Atualmente, a utilização de azo corantes pelas indústrias de tingimento constitui um problema ambiental e de saúde, considerando o lançamento de quantidades elevadas para o meio ambiente e a falta de dados toxicológicos dos corantes disponíveis para as indústrias. Vários estudos têm demonstrado o potencial genotóxico de diversos corantes azóicos, porém para os corantes Disperse Red 1, Disperse Orange 1 e o Disperse Red 13, não foram encontrados dados na literatura relativos à sua capacidade de dano ao material genético. Considerando que esses corantes são empregados em processos de tingimento no Brasil, esse trabalho teve como objetivo a avaliação de sua atividade mutagênica, utilizando o teste de micronúcleo (MN) em linfócitos humanos e em células HepG2. Os resultados obtidos no teste com linfócitos, demonstram que na menor concentração testada (0,2 µg/mL), o número de micronúcleos presentes foi semelhante ao controle negativo, mas esse número aumenta à medida que eleva-se a concentração. No entanto, a partir da concentração de 1,0 µg/mL, este valor começa a decair. Isso provavelmente se deve à citotoxidade dos corantes, levando à morte celular ou redução da divisão celular e, conseqüentemente, não há a formação de micronúcleo. Embora o perfil de mutagenicidade dos três corantes seja semelhante, o corante Disperse Red 13 parece ter maior potencial de dano sobre os linfócitos em relação aos demais, seguido pelo Disperse Red 1 e Disperse Orange 1, respectivamente. Os resultados obtidos para o teste de MN em células HepG2 foram semelhantes aos obtidos no teste feito em linfócitos. O aumento do número de micronúcleos em relação ao aumento da concentração dos corantes, ocorreu até o limite de 2,0 µg/mL em células HepG2, excetuando-se o corante Disperse Red 13, para o qual o limite foi de 1,0 µg/mL. E a partir desses pontos, considerados como limites, ocorreu uma redução no número de MN. Para este sistema celular, os três corantes parecem ter potencial mutagênico bastante semelhante. Portanto, a análise dos resultados mostrou que os corantes Disperse Red 13, Disperse Red 1 e Disperse Orange 1 são mutagênicos para sistemas celulares diferentes. Foi também avaliado Índice de Proliferação do Bloqueio de Citocinese (IPBC), que permite a avaliação de toxicidade celular ou atraso no ciclo celular por meio da determinação da proliferação celular nas culturas. Porém, neste estudo não foram observadas diferenças estatísticas entre o controle negativo e as concentrações testadas. Nossos resultados confirmam que os azo corantes constituem uma importante classe de contaminantes ambientais e devem ser avaliados e utilizados de forma cautelosa. / Currently, the use of azo dyes for the textile industries can causes direct and/or indirect effects on human health and on the environment, considering the discharge of industrial effluents that contain toxic dyes and the lack of reports in the literature about the toxic effects of these compounds. Several studies have been demonstrated the genotoxic effect of diverse azo dyes, however for the dyes Disperse Red 1, Disperse Orange 1 and Disperse Red 13 no information about their capacity to cause DNA damage was found in the literature. Considering that these dyes are used for dying processes in Brazil, the main of this work was the evaluation of the mutagenic activity of Disperse Red 1, Disperse Orange 1 and Disperse Red 13, using the micronucleus assay (MN) in human lymphocytes and HepG2 cells. For the lymphocytes assay, we observed that the number of micronucleus induced by the lowest concentration of each dye (0,2 µg/mL) was similar to the negative control. For the other concentrations we observed a dose response micronucleus formation, until 1,0 µg/mL. Above this concentration, the number of micronucleus has decreased, probably because of the cytotoxic effects of the dyes, which leads to cellular death or reduction of cellular division and, consequently, does not have the micronucleus formation. Although the mutagenicity profile of the three dyes is similar, Disperse Red 13 seems to be the strongest for the lymphocytes, followed by Disperse Red 1 and Disperse Orange 1, respectively. For the HepG2 cells the results were similar to the lymphocytes. For the three dyes we noted a dose dependent increase in the frequency of micronuclei. However, for the HepG2 the threshold for this increase was 2,0 µg/mL, except for Disperse Red 13, which the limit was at 1 µg/ml, after this point a reduction in the MN number occurred. For this cellular system, the three dyes seem to have similar mutagenic potential. Therefore, our results suggest that the dyes Disperse Red 13, Disperse Red 1 and Disperse Orange 1 are potentially mutagenic for different cellular systems. Besides, cytokinesis-block proliferation index (CBPI) was calculated, in order to evaluate cellular toxicity or delay in the cellular cycle through of the determination of the cellular proliferation in the cultures. No statistical difference was detected between the tested concentrations and the negative control. Our results confirmed that azo dyes constitute an important class of environmental contamination and they should be evaluated and used carefully.

Atualmente, a utilização de azo corantes por vários ramos industriais constitui um problema ambiental e de saúde pública, tendo em vista o lançamento de quantidades elevadas para o meio ambiente e a falta de dados toxicológicos a cerca dos corantes e de seus metabólitos gerados, principalmente, após os processos de oxidação e redução. Nosso grupo realizou ensaios com micronúcleos em linfócitos humanos e em células HepG2 e ensaio de mutagenicidade com Salmonella typhimurium, demonstrando que os azo corantes Disperse Red 1, Disperse Red 13 e Disperse Orange 1 são mutagênicos para os diferentes parâmetros. Dessa forma, neste trabalho foi avaliada a capacidade de ligação dos corantes originais com o DNA e a base nitrogenada guanosina, a fim de elucidar o mecanismo de ação mutagênica. Adicionalmente, foi realizada a análise do potencial mutagênico do corante Disperse Red 1 e de seus metabólitos por meio do teste de mutação gênica em células de linfomas de camundongo (Mouse Lymphoma Assay), e também foram avaliados os produtos de oxidação e redução dos azo corantes Disperse Red 1, Disperse Red 13 e Disperse Orange 1, por meio do teste de mutagenicidade com Salmonella typhimurium. Posteriormente, foi investigada a possível formação de aminas aromáticas e de outros compostos, após os ensaios eletroquímicos e reação com S9, utilizando CLAE/DAD e CG/EM. Nossos resultados mostraram que a formação de adutos com o DNA, especificamente com a base guanosina, não é o mecanismo de ação tóxica preferencial para os azo corantes estudados. O corante Disperse Red 1 e seus produtos de biotransformação apresentaram resultados negativos no teste de mutação gênica em células de linfoma de camundongos. No entanto, tanto os produtos de oxidação como os de redução dos três corantes estudados apresentaram potencial mutagênico ao serem testados no Ensaio Salmonella/microssoma. Os produtos identificados após a oxidação química e enzimática (utilizando S9) e redução química dos três corantes estudados foram: sulfato 2-[(4-aminofenil)etilamino]- etanol monohidratada, 2-cloro-4-nitro-benzamina, benzamina, nitrobenzeno, 4-nitro-benzamina, 2-(etilfenilamino)-etanol, N-fenilbenzamina, N-fenil-1,4-benzenodiamina. Portanto, nossos dados mostram que a exposição por via oral a esses corantes tem relevância toxicológica, visto que podem causar danos à saúde não somente pela exposição aos corantes inalterados, mas também devido à formação de produtos tóxicos após a biotransformação. Cabe ressaltar que os corantes estudados no presente trabalho são amplamente utilizados por indústrias têxteis no Brasil, o que pode levar à contaminação de águas e alimentos. / Currently, the use of azo dyes by various industries is an environmental problem and public health, considering the release of large quantities to the environment and the lack of toxicological data about the dyes and their metabolites generated, especially after the processes of oxidation and reduction. Our group carried out micronuclei assay in human lymphocytes and HepG2 cells and mutagenicity test with Salmonella typhimurium, indicating that the azo dyes Disperse Red 1, Disperse Red 13 and Disperse Orange 1 are mutagenic to the different parameters. Thus, this study evaluated the binding capacity of the original dyes with DNA and nitrogenous base guanosine in order to elucidate the mechanism of mutagenic action. Additionally, we performed the analysis of mutagenic potential of Disperse Red 1 dye and its metabolites using Mouse Lymphoma Assay, and also evaluated the products of oxidation and reduction of the azo dyes Disperse Red 1, Disperse Red 13 and Disperse Orange 1, using the mutagenicity test with Salmonella typhimurium. Also it was investigated the possible formation of aromatic amines and other compounds after the electrochemical assays and reaction with S9, using HPLC / DAD and GC / MS. Our results showed that the formation of adducts with DNA, specifically with the guanosine base is not the preferred mechanism of toxic action for the azo dyes studied. The dye Disperse Red 1 and its biotransformation products had negative results in the mouse lymphoma assay. However, both the products of oxidation and the reduction of three dyes studied showed mutagenic potential in the Salmonella/ microsome assay. The products identified after chemical and enzymatic oxidation (using S9) and chemical reduction of three dyes studied were: sulfate 2-[(4- aminophenyl)ethylamino]-Ethanol monohydrate, 2-chloro-4-nitro-benzamine, benzamine, nitrobenzene, 4-nitro-benzamine, 2-(ethylphenylamino)-Ethanol, Nphenyl- benzamine, N-phenyl-1,4-benzenediamine. Therefore, our data show that oral exposure to these dyes have toxicological significance, since it can cause damage to health not only by exposure to dyes unchanged, but also due to the formation of toxic products after the biotransformation. It is noteworthy that the dyes studied in this work are widely used by textile industries in Brazil, which can lead to contamination of food and water.

O processamento de citros se justifica pela dificuldade de descascamento destes frutos. Estudos de descascamento de laranja \'Pêra\' realizados na ESALQ-USP vêm mostrando o potencial do uso do tratamento hidrotérmico para facilitar o descascamento desta variedade. As laranjas são imersas em água a 50ºC por 8 minutos, isto facilita o descascamento e não afeta a qualidade da fruta. Entretanto, é necessário estudar outros tempos de imersão para flexibilizar o uso desta técnica em escala industrial, bem como estendê-la a outras variedades. O objetivo deste trabalho foi adequar a tecnologia de descascamento de frutas cítricas pelo uso do tratamento hidrotérmico, bem como avaliar sua influência na qualidade fisiológica, físico-química, microbiológica e sensorial de laranjas \'Pêra\', laranjas \'Valência\' e tangores \'Murcott\'. Também foram avaliados o tempo de descascamento, o rendimento em frutos comercializáveis e a temperatura interna dos frutos durante o tratamento. Os frutos foram lavados, sanitizados, resfriados a 5ºC por 12 horas, submetidos ao tratamento hidrotérmico e descascados. O tratamento hidrotérmico consistiu em colocar os frutos em banho-maria a 50ºC por 5 (somente para o tangor \'Murcott\'), 10, 15, 20, 25 e 30 minutos. Posteriormente, os frutos foram descascados retirando-se a parte peduncular com a faca e, em seguida, o flavedo foi retirado, manualmente, junto com o albedo. Os frutos sem tratamento hidrotérmico (controle) foram descascados retirando-se primeiramente o flavedo, e depois o albedo cuidadosamente para causar o mínimo de injúria possível. Os frutos foram analisados durante seis dias de armazenamento a 5ºC. Os experimentos foram conduzidos separadamente para cada fruta cítrica e de acordo com cada tipo de análise. Foram utilizados delineamentos inteiramente ao acaso e em blocos casualizados, adequados para cada variável analisada. O tratamento hidrotérmico provocou alterações na atividade respiratória dos frutos somente nas primeiras horas após o processamento. A temperatura interna dos frutos (medida a 2 cm de profundidade em relação ao epicarpo) após 30 minutos de tratamento atingiu aproximadamente 35ºC, temperatura comumente observada em algumas etapas da cadeia de comercialização dos frutos. A coloração externa das laranjas sem tratamento apresentou maior valor de luminosidade (L) quando comparadas às frutas tratadas. Não houve alterações nas outras características físico-químicas e nas características microbiológicas dos frutos. O tratamento não alterou o sabor e melhorou a aparência em relação aos frutos sem tratamento devido à ausência de resquícios de albedo nos frutos. Além disso, o tratamento diminuiu em até 78% o tempo de descascamento dos frutos tratados para a laranja \'Pêra\', em até 75% para a laranja \'Valência\' e em até 57% para o tangor \'Murcott\', quando comparados aos frutos sem tratamento, e aumentou o rendimento em frutos comercializáveis. O tratamento hidrotérmico realizado até 30 minutos a 50ºC pode ser utilizado como técnica de descascamento para laranja \'Pêra\', laranja \'Valência\' e tangor \'Murcott\'. / The citrus fruit processing is justified for the difficulty of peeling of these fruits. Studies of peeling of \'Pera\' sweet orange fruit accomplished at ESALQ-USP are showing the potential of the use of the hydrothermal treatment to facilitate the peeling of this variety. The oranges are immersed in hot water at 50ºC for 8 minutes. This process to make easy the peeling and it doesn\'t affect the quality of the fruit. However, it is necessary to study other immersion times to make flexible the use of this technique in industrial scale, as well as to extend it for other varieties. The purpose of this work was to adapt the technology of peeling of citric fruits for the use of the hydrothermal treatment, as well as to evaluate the influence of the hydrothermal treatment in the physiological, physicochemical, microbiologic and sensorial qualities of \'Pera\' sweet orange, \'Valencia\' sweet orange and \'Murcott\' tangor. The peeling time, the yield of marketable fruits and the internal fruit temperature were also evaluated during the treatment. Fruits were washed, sanitized, cooled at 5ºC for 12 hours, submitted to hydrothermal treatment and peeled. The hydrothermal treatment consisted of putting fruits in water-bath at 50ºC for 5 (only for \'Murcott\' tangor), 10, 15, 20, 25 and 30 minutes. Then, fruits were peeled by first opening a gap on the peduncular region with a knife and then, the flavedo was removed, manually, with the albedo. Fruits with no hydrothermal treatment (control) were peeled by first removing the flavedo and then, the albedo was removed carefully to cause the less of injuries possible. The fruits were analyzed for six days of storage at 5ºC. The experiments were carried out separately for each citric fruit and in agreement with each analysis type. The experimental designs used were completely randomized and in randomized blocks, appropriate for each analyzed variable. Hydrothermal treatment caused changes in respiratory activity just in first hours after treatment. Internal fruit temperature (evaluated at 2 cm depth in relation to the epicarp) after 30 minutes of treatment reached 35ºC approximately, temperature commonly observed in some stages of the commercialization\'s chain of the fruits. The external coloration of the oranges without treatment presented larger value of brightness (L) when compared to the treated fruits. There were no changes in the others physicochemicals and microbiologics characteristics of the fruits. The treatment did not change the flavor and it improved the appearance in relation to the fruits without treatment due to the lower albedo residue in the fruits. Besides, the treatment decreased in up to 78% the peeling time of the treated fruits for the \'Pera\' sweet orange, in up to 75% for the \'Valencia\' sweet orange and in up to 57% for the \'Murcott\' tangor, when compared to the fruits without treatment, and it increased the yield of marketable fruits. The hydrothermal treatment accomplished up to 30 minutes at 50ºC can be used as peeling technique for \'Pera\' sweet orange, \'Valencia\' sweet orange and \'Murcott\' tangor.

The largest known marine diamond placer, the Namibian mega-placer, lies along the Atlantic coast of south-western Africa from the Orange River mouth 1,000 km northwards to the Namibian-Angolan border. The most economically viable portion of the Namibian mega-placer (>75 million carats recovered at >95% gem quality) comprises onshore and offshore marine deposits that are developed within ∼100km of the Orange River outfall. For much of the Cainozoic, this long-lived fluvial system has been the main conduit transporting diamonds from kimberlitic and secondary sources in the cratonic hinterland of southern Africa to the Atlantic shelf that has been neutrally buoyant over this period. Highly energetic marine processes, driven in part, by southerly winds with an attendant northward-directed longshore drift, have generated terminal placers that are preserved both onshore and offshore. This study, through detailed field sedimentological and diamond analyses, investigates the development and mineralisation of gravel barrier deposits within the ancestral Orange River mouth area during a major ∼30 m regional transgression ('30 m Package') in the Late Pliocene. At that time, diamond supply from this fluvial conduit was minimal, yet the corresponding onshore marine deposits to the north of the Orange River mouth were significantly diamond enriched, enabling large-scale alluvial diamond mining to take place for over 75 years. Of the entire coastline of south-western Africa, the most complete accumulation of the '30 m Package' is preserved within the palaeo-Orange River mouth as barrier spit and barrier beach deposits. Arranged vertically and laterally in a 16m thick succession, these are deposits of: (1) intertidal beach, (2) lagoon and washover, (3) tidal inlet and spit recurve and (4) storm-dominated subtidal settings. These were parts of larger barrier features, the bulk of which are preserved as highstand deposits that are diamond-bearing with varying, but generally low grades (<13 stones (diamonds) per hundred tons, spht). Intertidal beach and spit recurve deposits have higher economic grades (12-13 spht) due to the energetic sieving and mobile trapping mechanisms associated with their emplacement. In contrast, the less reworked and more sandy subtidal, tidal inlet and washover deposits have un-economic grades (<2 spht). Despite these low grades, the barrier deposits have the largest average stone (diamond) size (1-2 carats/stone, cts/stn) of the entire Namibian mega-placer, given their proximity to the ancestral Orange River outfall. This study demonstrates that barrier shoreline evolution at the fluvial/marine interface was controlled by: (1) a strong and coarse fluvial sediment supply that sustained shoreline growth on a highly energetic coast, (2) accommodation space facilitating sediment preservation and (3) short-duration, high-frequency sea-level cycles superimposed on the∼30 m regional transgression, promoting hierarchal stacking of progradational deposits. During these sea-level fluctuations, diamonds were 'farmed' from older, shelf sequences in the offshore and driven landward to accumulate in '30 m Package' highstand barrier deposits. In spite of the large supply of diamonds, their retention in these deposits was poor due to an incompetent footwall of ancestral Orange River mouth sediment and the inherent cobble-boulder size of the barrier gravels. Thus the principal process controlling diamond entrapment in these barrier deposits was kinetic sieving in a coarse-grained framework. Consequently, at the marine/fluvial interface and down-drift for ∼5 km, larger diamonds (1-2 cts/stn) were retained in low-grade (<2 spht), coarse-gravel barrier shorelines. Smaller diamonds (mostly < I cts/stn) were rejected into the northward-driven littoral sediments and further size-sorted along ∼95 km of Namibian coast to accumulate in finer, high-grade beach placers (> 100 spht) where bedrock footwall promoted such high concentrations. The gravel-dominated palaeo-Orange River mouth is considered to be the ' heart' of the Namibian mega-placer, controlling sediment and diamond supply to the littoral zone further north. Although coarse gravel is retained at the river mouth, the incompetence of this highly energetic setting to trap diamonds renders it sub-economic. This ineffectiveness at the fluvial/marine interface is thus fundamental in enriching the coastal tract farther down-drift and developing highly economic coastal placers along the Atlantic coast of south-western Africa. / KMBT_363 / Adobe Acrobat 9.54 Paper Capture Plug-in

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Tesis por compendio / [ES] La industria alimentaria ha mostrado un enorme interés por desarrollar nuevos productos a base de fruta con el fin de satisfacer la demanda saludable y sostenible de productos alimentarios de los consumidores. En este sentido, un puré de naranja liofilizado podría representar una opción viable. La liofilización del puré da lugar a una torta que puede consumirse directamente como snack, o puede triturarse para obtener un polvo que puede utilizarse para una amplia gama de aplicaciones. Una optimización adecuada de las condiciones de liofilización podría ayudar a reducir su duración sin afectar a las características del producto final. Sin embargo, los alimentos deshidratados pueden presentar problemas de colapso estructural relacionados con su baja temperatura de transición vítrea. En este sentido, una técnica frecuente para la estabilización de estos productos deshidratados es la incorporación de biopolímeros de alto peso molecular. El objetivo de esta Tesis ha sido el diseño del proceso de liofilización para la obtención de un snack de naranja. Para ello se ha estudiado la influencia de diferentes combinaciones de biopolímeros en la estabilidad física del puré de naranja liofilizado (snack de naranja) y en la bioaccesibilidad in vitro de sus compuestos bioactivos. Asimismo, se ha evaluado su efecto en las propiedades de flujo en aire y de rehidratación del polvo de naranja. Se ha trabajado con diferentes combinaciones de goma Arábiga, maltodextrina, almidón sustituido por grupos octenil succínico, almidón nativo de maíz, fibras de guisante y de bambú. Los resultados mostraron la necesidad de incorporar estos biopolímeros para aumentar la actividad de agua crítica y el contenido de agua crítico para la transición vítrea, el cual se ha relacionado con la pérdida de la textura del snack. Si bien ninguna de las mezclas de biopolímeros fue mejor que las otras en higroscopicidad, carácter anti-plastificante, color y propiedades mecánicas del snack, la mezcla GA con FB fue la que mejoró la bioaccesibilidad de la vitamina C (VC) y de los compuestos fenólicos totales (TP). Además, esta misma combinación fue la que ofreció uno de los tiempos de mojado más cortos y una menor viscosidad del producto rehidratado, deseado para un producto tipo zumo. Por otra parte, se ha estudiado el impacto de las condiciones de liofilización en el consumo de energía del proceso y en la calidad del snack formulado con GA y FB. Las variables del proceso consideradas han sido la velocidad de congelación (convencional y abatidor), la temperatura de bandeja (30, 40, 50 ºC) y presión de trabajo (5, 100 Pa) durante el secado. Menor presión y mayor temperatura promovieron un ligero mayor secado de las muestras, obteniendo un producto más crujiente, con un color amarillo menos intenso, mejor preservación de VC y ß-caroteno (BC), y una reducción significativa, de hasta un 75%, en el consumo de energía total durante el secado, debido a la reducción del tiempo del proceso. La velocidad de congelación no tuvo impacto significativo sobre ninguna de las propiedades evaluadas. Por tanto, las condiciones recomendadas para el secado por liofilización son 5 Pa de presión y 50 ºC como temperatura de bandeja. Por último, se evaluó la estabilidad física y de los compuestos bioactivos y actividad antioxidante del snack almacenado en bolsas zip, a 4 y 20 ºC, durante 6 meses, simulando condiciones domésticas de almacenamiento. Como resultado, la muestra ganó cierta humedad, con la consecuente pérdida en porosidad y carácter crujiente a partir de los 2 meses. Asimismo, la luminosidad del snack almacenado a 20 ºC disminuyó pasados 2 meses, probablemente debido a las reacciones de pardeamiento, que incluyen la degradación de la VC (20%). BC sufrió una gran disminución, desde el inicio del almacenamiento y más cuanto mayor fue la temperatura. Por lo tanto, para este producto se recomienda un almacenamiento en refrigeración para una mejor preservación de los compuestos bioactivos. / [CA] La indústria alimentària ha mostrat un enorme interés per desenvolupar nous productes a base de fruita amb la finalitat de satisfer la demanda saludable i sostenible de productes alimentaris dels consumidors. En aquest sentit, un puré de taronja liofilitzat podria representar una opció viable. La liofilització del puré dona lloc a una coca que pot consumir-se directament com a snack, o pot triturar-se per a obtindre una pols que pot utilitzar-se per a una àmplia gamma d'aplicacions. Una optimització adequada de les condicions de liofilització podria ajudar a reduir la seua duració sense afectar les característiques del producte final. No obstant això, els aliments deshidratats poden presentar problemes de col·lapse estructural relacionats amb la seua baixa temperatura de transició vítria. En aquest sentit, una tècnica freqüent per a l'estabilització d'aquests productes deshidratats és la incorporació de biopolímers d'alt pes molecular. L'objectiu d'aquesta Tesi ha sigut el disseny del procés de liofilització per a l'obtenció d'un snack de taronja. Per a això s'ha estudiat la influència de diferents combinacions de biopolímers en l'estabilitat física del puré de taronja liofilitzat (snack de taronja) i en la bioaccessibilitat in vitro dels seus compostos bioactius. Així mateix, s'ha avaluat el seu efecte en les propietats de flux en aire i de rehidratació de la pols de taronja. S'ha treballat amb diferents combinacions de goma Aràbiga, maltodextrina, midó substituït per grups octenil succínic, midó natiu de dacsa, fibres de pésol i de bambú. Els resultats van mostrar la necessitat d'incorporar aquests biopolímers per a augmentar l'activitat d'aigua crítica i el contingut d'aigua crític per a la transició vítria, el qual s'ha relacionat amb la pèrdua de la textura del snack. Si bé cap de les mescles de biopolímers va ser millor que les altres en higroscopicitat, caràcter anti-plastificant, color i propietats mecàniques del snack, la mescla GA amb FB va ser la que va millorar la bioaccessibilitat de la vitamina C (VC) i dels compostos fenòlics totals (TP). A més, aquesta mateixa combinació va ser la que va oferir un dels temps de mullat més curts i una menor viscositat del producte rehidratat, desitjat per a un producte tipus suc. D'altra banda, s'ha estudiat l'impacte de les condicions de liofilització en el consum d'energia del procés i en la qualitat del snack formulat amb GA i FB. Les variables del procés considerades han sigut la velocitat de congelació (convencional i abatedor), la temperatura de safata (30, 40, 50 °C) i pressió de treball (5, 100 Pa) durant l'assecat. Menor pressió i major temperatura van promoure un lleuger major assecat de les mostres, obtenint un producte més cruixent, amb un color groc menys intens, millor preservació de VC i ß-caroté (BC), i una reducció significativa, de fins a un 75%, en el consum d'energia total durant l'assecat, a causa de la reducció del temps del procés. La velocitat de congelació no va tindre impacte significatiu sobre cap de les propietats avaluades. Per tant, les condicions recomanades per a l'assecat per liofilització són 5 Pa de pressió i 50 °C com a temperatura de safata. Finalment, es va avaluar l'estabilitat física i dels compostos bioactius i activitat antioxidant del snack emmagatzemat en bosses zip, a 4 i 20 °C, durant 6 mesos, simulant condicions domèstiques d'emmagatzematge. Com a resultat, la mostra va guanyar una certa humitat, amb la conseqüent pèrdua en porositat i caràcter cruixent a partir dels 2 mesos. Així mateix, la lluminositat del snack emmagatzemat a 20 °C va disminuir passats 2 mesos, probablement a causa de les reaccions de enfosquiment, que inclouen la degradació de la VC (20%). BC va patir una gran disminució, des de l'inici de l'emmagatzematge i més com més gran va ser la temperatura. Per tant, per a aquest producte es recomana un emmagatzematge en refrigeració per a una millor preservació dels compostos bioactius. / [EN] Food industries have showed a huge interest in developing new fruit-based products to satisfy the healthy and sustainable demand of food products by consumers. In this sense, offering a freeze-dried orange puree could represent a feasible option. Freeze-drying the puree results in a cake that can be consumed directly as a snack, or it can be crushed to obtain a powder that can be used for a wide range of applications. A suitable optimisation of the freeze-drying conditions could help to reduce its duration without affecting the characteristics of the final product. However, dehydrated foods may present problems of structural collapse related to its low glass transition temperature. In this sense, an approach for the stabilisation of dehydrated products is the incorporation of high molecular weight biopolymers. The aim of this Thesis has been the design of the freeze-drying process to obtain an orange snack. The influence of different combinations of biopolymers on the physical stability of the freeze-dried orange puree (orange snack) and on the in vitro bioaccessibility of its bioactive compounds has been studied. Their effect on the air flow and rehydration properties of an orange powder obtained after crushing the snack has also been evaluated. Different combinations of gum Arabic, maltodextrin, starch substituted with octenyl succinic groups, native corn starch, pea and bamboo fibres were used. The results showed the need to incorporate these biopolymers to increase the critical water activity and the critical water content for the glass transition, which has been related to the loss of snack texture. Although none of the biopolymer combinations was better than the others in terms of hygroscopicity, anti-plasticising character, colour, and mechanical properties of the snack, the GA mixed with FB was the one that improved the bioaccessibility of vitamin C (VC) and total phenolic compounds (TP). This same combination offered the shortest wetting times and a lower viscosity of the rehydrated product, which is desirable for a juice-type product. Also, the impact of the freeze-drying conditions on the energy consumption of the process and on the quality of the snack formulated with GA and FB has been studied. The process variables considered were freezing rate (conventional and blast freezer), shelf temperature (30, 40, 50 ºC) and working pressure (5, 100 Pa) during drying. Lower pressure and higher temperature promoted a slightly higher drying of the samples, which resulted in a crispier product, as well as a less intense yellow colour. However, at the sensory level, there was no significant preference for any of the samples processed under the different conditions studied. In addition, VC and ß-carotene (BC) were better preserved under these conditions, conditions which significantly reduced, up to 75%, the total energy consumption during drying, due to the reduction of the process time. The freezing rate had no significant impact on any of the properties evaluated. Therefore, the recommended conditions for freeze-drying to maximise the preservation of bioactive compounds, with a lower energy consumption, while providing a snack perceived as a crispy product by consumers, are 5 Pa pressure and 50 ºC as shelf temperature. Finally, the physical stability and the stability of bioactive compounds and antioxidant activity of the snack stored in zip bags at 4 and 20 ºC for 6 months, simulating domestic storage conditions, was evaluated. As a result, a certain moisture gain of the sample was observed, with a consequent loss in porosity and crispness after 2 months. Also, the luminosity of the snack stored at 20°C decreased after 2 months, probably due to browning reactions, including degradation of VC (20%). BC suffered a large decrease, from the beginning of storage and more so the higher the temperature. Therefore, refrigerated storage is recommended for better preservation of the bioactive compounds of this product. / Silva Espinoza, MA. (2021). Design of the process of obtaining a freeze-dried orange puree. Formulation, freeze-drying variables, and storage conditions [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/170354 / TESIS / Compendio

Les cyanobactéries sont des organismes procaryotes photosynthétiques. Si l’énergie leur est essentielle, elle peut également être délétère. Afin de se protéger, elles ont acquis plusieurs mécanismes de photoprotection. La thématique de ma thèse est l’étude de l’un d’entre eux par des approches combinées de biologie moléculaire, biochimie et biophysique.Les antennes collectrices de lumière des cyanobactéries sont des complexes extra-membranaires solubles appelés les phycobilisomes. Ils permettent de canaliser l'énergie vers les centres réactionnels des photosystèmes. Sous forte lumière, l'afflux d’énergie y parvenant crée notamment des espèces réactives de l’oxygène, ce qui est délétère pour la cellule. L’Orange Carotenoid Protein (OCP) est impliquée dans un mécanisme de photoprotection qui diminue l'énergie arrivant au niveau des centres réactionnels en augmentant la part d’énergie dissipée sous forme de chaleur. L’OCP est une caroténo-protéine composée de deux domaines globulaires N- et C-terminal qui lie un caroténoïde. Cette protéine photoactivable est à la fois le senseur, et l’acteur du mécanisme de photoprotection. Le mécanisme est désactivé par une seconde protéine, la FRP (Fluorescence Recovery Protein).Le premier chapitre de ce travail de thèse rapporte l’étude de la spécificité des OCPs isolées chez deux souches différentes pour différentes classes de phycobilisomes dont l’architecture du cœur diffère. Le second chapitre présente la méthode mise au point au laboratoire de production de l’OCP chez E.coli, ainsi que la caractérisation d’OCPs clonées depuis le génome de Synechocystis, A. variabilis et A. platensis et surexprimés chez E. coli. Le troisième présente la structure tridimensionnelle du domaine N-terminal, qui est le domaine effecteur de l’OCP. Dans ce chapitre, nous démontrons que le cofacteur caroténoïde se déplace de 12 angstrom au sein de l’OCP lors de la photoactivation. Le quatrième rapporte que le bras N-terminal de l’OCP est une structure singulière qui maintient la protéine fermée à l’obscurité, évitant que l’OCP ne s’active sous faible lumière, ou à l’obscurité. Le cinquième présente la résolution de la structure et l’identification du site actif de la FRP qui nous ont permis de prédire in silico le site d’attachement putatif de la FRP sur le domaine C-terminal de l’OCP. Dans le chapitre 6, je rapporte que deux résidus, l’aspartate 220 et la phénylalanine 299, sont requis pour que l’activité de la FRP soit maximale, confirmant le site d’interaction prédit. / Cyanobacteria, a photosynthetic prokaryote organism, harvest light for living. But harvesting too much light can be harmful. To protect themselves against this stress, cyanobacteria have developed several photoprotective mechanisms. This manuscript reports my work about one of them by combined technics of molecular biology, biochemistry and biophysics.Cyanobacterial light harvesting antennae are extra-membranous complexes called phycobilisomes. They funnel harvested energy into the photosynthetic reaction centers. Under high light, high energy input induces the formation of reactive oxygen species (ROS), which are harmful in excess. One of the existent photoprotective mechanisms helps to avoid ROS formation by decreasing the energy arriving at the reaction centers. The main actor of this mechanism is the photoactive Orange Carotenoid Protein (OCP) that binds to the phycobilisome, and induces an increase of the part of energy dissipated as heat. The OCP is a protein composed by two globular domains (called N- and C- terminal) and binds a carotenoid cofactor. High intensity of blue-green light triggers conformational changes in the inactive orange OCP, which turns red and is now able to binds the PBs. Under low light conditions, this mechanism is turned off by another protein, the Fluorescence Recovery Protein (FRP).The first chapter of this manuscript reports the study of the specificity of OCPs isolated from two strains for different classes of phycobilisomes with different core architecture. The second describe the development of a method to produce holoOCP in E. coli cells. Furthermore, it reports the characterization of the Synechocystis, A. variabilis and A. platensis OCPs isolated from E. coli. The third chapter presents the tridimensional structure of the active N-terminal domain of the OCP. In this chapter, we demonstrate that the carotenoid undergoes a 12anstrom movement upon photoactivation. The fourth chapter rapports that the N-terminal arm of the OCP helps to maintain closed the inactive orange OCP in darkness or low light, avoiding OCP activation and consequent unwanted PBs fluorescence quenching. The fifth presents the resolution of the structure and the identification of the active site of the FRP. These data allow to compute a predictionnal model of interaction between OCP and FRP. I assessed the validity of the model by isolating several modified OCPs. Results shown in chapter 6 report that the aspartate 220 end the phenylalanine 299 are required for effective FRP action.

La lumière est essentielle pour les organismes photosynthétiques qui convertissent l'énergie solaire en énergie chimique. Cependant, la lumière devient dangereuse lorsque l'énergie qui arrive aux centres réactionnels de l'appareil photosynthétique, est en excès par rapport à l’énergie consommée. Dans ce cas, la chaîne de transport d'électrons photosynthétiques se réduit et les espèces réactives de l'oxygène (ROS) sont accumulées, notamment au niveau des deux photosystèmes, PSI et PSII. Les cyanobactéries ont développé des mécanismes photoprotecteurs qui diminuent l'énergie transférée au PSII atténuant ainsi l'accumulation de ROS et les dommages cellulaires, comme l’extinction non-photochimique (NPQcya) induite par la lumière bleue-verte. La soluble Orange Caroténoïde Protéine (OCPo) est essentielle pour ce mécanisme de photoprotection. L'OCP agit comme un senseur de l’intensité lumineuse et un inducteur de la dissipation d'énergie des phycobilisomes (PBS), l'antenne extra-membranaire des cyanobactéries. L'OCP est la première protéine photo-active à caroténoïde connue comme senseur. Une forte lumière bleue-verte déclenche des changements structurels dans l'OCPo qui induisent une forme active, rouge (OCPr). Le domaine N-terminal de l’OCPr, en s’intercalant entre les trimères externes d’un des cylindres basaux du cœur du PBS, augmente la dissipation thermique de l'énergie au niveau de l'antenne. L'OCP possède aussi une autre fonction : l’extinction de l’oxygène singulet, qui protège les cellules du stress oxydatif. Pour récupérer pleinement la capacité de l’antenne en faible lumière, une deuxième protéine est nécessaire, la "Fluorescence Recovery Protein" (FRP), dont le rôle est de détacher l’OCPr des PBS et d’accélérer sa reconversion en OCPo inactive. Ce manuscrit est un état des lieux des connaissances et des dernières avancées sur le mécanisme de NPQ associé à l'OCP dans les cyanobactéries. / Photosynthetic organisms use light energy from the sun in order to perform photosynthesis and to convert solar energy into chemical energy. Absorbance of excess light energy beyond what can be consumed in photosynthesis is dangerous for these organisms. Reactive oxygen species (ROS) are formed at the reaction centers and collecting light antennas inducing photooxidative damage which can lead to cell death. In cyanobacteria, one of these photoprotective mechanisms consists to reduce the amount of energy arriving to the reaction centers by thermal dissipation of the excess absorbed energy. Energy dissipation is accompanied by a decrease of Photosystem II-related fluorescence emission called non-photochemical quenching (NPQ). The soluble Orange Carotenoid Protein (OCPo) is essential for this photoprotective mechanism. The OCP is the first photo-active protein with a carotenoid known as light intensity sensor and acts as energy quencher of the phycobilisome (PB), the extra-membrane antenna of cyanobacteria. Structural changes occur when the OCPo absorbs a strong blue-green light leading to a red active form (OCPr). The N-terminal domain of OCPr burrows into the two external trimers of the core basal APC cylinders of the PB and increases thermal energy dissipation at the level of antenna. The OCP has an additional function in photoprotection as oxygen singlet quencher protecting cells from oxidative stress. Under low light conditions, to recover the full antenna capacity, a second protein is needed, the "Fluorescence Recovery Protein" (FRP), whose role is to detach the OCPr from the PB and accelerate its conversion into an inactive OCPo. In this manuscript, I will review the knowledge about the OCP, since the discovery of the mechanism and its characterization to the latest advances on the OCP-related-NPQ mechanism in cyanobacteria.

Orientador: Margarida Maria Barros / Resumo: Foi avaliada a capacidade antioxidante do frit de laranja (OF) no desempenho produtivo, perfil hematológico e atividade das enzimas antioxidante na tilápia-do-Nilo submetidas ao estresse térmico. Um grupo de 440 tilápias-do-Nilo machos (31.7g ± 0.34) foram distribuidos em 40 aquários 250 – L (11 peixes/caixa) e arraçoados com cinco dietas práticas com diferentes níveis de OF 0, 0.2, 0.4, 0.6, 0.8% OF por 70 dias. As dietas foram formuladas para conter 26% de proteína digestível e 14.24 MJ de energia digestível kg-1. Após o período de alimentação foi determinado o desempenho produtivo, perfil hematológico, e atividade das enzimas antioxidante. Após, os peixes foram submetidos ao estresse térmico (32°C) durante três e o mesmo perfil hematológico e atividade das enzimas antioxidante foram determinados. Não houve diferença estatística no desempenho produtivo entre os tratamentos. Peixes alimentados com as dietas 0 Of demonstraram menores valores no perfil hematológico após o estresse térmico (P < 0.05). Os peixes alimentados com dietas que continham 0.6 OF apresentaram menor taxa de hematócrito e os alimentados com 0.8 OF menores taxas de hemoglobin e hematócrito após o desafio de estresse térmico. Peixes alimentados com dietas que continham 0.4 OF demonstraram valores mais baixos de volume corpuscular médio (VCM) e maiores valores de concentração de hemoglobina corpuscular media (CHCM) quando comparados antes e após estresse térmico. As dietas com inclusão de frit de laranja det... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The antioxidant capacity of dietary orange frit (OF) on growth, hematological profile and antioxidant enzymes activity of Nile tilapia subjected to heat-induced stress (HIS) was analyzed. A group of 440 male Nile tilapia (31.7g ± 0.34) was randomly distributed in 40 250-L aquaria (11 fish/tank) and fed five practical diets with graded levels of OF at 0, 0.2, 0.4, 0.6 and 0.8% OF orange frit kg-1 diet for 70 days. The diets were formulated to contain 26% digestible protein and 14.24 MJ digestible energy kg-1. After the feeding period, growth performance, hematological profile and antioxidant enzymes activity were determined. Then, fish were subjected to HIS (32°C) for three days and the same hematological profile and antioxidant enzymes activity were determined. There was no statistical difference for growth performance among treatments. Fish fed 0 OF diet showed lower hematological profile after HIS (P < 0.05). Fish fed 0.6OF presented lower hematocrit and fed 0.8 OF lower hemoglobin and hematocrit after HIS. Fish fed 0.4 OF showed the lowest mean corpuscular volume (MCV) and the highest mean corpuscular hemoglobin concentration (MCHC) comparing the vales before and after HIS. Dietary orange frit determined different activities (P < 0.05) for catalase before HIS. Fish fed 0 OF diet showed the highest activity and 0.2 OF the lowest. After HIS fish fed 0 OF and 0.2 OF showed the lowest activity and 0.6 OF the highest (P < 0.05). A comparison of the values before and after HIS s... (Complete abstract click electronic access below) / Mestre

This essay aims at analysing and understanding different choices made by Caj Lundgren when he translated Anthony Burgess’s novel A Clockwork Orange (1962), more specifically when he translated the fictive language called Nadsat, which is one of the features that made this novel famous. The method used to analyse Lundgren’s translation is based on Reiß and Veermer’s (1984/2014) concept of skopos and functional equivalence. Conclusions drawn are that Lundgren uses a compensatory translation strategy, and that a model can be created for identification of Russian words in the original text’s Nadsat and the transfer of these words to the target text as Swedish Nadsat. In a comparison between Lundgren and two Russian translators’ strategies, their strategies are shown to differ, which could be caused by different translation problems dependent on the target languages. / Denna uppsats mål är att analysera olika val som Caj Lundgren gjorde i sin översättning av Anthony Burgess roman A Clockwork Orange (1962), specifikt avseende originalets fiktiva språk, som Burgess döpte till nadsat. Metoden som använts för analysen baseras på Reiß och Veermers (1984/2014) term skopos och funktionell ekvivalens. Slutsatser som dras är dels att Lundgren använder sig av en kompensatorisk strategi vid översättning, dels att ett schema kan uppställas för identifiering av ryska ord i originalets nadsat och deras överföring till måltexten som svensk nadsat. I en jämförelse mellan Lundgrens och två ryska översättares strategier visas att deras strategier skiljer sig åt, vilket kan bero på översättningsproblematik avhängig målspråken.