A diagnosis of coeliac disease (CD) in adults relies on the presence of a structurally abnormal intestinal mucosa, followed by a clear clinical remission on a gluten-free diet. There is a clear need for a rapid, simple, safe and sensitive method to determine the type and intensity of inflammation in the gut mucosa in clinical practice. The overall aims of our studies were to develop and evaluate a new technique, “the mucosal patch technique”, to characterize rectal local inflammatory process after rectal food challenge in patients with CD<b>. In study 1</b> we evaluated the potential of the new technique. The technique was well tolerated and easily applied. Pronounced neutrophil and eosinophil involvement in ulcerative colitis (UC) was demonstrated. With the high sensitivity of the technique, low-degree mucosal neutrophil activation could also be quantified in patients with collagen colitis,UC in clinical remission and in patients with irritable bowel syndrome. <b>In study 2 and 3</b> the aim was to elucidate the dynamics of the rectal inflammatory response and nitric oxide (NO) production after rectal gluten challenge. We found a pronounced neutrophil activation in coeliac patients after rectal gluten challenge. This activation was apparent 4 hours after challenge and remains for at least 48 hours. A more modest eosinophil activation started 1-2 hours later and remained at least for 48 hours. The biphasic pattern of neutrophil and eosinonphil activation after challenge suggests a biphasic inflammatory reaction. The activation of neutrophils and eosinophils precedes a pronounced enhancement of mucosal NO production. Some of our coeliac patients displayed signs of an inflammatory reaction after rectal corn gluten challenge. <b>In study 4</b> the aim was to investigate the local inflammatory reaction to gluten and cow’s milk protein in CD patients in remission. The findings indicate that not only gluten sensitivity but also cow’s milk (CM) protein sensitivity is common in CD. The data support the hypothesis that CM sensitivity may contribute to persistent symptoms in coeliac patients on gluten-free diet.

The quality of food wheat is annually evaluated with all deliveries of agricultural primary production in the chosen farming enterprise. During the years 2008 - 2012 particular characteristics with files of 42 - 222 samples were determined, except for the year of 2009 when only one supply was technologically incorporated to food wheat. Countrywide observation of the quality of food wheat is executed in Agricultural research institute Kroměříž, Ltd and in Agrotest fyto, Ltd. It is co-financed by Ministry of Agriculture within a functional task which follows the project of Ministry of Agriculture number QG50041 Factors of quality and safety with food grains (2005-2010). Harvest samples, obtained from producers in time span 2008-2012, were represented by files of 500-1035 samples. Characteristics which influence the quality of the grains as well as criteria and methods of the evaluation of food wheat were described. Their applications in Agrotest fyto, Ltd and under operating conditions in ZZN Pelhřimov a.s. were given. Average values of observed parameters were calculated from the results of each testing in the laboratory of the detached post Záhoří and consequently compared with countrywide norm and the norm of South Bohemia. The quality of food wheat grains in years 2008-2012 showed a large-scale variability in particular parameters both at countrywide level and in South Bohemia as well as in chosen farming enterprise. Final quality was considerably influenced by course of weather annually.

Made available in DSpace on 2014-07-29T15:22:54Z (GMT). No. of bitstreams: 1 Dissertacao Thaisa Anders Carvalho Souza.pdf: 2450969 bytes, checksum: 96a1e35514fbf17aedfa8fc3705b6ae4 (MD5) Previous issue date: 2011-02-28 / This study aimed to verify the microbiological safety of solid waste from cassava starch (peel and bagasse), with different times of storage at room temperature with and without chemical treatment and apply peels and dried cassava bagasse in the formulation of gluten-free cake mixes substituting rice flour. The pH values ranged from 5.3 to 4.3 in fresh cassava peel, while the cassava peel sanitized between 5.6 and 4.4. The pH values ranged from 5.24 to 5.92 on fresh cassava bagasse, while in the acidified cassava bagasse these values were between 4.7 and 5.5. The time significantly influenced the pH values of all samples of peel and cassava bagasse during storage at room temperature. None of the samples of flour and cake analyzed was detected Salmonella, and counting of Bacillus cereus, Clostridium sp. and fecal coliform (45 °C). There were only counts of coliforms, yeasts and molds in peel flour and cassava bagasse flour. All instrumental parameters of color, pH and moisture of the samples of rice flour, peel flour and cassava bagasse flour, showed differences. The rice flour had become clearer, with less red and yellow color when compared to the peel flour and cassava bagasse flour. The pH differed among the three flours, being the pH of the rice flour larger than those observed in flours of peel and cassava bagasse. The moisture content of rice flour was lower than the peel flour and cassava bagasse flour. The flours of peel and cassava bagasse showed higher contents of ash, total dietary fiber, soluble and insoluble fiber and lower protein and carbohydrates, in addition to the total energy. The specific volumes of experimental cakes made with rice flour and cassava peel flour did not differ in any treatment, already in those made with cassava bagasse flour, treatments with 75% and 100% substitution of rice flour by cassava bagasse flour differed from other treatments. The crumbs of cakes tended to stain darker with increasing levels of replacement of rice flour by peel flour and cassava bagasse flour. In relation to moisture all the cakes made with experimental flours were different, except the cake without replacement and with 25% substitution of rice flour with cassava bagasse flour. The performance index also tended to increase proportionately to the increase levels of replacement of rice flour by flour of peel and cassava bagasse flour. The ash, lipids and total and insoluble dietary fibers tended to increase in the cakes with replacement of rice flour by peel flour and cassava bagasse flour, carbohydrates, protein and total energy were lower in cakes with increasing replacement of flour waste. All experimental cakes exceeded the cutoff point (score 6) for acceptance testing in sensorial analysis. Cassava peel sanitized and acidified cassava bagasse can be considered safe for use as an ingredient (flour) in the formulation of food products, especially those who suffer heat treatment. The results obtained in the formulations of the cakes were adequate, since they showed satisfactory results in relation to the physical, nutritional, microbiological safety and sensory characteristics. / Este trabalho objetivou verificar a segurança microbiológica dos resíduos sólidos de fecularia de mandioca (cascas e bagaço), com diferentes tempos de armazenamento a temperatura ambiente com e sem tratamento químico e aplicar as cascas e bagaço de mandioca desidratados na formulação de misturas de bolos sem glúten em substituição a farinha de arroz. Os valores de pH variaram entre 5,3 a 4,3 na casca de mandioca in natura, enquanto na casca de mandioca sanitizada entre 5,6 e 4,4. Os valores de pH variaram entre 5,24 a 5,92 no bagaço de mandioca in natura, enquanto no bagaço de mandioca acidificado estes valores ficaram entre 4,7 e 5,5. O tempo influenciou significativamente os valores de pH de todas as amostras de casca e de bagaço de mandioca durante o armazenamento a temperatura ambiente. Em nenhuma das amostras de farinhas e de bolos avaliadas foi detectada presença de Salmonella sp, e contagem de Bacilus cereus, Clostridium sp. e coliformes termotolerantes (45 °C). Apenas verificaram-se contagens de coliformes totais e bolores e leveduras nas farinhas de cascas e de bagaço de mandioca. Todos os parâmetros instrumentais de cor, o pH e a umidade das amostras de farinha de arroz, farinha de casca de mandioca e farinha de bagaço de mandioca, apresentaram diferenças. A farinha de arroz apresentou-se mais clara, com coloração menos avermelhada e amarelada, quando comparada às farinhas de casca e de bagaço de mandioca. O pH diferiu entre as três farinhas, sendo o pH da farinha de arroz maior que o observado nas farinhas de casca e de bagaço de mandioca. A umidade da farinha de arroz foi menor que as das farinhas de casca de mandioca e de bagaço de mandioca. As farinhas de casca e de bagaço de mandioca apresentaram teores superiores de cinzas, fibras alimentares total, solúvel e insolúvel e inferiores de proteínas e carboidratos, além do valor energético total. Os volumes específicos dos bolos experimentais formulados com farinha de arroz e farinha de casca de mandioca não diferiram em nenhum dos tratamentos, já nos formulados com farinha de bagaço de mandioca, os tratamentos com 75% e 100% de substituição da farinha de arroz pela farinha de bagaço de mandioca diferiram dos demais tratamentos. Os miolos dos bolos apresentaram tendência à coloração mais escura, com o aumento dos níveis de substituição da farinha de arroz pela farinha de casca e de bagaço de mandioca. Em relação à umidade todos os bolos experimentais formulados com as farinhas apresentaram diferença, exceto o bolo sem substituição e com 25% de substituição de farinha de arroz por farinha de bagaço de mandioca. O índice de rendimento também apresentou tendência a aumentar proporcionalmente ao aumento dos níveis de substituição de farinha de arroz por farinha de casca e de bagaço de mandioca. As cinzas, os lipídeos e as fibras alimentares total e insolúvel apresentaram tendência de aumentar nos bolos com substituição de farinha de arroz pela farinha de casca e de bagaço de mandioca, já os carboidratos, proteínas e o valor energético total apresentaram valores inferiores nos bolos com o aumento da substituição das farinhas dos resíduos. Todos os bolos experimentais superaram o ponto de corte (escore 6) para o teste de aceitação na análise sensorial. A casca de mandioca sanitizada e o bagaço de mandioca acidificado podem ser considerados seguros para utilização como ingrediente (farinha) em formulações de produtos alimentícios, principalmente aqueles que sofrerão tratamento térmico. Os resultados obtidos nas formulações dos bolos foram adequados, uma vez que estes apresentaram resultados satisfatórios em relação às características físicas, valor nutricional, segurança microbiológica e características sensoriais.

Este trabalho teve o objetivo de avaliar o efeito da adição da transglutaminase microbiana (MTGase) na fabricação de pão de forma, através do desenvolvimento de formulação ideal, com combinações de aditivos e enzima, e da avaliação do efeito da enzima nas proteínas, na massa crua, na massa após a fermentação e no produto final. Para comparar a qualidade dos pães produzidos com ou sem enzima, foram testadas três formulações: a básica, livre de aditivos (pão Zero); com a adição de emulsificante e ácido ascórbico (pão Controle); e a preparada com a formulação básica adicionada de enzima (pão MTGase). A avaliação da qualidade dos pães foi feita por meio de medidas físicas e instrumentais. A análise de textura foi realizada pelo método TPA (Texture Profíle Analysis), cujas respostas de firmeza, elasticidade, mastigabilidade e gomosidade podem ser correlacionadas com análises sensoriais. Paralelamente, de amostras de farinha, de massa e de pão foram obtidas as frações protéicas de gliadinas, gluteninas e os resíduos de extração. As gliadinas e as gluteninas foram analisadas por cromatografia líquida de alta eficiência em fase reversa e por eletroforese em gel de poliacrilamida contendo SDS. Os resultados de volume e de firmeza dos diferentes pães apresentaram diferenças significativas a nível de 5%, em que as respostas do pão MTGase foram melhores que as do pão Zero, porém ainda inferiores às do Controle. A melhor formulação foi obtida por meio de um planejamento composto central, com variações nas concentrações de emulsificante, ácido ascórbico e enzima, com os resultados avaliados pela metodologia de superfície de resposta. Exceto para a coesividade, todos os outros parâmetros de volume, dureza (TA), firmeza, elasticidade, mastigabilidade e gomosidade (TPA) apresentaram resultados positivos pela ação da transglutaminase a 0,6%, combinada com 0,2 % de emulsificante e 70 ppm de ácido ascórbico. Os resultados sugerem que a enzima foi capaz de modificar as propriedades químicas das proteínas, o comportamento reológico da massa e as propriedades funcionais do pão, melhorando a força da massa, a textura e o volume dos pães. As análises das frações gliadínicas apresentaram cerca de 3% de Nitrogênio total, em base seca, e as frações glutenínicas apresentaram entre 2 e 5% de Nitrogênio total. Os perfis cromatográficos e eletroforéticos dessas frações sugerem que as gliadinas não foram afetadas pela presença da enzima, que envolveram sobretudo as gluteninas. O conjunto de resultados indica que a aplicação de MTGase em associação com aditivos convencionais pode ser uma alternativa à panificação, embora os mecanismos de sua ação na massa não estejam completamente esclarecidos. / The application of microbial transglutaminase on weak gluten flour used in breadmaking was studied over the process. To verify the enzyme effects, three formulations were tested: Base formulation, characterized by the absence of enzyme and emulsifying agents; Control formulation, comprised by the presence of emulsifying agents and ascorbic acid and MTGase formulation, with the enzyme. Samples of flour, dough and bread were analyzed. The effect of enzyme on bread quality was estimated by parameters of Texture Analysis, Texture Profile Analysis and specific volume. The protein contents from those samples were determined by the total nitrogen in glutenin and gliadin fractions, that were also analyzed by RP-HPLC (reversed phase high performance liquid chromatography) and by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Although the MTGase bread did not reach the same quality parameters as those achieved by the Control samples, it showed as an alternative formulation to reduce the quantity of emulsifying agents and ascorbic acid as compared to the Control. The results indicate that the enzyme modified chemical and functional properties of glutenin fraction, improving dough strength and bread volume. Results of total nitrogen content, and electrophoretic and chromatographic profiles of the protein fractions suggest that while glutenin proteins were modified by enzyme, gliadin proteins were not affected.

Foram conduzidos dois experimentos no Departamento de Zootecnia da ESALQ/USP com o objetivo de estudar a substituição do milho moído fino pelos co-produtos farelo de glúten de milho e polpa cítrica peletizada em rações para bovinos terminados em confinamento. No Experimento 1 foram utilizados 88 machos cruzados (½ Braunvieh ¼ Angus ¼ Nelore) não castrados (407 kg), distribuídos em 24 baias por 57 dias. As rações continham 12% de bagaço de cana in natura e 88% de concentrado. Os tratamentos foram: (1) milho moído fino (M); (2) milho moído fino e farelo úmido de glúten de milho (MFUG); (3) polpa cítrica peletizada (P); (4) polpa cítrica peletizada e farelo úmido de glúten de milho (PFUG). O GPD e a EA dos animais não diferiram entre os tratamentos (P>0,05), porém houve efeito da fonte energética sobre a IMS, que foi maior para as rações com milho (P<0,05). Não foi observado efeito de fonte energética ou nível de farelo úmido de glúten de milho nos valores de ELm e ELg observados neste experimento (P>0,05), mas houve interação (P<0,05) com valores de energia mais altos para o tratamento PFUG em relação aos demais. No Experimento 2, foram utilizados 99 machos Nelore não castrados (348 kg), distribuídos em 20 baias por 85 dias. Os tratamentos foram: (1) milho moído fino (M); (2) milho moído fino e polpa cítrica peletizada (MPC); (3) milho moído fino e farelo úmido de glúten de milho (MFUG); (4) polpa cítrica peletizada e farelo úmido de glúten de milho (PFUG); (5) polpa cítrica peletizada e farelo seco de glúten de milho (PFSG). As rações continham de 5 a 11% de feno de gramínea como fonte de volumoso, formuladas para serem isoprotéicas. Não houve diferença (P>0,05) na IMS, GPD e EA para os tratamentos utilizados. A área de olho de lombo e espessura de gordura não foram afetadas pelos tratamentos, bem como as características qualitativas de carne (P>0,05). Acompanhando os resultados de desempenho, o valor energético das rações não diferiram entre si (P>0,05). Através dos resultados de ELm e ELg observados/esperados, concluiu-se que o milho brasileiro tem seu valor energético inferior ao milho descrito pelo NRC (1996). Os valores de energia do milho (ELm e ELg de 2,24 e 1,55 Mcal/kg) preditos através do amido fecal são semelhantes aos valores tabulados para o milho quebrado americano e superiores ao do milho moído. Com os resultados obtidos é possível afirmar que o farelo de glúten de milho e a polpa cítrica peletizada podem ser utilizados em substituição parcial ao milho moído fino e quando combinados, em substituição total ao milho moído fino em rações com altos teores de concentrado para bovinos em terminação, sem prejuízos ao desempenho e características de carcaça. / Two experiments were conducted at ESALQ/USPs Animal Science Department to evaluate corn gluten feed and dry citrus pulp in substitution for fine ground corn in high concentrate diets for finishing feedlot cattle. In Experiment 1, 88 croosbred (½ Braunvieh ¼ Angus ¼ Nelore) bulls (BW = 407 kg) were kept in 24 pens for 57 days. The rations contained 12% sugarcane bagass and 88% concentrate. The treatments were: (1) fine ground corn (M); (2) fine ground corn and wet corn gluten feed (MFUG); (3) citrus pulp pellets (P); (4) citrus pulp pellets and wet corn gluten feed (PFUG). ADG and G:F ratio did not differ among treatments (P>0,05), but there was an energetic source effect on DMI, higher for corn based diets (P<0,05). No differences were observed for energy souce or corn gluten feed level for NEm and NEg on this experiment (P>0,05), but an interaction (P<0,05) was observed for PFUG wich had higher energy value then the other treatments. In Experiment 2, 99 Nelore bulls (BW = 348 kg) were kept in 20 pens for 85 days. The treatments were: (1) fine ground corn (M); (2) fine ground corn and citrus pulp pellets (MPC); (3) fine ground corn and wet corn gluten feed (MFUG); (4) citrus pulp pellets and wet corn gluten feed (PFUG); (5) citrus pulp pellets and dry corn gluten feed (PFSG). The diets had 5 to 11% grass hay and 89 to 95% concentrate. No differences (P>0,05) were observed for DMI, ADG and G:F ratio among treatments. Diets energy value showed no differences among treatments (P>0,05). Observed/expected NEm and NEg values led to the conclusion that brazilian corn grain has lower energy value than what NRC (1996) tables show. Corn energy values (NEm and NEg was 2,24 and 1,55 Mcal/kg) predicted from fecal starch were similar to american cracked corn and higher than ground corn. Wet corn gluten feed and citrus pulp can be used in substitution for fine ground corn, alone or in combination, in high concentrate finishing feedlot diets, without depressing animal performance and carcass charateristics.

The effect of gluten-free diet on β-cell residual capacity, immune function and gut microbiome in children with newly diagnosed type 1. diabetes Abstract The pathophysiology of the onset and progression of type 1 diabetes (T1D) is not fully understood. Gluten has a proinflammatory effect on the immune system and is therefore considered as one of the factors affecting the onset and progression of T1D. The aim of the thesis is to allow a complex insight into the role of the GFD on the residual β-cell capacity, T1D control, gut microbiome, gut permeability, subtypes of immune cells and the effect of gut microbiome transfer into germ-free non-obese diabetic (NOD) mice on the incidence of diabetes. On the group of 45 children with T1D (26 intervention group, 19 control group) we proved the association of the GFD with slower decrease of β-cell residual capacity (the difference in the trend of C-peptide decrease 409 pmol/l/year; p = 0,04) and lower HbA1c (by 7,8 mmol/mol; p=0,02). We also described the changes in the gut bacteria that were differentially abundant after the administration of the GFD and the changes in abundance of the regulatory and effector immune cells. We showed there was no change in the gut permeability with respect to the study group. We also proved that the transfer of human gut microbiota...

Diploma thesis is focused on Slovak company Nutribak s.r.o. and its potential business activities on the Czech market, especially with gluten-free goods. The created methodology is based on a critical search of current literature related to the the field of marketing, management and trade. Suggested steps are applied to the company Nutribank s.r.o. according to results of the analysis of the current situation, there were created certain steps, that will provide the company with a plan for evolving their business activities and also supporting their position in the Czech market

[ES] La presente tesis doctoral titulada "New nanostructured suports with signal amplification features for the detection of molecules and biomolecules of interest" se centra en el diseño y preparación de nuevos materiales híbridos orgánicos-inorgánicos constituidos por puertas moleculares soportadas sobre alúmina mesoporosa con el objetivo de desarrollar nuevos sistemas sensores con aplicaciones potenciales en el campo de la diagnosis y del control alimentario. En el primer capítulo de la tesis se introducen los conceptos en los que están basados los estudios realizados y los materiales preparados. A continuación, en el segundo capítulo se describen los objetivos generales de la tesis que serán abordados en los siguientes apartados. En el tercer capítulo se presenta el diseño y optimización de un nanodispositivo para la detección de la bacteria Mycoplasma fermentans. En primer lugar, los poros de una placa de alúmina mesoporosa se cargan con un indicador fluorescente (rodamina B). Seguidamente, la superficie es funcionalizada con una secuencia de ADN complementaria a una región altamente conservada de la subunidad ribosomal 16S de la bacteria Mycoplasma fermentans. El impedimento estérico generado por las secuencias de ADN ancladas al exterior de los poros impide la salida del indicador encapsulado. Únicamente en presencia de DNA de la bacteria Mycoplasma fermentans, se produce la apertura de los poros permitiéndose la difusión de la carga (rodamina B) que es posteriormente medida mediante espectroscopía de fluorescencia. En el capítulo cuatro se diseña de un nanodispositivo capaz de detectar de forma rápida, sensible y selectiva la bacteria Staphylococcus aureus. Para la preparación del material sensor, un soporte de alúmina mesoporosa es, en primer lugar, cargado con el indicador fluorescente rodamina B. A continuación, los poros del soporte son tapados mediante el anclaje de un aptámero que reconoce de forma específica la bacteria. Solamente en presencia de Staphylococcus aureus se produce la liberación del indicador encapsulado, que es posteriormente medido mediante espectroscopía de fluorescencia. Además, la respuesta obtenida es específica para Staphylococcus aureus. Este sistema ha sido ensayado en muestras reales. En el sexto capítulo, diseña un nanodispositivo híbrido orgánico-inorgánico consistente en un material de alúmina mesoporosa cubierto con una secuencia de ADN específica para la detección de ADN del hongo Pneumocystis jirovecii. En este caso, el soporte de alúmina cargado con rodamina B se recubre con una secuencia de ADN específica para el reconocimiento de este hongo. En presencia del organismo, la horquilla hibrida con el ADN del hongo, lo que resulta en una conformación triplex con elevada afinidad y estabilidad que induce, al mismo tiempo, el desplazamiento de este complejo de la superficie. Como consecuencia de este reconocimiento la carga se libera y es cuantificada mediante espectroscopía de fluorescencia. El sistema ha sido satisfactoriamente validado. En el séptimo capítulo, se diseña un sistema sensor con la capacidad de detectar gluten de forma rápida y sencilla en extractos de alimentos procesados y no procesados. Para ello, un soporte de alúmina mesoporosa se carga con rodamina B y los poros se recubren con un aptámero específicamente diseñado para la detección de la proteína gliadina, que constituye el 50 % del total del clúster de elementos que forman el gluten. La elevada afinidad y especificidad entre el aptámero y la proteína en cuestión hacen que en presencia de ésta se produzca un desplazamiento de la puerta molecular que permite la difusión del colorante encapsulado que es finalmente monitorizado mediante espectroscopía de fluorescencia. Finalmente, en el capítulo octavo se discuten de forma conjunta los resultados obtenidos en los capítulos anteriores y la potencial aplicación de los sistemas desarrollados en el actual sistem / [CA] La present tesi doctoral, titulada "New nanostructured supports with signal amplification features for the detection of molecules and biomolecules of interest", es centra en el disseny i preparació de nous materials híbrids orgànics-inorgànics constituïts per portes moleculars suportades sobre alúmina mesoporosa amb l'objectiu de desenvolupar nous sistemes sensors amb potencials aplicacions en el camp de la diagnosi i del control alimentari. En el primer capítol de la tesi s'introdueixen els conceptes en què estan basats els estudis realitzats i els materials preparats. A continuació, en el segon capítol es descriuen els objectius generals de la tesi que seran abordats en els següents apartats. En el tercer capítol es presenta el disseny i optimització d'un nanodispositiu per a la detecció de la bactèria Mycoplasma fermentans. Primerament, els porus d'una placa d'alúmina mesoporosa són carregats amb un indicador fluorescent (rodamina B). Seguidament, la superfície és funcionalitzada amb una seqüència d'ADN complementaria a una regió altament conservada de la subunitat ribosomal 16S de la bactèria Mycoplasma fermentans. L'impediment estèric generat per les seqüències d'ADN ancorades a l'exterior dels porus impedeix l'alliberament de l'indicador encapsulat. Únicament en presencia d'ADN de la bactèria Mycoplasma fermentans, es produeix l'obertura dels porus permetent la difusió de la càrrega (rodamina B) que és posteriorment mesurada mitjançant fluorescència. En el capítol quatre es dissenya un nanodispositiu capaç de detectar de forma ràpida, sensible i selectiva la bactèria Staphylococcus aureus. Per a la preparació del material sensor, el suport d'alúmina mesoporosa és, primerament, carregat amb l'indicador fluorescent rodamina B. A continuació, els porus del suport són tapats mitjançant l'ancoratge d'un aptàmer que reconeix de forma específica a la bactèria. Solament en presència de Staphylococcus aureus es produeix l'alliberament de l'indicador encapsulat, que és posteriorment mesurat mitjançant espectroscòpia de fluorescència. A més a més, la resposta obtinguda és específica per Staphylococcus aureus. Aquest sistema ha sigut validat amb mostres reals de pacients. En el sisè capítol, es dissenya un nanodispositiu híbrid orgànic-inorgànic consistent en un material d'alúmina mesoporosa cobert amb una seqüència d'ADN específica per a la detecció de l'ADN del fong Pneumocystis jirovecii. En aquest cas, el suport d'alúmina carregat amb l'indicador fluorescent rodamina B és recobert amb una seqüència d'ADN específica per al reconeixement d'aquest fong. En presència de l'organisme, la forquilla hibrida amb l'ADN del fong, resultant en una conformació triplex amb elevada afinitat i estabilitat, que indueix, al mateix temps, el desplaçament d'aquest complex de la superfície. Com a conseqüència d'aquest reconeixement la càrrega és alliberada i quantificada mitjançant espectroscòpia de fluorescència. El sistema ha sigut validat com a mètode diagnòstic mitjançant l'anàlisi de mostres reals de pacients. En el seté capítol, es dissenya un sistema sensor amb la capacitat de detectar gluten de forma ràpida i senzilla en extractes d'aliments processats i no processats. Per a això, un suport d'alúmina mesoporosa es carrega amb indicador fluorescent rodamina B i posteriorment és recobert amb un aptàmer específicament dissenyat per a la detecció de la proteïna gliadina, que constitueix el 50 % del total del clúster d'elements que formen el gluten. L'elevada afinitat i especificitat entre l'aptàmer i la proteïna en qüestió fa que en presència d'aquesta es produesca un desplaçament de la porta molecular que permet la difusió de la càrrega encapsulada i que serà finalment monitoritzada mitjançant espectroscòpia de fluorescència. Finalment, en el capítol vuité es discuteixen de manera conjunta els result / [EN] The PhD thesis hereby presented and entitled "New nanostructured supports with signal amplification features for the detection of molecules and biomolecules of interest", focuses in the design and preparation of new hybrid organic-inorganic materials constituted by molecular gates supported over mesoporous alumina with the aim of developing new sensor probes of potential applications in the fields of diagnosis and food control. In the first chapter, the concepts in which studies and prepared materials are based, are introduced. Next, the second chapter describes the general objectives of this thesis, which will be approached in the following sections. In the third chapter, it is presented in detail the design and optimization process of a nanodevice applied for the detection of Mycoplasma fermentans bacterium. First of all, mesoporous alumina porous films are charged with a fluorescent indicator (rhodamine B). Then, the surface is functionalized with a DNA sequence complementary to a highly conserved region of the 16S ribosomal subunit of the bacterium Mycoplasma fermentans. Steric hindrance generated by DNA sequences on the surface inhibits the release of the encapsulated indicator. Only in the presence of bacterium Mycoplasma fermentans DNA, molecular gates open, allowing payload diffusion to the solution, which is measured by fluorescence spectroscopy. In chapter four, it is carried out the design and optimization of a nanodevice able to detect Staphylococcus aureus bacterium in a fast, sensitive and selective way. For the sensor preparation, alumina mesoporous support is, first, loaded with the rhodamine B fluorescent dye. Then, the mesoporous are blocked through the attachment of an aptamer that recognises specifically this bacterium. Exclusively in the presence of Staphylococcus aureus it is accomplished the release of the encapsulated dye, which is later monitored by fluorescence spectroscopy. The response obtained is specific for Staphylococcus aureus. This system has been validated in real samples. In the sixth chapter, it is detailed the design and optimization process of a hybrid organic-inorganic nanodevice based on a capped mesoporous alumina material for the detection of Pneumocystis jirovecii fungus DNA. In this case, the mesoporous alumina support is loaded with a fluorescent dye and decorated with a specific oligonucleotide sequence designed for the recognition of Pneumocystis fungus. In the presence of the target organism, the fork-like oligonucleotide hybridises with the DNA of the fungus, which results in the adoption of a triplex conformation with high affinity and stability that induces, at the same time, the displacement of this complex from the surface. Consequently, the payload diffused to the solution is quantified through fluorescence spectroscopy. The system has been successfully validated. In the seventh chapter, it was developed a sensor system for gluten detection, in a quick and easy way, in processed and non-processed food extracts. For this, a mesoporous alumina support is loaded with the fluorescent dye rhodamine B, and later was functionalized with an aptamer specifically designed for the detection of gliadin, a protein that constitutes 50 % of average cluster elements that forms gluten. The protein-aptamer high affinity and specificity induce the displacement of the capping aptamer and cargo delivery, which is monitored through fluorescence spectroscopy. Finally, in the eighth chapter, the results obtained in the previous chapters and the potential application of the systems developed as health and food control system are discussed. / We thank the Spanish Government projects MAT2015-64139-C4-1-R, AGL2015-70235-C2-2-R, and TEC2015-71324-R (MINECO/FEDER, UE), the Generalitat Valenciana (project PROMETEOII/2014/047), the Catalan authority (project AGAUR 2014SGR1344), and ICREA under the 2014 ICREA Academia Award for support. This study was supported by the Spanish Government projects RTI2018-100910-B-C41 and SAF2017-82251-R (MCUI/AEI/FEDER, UE), the Generalitat Valenciana (project PROMETEO/2018/024), the Universitat Politècnica de València−Instituto de Investigación Sanitaria La Fe (B02-MIRSA project), CIBER-BBN (NANOPATH and valorization project CANDI-EYE) and co-financed by the EU through the Valencian Community ERDF PO 2014-2020. This research was funded by the Spanish Government, projects RTI2018-100910-B-C41 (MCUI/AEI/FEDER, UE) and CTQ2017-84415-R / Pla Blasco, L. (2021). New nanostructured supports with signal amplification features for the detection of molecules and biomolecules of interest [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/166500

Tortilla sales are projected to exceed 9.5 billion by 2014. However, currently no wheat cultivars have been identified that possess the intrinsic quality attributes needed for the production of optimum quality tortillas. Tortillas made with refined wheat flour low in dietary fiber (DF) are popular in the United States due to their sensory properties. This study explored the use of wheat lines (WL) possessing variations in high molecular weight glutenin allele sub-units (HMW-GS) for production of tortillas and also investigated the use of corn based resistant starches (RS), type II (RS2) and wheat based RS type IV (RS4) to increase DF in tortillas. Tortillas were made with 0-15 percent RS and 100 percent whole white wheat (WW). Flour protein profiles, dough, and tortilla properties were evaluated to determine the effects of the allelic variations and RS substitution on tortilla quality. Sensory properties of tortillas with RS were determined. Variations in HMW-GS composition significantly affected the protein quality and tortilla properties. Flour from WL possessing allelic combinations (2*, 17+18, 7, 2+12), (1, 17+18, 5+10), (2*, 17, 2+12) and (1, 2*, 17+18, 2+12) had 12.8-13.3 percent protein. These WL had extensible doughs and produced large diameter tortillas with superior (greater than or equal to 3.0) flexibility after 16 days compared to control. However, WL with (17+18 and 5+10) and (2*, 17+7, 5) produced extensible doughs, large, but less flexible, tortillas compared to control. WL with (2*,17+18,5+10) and (1,2*,7+9,5+10) produced smaller diameter tortillas, but with superior flexibility compared to control. RS2, WW, and cross-linked-pre-gelatinized RS4 (FiberRite) produced hard, less-extensible doughs and thinner tortillas compared to control, due to high water absorption. Cross-linked RS4 (Fibersym) dough and tortillas were comparable to control. 15 percent of RS2 and RS4 increase DF in control to 6 and 14 percent respectively, compare to control (2.8 percent DF). WW tortillas were less acceptable than control in appearance, flavor and texture, while tortillas with 15 percent Fibersym had higher overall acceptability than control. RS2 negatively affected dough machinability and tortilla shelf stability. However, 15 percent RS4 improved the DF in refined flour tortillas to meet FDA's "good source of fiber claim," without negatively affecting dough/tortilla quality.

The Master's thesis focuses on international expansion process of a company. For the application part of the thesis I have chosen a company called Žatec brewery Ltd. which belongs into a category of small and medium enterprises. The main objectives of the thesis comprise selection of a convenient Western Balkans region market for expansion of the brewery and subsequent formulation of a specific market entry strategy. The market entry strategy consists of a market entry mode selection, competitive strategy and marketing strategy as well as marketing mix 4P. The minor objectives include characteristics of the brewery's business strategy and the role of international expansion in it and also the company's financial analysis whose results underpine its expansion possibilities. The other minor objectives comprise multi-criteria decision model creation in order to decide for the most attractive target market and the expansion key success factors identification. As for the methods used, analyzes like financial analysis, PEST analysis, Porter's model of five forces and SWOT analysis, and then comparisons dominate. Croatia was finally identified as the most suitable regional market for the brewery's expansion and the company was advised to enter it with its gluten-free lager product. It was suggested, at the same time, that the brewery use to its advantage the background provided by its current owner - beer brewing company Carlsberg.