Orientador: Fernanda Paula Collares / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-06T15:44:35Z (GMT). No. of bitstreams: 1 UclesSantos_JoseRicardo_M.pdf: 2719947 bytes, checksum: c27f8219ba712b28246c216abd73b478 (MD5) Previous issue date: 2006 / Resumo: O pão de queijo, um produto tipicamente brasileiro, é considerado um produto de panificação que não contém glúten, pois em sua formulação não é utilizada farinha de trigo e sim fécula nativa de mandioca ou fécula de mandioca fermentada naturalmente e/ou modificada. Devido ao seu alto teor de carboidratos, possui grande aceitabilidade e pode ser utilizado por pessoas portadoras de doença celíaca. O alto consumo deste produto no Brasil e a procura crescente dos consumidores por produtos alimentícios não somente nutritivos e saborosos, mas que também tragam benefício à saúde, estimula o estudo da incorporação de ingredientes nutricionais e funcionais à massa de pão de queijo que não afetem suas propriedades reológicas, físicas e sensoriais. O objetivo deste trabalho, portanto, foi avaliar o efeito da incorporação de componentes nutricionais e funcionais, como o isolado protéico de soja e a polidextrose, na qualidade da massa e do pão de queijo assado, com e sem adição de fécula de mandioca pré-gelatinizada, visando determinar quais dos teores adicionados de cada um destes ingredientes conferem as melhores características físicas e sensoriais ao pão de queijo. Para atingir este objetivo, o trabalho foi dividido em três etapas: (i) avaliação do teor adequado de fécula de mandioca e polvilho azedo adicionado à formulação de pão de queijo, através da determinação de suas propriedades físicas, reológicas e sensoriais; (ii) análise do efeito combinado dos ingredientes nutricionais e funcionais incorporados à massa de pão de queijo com diferentes teores de fécula de mandioca e polvilho azedo, com e sem a etapa de escaldamento, nas suas propriedades reológicas, físicas e sensoriais, utilizando a metodologia de superfície de resposta; (iii) caracterização física, química, reológica e sensorial das formulações de pão de queijo, com e sem a adição de fécula de mandioca pré-gelatinizada, escolhidas no item anterior. Na primeira etapa do trabalho, massas elaboradas com teores de 70% e 100% de fécula de mandioca apresentaram maiores consistências em U.F. em relação às massas elaboradas com altos teores de polvilho azedo, durante o processo de mistura similado no farinógrafo Brabender. Os pães de queijo elaborados com altos teores de polvilho azedo apresentam coeficientes de expansão de até 80,00%, sendo que os pães elaborados com fécula de mandioca apresentaram valor máximo de expansão de 66,67%. Devido a este poder de expansão apresentado pelos pães de queijo elaborados com polvilho azedo, a força de compressão (textura dos pães) obtida foi de 7,85% a 28,40% menor à reportada para pães de queijo elaborados com fécula de mandioca. Sensorialmente, os pães de queijo elaborados com altos teores de fécula de mandioca apresentaram as maiores médias em relação aos atributos de aceitação geral, textura, cor e sabor em comparação com os pães de queijo com altos teores de polvilho azedo, que segundo os provadores lembra o biscoito de polvilho. Na segunda etapa do trabalho, foi possível a determinação de um modelo matemático para o volume específico dos pães de queijo elaborados com adição de fécula de mandioca pré-gelatinizada, e para o ângulo de tonalidade do miolo (h) de pães de queijo elaborados sem adição de fécula de mandioca prégelatinizada, explicando a interação que existe entre as variáveis (fécula de mandioca, polvilho azedo, isolado protéico de soja e polidextrose). O modelo matemático do volume mostra que a utilização de teores entre 79,76% e 100% de polvilho azedo, 5,50% de isolado protéico de soja e teores abaixo de 1,03% de polidextrose confere grandes volumes ao pão de queijo, assim como teores de 0,00% de polvilho azedo, 5,50% de isolado protéico de soja e teores de polidextrose acima de 5,07%. Para o ângulo de tonalidade (h), o modelo matemático determinou que a utilização dos valores extremos, teores mínimo e máximo, de polvilho azedo (0,00% e 100,00%) com teores de 0,00% a 9,00% de isolado protéico de soja e teores médios de polidextrose (2,21%) na formulação de pão de queijo, conferiu altos ângulos de tonalidade do miolo. Os parâmetros de textura, luminosidade e saturação da cor da crosta e do miolo dos pães de queijo, com e sem a adição de fécula de mandioca prégelatinizada, não apresentaram diferença significativa (p=0,05), sendo que a elaboração de pães de queijo utilizando concentrações na faixa estudada não produz alterações na qualidade do produto. Na última etapa do trabalho, duas formulações de pão de queijo, selecionadas na 2a etapa considerando os escores sensoriais de aparência geral, textura, aroma e sabor, foram caracterizadas física, química, reológica e sensorialmente, sendo que as duas formulações apresentaram diferença significativa (p=0,05) no perfil reológico da massa, no volume, textura e cor dos pães de queijo assados, mas sensorialmente foram igualmente aceitas pelos consumidores, quanto à aparência geral, cor do miolo, textura, aroma e sabor / Abstract: ¿Pao de queijo¿ (cheese bread), a typically Brazilian product, is considered a gluten-free bakery product, as its formulation does not contain wheat flour but native cassava starch, naturally fermented and/or modified. Due to its high content of carbohydrates, it has great acceptance and can be consumed by individuals who suffer from celiac disease. The high consumption of this product in Brazil and the growing demand, by consumers, of food products that are not only nutritious and tasty, but also bring health benefits, stimulates the study of the incorporation of nutritional and functional ingredients to cheese bread doughs, which do not affect its rheological, physical and sensorial properties. The objective of this study, therefore, was to evaluate the effect of the incorporation of the nutritional and functional components, such as soy protein isolate and the polydextrose, on the quality of cheese bread dough and of baked cheese bread, with or without the addition of pre-gelatinized cassava starch, with the objective of determining the contents of each of these ingredients that confer the best physical and sensorial characteristics to cheese bread. In order to achieve this objective, the study was divided in three phases: (i) evaluation of the appropriate content of cassava starch and sour cassava starch added to the formulation, by determining physical, rheological and sensorial properties; (ii) analysis of the combined effect of functional and nutritional ingredients incorporated to the cheese bread dough with different contents of cassava starch and sour cassava starch, with and without the scalding process, on rheological, physical and sensorial properties, using the response surface methodology; (iii) physical, chemical and rheological characterization of the cheese bread formulations, with or without the addition of pre-gelatinized cassava starch, chosen in the previous stage/step. In the first phase of the study, doughs elaborated with 70% and 100% of cassava starch presented greater consistency profiles when compared to high contents of sour cassava starch, during the mixing process simulated in the Brabender farinograph. The cheese breads elaborated with high contents of sour cassava starch presented expansion coefficients of up to 80.00%, while the cheese breads elaborated with cassava starch presented a maximum value of expansion of 66.67%. Due to this power of expansion presented by the cheese breads elaborated with sour cassava starch, the force of compression (texture of breads) obtained was 7.85% to 28.40% lower than that of those reported for cheese breads elaborated with cassava starch. Sensorially, the cheese breads produced with high contents of cassava starch presented the highest scores with respect to the attributes general acceptance, texture, color and flavor, in comparison to the cheese breads with high contents of sour cassava starch, which according to the panelists resembled the sour cassava starch biscuits (¿biscoito de polvilho¿). In the second phase of the study, it was possible to determine a mathematical model for the specific volume of cheese breads elaborated with the addition of pre-gelatinized cassava starch, and for the shade angle (h) of cheese breads elaborated without the addition of pre-gelatinized cassava starch, explaining the interaction between the variables (cassava starch, sour cassava starch, soy protein isolate and polydextrose). The mathematical model showed that the use of contents between 79.76% and 100% of sour cassava starch, 5.50 % of soy protein isolate and contents below 1.03% of polydextrose confers great volumes to cheese bread, as well as contents of 0.00% of sour cassava starch, 5.50% of soy protein isolate and contents of polydextrose above 5.07%. For the shade angle (h), the mathematical model determined that the use of extreme values, minimum and maximum contents, of sour cassava starch (0.00% and 100.0%) with contents of 0.00% to 9.00% of soy protein isolate and intermediate contents of polydextrose (2.21%) in the formulation of cheese bread conferred high shade angles to the crumb. The parameters of texture, luminescence (L*) and color saturation (C*) of the crust and crumb of the cheese breads, with and without the addition of pregelatinized cassava starch, did not present significant difference (p=0.05), indicating that the elaboration of cheese breads using the concentrations in the range studied for the different variables does not produce alterations in the quality of the product. In the last phase of the study, two formulations of cheese bread, selected in the second phase considering the sensorial scores of general appearance, aroma and flavor, were characterized physically, sensorially, rheologically and chemically. These two formulations presented significant differences (p=0.05) in the rheological profile of the dough, specific volume, texture and color of the baked cheese breads, but were equally accepted by consumers, as to general appearance, color of the cr8umb, texture, aroma and flavor / Mestrado / Mestre em Tecnologia de Alimentos

Orientador: Celio Kenji Miyasaka / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-15T19:03:10Z (GMT). No. of bitstreams: 1 Abecia-Soria_MariaInes_D.pdf: 1397720 bytes, checksum: efabd9293a2a8d45841c5d131da10056 (MD5) Previous issue date: 2010 / Resumo: As proteínas do soro de leite são consideradas de alto valor nutritivo. Elas têm escore químico superior às de outras proteínas de origem animal, possuem elevadas concentrações de aminoácidos de cadeia ramificada (BCAA), com excelente balanço e biodisponibilidade de aminoácidos essenciais. As proteínas de soro de leite vêm sendo largamente utilizadas pela indústria de alimentos como suplemento na alimentação de esportistas, devido às suas características fisiológicas e funcionais, destacando-se o uso dos hidrolisados, que tem sido recomendado para situações de estresse metabólico como o exercício físico em que a reposição de proteínas no organismo se torna necessária. O presente trabalho teve como objetivo avaliar alterações metabólicas induzidas pelo consumo de proteínas do soro de leite [isolado (ISL) e seu hidrolisado (HSL) com grau de hidrólise 10%] em comparação com a dieta padrão [caseína (CAS)], utilizadas como única fonte protéica em ratos submetidos ao exercício físico. Foram utilizados 96 ratos machos Wistar (~100g) divididos em grupos segundo o tipo de dieta (CAS, ISL e HSL) e a intensidade do exercício físico [treinados (T), treinados exaustos (TEX), sedentários (S) e sedentários exaustos (SEX)] durante 35 dias. Os seguintes parâmetros foram analisados: tempo de exaustão, concentração de lactato no sangue, glicogênio hepático (GH) e muscular (GM), atividade das enzimas marcadoras de lesão tecidual, incluindo lactato desidrogenase (LDH), creatina quinase (CK), alanina e aspartato aminotranferase (ALT e AST, respectivamente). Os animais alimentados com HSL nos grupos TEX e SEX conseguiram correr por muito mais tempo até atingir a exaustão em relação aos que receberam as dietas com CAS e ISL (p<0.05). O tempo de exaustão em relação à dieta CAS foi 72 min maior no grupo TEX e 44 min maior no grupo SEX. A diferença no tempo de exaustão entre os animais alimentados com as dietas de HSL e ISL foi de 40 min. no grupo TEX e de 13 min no grupo SEX. A concentração de lactato dos animais alimentados com ISL e HSL foi significativamente menor em relação aos alimentados com CAS nos grupos T e S (p<0,05). A concentração de glicogênio hepático dos animais que receberam as dietas com ISL e HSL foi estatisticamente superior em comparação com os que consumiram CAS nos grupos T(46%) e S (61%). A concentração de glicogênio muscular não apresentou diferenças significativas entre as três dietas nos diferentes grupos de treinamento (TEX, SEX, T e S). A atividade da ALT dos animais que consumiram HSL nos grupos SEX e S foi estatisticamente menor que os que receberam CAS. A atividade da AST dos animais alimentados com HSL foi significativamente menor que a dos que consumiram a dieta com CAS nos diferentes grupos de treinamento (TEX, SEX, T e S). A atividade da CK e LDH dos ratos que receberam a dieta com HSL foi estatisticamente menor que dos alimentados com CAS em todos os grupos de treinamento exceto no grupo S. Dos resultados obtidos neste trabalho pode-se concluir que o uso das proteínas do soro de leite [principalmente o hidrolisado (10% grau de hidrólise)] em relação à proteína padrão (CAS) como única fonte protéica em ratos submetidos a exercício físico, promoveu a) menor ganho de peso em todos os grupos, b) maior resistência à exaustão tanto no grupo TEX como no SEX, c) maior concentração de glicogênio hepático nos grupos T e S e d) maior proteção contra possíveis lesões hepáticas e musculares nos diferentes grupos estudados (TEX, SEX, T e S) / Abstract: The milk whey proteins are considered to be of high nutritive value and superior chemical score in comparison to other proteins of animal origin. They have high concentrations of branched-chain amino acids (BCAA) with excellent balance and bioavailability of essential amino acids. The milk whey proteins are widely used in the food industry as supplements for sportsmen due to their physiological and functional properties, and their hydrolysates are thought to be more efficient in the recovery of debilitated organisms under pronounced catabolic state because of their great stimulation to protein synthesis. In view of the energy value of proteins in exhaustive exercise, several studies on exercise and protein metabolism have been carried out in the attempt to elucidate the proper amount of protein that must be consumed by sportsmen. The present work had the objective of evaluating the metabolic changes induced by consumption of milk whey proteins [isolate (WPI) and its hydrolysate (WPH) with hydrolysis degree 10%] used as the only protein source in comparison to the standard diet [casein (CAS)] in rats submitted to physical exercise. 96 male Wistar rats (~100g) were divided in groups according to the type of diet (CAS, ISL and HSL) and the intensity of physical exercise [trained (T), trained taken to exhaustion (TEX), sedentary (S) and sedentary exhausted (SEX)] during 35 days. The following parameters were analyzed: time of exhaustion (min.), blood lactate concentration (mmol/L), liver glycogen (LG) and muscular glycogen (MG) (g/100g of tissue), activity of tissue injury marker enzymes (U/L) including lactate desidrogenase (LDH), creatine kinase (CK) and alanine and aspartate aminotranferase (ALT and AST respectively). The results showed that the animals fed with HSL in the groups TEX and SEX ran for much longer times until exhaustion in relation to the ones that received the diets with CAS and ISL (p<0.05). The exhaustion time in relation to the CAS diet was 72 min longer for the TEX group and 44 min longer for the SEX group. The difference in exhaustion time between the animals fed with the WPH and WPI diets was of 40 min for the group TEX and 13 min for the SEX group. The lactate concentration in the animals fed with WPI and WPH was statistically lower than in the ones fed with CAS in the groups T and S (p99<0,05). The concentration of liver glycogen in the animals that received the diets with WPI and WPH was statistically higher than in the animals that consumed CAS in the groups T (46%) and S (61%). The concentration of muscular glycogen did not have significant differences between the three types of diet in the different groups of training (TEX, SEX, T and S). The activity of the ALT of the animals that consumed WPH in the groups SEX and S was statistically lower than that of the animals that received CAS. The activity of the AST of the animals fed with WPH was significantly lower than that of the animals that consumed the diet with CAS in all of the groups of training (TEX, SEX, T and S). The activity of CK and LDH in the rats fed with HSL diet was statistical lower than in the ones fed with CAS in all of the groups of training except in the group S. From the results obtained in this work it can be concluded that the use of milk whey proteins [mainly the whey protein hydrolysate (10% hydrolysis degree)] in relation to the standard protein (CAS) as the only protein source in rats submitted to physical exercise promoted a) lesser weight gain in all of the groups, b) higher resistance to exhaustion in groups TEX and SEX, c) higher hepatic glycogen concentration in groups T and S and d) higher protection against possible hepatic and muscular injuries in all of the studied groups (TEX, SEX, T and S) / Doutorado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Doutor em Alimentos e Nutrição

Os isolados protéicos de soja são utilizados como ingredientes em diversos alimentos e sua utilização vêm aumentando juntamente com o aumento das pesquisas sobre os metabólitos secundários da soja, as isoflavonas. Alguns efeitos benéficos vem sendo associados às isoflavonas, entre estes a sua ação antioxidante, a redução ao risco de câncer, doenças cardiovasculares e osteoporose. O objetivo deste estudo foi o de otimizar as condições de extração das isoflavonas e de suas formas conjugadas a partir da farinha desengordurada de soja, visando o preparo de isolado protéico de soja. Os resultados mostraram que a obtenção de isolados protéicos de soja com teor aumentado de isoflavonas depende da utilização de condições brandas de centrifugação para a separação do precipitado isoelétrico, assim como da utilização de água acidificada na sua lavagem. A presença de isoflavonas no isolado resulta de três fatores, o primeiro referindo-se à associação entre isoflavonas e proteínas através de interações hidrofóbicas, eletrostáticas, e pontes de hidrogênio; o segundo à menor solubilidade das isoflavonas presentes na farinha desengordurada de soja no pH isoelétrico; e o último ao processo de carreamento (físico) das isoflavonas pelas proteínas insolubilizadas. / Soy protein isolates are used as ingredients in several food products and their use is increasing together with the increase of the researches on the secondary metabolites of soy, the isoflavones. Some beneficial effects have been associated to the isoflavones, among these their antioxidant action, prevention of cancer, cardiovascular diseases and osteoporosis. The objective of this study was to optimize the extraction conditions of the isoflavones from the defatted soy flour, seeking the preparation of soy protein isolates. The results showed that the obtention of soy protein isolates with increased content of isoflavones depends on the use of mild conditions of centrifugation for the separation of the isoelectric precipitate, as well as on the use of water acidified in the washing step. The presence of isoflavones in the isolates resulted from three factors, the first refers to the association between isoflavones and proteins through hydrophobic; and electrostatic interactions, and hydrogen bonding; the second to the decreased solubility of the isoflavones extracted from the defatted soy flour in the isoelectric pH; and the last to the carrying process (physical) of isoflavones by the precipitating proteins.

La tristeza es la enfermedad viral más grave del cultivo de los cítricos y su agente causal es el virus de la tristeza de los cí­tricos (CTV). CTV induce uno o más de los siguientes sí­ndromes: I) decaimiento y muerte de los naranjos dulces (ND), pomelos y mandarinos injertados sobre el patrón naranjo amargo (NA), sí­ndrome conocido como "tristeza", II) enanismo, acanaladuras en la madera y fruta de pequeño calibre (stem pitting, SP), y III) enanismo y amarillamiento de plantas de semilla de limonero, pomelo y NA (seedling yellows, SY). La gama de huéspedes de CTV es muy restringida y hasta hace poco no se conocí­a ningún huésped herbáceo experimental. Actualmente, se sabe que la agroinfiltración de Nicotiana benthamiana, con clones de DNA complemantario (cDNA) del aislado T36 de CTV produce la infección sistémica de la planta, acompañada de sí­ntomas similares a los inducidos en cí­tricos, si bien la infección no queda limitada al floema. El aislado T36 induce SY y SP de lima Mejicana (LM), pero no en otros huéspedes como pomelo o ND. El estudio de los determinantes genéticos responsables de la inducción del sí­ndrome de SP requerí­a desarrollar un sistema genético basado en clones agroinfecciosos de un aislado inductor de estos sí­ntomas, como el aislado español T318A. Para ello, se partió de clones de cDNA de longitud completa de T318A previamente desarrollados en el laboratorio, capaces de replicarse en hojas de N. benthamiana pero incapaces de inducir infección sistémica y que presentaban varias mutaciones en su proteína de cápsida minoritaria p27. La corrección de dichas mutaciones y la construcción de nuevos clones de longitud completa de T318A marcados con el gen gfp, mostraron una correcta replicación en hojas agroinfiltradas de N. benthamiana, pero resultaron incapaces de inducir infección sistémica en este huésped experimental. La respuesta diferencial de N. benthamiana frente a distintas cepas de CTV permite estudiar los factores implicados en la interacción virus-huésped. Se analizó la interacción de las proteí­nas virales p20 y p25 de los aislados T36 y T318A con proteí­nas de N. benthamiana utilizando un abordaje consistente en: i) la expresión transitoria de p20/p25 marcadas con una etiqueta Strep-Tag en hojas de N. benthamiana, ii) purificación de los complejos proteí­na CTV-proteína huésped y análisis interactómico de los datos, y iii) estudio de la interacción directa entre p20/p25 y proteínas seleccionadas del huésped mediante análisis del doble hibrido en levadura y complementación bimolecular de fluorescencia (BIFC). Este abordaje proteómico mostró claras diferencias entre aislados que pueden explicar, en parte, el comportamiento diferencial de los aislados T36 y T318A en dicho huésped experimental. La inducción el síndrome de decaimiento por parte de CTV ha obligado a utilizar patrones tolerantes al decaimiento. Dichos patrones son menos adecuados. Las plantas de cí­tricos propagadas sobre NA e infectadas por CTV muestran necrosis en los tubos cribosos y disminución del floema funcional. Éstos desórdenes podrí­an ser consecuencia de la activación de los mecanismos de defensa como la reacción de hipersensibilidad desencadenada por la ruta del ácido salicí­lico o el silenciamiento génico mediado por RNA (post-transcriptional gene silencing, PTGS). Con el objetivo de avanzar en el mecanismo molecular de la resistencia del NA a la infección por CTV, se estudió el papel de diferentes genes de la planta implicados en las rutas mediante el uso de un vector viral basado en el genoma del virus del manchado foliar de los cítricos (citrus leaf blotch virus, CLBV). El silenciamiento génico de las rutas del AS o del PTGS en plantas NA y la inoculación de tres aislados de CTV patogénicamente diferentes mostró la implicación de ambas rutas en la defensa del NA frente a CTV. / Tristeza is the most important viral disease affecting citrus plants and Citrus tristeza virus (CTV) is the causal agent of this disease. CTV induces at least one of this syndromes: I) decline and death of sweet orange (SwO), grapefruits and mandarin trees grafted on sour orange (SO) rootstock, this syndromes is known as "tristeza", II) stunting, stem pitting (SP) and small fruits, and III) stunting and leaf chlorosis of lemon, grapefruit and SO seedlings (seedling yellows, SY). The host range of CTV is restricted and until recently no experimental herbaceous host was known. The agroinoculation Nicotiana benthamiana with clones of complementary DNA (cDNA) from the CTV isolate T36 cause the systemic infection of the plant and similar symptoms to those observed in citrus, although the infection is not limited to the phloem. T36 isolate induces SY and SP of Mexican lime (ML), but not in other hosts such as grapefruit and SwO. Therefore, to study the genetic determinants responsible of the SP syndrome induction was necessary to develop a genetic system based on agroinoculated clones from an isolate able to induce these symptoms, such as the Spanish isolate T318A. To do this, full length cDNA clones from T318A were obtained. They are able to replicate in N. benthamiana leaves but unable to induce systemic infection and showed several mutations in their protein of the minor coat, p27. The correction of these mutations and the construction of new clones of complete length from T318A labeled with the gfp gene, showed a proper replication in agroinoculated leaves of N. benthamiana, but they were still unable to induce systemic infection in this experimental host. The differential response of N. benthamiana to different CTV strains allows the study of the potential factors involved in the virus-host interaction. The aim of this work was study the interaction between the viral proteins p20 and p25 from the isolates T36 and T318A with N. benthamiana proteins with an analysis consisted in: I) the transitory expression of p20/p25 fused to Strep-Tag in N. benthamiana leaves, II) purification of the CTV protein-host protein complex and interatomic analysis of the data, and III) the study of the direct interaction between p20/p25 and selected plant proteins by the analysis of the double hybrid in yeast and bimolecular complementation of fluorescence (BIFC). The proteomic analysis showed strong differences between isolates that may partially explain the differential behavior of the T36 and T318A isolates in this experimental host. The induction of decline syndrome by CTV in citrus has leaded the use of tolerant rootstocks to decline. However, the use of such rootstocks is less suitable. Citrus plants propagated on SO rootstock and infected by CTV show phloem necrosis below the bud union that reduces the flow of carbohydrates to the roots. These symptoms may be a consequence of the activation of defense pathways in the plant, such as the hypersensitive reaction, hormone salicylic acid (SA) pathways or the RNA mediated post-transcriptional gene silencing (PTGS). Their relation is essential to know their implication in the decline. Therefore, the role of different genes involved in SA and PTGS has been studied by the silencing of plant genes using a viral vector (VIGS) based in the genome of the citrus leaf blotch virus (CLBV). The gene silencing of the SA and PTGS in SO and the inoculation of three different pathogenicity CTV isolates showed that both pathways are involved in the SO defense against CTV. The analysis of the proteins p20, p23 and p25 as possible suppressors of the AS indicating that the more virulent CTV isolates possess the more powerful suppressors. / La Tristesa és la malaltia viral més greu del cultiu dels cítrics. CTV induïx un o més de les sí­ndromes següents: I) decaïment i mort de taronger dolç§ (ND), pomelo i mandariner empeltats sobre el patró taronger amarg (NA), sí­ndrome conegut com "Tristesa", II) nanisme, estries en la fusta i fruita de xicotet calibre (SP) i III) nanisme i tonalitat groguenta de plantes de llavor de llimera, pomelo i taronger amarg (SY). El rang d'hostes de CTV és molt restringit i fins fa poc no es coneixia cap hoste herbaci experimental. Actualment es sap que la infecciò sistèmica en Nicotiana benthamiana amb clons de DNA complementari (cDNA) de l`aïllat de T36 provoca la infecció sistemàtica de la planta, acompanyada de síntomes similars als induïts en cí­trics, si be la infecció no queda llimitada al floema. L' aïllat T36 induïx SY i estries en la fusta de Llima Mexicana (LM), però no en altres hostes com a pomelo, ND o NA, l'estudi dels determinants genètics responsables de la inducció de la síndrome de SP requeria desenvolupar un sistema genètic basat en clons agroinfecciosos d'un aïllat inductor d'estos sí­mptomes, com l'aïllat espanyol T318A. Per a això, es va partir de clons de cDNA longitud completa de T318A prèviament desenvolupats al laboratori, capaços de replicar-se en fulls de N. benthamiana però incapaços d'induir infecció sistèmica i que presentaven varies mutacions en la seua proteïna de càpsida minoritatia p27. La correcció d`aquestes mutacions i la construcció de nous clons T318A de longitud completa marcats amb el gen gfp, van mostrar una correcta replicació en fulls agroinfiltradas de N. benthamiana però van resultar incapaços d'induir infecció sistèmica en aquest hoste experimental. La resposta diferencial dependent d'aïllat en N. benthamiana front CTV permet estudiar els possibles factors de la interacció virus- hoste. Es va dur a terme l'estudi de la funció de les proteínes virals p20 i p25 dels aïllats T36 i T318A amb proteïnes de N. benthamiana utilitzant un abordatge consistent en: i) l' expressió transitòria de les dues proteïnes p20/p25 marcades amb una etiqueta Strep-Tag en fulls de N. benthamiana, ii) purificació dels complexos proteïna CTV-proteïna hoste i anàlisi interactómic de les dades, i iii) estudi de la interacció directa per mitjà  de doble híbrid en llevat i complementació bimolecular de fluorescència (BIFC) de les proteïnes virals i determinades proteïnes de N. benthamiana. Aquest abordatge proteòmic va mostrar clares diferències entre aïllats que poden explicar el comportament diferencial dels aïllats T36 i T318A en aquest hoste experimental. La inducció de la sí­ndrome de decaïment per part de CTV en cí­trics ha obligat la utilització de patrons tolerants al decaïment. No obstant, aquestos patrons són agronòmicament menys adequats. Les plantes de cítrics propagades sobre NA i infectades por CTV mostren necrosi als tubs cribosos i disminució del floema funcional. Aquestos sí­mptomes poden ser conseqüència de l'activació de les rutes de defensa de la planta com la reacció d'hipersensibilitat, desencadenada per la ruta de l'àcid salicí­lic o el silenciamient gènic mediat per RNA (PTGS). Amb l'objectiu d'analitzar la implicació d¿aquestes rutes en la defensa, es va estudiar el paper de diferents gens implicats en la ruta de l'AS i del PTGS per mitjà  del silenciamient gènic induït per virus basat en el genoma del tacat foliar dels cítrics (CLBV). El silenciamient gènic de les rutes AS o PTGS en plantes NA i la inoculació de tres aïllats de CTV patogènicament diferents va mostrar la implicació de les dues rutes en la defensa del NA front CTV. L'analisis de les proteïnes p20, p23 i p25 com a possibles supressors de la ruta de l'AS va indicar que els aïllats més virulents de CTV posseïxen supressors més potents. / Gómez Muñoz, N. (2017). ESTUDIO DE LA INTERACCIÓN DIFERENCIAL ENTRE AISLADOS DEL VIRUS DE LA TRISTEZA DE LOS CÍTRICOS (CTV) Y SUS HUÉSPEDES [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/94624 / TESIS

Bordetella pertussis is a Gram-negative strictly human pathogen and the major causative agent of whooping cough or pertussis. The incidence of this highly contagious respiratory disease in developed countries has increased in the last decades. One of the less characterized virulence factors of B. pertussis is the type three secretion system (TTSS) which is responsible for the secretion of the effector proteins into host eukaryotic cells. This diploma thesis sheds light onto factors influencing TTSS in vitro activity. Although TTSS of laboratory strain Tohama I was induced by biologically active compounds present in blood (e. g. complement proteins), TTSS of recent clinical isolate B1917 seems to be induced permanently. Furthermore, BB0302 encoding a GntR family transcription regulator in B. bronchiseptica RB50 (homologous to BP0209 of Tohama I) was studied, however, the deletion of this gene did not affect the TTSS functionality. Serum resistance is a factor that plays a key role in the pathogenesis of B pertussis. We show that Czech recent isolates (2008-2015) are significantly more resistant to serum killing in vitro than the original vaccine strains (1954-1965). This phenomenon seems to result from the adaptation of global B. pertussis population to its human host. In addition, this diploma...

Bovine trypanosomosis is a serious constraint to livestock development in large parts of Mozambique. In most areas where tsetse flies are present, the disease in livestock is controlled using curative and prophylactic trypanocidal drugs. Those drugs have been used for many years and new drugs are unlikely to become available in the near future. As a result, trypanosomes have developed resistance against the currently available trypanocidal compounds. Drug resistance has been detected in various African countries and is a serious impediment to the control of livestock trypanosomosis. A study was initiated to determine whether drug resistant trypanosome strains are present in Zambézia Province of Mozambique. The aim of this study was to determine the sensitivity of Trypanosoma congolense isolates from Chinde, Nicoadala and Maganja da Costa Districts to diminazene aceturate, isometamidium chloride and homidium chloride. To assess the effect of the farming system and the intensity of drug regimens on the development of drug resistance, trypanosome isolates were collected from cattle from subsistence, semisubsistence and commercial livestock production systems. Drug-use practices in each of the production systems were determined using a questionnaire. The methodology used to assess the level of drugs resistance in the trypanosome isolates was the standardized method described by Eisler et al. (2001). Seven isolates were selected for resistance testing. For each of the seven isolates, five different doses varying between 0.01-20 mg/kg body weight for isometamidium chloride, 0.01-10 mg/kg body weight for homidium chloride and 1-30 mg/kg body weight for diminazene aceturate were used. For each dose rate six mice were treated intraperitoneally with the appropriate quantity of the drug dissolved in 0.2 ml of sterile distilled water 24 hours after the inoculation of the blood containing the trypanosomes. The control mice (six mice per trypanocidal drug) received the same amount of water without the drug. In four of the seven isolates high levels of multiple drug resistance (diminazene aceturate and isometamidium chloride) were detected. One isolate had a low level of multiple (diminazene aceturate and isometamidium chloride) drug resistance. Two isolates were susceptible to both diminazene aceturate and isometamidium chloride. One of those was highly susceptible to isometamidium chloride even at the lowest dose rate. The observed levels of drug resistance could in most cases be correlated to the drug-use practices in the particular livestock production system. The results obtained from homidium chloride treatment are not conclusive, because most the mice cured after receiving 10 mg/kg body weight of the drug. Hence more research is required to establish the homidium threshold in mice. The results of this study should be useful to define the strategy of disease control in places where resistance of trypanocide were been reported. / Dissertation (MSc (Veterinary Science))--University of Pretoria, 2005. / Veterinary Tropical Diseases / unrestricted

<p>  </p> <p>  In this study, whey protein isolate (WPI) and high gellan gum (HAGG) were selected as natural ingredients to produce food-grade biopolymer particles for stabilizing the air-water interface. To achieve this, different mixing ratios of WPI and HAGG were employed, and heat treatment was implemented at different pH levels under the same concentration based on investigations of the pH-driven phase behavior of the WPI/HAGG complex system. The resulting WPI/HAGG complex particles were then evaluated for their ability to stabilize air-water interfaces by measuring their foaming properties.</p> <p><br></p> <p>  Foams generated using 0.1% (wt/wt) WPI/HAGG complex particles, heated at pH 5 with the mixing ratio 2:1 has demonstrated enhanced stability over a period of 30 hours compared to the WPI alone. The unique properties of these complex particles, including their smaller size (around 500nm), greater negative charge (more negative than -30 mV), and compact spherical core-shell structure, along with the higher viscosity in the continuous phase as well as the presence of small protein particles and gellan chains at the interface, collectively contribute to their superior performance as foam stabilizers. This allows for the creation of aerated food products with desirable characteristics, including product handling, enhanced texture, and prolonged shelf life in food industry.</p>

Under det här kandidatexamensarbetet har protein-nanofibers påverkan på material undersökts genom att jämföra fibrillerade filmer med ofibrillerade. Sojaproteinisolat fibrillerades under förhållandena pH 2 och 85 ◦C under minst ett dygn och de syntetiserade nanofibrerna analyserades med Thioflavin T (ThT) fluorescens och atomkraftsmikroskopi (AFM). Spektra från analysmetoden ThT fluorescens indikerade på förekomsten av β-flak och analyserna med AFM visade på att fibrerna hade en morfologi som är karakteristisk för protein-nanofibrer. Resultaten antyder att de parametrar som påverkar morfologin hos fibrerna är fibrilleringstid och typ av protein. De gjutna filmerna från fibrillära respektive ofibrillära proteiner var sammanhängande bortsett från vissa sprickor. Värdena på E-modulen från AFM visade att det fibrillerade materialet var mer heterogent än det ofibrillerade. Filmer med sammanhängande yta erhölls vid tillsats av det mjukgörande additivet glycerol. Slutligen, material av både fibrillär och ofibrillär form kan framställas, däremot krävs vidare forskning för att optimera materialens egenskaper. / During this bachelor thesis project, the impact of protein nanofibers on materials has been analysed by comparing films made from fibrillar and non-fibrillar protein. Fibrillation of soy protein isolate was performed during at least 24 hours at pH 2 and a temperature of 85 ◦C. Analysis of the nanofibers was made with Thioflavin T (ThT) fluorescence and atomic force microscopy (AFM). The spectra from ThT Fluorescens indicated the presence of β-sheets and AFM confirmed that the fibrils had a morphology that is characteristic of protein nanofibers. The results indicated that heating time and protein type were the parameters which had the largest impact on the morphology of the fibrils. The synthesised films from both fibrillar and non-fibrillar protein were coherent with exception of some cracks. The elastic modulus from AFM indicated that the fibrillar film was more heterogeneous compared to the non-fibrillar film. To attain coherent films, the plasticising agent glycerol was added. To summarise, both fibrillar as well as non-fibrillar materials were successfully synthesised, however, further research is necessary to optimise the properties of the material.

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