京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第20430号 / 農博第2215号 / 新制||農||1048(附属図書館) / 学位論文||H29||N5051(農学部図書室) / 京都大学大学院農学研究科食品生物科学専攻 / (主査)教授 谷 史人, 教授 安達 修二, 教授 松村 康生 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

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Introduction Several studies have found celiac disease may be associated with a variety of cardiac manifestations. Atrial fibrillation (AF) is one of the most common arrhythmias that can cause significant morbidity. However, the risk of atrial fibrillation in patients with celiac disease according to epidemiological studies remains unclear. The aim of this meta-analysis study is to assess the risk of atrial fibrillation in patients diagnosed with celiac disease compared to controls. Methods A systematic literature review was conducted in MEDLINE, EMBASE, Cochrane databases from inception through December 2017 to identify studies that evaluated the risk of atrial fibrillation in patients with celiac disease. We included randomized controlled trial, cross sectional and cohort studies that reported the odds ratio, relative risk, hazard ratio, and standardized incidence ratio comparing the risk of developing atrial fibrillation among patients with celiac disease, versus patients without celiac disease as control. The Newcastle-Ottawa scale was used to determine the quality of the studies. Effect estimates from individual studies were extracted and combined using random-effect, generic inverse variance method of DerSimonian and Laird. Results Celiac disease is an autoimmune condition. This inflammatory state predisposes patients to develop AF. After a review of the literature, four observational studies with a total of 64,397 participants were enrolled. The association between celiac disease and increased risk of atrial fibrillation was significant, with a pooled OR of 1.38 (95% CI: 1.01-1.88). No publication bias as assessed by the funnel plots and Egger's regression asymmetry test with p = 0.54. However, the heterogeneity of the included studies was high (I2 = 96). Conclusion A significant association between celiac disease and risk of atrial fibrillation was reported in this study. There is a 38% increased risk of atrial fibrillation. Additional studies are needed to clarify the mechanistic link between atrial fibrillation and celiac disease. Some of the limitations of this study are that all were observational studies, some were medical registry-based and there was high heterogeneity between studies.

Because of the nutritious value of sodium glutamate and its meat-Like flavor, won 18 very pleasing to most people, and hence Its demand if it were properly promoted, and because of the abundance of protein in accessible form for its manufacture, it is only reasonable to assume that if too optimum continuous of protein hydrolysis were found, tho manufacturing metnous could be made a.highly successful enterprise.

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El contenido de proteína en grano y la fuerza de gluten son objetivos importantes en los programas de mejoramiento de trigo candeal (Triticum turgidum ssp. durum). Una serie de experimentos fue conducida en el sur de la provincia de Buenos Aires para determinar los días a espigazón, rendimiento, contenido de proteína en grano, peso de mil granos y fuerza de gluten en una colección de germoplasma de trigo candeal de base amplia (n=170). La colección fue genotipada con el array 35K para trigo y marcadores STS. Se determinó la estructura poblacional con un subconjunto de 1000 SNPs en bajo desequilibrio de ligamiento y se realizó el mapeo por asociación (MA) para los caracteres fenotípicos evaluados con 3745 marcadores. Fue posible encontrar asociaciones entre los cinco caracteres fenotípicos y los marcadores moleculares utilizados, confirmando regiones cromosómicas previamente informadas y en algunos casos, se encontraron QTLs novedosos. Fueron hallados cinco loci asociados a los días a espigazón en los cromosomas 2AS, 2B, 5A, 7A y 7B. Los SNPs asociados a peso de mil granos en el presente trabajo tendrían relación con QTLs/ genes encontrados por otros autores en regiones del genoma similares tanto en trigo pan como en candeal. Los marcadores hallados para contenido de proteína y rendimiento fueron considerados insuficientes al momento de buscar asociaciones genotipo-fenotipo en varios ambientes. Existe una región comprendida entre 7,46 - 36,42 cM del cromosoma 1B que sería responsable por la variabilidad en la fuerza de gluten observada en esta tesis. El MA realizado con variedades y líneas elite fue efectivo al momento de detectar varios de los QTL informados previamente en las mismas regiones cromosómicas a través del mapeo biparental. Los resultados sugieren que el MA podría ser una herramienta valiosa para identificar regiones genómicas para caracteres que se evalúan de manera rutinaria en los programas de mejoramiento de trigo. Las distintas configuraciones alélicas para los días a espigazón, peso de mil granos y fuerza de gluten serían el punto de partida para la selección asistida mediante varios marcadores moleculares. La eficiencia de selección sería similar a la selección fenotípica, con la ventaja de no requerir ensayos a campo ni análisis de calidad. Además, la información generada por esta tesis podrá ser utilizada holísticamente para la planificación de cruzamientos que aporten variabilidad en los programas de mejoramiento nacionales. / Grain protein content and gluten strength are major targets in wheat breeding programs (Triticum turgidum ssp. durum). A series of experiments was conducted in the south of Buenos Aires province to determine days to heading, yield, grain protein content, thousand kernel weight and gluten strength in a world-wide durum wheat germplasm collection (n=170). This collection was genotyped with the 35K array for wheat and STS markers. The population structure was determined using a subset of 1000 SNPs with low linkage disequilibrium and association mapping (AM) was performed for all the phenotypic characters using 3745 markers. It was possible to find associations between the five phenotypic characters and the molecular markers used, confirming previously reported chromosomal regions and in some cases, novel QTLs were found. Five loci associated with days to heading on chromosomes 2AS, 2B, 5A, 7A and 7B were found. The SNPs associated with thousand kernel weight found in this work would be related to other QTLs / genes previously reported in similar genome regions in bread and durum wheat. Markers found for grain protein content and yield were considered unsatisfactory when looking for genotype-phenotype associations in several environments. There is a region between 7.46 - 36.42 cM of chromosome 1B that would be responsible for the variability in gluten strength observed in this thesis. AM performed with varieties and elite lines was effective at the time of detecting several of the QTLs previously reported in the same chromosomal regions through biparental mapping. The results suggest that AM could be a suitable tool for identifying genomic regions for characters that are routinely measured in wheat breeding programs. The different allelic configurations for days to heading, thousand kernel weight and gluten strength would be the starting point for the assisted selection by several molecular markers in breeding programs. Selection efficiency would be similar to phenotypic selection, with the advantage of avoiding field trials or quality analysis. In addition, the information generated by this thesis could be used for planning crosses that contribute variability to national breeding programs.

The gluten-free diet is the only treatment available for gluten-related disorders, such as celiac disease, an autoimmune reaction to gluten, or non-celiac gluten or wheat sensitivity, a symptomatic reaction to wheat or gluten. However, gluten may not be the only culprit, and patients on a gluten-free diet have been suggested to symptomatically improve through the placebo effect, alterations in immune activity, and alterations in gut microbiota composition. It is unclear which of these mechanisms underlie symptoms in gluten-related disorders and well-designed clinical studies are needed to better understand them. This thesis aims to understand the mechanisms and symptomatic responses by which wheat and gluten affect individuals with gluten-related disorders. I hypothesize that patients with gluten-related disorders have increased psychological symptoms and immune reactivity which may be modulated by the gut microbiota. To test this, I conducted a clinical crossover trial to investigate whether whole wheat or gluten triggered symptoms versus gluten-free control, or nocebo, in irritable bowel syndrome patients adopting a gluten-free diet. Participants reacted similarly to each intervention, suggesting a strong 'nocebo effect' to be the main trigger of their symptoms. However, several participants did not comply to the protocol, muddying the results. Subsequent follow-up visits after disclosing personalized study results found no changes in participant beliefs, behaviours, and symptoms, and most remained on a gluten-free diet. Next, a systematic review of 65 observational studies found an elevated risk of IBD in celiac disease and vice versa. Finally, a systematic review of 6 RCTs found limited evidence that probiotics are safe and possibly therapeutic for ameliorating symptoms in celiac disease. Overall, the work presented in this thesis critically assesses the mechanisms by which gluten and wheat trigger symptoms in gluten-related disorders and highlights the importance of rigorous clinical trial design to control for psychological factors and patient compliance. / Thesis / Doctor of Philosophy (PhD) / Gluten, a wheat protein, is commonly associated with the autoimmune condition celiac disease, symptomatic worsening from gluten or wheat in non-celiac gluten/wheat sensitivity, and irritable bowel syndrome. This thesis strove to understand how gluten and other wheat proteins impact symptoms via psychological, immune, and/or bacteria-mediated pathways in gluten-related disorders. A clinical trial tested the effects of whole wheat, gluten, and gluten-free control on symptoms in irritable bowel syndrome patients on a gluten-free diet. We found no differences between interventions but discovered widespread diet non-compliance and that patient fears triggered symptoms. Informing patients of whether wheat, gluten, or gluten-free control triggered their symptoms did not change their dietary beliefs or behaviours. Additionally, two systematic reviews found a relationship between celiac disease and inflammatory bowel disease, and a possible therapeutic effect of probiotics in celiac patients. Our findings provided insights into the content and quality of the clinical evidence for gluten-related disorders.

This study was designed to examine if a sphingolipid-enriched lipid fraction from wheat gluten could affect the incidence of type I diabetes in BioBreeding diabetes prone (BBdp) rats. Wheat gluten was extracted with a chloroform-methanol (CM) mixture to isolate most of the lipids. Isolated lipids were subjected to silica gel column chromatography and saponification to remove most of neutral lipids and phospholipids, leaving behind a lipid fraction enriched in sphingolipids. This sphingolipid-enriched lipid fraction was used in a BBdp rat feeding study. BBdp rats were fed with one of five diets from weaning at 23 days of age until 125 days of age: a hydrolyzed casein based diet (HC), a NTP-2000 standard rodent diet (NTP-2000), a wheat gluten based diet (WG), a sphingolipid-free wheat gluten based diet (WGSLF), and a hydrolyzed casein plus sphingolipid-enriched lipid fraction diet (HC+SL). The yield of sphingolipid-enriched lipid fraction was about 0.62% of wheat gluten. The content of glycosylceramide in sphingolipid-enriched lipid fraction was increased more than five fold compared to that in total isolated lipids. Rats fed the NTP-2000 diet had the highest incidence of diabetes; while rats on the HC diet had the lowest diabetes incidence. There was no significant difference with regard to the onset age of diabetes among rats in the five diet groups. The addition of sphingolipid-enriched fraction to the HC diet caused a significant increase in the incidence of diabetes in BBdp rats in the first 80 days of the study. However, the ultimate diabetes incidence at day 125 was not changed. The removal of lipids from wheat gluten did not change the diabetes incidence in BBdp rats at any stages of the feeding study. These findings suggest that the sphingolipid-enriched fraction from wheat gluten acted as a possible promoter but not as a trigger of the development of type I diabetes in BBdp rats. There must be something that remains in wheat gluten after chloroform-methanol extraction that serves as a trigger for type I diabetes in these rodents. Type I diabetes in this animal model for the human disease seems to be caused by multiple factors, most likely, by the interaction of sphingolipids and some other unknown substances in wheat gluten. / Master of Science

Soy peroxidase enzyme obtained from isoelectic precipitation procedures was added to all-purpose flour (APF) to assess its effects on the rheological properties and consumer acceptability of yeast bread. A pH 4.8 isoelectrically precipitated fraction from soybeans was used because it produced the most precipitate and had about the same peroxidase activity as the other fractions. Gluten strength was determined using a farinograph for seven treatment groups: control (all-purpose flour), bread flour, all-purpose flour + soy flour, bread flour + soy flour, all purpose flour + pH 4.8 precipitate, all-purpose flour + 15 mg soybean peroxidase, and all-purpose flour + 25 mg soybean peroxidase. Four types of yeast bread were baked for loaf volume determination, texture analysis, and consumer acceptability: a control loaf using only all-purpose flour, a reference loaf using all bread flour, a loaf with all purpose flour + whole soy flour, and a loaf with all-purpose flour + pH 4.8 soy precipitate. The APF+soy flour, bread flour, bread flour + soy flour, and the APF + pH 4.8 precipitate produced an improvement in the gluten strength and mixing tolerance compared to the control (p<0.05). However, the improvement by the addition of the pH 4.8 precipitate cannot be attributed to the peroxidase enzyme because peroxidase needs hydrogen peroxide as a substrate and no hydrogen peroxidase could be added to the farinogragh; therefore, it was concluded that the increase in gluten strength produced by the pH 4.8 soy precipitate was due to an unknown component present in the pH 4.8 fraction. No significant differences (p<0.05) were found in crumb or crust texture for any of the treatment groups. The addition of pH 4.8 precipitate to APF significantly decreased (p<0.05) loaf volume compared to bread made from bread flour. The results from sensory analysis showed there was no difference in preference for any of the breads. This study showed no conclusive evidence that peroxidase enzyme improved gluten strength or loaf volume of yeast bread, but further research is warranted. / Master of Science