of Proteomics, considerable increase in the knowledge of the protein profile of the AP formed on tooth enamel, under different circunstances, has been observed. However, so far the proteomic profile of the AP formed on dentin has not been described. This is the first study to compare the proteomic profile of APs formed in situ for 10 minutes and 2 hours, on enamel and dentin, using quantitative label-free proteomics. The experiments were conducted for 3 consecutive days. Each day, 9 volunteers were submitted to dental prophylaxis and in sequence wore a vestibular device containing 6 human enamel and 6 human dentin blocks for 10 minutes or 2 hours. After these periods, the PA formed was collected with an electrode filter paper soaked in 3% citric acid. The papers from the 9 volunteers, for each substrate and time of pellicle formation were pooled and used for analysis. After protein extraction and digestion, peptides were separated by nano-HPLC (High-performance liquid chromatography) coupled to a mass spectrometer (nLC-ESI- MS/MS). The obtained MS/MS spectra were searched against human protein databases (UniProt and TrEMBL) using SEQUEST algorithm in Proteome Discoverer 1.3 software. For the AP formed on enamel, 160 and 64 proteins were identified for the times of pellicle formation of 10 minutes and 2 hours, respectively. The respective numbers of identified proteins for dentin were 86 and 52, respectively. For the times of 10 minutes and 2 hours, respectively, 25 and 11 proteins were common to both substrates. They were submitted to label-free quantification, which revealed that most of the proteins with differential expression were overexpressed in the dentin. For APs formed for 10 minutes, 135 and 61 proteins were identified exclusively for enamel or dentin, respectively. The corresponding number for the 2-hour APs was 53 and 41 proteins, respectively. Among the proteins identified exclusively in dentin, many proteins related with calcium/calmodulin complex, as well as proteins associated with tumorigenesis and protein phosphorylation/dephosphorylation were found. In addition, many of the identified proteins, both for enamel and dentin, remain uncharacterized and, therefore have no described function in the AP. In the future, their characterization and functional studies might open new avenues for the understanding of the importance of the AP for the protection of the dental structure, as well as for the use of the AP as a site for biomarkers of oral and systemic diseases.

Uma das características de qualidade dos vinhos espumantes, e que também impõe a sua identidade, é a aparência das borbulhas. Tradicionalmente, acredita-se que a capacidade de formação e estabilização dessas borbulhas depende de macromoléculas do vinho, em especial das proteínas, devido a sua ação tensoativa. Este trabalho de doutorado visou o estudo proteômico do vinho espumante brasileiro a fim de identificar quais proteínas estão presentes nesses vinhos para entender melhor a influência dessas na estabilização da espuma (perlage e colarinho), e com o intuito de potencializar essa característica em nossos produtos. Foram utilizados os métodos de extração de proteínas clássico, ácido tricloroacético/acetona e de última geração, biblioteca combinatória de ligantes peptídicos, sendo estas separadas por SDS-PAGE, 2DE e OFFGEL. As proteínas extraídas foram digeridas com tripsina e a mistura de peptídeos analisada por nLC-MS/MS com metodologia shotgun. Os resultados iniciais obtidos por eletroforese 2DE e OFFGEL, mostraram a presença de três grupos de proteínas de massa molecular distintas, sendo duas próximas a 25 kDa e uma próxima a 70 kDa. Estas proteínas parecem estar presentes nos vinhos em mais de uma isoforma evidenciado pelo espalhamento de todas as bandas de mesma massa molecular em diferentes pH. Foram identificadas 40 proteínas, sendo 17 proteínas de organismos do sub-reino Viridiplantae e 23 proteínas pertencentes ao gênero Saccharomyces, onde 10 e 6 proteínas, respectivamente, estão presentes em pelo menos duas amostras de espumantes nacionais. Dessas, seis proteínas foram identificadas pela primeira vez em vinhos. Três proteínas originárias da levedura Saccharomyces cerevisiae estão presentes em todos os produtos analisados, podendo essas proteínas serem as responsáveis pela melhor formação de espuma observada em nossos produtos em relação ao Champagne (vinho espumante tradicional da França). / The type of fizzy bubbles is one of the aspects that characterizes the quality of sparkling wines and also helps defining their identity. Traditionally, it is believed that the ability of these bubbles to form and stabilize depends on the macromolecules found in the wine, particularly proteins, due to their surfactant action. The aim of this work is the proteomic study of the brazilian sparkling wines in order to identify which proteins are present to better understand the influence of these molecules in the foam formation (perlage and collar), in order to improve our products. The protein extraction methods used were the classical TCA/acetone precipitation and the modern combinatory peptide ligand library. Then, proteins were separated by SDS-PAGE, 2DE and OFFGEL. The protein extracted were digested with trypsin and the peptide mixture were analyzed with nLC-MS/MS using the shotgun method. The first results obtained by electrophoresis 2DE and OFFGEL showed the presence of three groups of proteins with different molecular mass, two of them close to 25 kDa and the other one close to 70 kDa. These proteins appear to be present in wine in more than one isoform evidenced by spreading in all bands of similar molecular weight at different pH. In total, 40 proteins were identified, 17 protein from Viridiplantae sub-kingdom organisms and 23 proteins belonging to Saccharomyces genus, where 10 and 6 proteins, respectively, are present in at least two samples of domestic sparkling wines. Six of those proteins were identified in wine for the first time. Three proteins originating from Saccharomyces cerevisiae are present in all analyzed products, and those may be responsible for a better foam formation observed in our products in comparison to Champagne (traditional French sparkling wine).

Uma das características de qualidade dos vinhos espumantes, e que também impõe a sua identidade, é a aparência das borbulhas. Tradicionalmente, acredita-se que a capacidade de formação e estabilização dessas borbulhas depende de macromoléculas do vinho, em especial das proteínas, devido a sua ação tensoativa. Este trabalho de doutorado visou o estudo proteômico do vinho espumante brasileiro a fim de identificar quais proteínas estão presentes nesses vinhos para entender melhor a influência dessas na estabilização da espuma (perlage e colarinho), e com o intuito de potencializar essa característica em nossos produtos. Foram utilizados os métodos de extração de proteínas clássico, ácido tricloroacético/acetona e de última geração, biblioteca combinatória de ligantes peptídicos, sendo estas separadas por SDS-PAGE, 2DE e OFFGEL. As proteínas extraídas foram digeridas com tripsina e a mistura de peptídeos analisada por nLC-MS/MS com metodologia shotgun. Os resultados iniciais obtidos por eletroforese 2DE e OFFGEL, mostraram a presença de três grupos de proteínas de massa molecular distintas, sendo duas próximas a 25 kDa e uma próxima a 70 kDa. Estas proteínas parecem estar presentes nos vinhos em mais de uma isoforma evidenciado pelo espalhamento de todas as bandas de mesma massa molecular em diferentes pH. Foram identificadas 40 proteínas, sendo 17 proteínas de organismos do sub-reino Viridiplantae e 23 proteínas pertencentes ao gênero Saccharomyces, onde 10 e 6 proteínas, respectivamente, estão presentes em pelo menos duas amostras de espumantes nacionais. Dessas, seis proteínas foram identificadas pela primeira vez em vinhos. Três proteínas originárias da levedura Saccharomyces cerevisiae estão presentes em todos os produtos analisados, podendo essas proteínas serem as responsáveis pela melhor formação de espuma observada em nossos produtos em relação ao Champagne (vinho espumante tradicional da França). / The type of fizzy bubbles is one of the aspects that characterizes the quality of sparkling wines and also helps defining their identity. Traditionally, it is believed that the ability of these bubbles to form and stabilize depends on the macromolecules found in the wine, particularly proteins, due to their surfactant action. The aim of this work is the proteomic study of the brazilian sparkling wines in order to identify which proteins are present to better understand the influence of these molecules in the foam formation (perlage and collar), in order to improve our products. The protein extraction methods used were the classical TCA/acetone precipitation and the modern combinatory peptide ligand library. Then, proteins were separated by SDS-PAGE, 2DE and OFFGEL. The protein extracted were digested with trypsin and the peptide mixture were analyzed with nLC-MS/MS using the shotgun method. The first results obtained by electrophoresis 2DE and OFFGEL showed the presence of three groups of proteins with different molecular mass, two of them close to 25 kDa and the other one close to 70 kDa. These proteins appear to be present in wine in more than one isoform evidenced by spreading in all bands of similar molecular weight at different pH. In total, 40 proteins were identified, 17 protein from Viridiplantae sub-kingdom organisms and 23 proteins belonging to Saccharomyces genus, where 10 and 6 proteins, respectively, are present in at least two samples of domestic sparkling wines. Six of those proteins were identified in wine for the first time. Three proteins originating from Saccharomyces cerevisiae are present in all analyzed products, and those may be responsible for a better foam formation observed in our products in comparison to Champagne (traditional French sparkling wine).