The diploma thesis deals with the relationships and opinions of students of selected secondary schools, which are connected with alternative ways of eating. The theoretical part contains basic information about rational nutrition, individual nutrients, there is a comparison of the food pyramid and a healthy plate. Last but not least, alternative diets are also clarified and then information on five specific types of alternative diets is given. In the individual subchapters, vegetarianism, veganism, raw food, paleo diet and a gluten-free diet are mentioned. The aim of the practical part is to find out the opinion of students of selected secondary schools on alternative ways of eating. These are my chosen secondary schools in the Rakovnicko region. A questionnaire survey is used as a research tool. Based on the data obtained, the questionnaire was evaluated and the results were converted into tables and graphs, which are always commented. 187 respondents answered the questionnaire. It was found that 19 of all respondents have nutrition-related health restrictions. The surprising result is that students did not learn about alternative meals primarily at school, but from their own resources (friends, family, etc.). The results of the questionnaire survey point to the fact that secondary school students...

Weißbüschelaffen (WBA) sind wissenschaftlich häufig genutzte Modelltiere für diverse Humanerkrankungen. Zur Gesunderhaltung dieser Primaten sind grundlegende diagnostische Blutparameter unverzichtbar. Bisher erhobene Daten zeichneten sich jedoch durch große Divergenz aus. Ob Veränderungen in Haltungsbedingungen einen Einfluss auf diese Blutparameter nehmen, ist bis heute unklar. Somit war ein Ziel dieser Arbeit die Erhebung aktueller hämatologischer und klinisch-chemischer Blutparameter von WBA. Zudem wurde der Einfluss der routinemäßigen Umsetzung in eine neue Behausung auf die erhobenen Parameter sowie den Kortisolspiegel im Kot untersucht. Des Weiteren leiden WBA in menschlicher Obhut rezidivierend an gastrointestinalen Erkrankungen, die mittels klinischer Standardparameter allein nicht diagnostizierbar sind. Dabei spielt vor allem die Sensitivität gegenüber Futtermittelinhaltsstoffen (z. B. Gluten) eine Rolle, welche ursächlich im Zusammenhang mit dem Wasting Marmoset Syndrome (WMS) diskutiert wird. Im zweiten Teil der vorliegenden Arbeit sollten deshalb die gastrointestinalen Erkrankungen von in menschlicher Obhut lebenden WBA ätiologisch beleuchtet werden, vor allem hinsichtlich einer möglichen Sensitivität gegenüber Gluten. Im ersten Teil dieser Studie wurden von 54 WBA hämatologische und klinischchemische Richtwerte erhoben. Die ermittelten hämatologischen Blutrichtwerte ähneln denen aus den achtziger Jahren, die Daten der klinischen Chemie nur bedingt: Die Richtwertbereiche von Laktatdehydrogenase, Alaninaminotransferase, Lipase sowie Alkalische Phosphatase und Gesamtbilirubin weichen von den ehemals erhobenen Daten ab. Zudem wurden in der vorliegenden Arbeit geschlechtsabhängige Unterschiede ermittelt: Weibliche Tiere wiesen signifikant höheres mittleres Erythrozytenvolumen und mittleren Hämoglobingehalt des Einzelerythrozyten auf als männliche Tiere, wohingegen bei diesen ein signifikant höheres Gesamt- und Low density lipoprotein- Cholesterol im Vergleich zu weiblichen Affen messbar war. Des Weiteren wurden 16 Tiere über einen vierwöchigen Zeitraum in eine neue Umgebung verbracht, bevor sie in ihre Heimatbehausung zurückkehrten. Durch diese Umsetzung war bei den untersuchten Tieren die Leuko- und Lymphozytenzahl auch vier Wochen nach der Umsetzung erniedrigt. Zeitgleich lag ein erhöhter Kortisolspiegel vor, der im Kot bestimmt wurde. Im zweiten Teil der Studie wurden anhand humandiagnostischer Standards IgAAntikörper (AK) gegen Gliadin (AGA), Gewebstransglutaminase (tTG), deamidiertes Gliadin (ADGA) sowie Glykoprotein 2 (AGP2A) im Plasma von 24 WBA mittels eines ELISAs während glutenhaltiger (Diät 1) und glutenfreier Ernährung (Diät 2) bestimmt. Dabei wurden die klinische Symptomatik von WMS und das Körpergewicht der Tiere ebenfalls untersucht. Zudem erfolgte die Analyse von Kotproben antikörperpositiver Tiere hinsichtlich Qualität und Trockenmassegehalt während Diät 2 und einer darauf folgenden glutenhaltigen Provokationsdiät. Die serologische Diagnostik ergab einen signifikanten Rückgang von AGA, AK gegen tTG und AGP2A während Diät 2 bei Tieren, die nach Diät 1 erhöhte Werte aufwiesen. Diät 2 führte zu einem Rückgang der klinischen Symptome und einer signifikanten Gewichtszunahme bei antikörperpositiven WBA. Die glutenhaltige Provokationsdiät ergab eine verminderte Kotqualität mit einem niedrigeren Trockenmassegehalt. Es wurden im Rahmen dieser Arbeit aktuelle, hämatologische und klinisch-chemische Blutrichtwerte des WBA erhoben. Der durch Umsetzung in eine neue Behausung bedingte Stress ist bei WBA bis vier Wochen lang nachweisbar. Es ist sinnvoll, dies in der zeitlichen Planung wissenschaftlicher Studien zu berücksichtigen, um das Wohlbefinden der Tiere vor Versuchsbeginn sicherzustellen und den Einfluss von Stress auf experimentelle Ergebnisse zu minimieren. Der Nachweis grundlegender, an der Pathogenese der Zöliakie beteiligter Antikörper, in Kombination mit den klinischen Symptomen, deutet auf Glutensensitivität mit ätiologischer Beteiligung an WMS bei WBA hin. Die glutenfreie Ernährung von WBA in menschlicher Obhut ist daher sinnvoll und empfehlenswert. / Common marmosets are often used as animal models for human diseases. For their health maintenance, diagnostic blood values are absolutely essential. Previously obtained reference values are characterized by great value-specific differences. Moreover, the influence of routine measures on these blood parameters, e. g. changes in housing conditions, has not been examined yet. Therefore, the first aim of the present study was to update haematological and clinical chemical blood parameters of common marmosets. Further, the influence of stress, caused by relocation to a new housing, on these parameters and the cortisol level in feces was examined. In addition to that, common marmosets under human management are often affected by gastrointestinal diseases, which are difficult to diagnose with basic standard blood values. In this context, sensitivity to nutritional elements, e. g. gluten, plays an important role and is discussed as a potential cause of wasting marmoset syndrome (WMS). In the second part of this study, the recurrent gastrointestinal diseases of common marmosets under human management were aetiologically investigated, with special regard to possible gluten sensitivity. In the first part of this study, blood samples were obtained from 54 female and male common marmosets to evaluate standard values of haematology and clinical chemistry. The determined haematological parameters are similar to the already obtained data, the clinical chemistry values differ somewhat: The enzyme activities of lactate dehydrogenase, alanine aminotransferase and lipase in addition to the ranges of alkaline phosphatase and total bilirubin diverge from the data ascertained in this study. Moreover, female animals presented significantly higher mean corpuscular volume and mean corpuscular haemoglobin than males, whereas male common marmosets showed significantly higher total- and low density lipoprotein-cholesterol, compared to females. Further, 16 animals were relocated to a new environment for a time period of four weeks, before they returned to their home cages. The change of housing caused a decreased leuko- and lymphocyte count in all examined animals that was still measurable four weeks after the relocation. At the same time, an increased fecal cortisol level was determined. The aim of the second study was to investigate the modification of plasma antibodies to gliadin (AGA), tissue transglutaminase (tTG), deamidated gliadin (ADGA) and glycoprotein 2 (AGP2A) during two successive diets in 24 animals: A gluten-containing diet (diet 1) and a gluten-free diet (diet 2). Further, clinical symptoms of WMS and the animals’ body weight were also examined. An analysis of the feces of antibody-positive animals regarding changes in quality and dry matter content was carried out with samples collected during diet 2 and a successive gluten challenge diet of two months duration. The serological diagnostics resulted in a significant decline of AGA, antibodies to tTG and AGP2A during diet 2 in animals that had shown increased antibody concentrations during diet 1. Diet 2 also caused an amelioration of clinical symptoms and an increased body weight in antibody-positive animals. The gluten challenge resulted in a decreased feces quality and a lower fecal dry matter, compared to fecal samples of diet 2. In the context of this dissertation, parameters of haematology and clinical chemistry of the common marmoset were updated. Stress caused by relocation to a new housing was still measurable for a period of four weeks. It is therefore essential to consider this time span in the design of scientific studies to secure animal welfare prior to the study and to reduce the influence of stress on experimental results. In combination with the clinical symptoms, the detection of antibodies that are part of the pathogenesis of coeliac disease in humans strongly suggests gluten sensitivity with an aetiological connection to WMS in common marmosets. Therefore, gluten-free nutrition of common marmosets under human management is highly recommendable.

<p>Sagledavajući značaj nutritivne vrednosti bezglutenskog hleba u prevenciji dodatnih zdravstvenih poremećaja kod obolelih od celijakije, kao i prisutnog trenda bezglutenske ishrane, u okviru ove disertacije analizirana je mogućnost obogaćenja bezglutenskog hleba prehrambenim vlaknima. Ispitan je uticaj različite vrste i količine (3, 5, 7%) prehrambenih vlakana (vlakna &scaron;ećerne repe i vlakna jabuke), količine hidroksipropil metil celuloze - HPMC (2, 3, 4%) i količine vode (180&ndash;230%) na reolo&scaron;ke osobine bezglutenskog testa (svojstva proticanja i viskoelastične osobine), kvalitet i nutritivnu vrednost bezglutenskog hleba. Rezultati ove disertacije ukazuju da dodatak vlakana &scaron;ećerne repe i jabuke ne umanjuje pozitivan uticaj HPMC na reolo&scaron;ke osobine bezglutenskog testa, formiranje i ojačavanje njegove strukture. Bezglutenski hleb privlačne boje, veće zapremine, manje tvrdoće sredine i odličnih senzorskih karakteristika dobijen je kod uzoraka sa 4% HPMC, 3, 5 ili 7% vlakana &scaron;ećerne repe/jabuke i 220%/190% vode. Sadržaj ukupnih prehrambenih vlakana kod uzoraka sa 3% vlakana je iznad 4,5 g/100 g, a kod uzoraka sa 7% vlakana čak oko 6 g/100 g bezglutenskog hleba. Postignut sadržaj ukupnih prehrambenih vlakana je iznad propisane vrednosti za proizvode koji mogu biti nosioci nutritivne izjave &bdquo;izvor vlakana&ldquo;, čime je ostvaren cilj ove disertacije.</p> / <p>Considering the importance of the gluten-free bread nutritional value in the prevention of additional health disorders in patients with celiac disease, as well as the arising gluten-free diet trend, the possibility of enriching gluten-free bread with dietary fibers was analyzed within this dissertation. The influence of different types and amounts (3, 5, 7%) of dietary fibers (sugar beet and apple fibers), the amount of hydroxypropylmethylcellulose - HPMC (2, 3, 4%) and the amount of water (180&ndash;230%) on rheological properties of gluten-free batter (flow and viscoelastic properties), quality and nutritional value of gluten-free bread. The results of this dissertation indicate that the addition of sugar beet and apple fibers does not diminish the positive influence of HPMC on the rheological properties of the gluten-free batter, the formation and strengthening of its structure. Gluten-free bread with appealing color, higher volume, lower hardness and excellent sensory characteristics was obtained in samples with 4% HPMC, 3, 5 or 7% sugar beet/apple fibers and 220%/190% water. The total dietary fibers content in samples with 3% fibers is above 4.5 g/100 g, and in samples with 7% fibers approximately 6 g/100 g of gluten-free bread. The accomplished total dietary fibers content is above the prescribed value for products that may bear the nutritional statement &quot;source of fiber&quot;, thus achieving the goal of this dissertation.</p>

[ES] La inspección visual es un parámetro de calidad que describe tanto el aspecto externo como la estructura interna en los productos derivados de cereales. El análisis digital de imagen (ADI) es una herramienta ampliamente utilizada en diversos campos para procesar y analizar los objetos que observamos visualmente. Sin embargo, a pesar de su potencial, ha sido escasamente explotado en el ámbito de alimentos. La estructura de la miga de los productos derivados de cereales es uno de los parámetros característicos del pan y en este contexto es fundamental realizar el seguimiento de los cambios estructurales que se producen durante el proceso o debido a las modificaciones en la formulación. Por ello, el objetivo principal de la presente tesis doctoral fue definir la interrelación entre la estructura y parámetros de calidad de masas y productos horneados, así como extender la utilidad del ADI y su uso para optimizar los procesos y las formulaciones en productos horneados. Para ello, en primer lugar, se establecieron las condiciones óptimas de captura y se estudió la forma de mejorar la detección de bordes. Posteriormente se evaluó el potencial del análisis de imagen para cuantificar los cambios estructurales ocurridos en la formación de geles de almidón en un analizador rápido de fuerza. Seguidamente, se estudió el impacto en la estructura de la adición o sustitución de distintos ingredientes o aditivos en panes con y sin gluten. Y finalmente se investigaron las posibles correlaciones entre los resultados obtenidos de la caracterización tecnológica (perfil de textura, estructura de la miga, humedad) frente a la percepción sensorial (propiedades del bolo, masticación y deglución). El ADI permitió cuantificar las diferentes estructuras obtenidas con los distintos almidones. Además, el estudio reveló las diferencias en el tiempo necesario para formar una estructura homogénea y bien distribuida, que fue dependiente del origen del almidón. La modificación de la formulación de panes con o sin gluten requirió de la adaptación del proceso y permitió obtener diferentes estructuras atendiendo a los ingredientes /aditivos estudiados. El ADI resultó ser una herramienta eficaz para detectar y cuantificar los cambios originados a nivel estructural, en masa y producto final. Además, los cambios cuantificados mediante ADI obtuvieron correlaciones estadísticamente significativas con varios de los parámetros estudiados de percepción sensorial, como son la adhesividad del bolo y el tiempo total para masticar y deglutir una muestra. Por lo tanto, el ADI resultó una herramienta suficientemente específica y no destructiva, que permitió caracterizar el impacto del cambio de la formulación en la distribución alveolar de panes con y sin gluten. Existiendo relación entre este y los parámetros utilizados habitualmente en la caracterización de productos derivados de cereales, así como con la percepción sensorial. / [CA] La inspecció visual és un paràmetre de qualitat que descriu tant l'aspecte extern com l'estructura interna en els productes derivats de cereals. L'anàlisi digital d'imatge (ADI) és una eina àmpliament utilitzada en diversos camps per a processar i analitzar els objectes que observem visualment. No obstant això, malgrat el seu potencial, ha sigut escassament explotat en l'àmbit d'aliments. L'estructura de la molla dels productes derivats de cereals és un dels paràmetres característics del pa i en aquest context és fonamental realitzar el seguiment dels canvis estructurals que es produeixen durant el procés o a causa de les modificacions en la formulació. Per això, l'objectiu principal de la present tesi doctoral va ser definir la interrelació entre l'estructura i paràmetres de qualitat dels diferents tipus de masses i productes enfornats, així com estendre la utilitat de l'ADI i el seu ús per a optimitzar els processos i les formulacions en productes enfornats. Per a això, en primer lloc, es van establir les condicions òptimes de captura i es va estudiar la manera de millorar la detecció de les vores. Posteriorment es va avaluar el potencial de l'anàlisi d'imatge per a quantificar els canvis estructurals ocorreguts en la formació de gels de midó en un analitzador ràpid de força. Seguidament, es va estudiar l'impacte en l'estructura de l'addició o substitució de diferents ingredients o additius en pans amb glútens i sense glútens. I finalment es van investigar les possibles correlacions entre els resultats obtinguts de la caracterització tecnològica (perfil de textura, estructura de la molla, humitat) enfront de la percepció sensorial (propietats del bol, masticació i deglució). L'ADI va permetre quantificar les diferents estructures obtingudes amb els diferents midons. A més, l'estudi va revelar les diferències en el temps necessari per a formar una estructura homogènia i ben distribuïda, que fou dependent de l'origen del midó. La modificació de la formulació de pans amb gluten o sense va requerir de l'adaptació del procés i va permetre obtindre diferents estructures atesos els ingredients /additius estudiats. L'ADI va resultar ser una eina eficaç per a detectar i quantificar els canvis originats a l'entorn estructural, en massa i producte final. A més, els canvis quantificats mitjançant ADI van obtindre correlacions estadísticament significatives amb els diversos paràmetres estudiats de percepció sensorial, com són l'adhesivitat del bol i el temps total per a mastegar i deglutir una mostra. Per tant, l'ADI va resultar una eina prou específica i no destructiva, que va permetre caracteritzar l'impacte del canvi de la formulació en la distribució alveolar de pans amb glutens i sense. Tot i existint relació entre aquest i els paràmetres utilitzats habitualment en la caracterització de productes derivats de cereals, així com amb la percepció sensorial. / [EN] Visual inspection is a quality parameter that describes both the external appearance as the internal structure in cereal-based products. Digital image analysis (DIA) is a tool widely used in several fields to process and analyse objects visually observed by the human eye. However, despite of its potential, it has been barely explored for food analysis purposes. Crumb structure in cereal-derived products is one of the most characteristic parameters in bread. Due to this, following up the structural changes caused by the process or by the formulation changes, is of fundamental importance. For that reason, the main target of this thesis was the definition of the relationship between the structure and the quality parameters of dough and baked products. Moreover, extending the DIA utility and its use to improve process and mixes in baked products represented other main objective of this work. To do that, first, the optimal capture conditions were established and the way to improve the edge detection was studied. Afterwards, image analysis was evaluated to quantify the structural changes that happened during the starch gels formation in a fast force analyser. Then, the impact on the structure by adding or replacing different ingredients or additives in gluten-free breads was studied. Finally, the possible correlations between the results obtained from technological characterization (texture profile, crumb structure, moisture content) versus sensory perception (bolus properties, chewing and swallowing) were also investigated. DIA allowed the quantification of the different structures obtained from different starches. In addition, the study revealed the differences in time needed to form a homogeneous and well-distributed structure, which was dependent on the origin of the starch. Formulation modification in bread with or without gluten required a process adjustment that allowed to obtain different structures depending on the ingredients / additives studied. DIA has been proved to be an effective tool for detecting and quantify changes originated at the structural, dough and final product levels. In addition, the changes quantified by DIA obtained statistically significant correlations with several parameters studied for the sensory perception, such as bolus adhesion and total time to chew and swallow the sample. Therefore, DIA was a sufficiently specific and non-destructive tool, which allowed to characterize the impact of the formulation change on the bread cell distribution with and without gluten. There being a relationship between this, and the parameters usually used in cereal products characterization, as well as sensory perception. / Authors acknowledge the financial support of the Spanish Ministry of Economy and Competitiveness (Projects AGL2011-23802 and AGL2014-52928-C2-1-R), the European Regional Development Fund (FEDER) and Generalitat Valenciana (Projects Prometeo 2012/064 and Prometeo 2017/189), the Spanish Ministry of Science, Innovation and Universities (RTI2018-095919-B-C21), and the European Regional Development Fund. This work is based upon the work from COST Action 18101 SOURDOMICS – Sourdough biotechnology network towards novel, healthier and sustainable food and bioprocesses, the European Regional Development Fund and the joint project between research group from IATA-CSIC and the research groups of Nutriomics and CIDPRO from Tecnologico de Monterrey, Mexico. / Garzón Lloría, R. (2021). Análisis estructural de los productos derivados de cereales y su aplicación en la optimización de procesos y productos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/166856

Yes / Objectives. Coeliac disease (CD) is an autoimmune condition managed by a lifelong therapeutic gluten-free diet. Previous research suggests that the chronicity of CD, the limitations imposed by the gluten-free diet, and the risk of other associated diseases can have a negative impact on health-related quality of life (HRQoL) and psychological well-being. The aim of this study was to explore the illness perceptions and self-efficacy beliefs of adults with CD in the United Kingdom and to report their subjective levels of HRQoL and psychological well-being. Design. The study employed a cross-sectional postal questionnaire design. Method. Participants (n= 288) were adults with CD recruited via Coeliac UK. Measures of well-being, HRQoL, self-efficacy, illness perceptions, and dietary self-management were analysed. Preliminary descriptive and univariate procedures were employed before bivariate tests of association or difference were carried out. Backward stepwise multiple regression analysis was used to investigate the predictive strength of variables on well-being, quality of life, and self-efficacy. Logistic regression was used to look at the influence of variables on adherence. Results. Results indicate that HRQoL and psychological well-being were comparable to those found in previous related studies. Participants with weak beliefs in the serious consequences of CD and poorer emotional reactions to the condition had a greater likelihood of having enhanced HRQoL, improved psychological well-being, and higher self-efficacy. Strong beliefs in personal control and a greater perceived understanding of CD were associated with greater self-efficacy. Conclusions. Perceived self-efficacy and illness perceptions could play a role in informing psychological interventions for individuals with CD.

La maladie cœliaque est une entéropathie inflammatoire chronique se développant chez des individus génétiquement prédisposés par l’expression des haplotypes HLA-DQ2 ou HLA-DQ8 et survenant suite à la consommation de gluten. Elle se caractérise par le développement d’une atrophie des villosités de la muqueuse intestinale débouchant sur un syndrome de malabsorption alimentaire. La seule thérapie actuelle est le suivi d’une diète sans gluten mais cette éviction totale du gluten n’est pas toujours efficace et est lourde en concessions. Il est par conséquent urgent de développer des thérapies alternatives mais ce domaine constitue un pipeline évoluant lentement, notamment suite à l’absence d’un modèle animal pertinent et complet sur le plan physiologique. L’objectif de cette thèse doctorale est de répondre à ce besoin crucial en développant un modèle murin capable de récapituler les caractéristiques de la maladie. Le chapitre 1 dresse le portrait de la maladie en quatre parties amenant progressivement le lecteur dans les détails de sa pathogenèse. Cette introduction débute par un rappel sur la physiologie et l’immunité intestinale puis elle définit la face clinique de la maladie. Ensuite, le lecteur évolue dans une partie plus détaillée de la pathogenèse aidant au discernement de ses acteurs cellulaires et moléculaires. Finalement, elle se termine par une revue de la littérature sur les actuels modèles animaux. Le chapitre 2 brossent les objectifs de la thèse sur base de données clés de la littérature, notamment, les patients présentent au minimum une copie de l’halplotype HLA-DQ2 ou HLA-DQ8 et plus des deux-tiers sur-expriment la cytokine pro-inflammatoire interleukine-15 au niveau de leur muqueuse intestinale. Il est donc raisonnable de penser qu’ensemble, le gluten, l’haplotype HLA et l’interleukine-15 contribuent activement à la pathogenèse. Bien que soupçonnés, leurs rôles et interactions nécessitent l’apport de preuves tangibles in vivo. Le chapitre 3 détaille ainsi ces interactions démontrées à l’aide du développement de notre nouveau modèle murin. Ce dernier est caractérisé par la surexpression de l’interleukine-15 dans l’épithélium et dans la lamina propria intestinale et par l’expression de l’haplotype HLA-DQ8. Ce travail démontre que l’exposition de cette souris au gluten s’accompagne d’une atrophie villositaire et de la signature complète de la maladie, tant sur le plan sérologique, cellulaire que transcriptionnel. Nous démontrons que la surexpression simultanée de l’interleukine-15 dans les deux compartiments de la muqueuse intestinale que sont la lamina propria et l’épithélium est une condition sine qua non au développement de l’atrophie. Aussi, cette étude permet de mettre en lumière la nécessité des cellules T CD4+ et de l’interféron-gamma dans l’activation des lymphocytes intraépithéliaux et le développement de l’atrophie. Finalement, cette recherche établit le rôle central joué par l’haplotype HLA-DQ8 et par l’enzyme transglutaminase II tissulaire dans la survenue de ces lésions. De manière générale, les résultats issus de ce modèle et présentés au chapitre 3 reflètent toute la complexité des interactions entre le gluten, la génétique et l’IL-15 dans le développement de la maladie cœliaque. Enfin, le chapitre 4 apporte une conclusion à ce travail et le chapitre 5 discute des futures directions envisagées pour ce modèle préclinique. Ce dernier va sans doute contribuer à une meilleure compréhension de la maladie cœliaque et permettre l’identification de potentielles cibles thérapeutiques. / Coeliac disease is a chronic inflammatory enteropathy characterized by autoimmune features. This disease occurs in genetically predisposed individuals expressing HLA-DQ2 or HLA-DQ8 haplotypes and is triggered following gluten consumption. The disease is characterized by the development of intestinal villous atrophy leading to malabsorption. The only current therapy is the adherence to a gluten-free diet, but the diet is not always effective and is heavy in concessions. Therefore, the development of alternative therapies is urgent but is a slowly evolving pipeline, mainly due to the absence of a physiologically relevant animal model. The aim of this thesis is to answer this unmet need by developing an animal model capable of recapitulating the main characteristics of the disease. Chapter 1 depicts a portrait of the disease in four points, gradually leading the reader into the details of its pathogenesis. This introduction begins with a review of the physiology and intestinal immunity and then draws a clinical portrait of the disease. Third, the reader evolves in a more detailed part of the pathogenesis helping him to discern its cellular and molecular actors. Finally, the introduction ends with a review of the literature on current animal models. Chapter 2 outlines the thesis objectives based on key data from the literature, in particular, patients present at least one copy of the HLA-DQ2 or HLA-DQ8 haplotype and more than two-thirds over-express the proinflammatory cytokine interleukin-15 at the level of their intestinal mucosa. It is therefore reasonable to hypothesize that gluten, HLA haplotype and interleukin-15 together contribute to the pathogenesis. Although suspected, their roles and interactions still require the provision of tangible evidence in vivo. Chapter 3 details these interactions based on the proposed new mouse model. This model is characterized by the overexpression of interleukin-15 in the intestinal epithelium and lamina propria and by the expression of the HLA-DQ8 haplotype. This work demonstrates that the exposure of this mouse to gluten is accompanied by villous atrophy and the complete serological, cellular and transcriptional signature of the disease. We also demonstrate that simultaneous overexpression of interleukin-15 in both mucosal compartments is a prerequisite for the development of atrophy. This study also highlights the need for CD4+ T cells and interferon-gamma in the activation of intraepithelial lymphocytes and the development of villous atrophy. Finally, this research establishes the central role played by the HLA-DQ8 haplotype and the enzyme tissue transglutaminase II in the occurrence of these lesions. In general, the results from this model presented in Chapter 3 reflects the complexity of the interactions between gluten, genetics and IL-15 in the development of coeliac disease. Finally, chapter 4 concludes this work and chapter 5 discusses future directions for this powerful preclinical model that will undoubtedly contribute to a better understanding of coeliac disease and will allow the identification of new potential therapeutic targets.

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.

In order to better assist practitioners and better serve persons with autism spectrum disorders (ASD) and their families, it is vital for professionals to systematically evaluate the existing body of literature and synthesize its scientific evidence, so that the efficacy of research can be translated to evidence-based practices (EBPs) (Wheeler, 2007; Zhang & Wheeler, 2011). This research synthesis evaluated adherence to EBP standards and analyzed the effectiveness of gluten-free and casein-free (GFCF) diets for individuals with ASD. Four hundred and seventy articles were screened among peer-reviewed journals in English language published through 2010 using the Academic Search Complete search database. Twenty-three studies were selected, and the researchers used a systematic analysis model developed by Mayton, Wheeler, Menendez, and Zhang (2010) to investigate the degree of adherence to specific evidence-based practice standards. In addition, the study utilized quality indicators proposed by (a) Horner et al. (2005) for single-subject design studies and (b) Gersten et al. (2005) for group experimental design, to evaluate the efficacy of GFCF diet interventions. Results of this synthesis indicated that the efficacy of GFCF diet interventions for individuals with ASD is inconclusive, and the field needs better controlled studies to provide the scientific evidence base for the intervention.

<p>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.</p>