Nine Virginia soybeans grown in a single location were compared for their antioxidant properties and isoflavone profiles. The extracts were evaluated for their total phenolic contents (TPC), Oxygen Radical Absorbance Capacity (ORAC), and DPPH™ radical scavenging activities. In order to evaluate efficient preparation methods for soybean antioxidants, three Virginia-grown soybeans were extracted using different extraction strategies. The extraction techniques included soxhlet extraction, conventional solvent extraction, and ultrasonic-assisted extraction (UAE) with 5 different common solvent systems including 50% and 80% aqueous acetone, 50 and 70% aqueous ethanol, and 80% aqueous methanol. The TPC in the soybean extracts and isoflavone compositions were significantly different among cultivars. Malonylgenistin was the major isoflavone in all soybean seeds, accounting for 75-83% of the total measured isoflavones. The V01-4937 variety had the highest total isoflavone and malonylgenistin contents, followed by V03-5794. The antioxidant activities of the soybean extracts were also significantly different. Overall, the V01-4937 soybean was the variety that stood out from the other tested Virginia soybeans because it had the highest TPC, ORAC value, and isoflavone contents as well as the second highest DPPH™ scavenging activity. Ultrasonic treatment improved the extraction of soybean phenolics by more than 50% compared to solvent alone. The UAE with 50% aqueous acetone was the most efficient for extraction of phenolic compounds in the soybean seeds. The conventional and UAE with 70% aqueous ethanol extracts had the highest ORAC values, while the soxhlet methanol extracts had the highest DPPH™ radical scavenging activities. Our results suggest that different extraction technologies have a remarkable effect on soybean antioxidant estimation and the UAE is more appropriate for soybean phenolic extraction because it is less time and solvent consuming than the conventional solvent and soxhlet extractions. The V01-4937 soybean with the highest TPC was evaluated for its antioxidant activity and isoflavone contents in an in vitro digestive system. After gastrointestinal digestion, soybean extracts contained higher TPC and ORAC values than cooked soybean (before digestion) but they were relatively low in DPPH™ radical scavenging capacity. The glucosides, daidzin, genistin, and malonylgenistin showed stability during simulated digestion with 83.3 %, 59.4 %, and 10.7 % recovery, respectively. Aglycones, including daidzein and genistein, were recovered at 37 % and 73.7 %, respectively, after in vitro digestion. In this study, daidzin was the most stable and bioaccessible isoflavone determined using the in vitro digestive system. Among the aglycones, genistein was more stable and bioaccessible than daidzein after digestion. In conclusion, soybean antioxidant activities were different among cultivars and efficient extraction for TPC was found using UAE with 50% aqueous acetone. Furthermore, antioxidant activities were stable during digestion and genistein, within aglycones tested, was the most stable and bioaccessible compound following in vitro digestion. This information may provide manufacturers or researchers information required to develop food or nutraceutical products processed for better bioaccessibility of soybean bioactive components. / Ph. D.

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-09-04T13:08:22Z No. of bitstreams: 1 arquivototal.pdf: 2019647 bytes, checksum: 8311c23a2b59a1fe2d552ea303954693 (MD5) / Made available in DSpace on 2017-09-04T13:08:22Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2019647 bytes, checksum: 8311c23a2b59a1fe2d552ea303954693 (MD5) Previous issue date: 2017-03-21 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The fruit of the caatinga has aroused growing interest on the part of the food industry because of its potential health benefits. Exotic fruits such as seriguela (Spondias purpurea), umbu-cajá (Spondias spp.) and mangaba (Hancornia speciosa) are important sources of bioactive compounds, such as phenolics. However, the high perishability of these fruits is a factor that hampers their in natura supply, which leads to their processing, which in turn has a direct impact on the content of bioactive present due to the process of structural modification through reactions that may interfere in the bioaccessibility of bioactive compounds, causing nutritional losses. For these reasons, the objective of this work was to determine the phenolic profile of mango, seriguela and umbu-cajá fruits and their respective pulps, evaluating the antioxidant capacity and checking the behavior of phenolic compounds when exposed to simulated gastrointestinal digestion and post-digestion bioaccessibility . In this study, phenolic compounds were identified and quantified by High Performance Liquid Chromatography (HPLC) on fruits of seriguela (Spondias purpurea), umbu-cajá (Spondias spp.) and mangaba (Hancornia speciosa), and their respective pulps and Fractions from gastrointestinal digestion. The antioxidant activity of the bioacessible fraction after in vitro simulated gastrointestinal digestion was quantified by the DPPH and FRAP assays. From the determined phenolic profile of fruits compared to their respective frozen pulps, a reduction in the phenolic compounds content of fruits of the genus Spondias spp. In the mangaba fruit there was an increase in some increased free phenolic contents, such as 3,4-dihydroxybenzoic acid (+ 50%), and p-coumaric acid (+ 50%). In the fruit pulp the reduction of the phenolic compounds was observed in front of the antioxidant tests, in which the free radical scavenging capacity reduced after the fruit processing. From the simulated gastrointestinal digestion it was observed that fruits of the genus Spondias have a high fraction of their bioaccessible phenolic compounds after digestion, with a bioaccessibility of 73.92% (gallic acid, umbu-cajá pulp), unlike mangaba 5 , 60% (ferulic acid). The results obtained in this work demonstrate that the pulp consumption of these fruits, especially seriguela and umbu-cajá can contribute significantly to the ingestion of antioxidant compounds, since they have high concentration of bioaccessible phenolic compounds. / As frutas da caatinga tem despertado crescente interesse por parte da indústria de alimentos devido aos seus potenciais benefícios à saúde. Frutas exóticas como a seriguela (Spondias purpurea), umbu-cajá (Spondias spp.) e mangaba (Hancornia speciosa) são importantes fontes de compostos bioativos, como os fenólicos. Entretanto, a alta perecibilidade dessas frutas é um fator que dificulta sua oferta in natura, o que leva ao processamento das mesmas, que por vez tem impacto direto no conteúdo de bioativos presentes devido ao processo de modificação estrutural mediante reações que podem interferir na bioacessibilidade dos compostos bioativos, ocasionando perdas nutricionais. Por estas razões o presente trabalho objetivou determinar o perfil fenólico das frutas de mangaba, seriguela e umbu-cajá e de suas respectivas polpas, avaliando a capacidade antioxidante e verificando o comportamento dos compostos fenólicos quando expostos a digestão gastrointestinal simulada e sua bioacessibilidade pós-digestão. Neste estudo, os compostos fenólicos foram identificados e quantificados por Cromatografia Liquida de Alta Eficiência (CLAE) nas frutas de seriguela (Spondias purpurea), umbu-cajá (Spondias spp.) e mangaba (Hancornia speciosa), e nas suas respectivas polpas e nas frações oriundas da digestão gastrointestinal. A aatividade antioxidante da fração bioacessível após a digestão gastrointestinal simulada in vitro foi quantificada através dos ensaios DPPH e FRAP. A partir do perfil fenólico determinado das frutas em comparação com a suas respectivas polpas congeladas, pôde-se identificar uma redução no conteúdo de compostos fenólicos das frutas do gênero Spondias spp. Na fruta mangaba houve aumento de alguns teores de fenólicos livres aumentados, como ácido 3,4-dihidroxibenzoico (+50%), e ácido p-cumarico (+50%). Na polpa em relação a fruta a redução dos compostos fenólicos foi observada frente aos ensaios antioxidantes, em que a capacidade sequestradora de radicais livres reduziu após o processamento das frutas. A partir da digestão gastrointestinal simulada observou-se que as frutas do gênero Spondias possuem uma elevada fração de seus compostos fenólicos bioacessíveis após a digestão, com uma bioacessibilidade de 73,92% (ácido gálico, polpa umbu-cajá), diferentemente da mangaba 5,60% (ácido ferúlico). Os resultados obtidos neste trabalho demonstram que o consumo da polpa destas frutas, especialmente seriguela e umbu-cajá podem contribuir significativamente para a ingestão de compostos antioxidantes, ja que os mesmos possuem alta concentração de compostos fenólicos bioacessiveis.

Orientadores: Flavia Maria Netto, Ricardo de Lima Zollner / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-21T22:30:18Z (GMT). No. of bitstreams: 1 Morais_JanainaMadruga_M.pdf: 1775595 bytes, checksum: 5ee1d5b4bacec45d3dfff615d49f7ca8 (MD5) Previous issue date: 2013 / Resumo: O leite bovino contém várias proteínas capazes de induzir resposta alérgica, sendo que a ß-lactoglobulina (ß-Lg) e a a-lactalbumina (a-La), proteínas mais abundantes do soro, estão entre as principais proteínas antigênicas. A homogeneização à alta pressão (HAP) é uma tecnologia emergente que pode alterar a estrutura das proteínas do soro de leite (IPS) e, portanto, seu caráter alergênico. Estudos anteriores apontam que a polimerização com a enzima transglutaminase (TG) reduziu o potencial antigênico da ß-Lg. O presente estudo teve como objetivo avaliar o efeito da HAP associada à polimerização por TG na suscetibilidade à digestão gástrica e gastrointestinal e na resposta antigênica da ß-Lg e a-La antes e após a digestão gástrica. Soluções de IPS (1% m/v), tratadas termicamente (72 °C/22 min) ou não, foram homogeneizadas a diferentes níveis de pressão (0, 100, 180 MPa) ou multiprocessadas por 3 ciclos consecutivos a 180 MPa. As amostras foram caracterizadas por turbidez, pelo teor de grupos sulfidrila reativos e livres e por eletroforese em sistema nativo e em SDS-PAGE em condições redutoras e não-redutoras. As amostras processadas foram polimerizadas com TG (25 U g-1 de proteína) e caracterizadas por eletroforese SDS-PAGE em sistema redutor e densitometria dos géis. Foi avaliada a suscetibilidade da ß-Lg e da a-La à digestão gástrica e gastrointestinal após tratamento térmico, HAP e polimerização, associados ou não. As condições de digestão foram: 182 U de pepsina g-1 de proteína e a relação 1:25 (enzima:proteína) para a pancreatina. A caracterização dos digeridos foi realizada por eletroforese SDS-PAGE/Tricina e cromatografia líquida de alta eficiência de fase reversa (CLAE-FR). A antigenicidade das proteínas foi avaliada pelo método ELISA, utilizando soro anti-ß-Lg ou anti-a-La de camundongos BALB/c imunizados com as proteínas nativas. A homogeneização das soluções de IPS, previamente tratadas termicamente ou não, resultou na diminuição da turbidez e do teor de grupos sulfidrila reativos e livres com o aumento dos níveis de pressão. A polimerização com TG resultou na formação de polímeros com massa molecular (MM) acima de 97,4 kDa, principalmente nas amostras tratadas termicamente e homogeneizadas a 180 MPa por 1 e 3 ciclos, alcançando aproximadamente 20% de polímeros de alta MM. As amostras pré-tratadas e polimerizadas foram mais suscetíveis à digestão gástrica e gastrointestinal, resultando em menor residual de proteína intacta em relação às amostras homogeneizadas pré-tratadas termicamente ou não. Foi observado aumento (P< 0,05) da capacidade de ligação à IgE anti-a-La para a amostra de IPS tratado termicamente e homogeneizado a 180 MPa (IPS-TT-180, 27,88 µg IgE mL-1), em relação ao IPS nativo (IPS-N, 12,80 µg IgE mL-1), sugerindo que a associação do tratamento térmico e HAP aumentou a exposição de seus epítopos. Após a digestão gástrica, não foi observada redução da atividade antigênica da a-La em nenhuma das amostras analisadas. Já para a ß-Lg verificou-se que após a digestão gástrica a amostra de IPS tratado termicamente, homogeneizado a 180 MPa e polimerizado (IPS-TT-180 TG) apresentou diminuição (P< 0,05) da resposta antigênica para IgE anti-ß-Lg (13,59 µg IgE mL-1) em relação ao IPS-N (34,21 µg IgE mL-1). Em conclusão, os tratamentos utilizados alteraram a estrutura das principais proteínas do IPS, resultando na diminuição moderada da resposta antigênica da ß-Lg e no aumento da antigenicidade da a-La / Abstract: Bovine milk contains several proteins, which can induce allergic response. The most abundant whey proteins, â-lactoglobulin (â-Lg) and á-lactalbumin (á-La) are among the major antigenic proteins of cow¿s milk. High pressure homogenization (HPH) is an emergent technology which can alter whey proteins (WPI) structures and possibly WPI antigenicity. Previous studies indicated that polymerization with transglutaminase enzyme (TG) reduced the antigenic potential of â-Lg. The objective of the present study was to evaluate the effect of HPH associated to polymerization by TG on the susceptibility to simulated gastric and gastrointestinal digestion and on antigenic response of â-Lg and á-La before and after gastric digestion. Solutions of WPI (1% w/v) previously heat treated (72 °C/22 min) or without heat treatment were homogenized at several pressure levels (0, 100 and 180 MPa) or at 180 MPa for three consecutive cycles. The samples were characterized by turbidity, content of exposed and free sulfhydryl groups and by electrophoresis on native system and SDS-PAGE under non-reducing and reducing conditions. The processed samples were polymerized with TG enzyme (25 U g-1 of protein) and characterized by SDS-PAGE under reducing conditions followed by densitometry of gels. The susceptibility of â-Lg and á-La to gastric and gastrointestinal digestion was evaluated after heat treatment, HPH and polymerization, associated or not with each other. The digestion conditions were: 182 U pepsin g-1 protein and pancreatin:protein ratio of 1:25. Characterization of the digests was carried out by SDS-PAGE/Tricine and reversed-phase high performance liquid chromatography (RP-HPLC). Protein antigenicity was evaluated by ELISA assay, using serum from BALB/c mouse immunized with native â-Lg and á-La. The homogenization of the WPI solutions, previously heat treated or without heat treatment, resulted in reducing both turbidity and exposed and free sulfhydryl group content as the pressure levels increased. The polymerization with TG enzyme resulted in polymers with molecular mass (MM) above 97.4 kDa, mainly in the samples heat treated and homogenized at 180 MPa for 1 or 3 cycles, forming around 20% of high MM polymers. Either polymerized samples previously treated with heat and/or HPH were more susceptible to gastric and gastrointestinal digestion, resulting in less residual intact protein as compared to the homogenized samples previously heat treated or without heat treatment. It was observed higher (P< 0.05) binding capacity to anti-á-La IgE for the sample heat treated and homogenized at 180 MPa WPI (WPI-TT-180, 27.88 ìg IgE mL-1) than for native WPI (WPI-N, 12.80 ìg IgE mL-1) suggesting that the association of heat treatment and HPH increased the exposition of á-La epitopes. After gastric digestion it was not observed a reduction of á-La antigenic activity. In relation to â-Lg antigenicity, after gastric digestion, the WPI sample heat treated, homogenized at 180 MPa and polymerized (WPI-TT-180 TG) showed reduction (P< 0.05) of anti-â-Lg IgE response (13.59 ìg IgE mL-1) as compared to WPI-N (34.21 ìg IgE mL-1). In conclusion, the treatments studied altered the structure of major protein of WPI, resulting in moderate reduction of â-Lg antigenic response and increased á-La antigenicity / Mestrado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Mestra em Alimentos e Nutrição

Tesis por compendio / [ES] De entre las metodologías disponibles para estudiar la digestión de alimentos, los modelos de digestión in vitro se plantean como procedimientos válidos para este propósito. La digestión in vitro consiste en simular el proceso de digestión en el laboratorio, reproduciendo las condiciones fisiológicas en cuanto a composición de los fluidos digestivos (electrolitos y enzimas), pH, temperatura, fuerzas mecánicas y duración de las etapas oral, gástrica e intestinal. Abordar el estudio de la digestión de nutrientes es de especial relevancia en patologías que cursan con alteraciones pancreáticas o hepáticas, asociadas a una digestión de lípidos comprometida en la etapa intestinal, debido a la disminución de secreción de pancreatina, bicarbonato y sales biliares. Este es el caso de la fibrosis quística con insuficiencia pancreática, y los pacientes que padecen esta afección deben seguir la terapia de sustitución de enzimas pancreáticas, que consiste en el suministro exógeno de pancreatina encapsulada. Sin embargo, la dosis de este suplemento debe ajustarse a las características de los alimentos y no se dispone de ningún método válido para tal fin. Para hacer frente a este reto, en el proyecto financiado con fondos europeos MyCyFAPP se ha logrado desarrollar un método para ajustar la dosis óptima de los suplementos enzimáticos utilizados en la terapia. La presente tesis doctoral se realizó en el marco de dicho proyecto. Concretamente, esta tesis tiene como objetivo abordar el estudio de la digestión de lípidos en los alimentos de origen animal (carne y productos cárnicos, huevos, queso y pescado) en el contexto de la fibrosis quística. Para abordar este objetivo se aplicó un modelo de digestión in vitro estático con el fin de explorar el papel de las características inherentes a los alimentos (estructura de la matriz alimentaria como resultado del procesado) y los factores relacionados con el individuo (pH, concentración de sales biliares y concentración de pancreatina) como factores determinantes de la lipólisis en alimentos de origen animal. A lo largo de los cuatro capítulos presentados, centrados en el huevo, la carne, el queso y el pescado, se presenta un diseño experimental común para estudiar la lipólisis, la proteólisis y la degradación de la matriz. En cada estudio, las diferentes técnicas de procesado aplicadas a los alimentos evaluados también permitieron evaluar el efecto de las propiedades inherentes a los alimentos en los resultados del estudio. Los resultados han contribuido al desarrollo de un nuevo método basado en la evidencia para optimizar la terapia de reemplazo de enzimas pancreáticas e informan a la comunidad científica sobre nuevos conocimientos en el comportamiento de diferentes alimentos sometidos al proceso de digestión. / [CA] De les metodologies disponibles per estudiar la digestió d'aliments, els models de digestió in vitro es plantegen com a procediments vàlids per a aquest propòsit. La digestió in vitro consisteix en simular el procés de digestió al laboratori, reproduint les condicions fisiològiques pel que fa a la composició dels fluids digestius (electròlits i enzims), pH, temperatura, forces mecàniques i durada de les etapes oral, gàstrica i intestinal. Abordar l'estudi de la digestió de nutrients és d'especial rellevància en patologies que cursen amb alteracions pancreàtiques o hepàtiques, associades a una digestió de lípids compromesa en l'etapa intestinal, a causa de la disminució de secreció de pancreatina, bicarbonat i sals biliars. Aquest és el cas de la fibrosi quística amb insuficiència pancreàtica. Els pacients que pateixen aquesta afecció han de seguir la teràpia de substitució d'enzims pancreàtics, que consisteix en el subministrament exogen de pancreatina encapsulada. No obstant això, la dosi d'aquest suplement ha d'ajustar-se a les característiques dels aliments i actualment no es disposa de cap mètode vàlid per a tal fi. Per enfrontar a aquest repte, en el projecte finançat amb fons europeus, MyCyFAPP, s'ha aconseguit desenvolupar un mètode per a ajustar la dosi òptima dels suplements enzimàtics utilitzats en la teràpia. La present tesi doctoral es va realitzar en el marc d'aquest projecte. Concretament, aquesta tesi té com a objectiu abordar l'estudi de la digestió de lípids en els aliments d'origen animal (ous, carn i productes carnis, formatge i peix) en el context de la fibrosi quística. Per a abordar aquest objectiu es va aplicar un model de digestió in vitro estàtic amb la finalitat d'explorar el paper de les característiques inherents als aliments (estructura de la matriu alimentària com a resultat del processament) i els factors relacionats amb l'individu (pH, concentració de sals biliars i concentració de pancreatina) com a factors determinants de la lipòlisi en aliments d'origen animal. Als quatre capítols presentats, centrats en l'ou, carn, formatge i peix, es presenta un disseny experimental comú per a estudiar la lipòlisi, la proteòlisi i la degradació de la matriu. En cada estudi, les diferents tècniques de processament aplicades als aliments avaluats també van permetre avaluar l'efecte de les propietats inherents als aliments en els resultats de l'estudi. Els resultats han contribuït al desenvolupament d'un nou mètode basat en l'evidència per a optimitzar la teràpia de substitució d'enzims pancreàtics i informen la comunitat científica sobre nous coneixements en el comportament de diferents aliments sotmesos al procés de digestió. / [EN] Among the available methodologies to study food digestion, in vitro digestion models have raised as a valid procedure. In vitro digestion consists of simulating the digestion process in the laboratory, by reproducing the physiological conditions in terms of digestive fluids composition (electrolytes and enzymes), pH, temperature, mechanical forces and duration of the oral, gastric and intestinal stages. Addressing the study of nutrient digestion is of special relevance in pathologies coursing with pancreatic or hepatic alterations, which are associated with compromised intestinal lipid digestion due to reduced secretion of pancreatin, bicarbonate and bile salts. This is the case of Cystic Fibrosis along with pancreatic insufficiency, and the patients suffering this condition have to follow pancreatic enzyme replacement therapy, the exogenous supply of encapsulated pancreatin. However, the dose of this supplement should be adjusted to the specific characteristics of foods, and no valid method was available for such purpose. To tackle this challenge, the EU-funded project MyCyFAPP succeed to develop a method to adjust the optimal dose the enzyme supplements used in the therapy. The present doctoral thesis was conducted as a relevant part of this project. Concretely, this thesis aims at addressing the study of lipid digestion in foods to generate new knowledge regarding nutrient digestion in animal origin dietary sources (egg, meat and meat products, cheese and fish) in the context of Cystic Fibrosis. To address this goal a static in vitro digestion model was applied. The role of inherent-to-food characteristics (resulting food matrix structure from processing) and host related factors (pH and bile salts concentration and pancreatin concentration) were explored as determinants of lipolysis in animal-origin foods. Along the four chapters presented, focused on egg, meat, cheese and fish, a common experimental design was applied to study lipolysis, proteolysis and matrix degradation. In each study, different processing techniques applied to the assessed foods allowed for evaluating the effect of inherent-to-food properties on the study outcomes as well. The results have contributed to the development of a new evidence-based method to optimise pancreatic enzyme replacement therapy, and inform the scientific community about new insights on the behaviour of different foods undertaking the digestion process. / Authors of this paper acknowledge the European Union and the Horizon 2020 Research and Innovation Framework Programme (PHC-26-2014 call Self-management of health and disease: citizen engagement and mHealth) for fully funding this research in the context of MyCyFAPP Project, under grant agreement number 643806. The authors would like to thank the Conselleria de Educació i Investigació de la Generalitat Valenciana and also the European Union (“El Fondo Social Europeo (FSE) invierte en tu futuro”) for the PhD scholarship given to Andrea Asensio Grau (ACIF/2017/008). This study was developed thanks to the equipment funded with the support from the Generalitat Valenciana IDIFEDER/2018/041 (PO FEDER Comunitat Valenciana 2014-2020). Finally, we thank Antonio Martínez Cañada, from the Data Science and Biostatistics Unit of Instituto de Investigación Sanitaria La Fe, and Arash Javanidejad for the English corrections. / Asensio Grau, A. (2021). In vitro approach of dietary and host related factors affecting digestion of animal-origin foods under cystic fibrosis disease [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/171512 / TESIS / Compendio

Tesis por compendio / [ES] La presente tesis plantea distintas estrategias para el diseño de alimentos a partir del aprovechamiento del bagazo que se genera como subproducto en la industria de elaboración de zumos de frutos rojos. De este modo, se puede aportar un valor añadido al subproducto, al mismo tiempo que se desarrollan alimentos capaces de cubrir las necesidades y exigencias nutritivas que demanda el consumidor actual. Uno de los aspectos más interesantes que tiene el uso de estos subproductos, es su alto contenido en polifenoles y fibra, de gran importancia en la industria alimentaria por su efecto en las propiedades de los alimentos y sobre la salud. Se realizó un primer estudio para conocer el efecto de la sustitución de grasa en bizcochos por diferentes tipos de fibra: soluble, insoluble y una mezcla de ambas. En este trabajo se observó que las características texturales, estructurales y sensoriales se vieron afectadas con la incorporación de fibra, principalmente en el caso de la fibra insoluble. Por todo esto, se establecieron dos estrategias de mejora. Por una parte, se incorporaron diferentes impulsores químicos en combinación con bicarbonato (normal o encapsulado) en bizcochos formulados con bagazo y se caracterizaron desde un punto de vista físico químico, estructural y sensorial. Por otra parte, se estudió cómo afecta a la textura, estructura y a la digestibilidad in vitro del almidón la sustitución parcial de harina de trigo por harina extrusionada en magdalenas formuladas con bagazo. En ambos estudios se obtuvieron resultados satisfactorios en cuanto a la mejora de las propiedades del producto. Además de las estrategias de mejora en productos horneados, se decidió incorporar bagazo en matrices lácteas para desarrollar nuevas formulaciones de batidos. Se estudió el efecto de un tratamiento no térmico, las altas presiones hidrostáticas, sobre el contenido fenólico y la capacidad antioxidante de los batidos, y sobre la capacidad antimicrobiana de los fenoles presentes en el bagazo. Se observó que la mayor retención de compuestos fenólicos y capacidad antioxidante con la mínima supervivencia microbiológica se obtuvo para las presiones y tiempos más altos (500 MPa durante 10 min) en los batidos con las concentraciones más altas de bagazo estudiadas (10%). Por último, se llevó a cabo un estudio con sistemas modelo para conocer las interacciones que tienen los polifenoles, cuando forman parte del bagazo o cuando se encuentran en forma de extracto, con los principales macronutrientes de los alimentos y sus efectos sobre la bioaccesibilidad de los compuestos fenólicos tras la digestión in vitro. La bioaccesibilidad de los polifenoles cuando se incorporaron en los sistemas modelo en forma de bagazo aumentó en comparación con cuando se adicionaron como extracto. Por otro lado, los sistemas modelo formulados con bagazo y un solo nutriente, presentaron una mayor bioaccesibilidad de los compuestos fenólicos que cuando se encontraron todos los nutrientes en el sistema modelo. / [CAT] La present tesi planteja diferents estratègies per al disseny d'aliments a partir de l'aprofitament del bagàs que es genera com a subproducte en la indústria d'elaboració de sucs de fruits rojos. D'aquesta manera, es pot aportar un valor afegit al subproducte, al mateix temps que es desenvolupen aliments capaços de cobrir les necessitats i exigències nutritives que demanda el consumidor actual. Un dels aspectes més interessants que té l'ús d'aquests subproductes, és el seu alt contingut en polifenols i fibra, de gran importància en la indústria alimentària pel seu efecte en les propietats dels aliments i sobre la salut. Es va realitzar un primer estudi per a conéixer l'efecte de la substitució de greix en bescuits per diferents tipus de fibra: soluble, insoluble i una mescla d'ambdues. En aquest treball es va observar que les característiques texturals, estructurals i sensorials es van veure afectades amb la incorporació de fibra, principalment en el cas de la fibra insoluble. Per tot això, es van establir dues estratègies de millora. D'una banda, es van incorporar diferents impulsors químics en combinació amb bicarbonat (normal o encapsulat) en bescuits formulats amb bagàs i es van caracteritzar des d'un punt de vista físic químic, estructural i sensorial. D'altra banda, es va estudiar com afecta a la textura, estructura i a la digestibilitat in vitro del midó la substitució parcial de farina de blat per farina extrusionada en magdalenes formulades amb bagàs. En tots dos estudis es van obtindre resultats satisfactoris quant a la millora de les propietats del producte. A més de les estratègies de millora en productes enfornats, es va decidir incorporar bagàs en matrius làcties per a desenvolupar noves formulacions de batuts. Es va estudiar l'efecte d'un tractament no tèrmic, les altes pressions hidroestàtiques, sobre el contingut fenòlic i la capacitat antioxidant dels batuts, i sobre la capacitat antimicrobiana dels fenols presents en el bagàs. Es va observar que la major retenció de compostos fenòlics i capacitat antioxidant amb la mínima supervivència microbiològica es va obtindre per a les pressions i temps més alts (500 MPa durant 10 min) en els batuts amb les concentracions més altes de bagàs estudiades (10%). Finalment, es va dur a terme un estudi amb sistemes model per a conéixer les interaccions que tenen els polifenols, quan formen part del bagàs o quan es troben en forma d'extracte, amb els principals macronutrients dels aliments i els seus efectes sobre la bioaccessibilitat dels compostos fenòlics després de la digestió in vitro. La bioaccessibilitat dels polifenols quan es van incorporar en els sistemes model en forma de bagàs va augmentar en comparació amb quan es van addicionar com a extracte. D'altra banda, els sistemes model formulats amb bagàs i un sol nutrient, van presentar una major bioaccessibilitat dels compostos fenòlics que quan es van trobar tots els nutrients en el sistema model. / [EN] This thesis proposes different strategies for the design of foods based on the use of pomace generated as a by-product in the berry juice processing industry. In this way, it is possible to add value to the by-product while developing foods capable of meeting the needs and nutritional requirements demanded by today's consumers. One of the most interesting aspects of the use of these by-products is their high content of polyphenols and fiber, of great importance in the food industry due to their effect on food properties and health. A first study was carried out to determine the effect of replacing fat in sponge cakes with different types of fiber: soluble, insoluble and a mixture of both. In this work it was observed that the textural, structural and sensory characteristics were affected by the incorporation of fiber, mainly in the case of insoluble fiber. For all these reasons, two improvement strategies were determined. On the one hand, different chemical leavenings agents were incorporated in combination with bicarbonate (normal or encapsulated) in sponge cakes formulated with pomace and characterized from a physical-chemical, structural and sensory point of view. On the other hand, it was studied how the partial substitution of wheat flour by extruded flour in pomace-formulated muffins affects texture, structure and in vitro starch digestibility. In both studies satisfactory results were obtained in terms of improved product properties. In addition to the improvement strategies in baked products, it was decided to incorporate pomace in dairy matrices to develop new milkshake formulations. The effect of a non-thermal treatment, such as high hydrostatic pressures, on the phenolic content and antioxidant capacity of the milkshakes, and on the antimicrobial capacity of the phenols present in pomace was studied. It was observed that the highest retention of phenolic compounds and antioxidant capacity with the lowest microbiological survival was obtained for the highest pressures and times (500 MPa for 10 min) in the milkshakes with the highest pomace concentrations studied (10%). Finally, a study was carried out with model systems to learn about the interactions that polyphenols have when they are part of the pomace or when they are in extract form, with the main macronutrients of food and their effects on the bioaccessibility of phenolic compounds after in vitro digestion. The bioaccessibility of polyphenols increased when it is incorporated into the model systems as pomace compared to when it is added as extract. On the other hand, model systems formulated with pomace and a single nutrient presented higher bioaccessibility of phenolic compounds than when all nutrients were found in the model system. / The authors are grateful to Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA-Spain) for financial support through the BERRYPOM. Adding value to processing waste: innovative ways to incorporate fibers from berry pomace in baked and extruded cereal-based foods project included in the ERA-NET-SUSFOOD program, to Spanish Ministry of Science, Innovation and Universities through project AGL2017-86840-C2-2-R, and to FEDER funds from EU. / Díez Sánchez, E. (2021). Aprovechamiento de subproductos de zumos de frutos rojos para el diseño de alimentos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/172372 / TESIS / Compendio

Tesis por compendio / [ES] La presente tesis tiene como objetivo ofrecer estrategias que permitan incrementar la rentabilidad del cultivo de caqui "Rojo Brillante" mediante la valorización de los excedentes y destríos generados en los almacenes. La tesis aborda como objetivos principales la obtención de productos e ingredientes deshidratados con alto contenido en compuestos bioactivos y la evaluación de la digestibilidad in vitro de los mismos. La primera estrategia consistió en obtener caquis semisecos mediante secado "natural" evaluando la necesidad de aplicar un tratamiento previo de desastringencia. El tratamiento de secado en los caquis astringentes disminuyó significativamente el contenido en taninos solubles responsables de la astringencia de la fruta, alcanzando valores similares a los presentes en el caqui no astringente. Además, se observó que, el color de la pulpa permaneció con un tono anaranjado y poco pardeado cuando se partió de caqui astringente, por lo que no sería recomendable la aplicación de un tratamiento previo de desastringencia. Una segunda estrategia se centró en la liofilización para obtener snacks de caqui de alta calidad, al ser éste un tratamiento que permite obtener productos o ingredientes con alto valor añadido. Se establecieron las condiciones óptimas de procesado y almacenamiento mediante la obtención de los valores de humedad y actividad del agua críticos que garantizasen el estado vítreo de la matriz, evitando un aumento en la tasa de reacciones de deterioro, cambios de textura y color, y la pérdida de los compuestos bioactivos de la fruta. Como última estrategia se utilizó el tratamiento de secado por aire caliente en caqui astringente y no astringente, en distintos estados de madurez, para obtener snacks de caqui. Los snacks desarrollados presentaron un tono más anaranjado, en comparación con la fruta fresca, y menor contenido en taninos solubles. Se observó una alta correlación entre el nivel de astringencia percibido por los consumidores y la disminución del contenido en taninos solubles. Los snacks obtenidos a partir de caqui astringente en etapas de madurez avanzadas fueron igualmente aceptados por los consumidores que los obtenidos a partir de caqui no astringente. Seguidamente se estudió el efecto del secado con aire caliente y del estado de madurez sobre la fracción de carotenoides mediante diferentes técnicas cuantitativas y cualitativas como la fotoluminiscencia. El secado no afectó al contenido en carotenoides, pero disminuyó la capacidad antioxidante de los snacks. La fotoluminiscencia evidenció la isomerización de carotenoides y la formación de productos de degradación térmica, por lo que ambos hechos podrían explicar la pérdida de capacidad antioxidante en el caqui sometido a tratamientos de secado por aire caliente. Por último, se llevaron a cabo estudios de digestión in vitro. La recuperación de taninos solubles en la fase del intestino delgado fue mayor en los snacks obtenidos mediante secado por aire caliente que en la fruta fresca y en los snacks liofilizados. Los snacks obtenidos a partir de caqui no astringente presentaron un índice de recuperación de taninos solubles mayor que los obtenidos a partir de caqui astringente. Los taninos insolubles llegaron intactos a la fracción no absorbida, por lo que podrían llegar al colon y ejercer su potencial antioxidante. / [CA] La present tesi té com a objectiu oferir estratègies que permeten incrementar la rendibilitat del cultiu de caqui "Roig Brillant" mitjançant la valorització dels excedents i destriaments generats en els magatzems. La tesi aborda com a objectius principals l'obtenció de productes i ingredients deshidratats amb alt contingut en compostos bioactius i l'avaluació de la digestibilitat in vitro d'aquests. La primera estratègia va consistir a obtindre caquis semisecs mitjançant assecat "natural" avaluant la necessitat d'aplicar un tractament previ de desastringència. El tractament d'assecat en els caquis astringents va disminuir significativament el contingut en tanins solubles responsables de la astringència de la fruita, aconseguint valors similars als presents en el caqui no astringent. A més, es va observar que, el color de la polpa va romandre amb un to ataronjat i poc marró quan es va partir de caqui astringent, per la qual cosa no seria recomanable l'aplicació d'un tractament previ de desastringència. Una segona estratègia es va centrar en la liofilització per a obtindre snacks de caqui d'alta qualitat, en ser aquest un tractament que permet obtindre productes o ingredients amb alt valor afegit. Es van establir les condicions òptimes de processament i emmagatzematge mitjançant l'obtenció dels valors d'humitat i activitat de l'aigua crítics que garantiren l'estat vitri de la matriu, evitant un augment en la taxa de reaccions de deterioració, canvis de textura i color, i la pèrdua dels compostos bioactius de la fruita. Com a última estratègia es va utilitzar el tractament d'assecat per aire calent en caqui astringent i no astringent, en diferents estats de maduresa, per a obtindre snacks de caqui. Els snacks desenvolupats van presentar un to més ataronjat, en comparació amb la fruita fresca, i menor contingut en tanins solubles. Es va observar una alta correlació entre el nivell de astringència percebut pels consumidors i la disminució del contingut en tanins solubles. Els snacks obtinguts a partir de caqui astringent en etapes de maduresa avançades van ser igualment acceptats pels consumidors que els obtinguts a partir de caqui no astringent. Seguidament es va estudiar l'efecte de l'assecat amb aire calent i de l'estat de maduresa sobre la fracció de carotenoids mitjançant diferents tècniques quantitatives i qualitatives com la fotoluminiscència. L'assecat no va afectar el contingut en carotenoids, però va disminuir la capacitat antioxidant dels snacks. La fotoluminiscència va evidenciar la isomerització de carotenoids i la formació de productes de degradació tèrmica, per la qual cosa tots dos fets podrien explicar la pèrdua de capacitat antioxidant en el caqui sotmés a tractaments d'assecat per aire calent. Finalment, es van dur a terme estudis de digestió in vitro. La recuperació de tanins solubles en la fase de l'intestí prim va ser major en els snacks obtinguts mitjançant assecat per aire calent que en la fruita fresca i en els snacks liofilitzats. Els snacks obtinguts a partir de caqui no astringent van presentar un índex de recuperació de tanins solubles major que els obtinguts a partir de caqui astringent. Els tanins insolubles van arribar intactes a la fracció no absorbida, per la qual cosa podrien arribar al còlon i exercir el seu potencial antioxidant. / [EN] This thesis proposes different strategies to increase the profitability of "Rojo Brillante" persimmon by valuing the surpluses and discarded fruit generated in the warehouses. This thesis addresses the development of dehydrated products and ingredients with a high content of bioactive compounds and the evaluation of their in vitro digestibility. The first strategy was to obtain semidry persimmons by "natural" drying, evaluating the need to apply a previous desastringency treatment. The drying treatment in the astringent persimmons significantly decreased the soluble tannin content responsible for the astringency of the fruit, reaching values similar to those present in the non-astringent persimmon. In addition, the color of the pulp remained with an orange hue angle and moderate browning, when astringent persimmon was used, thus the application of a previous desastringency treatment would not be recommended. A second strategy focused on freeze-drying to obtain high-quality persimmon snacks, as a treatment that allows obtaining products or ingredients with high added value. Optimum processing and storage conditions were established by obtaining the critical water content and water activity values that would guarantee the glassy state, avoiding an increase in the rate of deterioration reactions, changes in texture and color, and the loss of bioactive compounds in the fruit. As a last strategy, the hot air-drying treatment was used in astringent and non-astringent persimmon, in different ripening stages, to obtain persimmon snacks. The snacks had a more orange hue angle, compared to fresh fruit, and lower soluble tannin content. A high correlation was observed between the level of astringency perceived by consumers and the decrease in soluble tannin content. Snacks obtained from astringent persimmon in advanced ripening stages were equally accepted by consumers as non-astringent ones. Then, the effect of hot air drying and the ripening stage on the carotenoid fraction was studied using different quantitative and qualitative techniques such as photoluminescence. Hot air-drying did not affect the carotenoid content but decreased the antioxidant capacity of the snacks. The photoluminescence evidenced the isomerization of carotenoids and the formation of thermal degradation products. Both facts could explain the loss of antioxidant capacity in persimmon subjected to hot air-drying treatments. Lastly, in vitro digestion studies were carried out. The recovery of soluble tannins in the small intestine phase was higher in the snacks obtained by hot air drying than in the fresh fruit and the freeze-dried snacks. The snacks obtained from non-astringent persimmon had a higher recovery index of soluble tannin than those obtained from astringent persimmon. The insoluble tannins reached the unabsorbed fraction intact, so that they could reach the colon and exert their potential antioxidant capacity. / The authors thank the Ministerio de Ciencia, Innovación y Universidades for the financial support given throughout Project RTA2017-00045-C02-02 / Martínez González, C. (2021). Tratamientos de secado para la obtención de ingredientes de alto valor nutritivo a partir del destrío postcosecha de caqui [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/180348 / TESIS / Compendio

Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. The ability of biofilm forming pathogens, such as Salmonella, to survive thermal and chemical processes is improved; it is unclear if biofilms will also improve survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation on Salmonella survival and gene expression after passage through an in-vitro digestion model. Cells of Salmonella enterica serotype Tennessee were deposited onto membranes for planktonic cells or on glass beads to create biofilms. The cells were subsequently dried at room temperature and stored in dried milk powder (aw = 0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto brilliant green agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and relative gene expression determined for genes associated with stress response (rpoS, otsB), virulence (hilA, hilD, invA, sipC) and a housekeeping gene 16S rRNA using quantitative real-time PCR. The physiological state and length of storage effected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in-vitro digestion system (p<0.05). Larger numbers of S. Tennessee were recovered by plate counts for biofilm cells, compared to planktonic cells. However, the numbers of 16S rRNA gene copies were not significantly different suggesting entry of S. Tennessee into a viable but non-culturable state. Prolonged storage in dry milk powder was not associated with increased cross-protection to gastric stress. Increased expression of stress response genes rpoS and otsB correlated with survival, indicating cross protection of low water activity and acid stress. Increased expression of virulence-associated genes was seen in cells exposed to short periods of dry storage, suggesting an increased virulence potential. / Master of Science in Life Sciences

Chronic conditions such as obesity, inflammatory bowel disease (IBD), and colitis are associated with gastrointestinal (GI) inflammation and compromised GI barrier integrity. Cocoa may be a potential dietary strategy to mitigate gut-related conditions and been shown to elicit anti-inflammatory, antioxidant, and prebiotic effects. Alkali treatment of cocoa was once thought to reduce its bioactivity, but new evidence suggests it may enhance cocoa's health properties, through the formation of new, potentially bioactive high molecular weight compounds. Inulin, a fructose-containing plant polymer, exerts prebiotic effects and has also been investigated in the mitigation of IBD. This study aims to 1) investigate effects of alkali processing on gut health related bioactivity and phytochemical composition of cocoa and 2) evaluate potential additive benefits of combining cocoa and inulin. Polyphenolic and flavanol compounds in natural cocoa, alkalized cocoa, and inulin powders were characterized using Folin-Ciocalteu (total polyphenols) and 4-dimethylaminocinnamaldehyde (total flavanols) assays, thiolysis , and HILIC UPLC-MS/MS. Treatments of cocoa and inulin were made in 1:2 cocoa:inulin and 1:4 cocoa:inulin mixtures for both natural and alkalized cocoas. Cocoa mixtures, in addition to both cocoa powders and inulin alone, were subjected to an in-vitro digestion to generate material for an in-vitro fecal fermentation. Samples collected from the fermentation at 0, 6, 12, and 24 hours were analyzed via HPLC-MS for microbial metabolites, applied to HT-29 colon cancer cells to assess anti-inflammatory activity, and applied to a florescence assay measuring PLA2 inhibitory activity. The alkalized cocoa powder was found to have a significantly lower concentration of total polyphenols and total flavanols, as well as a lower mDP, suggesting that alkalization may affect larger procyanidins more than smaller flavanol compounds. Inulin enhanced the inhibition of the PLA2 enzyme and enhanced the IL-8 anti-inflammatory properties of cocoa, although the trends were weak. Overall, we did not see any clear, significant effects of alkalization or the addition of inulin to cocoa's colonic metabolite formation or its gut bioactivity in vitro. However, we have demonstrated that colonic fermentation of cocoa may have a negative effect on its bioactivity in vitro. Future research should further explore flavanol DP and bioactivity, fiber's interaction with polyphenols, colonic metabolism of cocoa, and cocoa's gut health effects in vivo. / Master of Science in Life Sciences / Gut conditions like obesity-associated inflammation and inflammatory bowel disease are highly prevalent, debilitating, and currently have no cure. Cocoa has been investigated as a possible dietary strategy for the mitigation and prevention of chronic inflammatory gut conditions due to its anti-inflammatory and enzyme inhibiting properties. Most attribute these effects of cocoa to its abundance of compounds called polyphenols. It is widely thought that the ability of cocoa to promote health is lost when cocoa beans are processed, because of the loss of polyphenols. Alkalization, or "Dutching", is an optional step in cocoa processing that some manufacturers perform to enhance flavor and color formation. Dutching cocoa can promote the polymerization of many smaller, flavanol, protein, and other compounds into larger, indigestible compounds. These indigestible compounds will not be absorbed in the small intestine and may be broken down in the large intestine by colonic bacteria, forming new metabolites. We obtained cocoa powders, one natural (not alkalized) and one alkalized and compared them in terms of content of polyphenols, bioactivities, and anti-inflammatory abilities. Additionally, we added a known prebiotic, inulin, to our cocoa formulations to determine if there are additive benefits of cocoa and inulin together. Ultimately, we found that alkalized cocoa contained lower concentrations of all polyphenolic compounds, even the larger compounds. Inulin enhanced the inhibition of digestive enzymes and the anti-inflammatory properties of cocoa, though not significantly. Inulin also reduced the pH (i.e. increased the acidity) of a simulated gut environment, which may be beneficial. Alkalization did not significantly affect cocoa's enzyme inhibitory activity or anti-inflammatory activity. Overall, the addition of inulin to cocoa does not seem to be effective in increasing cocoa's ability to treat and prevent gut diseases, but more information is needed.

According to the American Heart Association, cardiovascular disease (CVD) is the leading cause of death in the U.S., representing about 20-30% of all deaths every year in the U.S. Major risk factors for developing CVD include high blood lipid and LDL-cholesterol levels. A large number of heart attacks and strokes could be prevented by controlling these factors through lifestyle modifications and diet interventions. Epidemiological evidence shows that consumption of dry or common beans (Phaseolus vulgaris L.) has positive effects on reducing blood LDL-cholesterol and lipid levels. These health benefits are mainly attributed to the high content of dietary fiber (DF) of beans, including soluble and insoluble DF (SDF and IDF). Some proposed mechanisms to explain the cholesterol and lipid-lowering effects of DF are related to the physico-chemical properties (e.g. viscosity) of DF, and involve binding to bile salts (BS) in the small intestinal to prevent BS re-absorption which further promote cholesterol catabolism and delay lipid digestion. Nevertheless, the precise mechanisms are not fully understood yet. In addition, cooking and processing operations, and in particular high-hydrostatic pressure (HHP) processing, can modify the composition, structure and functional properties of foods; however, whether HHP affects the ability of beans to interfere with different aspects of lipid digestion remains unknown. The overall goal of this research is to understand how common beans and HHP processing impact the ability of beans to bind BS and influence lipid digestion in vitro. The specific objectives are 1) to evaluate the effect of HHP treatments (and compared it with conventional cooking (HT)) on the thermo-rheological and functional properties of dry beans; 2) to identify the impact of major bean components on the in vitro BS-binding ability of beans, the role played by the bean matrix and how this is affected by HHP processing; 3) to investigate how bean (micro)structure and fiber fractions, as well as HHP processing of dry beans, influence lipid digestion in vitro. Results showed that HT caused complete starch gelatinization and protein denaturation of beans, while HHP treatments induced partial or no starch gelatinization and a lower degree of protein denaturation, which resulted in enhanced protein solubility and emulsifying activity/stability. It was observed that, while HT treatment reduced the capacity of bean flours to retain BS because of severe disruption of the bean cell wall integrity, protein matrices, and starch granules, HHP treatments maintained or enhanced BS retention, possibly by promoting the formation of starch/protein/fiber networks able to entrap BS. Furthermore, by using an in vitro dialysis-based digestion model combined with viscosity measurements and thermal analysis, it was shown that the interaction between bean tissue materials and primary BS was not only related to viscosity but also involved hydrophobic linkages. The contribution of IDF and proteins (other than SDF) to retain BS was also significant. There was a different binding preference of beans to four primary BS with sodium glycochenodeoxycholate, the more hydrophobic BS, showing the largest retention levels while sodium taurocholate being the least effectively retained BS by beans. Diverse sequences of the same processing operations showed distinct impacts on BS-retention by dry beans. By means of an in vitro digestion model simulating conditions in the upper gastrointestinal tract, bean flours delayed the digestion of extrinsic lipids to a higher extent, compared to isolated IDF and SDF. Furthermore, HHP treatment and less severe mechanical disintegration maintained the ability of beans to modulate lipid digestion, which suggests the importance of bean structural integrity in reducing the lipolysis rate and extent by beans. Overall, this research work shows that HHP processing is a promising minimal processing technology to produce bean flours with improved functionality. It also highlights the importance of considering the structure of foods, and not just their nutrient content, when evaluating potential health impacts. This knowledge could be applied to develop a range of bean-based ingredients and formulations with desirable health benefits. This work can be extended to research the influence of beans on the gut microbiota and profile of secondary BS and short-chain fatty acids, which are also closely related to cholesterol and lipid metabolism. / Doctor of Philosophy / According to the American Heart Association, cardiovascular disease (CVD) is the leading cause of death in the U.S., representing about 20-30% of all deaths every year in the U.S. Around the world, millions of people are struggling to control the risk of CVD. Major risk factors for developing CVD include high blood lipid and LDL-cholesterol levels. A large number of heart attacks and strokes can be prevented by controlling the major risk factors through lifestyle modifications and diet interventions. Epidemiological evidence shows that consumption of dry beans (Phaseolus vulgaris L.) has positive effects on reducing blood LDL-cholesterol and lipid levels. These health benefits are mainly attributed to the high content of dietary fiber (DF) in beans. DF is carbohydrate polymers that are not hydrolyzed by the endogenous enzymes in humans. However, some of them (water-soluble DF) could increase viscosity and retain the absorption of bile salts (BS) in the small intestinal. The BS retention or the binding of BS could promote more cholesterol convert to BS (thus reduce cholesterol levels) and decrease lipid digestion. Therefore, due to the increased viscosity and BS retention ability of DF, dry beans could help to reduce the blood cholesterol and lipid levels and further help to prevent CVD. Moreover, different cooking and processing method could also affect the composition, microstructure and functional properties of foods. The purpose of this research was to determine how common beans and high hydrostatic pressure (HHP) (compared with hydrothermal (HT)) processing, a non-thermal processing, influence the ability of dry beans to retain bile salts and modulate lipid digestion in vitro. This study showed that severe HT treatment disrupted the bean cell wall integrity severally and reduced the BS retaining the efficiency of dry beans, while HHP treatment, produced minimally processed beans, improved the application properties of dry beans and maintained/enhanced BS-retention by dry beans. It also showed that the whole bean matrix (other than soluble DF) also contributes to retain BS and modulate lipid digestion, indicating the importance of retaining intact food structures. The integrity of bean structures through HHP treatment and less severe mechanical treatment could help to retain the ability of dry beans to reduce lipid digestion. These findings suggest that dry beans, with a high content of dietary fiber and resistant starch, have significant health benefits related to lowering cholesterol and lipid levels. Increasing the consumption of dry beans would definitely help to improve overall health. HHP, as a non-thermal processing technology, showed the potential to produce minimally processed bean products with enhanced health benefits and diverse application properties. This study could be extended through continuing research into the influence of beans on the gut microbiota, which are also closely related to the cholesterol and lipid metabolism regulation.

Subprodutos de frutas são rotineiramente descartados pelas indústrias. Porém, são ricos em compostos bioativos, podendo ser utilizados como ingredientes em produtos funcionais, promovendo a saúde e minimizando o impacto ambiental. O objetivo deste trabalho foi estudar o potencial funcional de subprodutos desidratados de acerola e de laranja, como fontes de compostos fenólicos, e desenvolver leites fermentados probióticos adicionados deste resíduo, avaliando suas características físico-químicas durante o armazenamento sob refrigeração (28 dias; 4 ± 1 °C), bem como o impacto das condições gastrointestinais sobre os flavonoides e as cepas probióticas. Os subprodutos foram obtidos em indústrias de processamento de frutas do estado de São Paulo, e foram realizadas as seguintes análises para caracterizá-los: composição centesimal, teores de vitamina C, minerais, fibras alimentares, compostos fenólicos totais e proantocianidinas, capacidade antioxidante in vitro e perfil cromatográfico de flavonoides (CLAE). Foram elaboradas quatro formulações de leites fermentados: F0 (controle), sem adição de resíduo de acerola (RA); F2, com 2% de RA; F5, com 5% de RA; F10, com 10% de RA. Adicionou-se a cultura probiótica ABT-4 nos produtos, constituída de duas cepas probióticas: Bifidobacterium animalis subsp lactis Bb-12 e Lactobacillus acidophilus La-5, além da cultura starter Streptococcus thermophilus. As seguintes análises foram realizadas com as formulações de leites fermentados, durante o armazenamento sob refrigeração (28 dias, 4 ± 1 °C): composição centesimal, pH, acidez, viabilidade dos microrganismos, teor de compostos fenólicos totais (CF), cor e textura instrumentais. Além disso, os leites fermentados foram submetidos a condições gastrointestinais simuladas in vitro, para avaliação do impacto na viabilidade das cepas probióticas e nos compostos fenólicos. O RA mostrou-se excelente fonte de vitamina C (605 mg/100 g b.u.), além de apresentar melhor capacidade antioxidante in vitro do que o RL. Proantocianidinas foram encontradas apenas no RA, na concentração de 617 &#181;g EC/g b.s. O teor de compostos fenólicos totais do RA (3240 &#181;g EAG/100 g b.s.) foi 3,6 vezes maior que o do RL. Os principais compostos fenólicos encontrados no RA foram: derivados de quercetina, procianidina B1, rutina, e derivados de caempferol. No RL, foram identificados: naringenina, sinensetina, homorientina, isovitexina e derivados de ácido clorogênico. Os subprodutos estudados apresentaram elevados teores de fibras totais (acima de 60%) e proteínas totais (RA: 10,4%; RL: 9,9%), além de reduzido teor de lipídeos totais (RA: 1,6%; RL: 2,6%). Os principais minerais identificados em ambos os resíduos foram: potássio, magnésio, cálcio e fósforo. Quanto às quatro formulações de leites fermentados, estas apresentaram baixo teor de lipídeos totais (menor que 1%), e o teor de proteínas totais variou entre 3,9 e 5,1 g/100 g, estando de acordo com a legislação vigente para este tipo de produto. O pH das formulações F0 (controle) e F2 manteve-se estável significativamente (p > 0,05) ao longo do período de armazenamento sob refrigeração (28 dias; 4 ± 1 °C), e das outras formulações sofreu pequena queda, mesmo assim mantendo-se acima de 4,5. A acidez das formulações, que variou entre 0,92 a 1,28 mg de ácido lático/g, aumentou entre os dias 1 e 14 de armazenamento, depois se manteve até o final da vida de prateleira. O RA não interferiu de maneira negativa nas populações de microrganismos analisadas durante o armazenamento, já que as formulações F2, F5 e F10 mantiveram suas populações em torno de 8 log UFC/g. Quanto ao teor de CF, as amostras diferiram significativamente entre si (p < 0,05), sendo que F0 apresentou teor em torno de 5 vezes inferior a F10 (21,13 e 101,13 &#181;g EAG/100 g, respectivamente, no dia 1). A cor dos produtos se manteve até o final da vida de prateleira, e diferiram significativamente (p < 0,05) entre si. O RA influenciou pouco nos parâmetros de textura dos leites fermentados, mas a formulação controle foi a única que perdeu adesividade. Após a fase gástrica da digestão simulada in vitro, no 7° dia de armazenamento, as populações de bactérias probióticas diminuíram drasticamente (quedas em torno de 3 a 5 log UFC/g), e após a fase entérica não foram detectadas contagens. Por outro lado, os flavonoides encontrados nos leites fermentados adicionados de RA aumentaram em torno de 2 a 5 vezes, após a fase gástrica, mantendo-se ou sofrendo pequena queda após fase entérica. Estes resultados mostram que o pó de subprodutos de acerola é um valioso ingrediente a ser utilizado em alimentos funcionais, pois é rico em vitamina C, fibras e compostos fenólicos, agregando valor nutricional, além de servir como antioxidante natural. Seus flavonoides parecem ser altamente resistentes aos ácidos e sais da digestão, podendo, assim, exercer efeitos positivos sobre a saúde. / Fruits by-products are routinely discarded by industries. However, they are rich in bioactive compounds, and can be used as ingredients in functional foods, promoting health and minimizing environmental impact. The objective of this study was to investigate the functional potential of acerola and orange dehydrated by-products, as sources of phenolic compounds, and to develop probiotic fermented milks suplemented with this residues, evaluating its physico-chemical characteristics during refrigerated storage (28 days, 4 ± 1 °C), as well as the impact of gastrointestinal conditions on flavonoids and probiotic strains. The by-products were obtained from fruit processing industries of São Paulo, and the following analyzes were performed to characterize them: contents of moisture, ash, lipids, proteins, vitamin C, minerals, dietary fibers, total phenolic compounds and proanthocyanidins, antioxidant capacity in vitro and flavonoids chromatographic profile (HPLC). Were elaborated four formulations of fermented milks: F0 (control), without addition of acerola residue (AR); F2, with 2% AR; F5, with 5% AR; F10, with 10% AR. Was used the probiotic culture ABT-4, composed of two probiotic strains, Bifidobacterium animalis subsp lactis Bb-12 and Lactobacillus acidophilus La-5, in addition to the starter culture Streptococcus thermophilus. During the refrigerated storage (28 days, 4 ± 1 °C), the following analyzes were performed with the fermented milks: contents of moisture, ash, lipids and proteins, pH, acidity, viability of microorganisms, total phenolic compounds (PC), instrumental color and texture. In addition, the fermented milks were submitted to in vitro simulated gastrointestinal conditions to evaluate the impact on the viability of probiotic strains and phenolic compounds. AR presented excellent content of vitamin C (605 mg/100 g), in addition to presenting better antioxidant capacity in vitro than orange residue (OR). Proanthocyanidins were found only in AR (617 &#181;g CE/g). The PC content of AR (3240 &#181;g GAE/100 g) was 3.6 higher than in OR. The phenolic compounds identified in AR were quercetin-3-rhamnoside, rutin and others quercetin derivatives, procyanidin B1 and kaempferol derivatives. In OR, were identified naringenin, sinensetin, homorientin, isovitexin and chlorogenic acid derivatives. The by-products studied showed high total fibers content (above 60%) and total proteins (AR: 10.4%, OR: 9.9%), as well as reduced total lipids content (AR: 1.6%; OR: 2.6%). Both residues showed high levels of potassium, calcium, magnesium and phosphorus. The four formulations of fermented milks presented low total lipids content (below 1%), and the total proteins content ranged from 3.9 to 5.1 g/100 g, being in agreement with the legislation. The pH of F0 (control) and F2 formulations remained stable (p > 0.05) throughout the refrigerated storage period (28 days, 4 ± 1 °C), and the other formulations showed a small decreased, even thus remaining above 4.5. The acidity of the formulations, ranging from 0.92 to 1.28 mg of lactic acid/g, increased between days 1 and 14 of storage, then remained until the end of shelf life. The AR did not negatively interfere in the populations of microorganisms analyzed during storage, since the formulations F2, F5 and F10 maintained their populations around 8 log CFU/g. Regarding PC content, the samples differed significantly (p < 0.05), with F0 being about 5 lower than F10 (21.13 and 101.13 &#181;g GAE/100 g, respectively, in the day 1). The instrumental color of the products remained until the end of shelf life, and differed significantly (p < 0.05) from each other. The AR influenced a little in the texture parameters of the fermented milks, but the control formulation was the only one that lost adhesiveness. After the gastric phase of the simulated digestion in vitro, on the 7th day of storage, the populations of probiotic bacteria decreased dramatically (of 3 to 5 log CFU/g), and after the enteric phase no colonies were detected. On the other hand, the flavonoids found in the fermented milks that were suplemented with AR increased from 2 to 5 times, after the gastric phase, maintaining or suffering small decreased after enteric phase. These results show that acerola by-products powder is a valuable ingredient to be used in functional foods because it is rich in vitamin C, dietary fibers and phenolic compounds, adding nutritional value, and serving as a natural antioxidant. Its flavonoids appear to be highly resistant to the acids and salts of digestion and can thus have positive effects on health.