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Gut microbiome and its changes related to therapy of chronic diseases
Hurych, Jakub ; Cinek, Ondřej (advisor) ; Kolář, Milan (referee) ; Falt, Přemysl (referee)
This work examines the response of the gut microbiome to therapeutic interventions in three longitudinal studies of chronic gastrointestinal diseases: Crohn's disease, celiac autoimmunity and irritable bowel syndrome. Multiple methods of stool microbiome analysis (especially massively parallel 16S rDNA or 18S rDNA sequencing and metagenomic sequencing) followed by bioinformatic and statistical analysis were used. In Crohn's disease, we detected a previously undescribed secondary nature of changes in the gut bacteriome after anti-TNF treatment. In celiac disease autoimmunity, where previous works described an effect of probiotic intervention on serological markers of the disease, the gut bacteriome and metabolome, we described the absence of significant changes in beneficial gut protozoa. In irritable bowel syndrome, we observed a significant response of the bacteriome after administering four doses of mixed microbiota transplantation but no response in the reduction of clinical symptoms. The results of these studies could contribute to a better understanding of the gut microbiome's role in the pathogenesis of these serious diseases. Keywords: microbiome, Crohn's disease, celiac disease, irritable bowel syndrome
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Monitoring of cereal products uptake in a sample of Czech population
Vlachová, Martina ; Hurtová, Jana (referee) ; Márová, Ivana (advisor)
This bachelor’s thesis was focused on monitoring of consumption of cereal products and analysis of some of their active components. In theoretical part an overwiev of cereals and their composition was introduced. In experimental part a set of analytical parameters was obtained in selected cereal products: total phenolics, total flavonoids and antioxidant activity. The highest level of these active substances was detected in products containing chocolate and forest fruit. Further, 161 completed consumer questionnaires were collected. From these data common knowledge in consumers, their shopping conventions and individual products preferences were evaluated.
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Enzyme hydrolyses of wheat gluten for the gluten free application
Svobodová, Dominika ; Vránová, Dana (referee) ; Ing.Silvia Mošovská,Ph.D. (advisor)
Celiac disease is attributable to autoimmune diseases, where treatment is based on absolute respect for gluten-free diet. As the gluten used widely in all areas of food production, strict adherence to a gluten-free diet really can be difficult for people with celiac disease. Products suitable also for those suffering from the disease are usually expensive. Finally, patients that undergo exclusively on a gluten-free diet are robbing the important nutritional components. Therefore, interest in this type of products innovation enhance as to the nutritional, technological, as well as organoleptic properties. The aim of this final thesis was to draw up literature review of the celiac disease and gluten allergen, or his particular part of gliadin. In the experimental part, attention was focused on the first hydrolytic cleavage of starch, the substrate was sterilized wheat flour and water, where is incorporated spiral just gluten. The best ability of degrading the starch grains, the combination of enzymes and MT3K and GLUAMK concentration of 0, 1% over 4 hours of action, as indicated by a reduction in the starch content of the original amount of 72,65 % at a concentration of 29,00 %. The next item of work distribution used different proteases, to reduce the amount of gliadin to below 20 ppm per kilogram of final product. Low levels investigated allergen (10,79 ppm/kg) recorded by the enzyme combination exoprotease Flavourzyme (0,1 %) and endoprotease Neutral Protease (0,1 %) at 7 hours hydrolytic action. The conclusion of the thesis was prepared several alternatives gluten-free breads and through sensory evaluation assessed the characteristics of color, aroma and taste to predict the location of the products on the commercial market.
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Changes of gluten proteins during beer processing
Porubiaková, Otília ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The aim of thesis was monitoring of changes in the content of gluten proteins in the biotechnological process of beer production. During the production process of wheat and barley beer, the samples were collected and analysed using the electrophoresis and immunoassay method. The results of the analyses were compared with commercial Czech beers. The theoretical part contains description and composition of gluten proteins, malt and beer technology, the changes that occur in this process, and methods of gluten proteins analysis. The experimental part contains procedures for laboratory production of barley and wheat beer and analyses of gluten proteins. To identify the individual gluteal protein fraction acid and SDS electrophoresis methods were used. For quantification, enzyme immunoassay was used and evaluated spectrophotometrically. The identification of the gluten‘s fractions by electrophoretic methods has been shown to be less specific for samples with lower content of gluten proteins and for barley specimens. A decrease in the concentration of gliadins and glutenins in the beer production process was demonstrated. A significant change was found during wort production with 98% decrease of gluten content compared to the feedstock and during the fermentation, when the gluten concentration dropped below 10 mg/kg. This value is acceptable from the legislation for products labelled „gluten-free“.
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Speciální druhy pečiva a možnosti inovace receptur
Janečková, Šárka
Thesis is focused on the production of baked goods for population groups suffering from diseases that require special dietary therapy. Subject of the literature research was to get acquainted with three of the most serious of them - celiac disease, Crohn's disease and diabetes and a consequent summary of the individual dietary requirements related to the diseases. The aim was to create an overview of basic, innovative and non-traditional ingredients suitable for the production of special baked goods for people suffering from these diseases. Practical part of the work was based on the development of six recipes for the production of bakery products using non-traditional ingredients, followed by successful execution of baking expe-riment, instrumental analysis of samples and sensory evaluation of the products. The baking experiment resulted in 3 groups of baked goods - gluten-free, non-residual and low-carbohydrate. The best volume, firmness and sensory properties were achieved in the gluten-free bakery products, which were very similar to the conventional bakery products and were also judged to be the most sensory ac-ceptable based on the sensory questionnaires. For the other two groups of bakery products, the results of the measurements and sensory evaluation were not very sa-tisfactory, although very interesting crust and crumb colours were achieved in the-se samples due to the used ingeridents.
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Determination of selected elements in cereal flours and their gluten-free alternatives
Venclová, Veronika ; Hraníček, Jakub (advisor) ; Nováková, Eliška (referee)
This study deals with the determination of calcium, magnesium, phosphorus, potassium, sodium, iron, zinc, copper, manganese and molybdenum in cereal gluten flours and its gluten-free alternatives. The flour samples were digested using a microwave device and the elements in solution were subsequently determined using an inductively coupled plasma mass spectrometer. Optimization of the digestion of the selected flours was performer prior to the actual determination of the elements. The aim of the work was to determine the above listed elements in the flour samples and to establish a basic overview of the elemental composition of the different flours. This overview and comparison of the different flours is intended to help the people with celiac disease to choose gluten-free alternatives to wheat flour. Keywords Inductively coupled plasma mass spectrometry, microwave digestion, calcium, magnesium, phosphorus, potassium, sodium, iron, zinc, copper, manganese, molybdenum, wheat flour, gluten-free flours, celiac disease
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Mechanisms of antigen presentation in the etiopathogenesis of celiac disease
Hudec, Michael ; Černá, Marie (advisor) ; Hrdý, Jiří (referee) ; Slavčev, Antonij (referee)
1 ABSTRACT Celiac disease (CeD) is a chronic autoimmune disease that develops as a response of the immune system to the presence of gluten in the small intestine. CeD is manifested not only by classic intestinal symptoms: abdominal pain, constipation or diarrhea, as well as complex less common symptoms: anemia, osteoporosis, psychiatric disorders or menstrual cycle disorders. HLA risk alleles predisposing to origin of celiac disease are HLA-DQ2 (DQA1*05:01 / DQB1*02:01) and HLA-DQ8 (DQA1*03:01 / DQB1*03:02). There are other celiac disease-associated polymorphisms outside of HLA locus (6p21.3) that are located in 5q32 and 19p13 regions with unclear connection to CeD development. HLA class II glycoproteins are expressed on antigen presenting cells (APC) that include dendritic cells, macrophages and B cells. Monocytes are one of several possible dendritic cell precursors that circulate in the bloodstream. Deviations in the frequency of intermediate monocytes are directly associated with autoimmune disorders such as Crohn's disease or rheumatoid arthritis. It is known that the monocytes of CeD patients show pro-inflammatory reaction in the presence of gluten. It means that, in the context of CeD, the response to gluten arises earlier than the activation of gluten-specific T cells. The conventional way of direct...
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Monitoring of gluten content in barley varieties recommended for the production of Czech Beer
Barčinová, Nikola ; Boško, Rastislav (referee) ; Svoboda, Zdeněk (advisor)
Sandwich enzyme-linked immunosorbent assay ELISA was used for determination of gluten in barley varieties recommended for the production of Czech beer. Currently, it is the most widely used method for determining gluten in the food industry. Monitoring the gluten content in raw materials and foods made from them is especially important for people suffering from celiac disease. Gluten intolerance is an autoimmune disorder that causes morphological and functional changes in the small intestine. Currently, the most effective treatment is following a gluten-free diet. In this bachelor thesis, content of gluten in 14 barley samples was monitored. The RIDASCREEN® Gliadin set was used for the measurement. Results were evaluated with RIDA SOFT Win.NET software. Content of gluten in the barley samples varied from 36,3 g/kg to 82,5 g/kg.
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Screening of probiotics bacterial strains aimed at proof of gluten degradation
Korvasová, Lucie ; Obruča, Stanislav (referee) ; Ryšávka, Petr (advisor)
No other treatment for celiac disease and other gluten-related disorders has been found so far except for a lifelong diet, which is significantly restrictive for the patient. Moreover, it is not always possible to adhere strictly to this diet, mainly due to random gluten ingestion, which leads to health complications associated with these diseases. Currently, vaccination is being tested as an alternative treatment, and there is also increasing attention paid to enzymes that could break down the immunogenic part of gluten, specifically in its gliadin component. This thesis examined bacteria that could potentially break down this sequence in gliadin and thereby reduce its concentration during detection. A considerable proportion of the tested bacteria belong to probiotics to ensure their future use for humans is safe. To obtain data, the fact was utilized that as bacteria grow, the turbidity increases proportionally, which was subsequently detected to determine the amount of bacterial growth. The RIDASCREEN® Gliadin competitive kit was used for the gliadin detection test, which works on the principle of a competitive ELISA method with R5 antibodies. The result of the study was the finding that only the bacteria Bacillus subtilis was able to break down gliadin among the examined bacteria. It was also measured how much gliadin the bacteria could break down at different concentrations, and according to the data, it was evaluated that it is appropriate to adapt the culture to the presence of gliadin for a greater amount of broken gliadin.
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Molecular detection of selected gene polymorphisms related to nutrition (nutrichip validation)
TURKOVÁ, Kateřina
Lactose intolerance is the most common food intolerance in the world. Individuals with lactose intolerance are unable to produce the enzyme lactase in the small intestine, which makes it possible to break down the lactose contained in dairy products. Insufficient lactase production may be genetically determined. Two single nucleotide polymorphisms responsible for the persistence of lactase activity in adulthood have been found in the European population. Celiac disease is one of the autoimmune diseases that mainly affects the mucous membrane of the small intestine. The disease is characterized by intolerance to gliadin, which is part of gluten. Intolerance leads to chronic inflammation of the small intestinal mucosa, leading to chronic diarrhea, fatty stools, vomiting and fatigue. The development of celiac disease is conditioned by the presence of a genetic predisposition. Genetic predisposition is linked to HLA system alleles. Specifically, these are the HLA-DQ2 and HLA-DQ8 haplotypes.
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