Gluten Immunogenic Peptides as Standard for the Evaluation of Potential Harmful Prolamin Content in Food and Human Specimen

Gluten is a complex mixture of storage proteins in cereals like wheat, barley, and rye. Prolamins are the main components of gluten. Their high content in proline and glutamine makes them water-insoluble and difficult to digest in the gastrointestinal tract. Partial digestion generates peptide sequences which trigger immune responses in celiac and gluten-sensitive patients. Gluten detection in food is challenging because of the diversity, in various food matrices, of protein proportions or modifications and the huge number of immunogenic sequences with differential potential immunoactivity. Attempts to develop standard reference materials have been unsuccessful. Recent studies have reported the detection of a limited number of dominant Gluten Immunogenic Peptides (GIP) that share similarities to epitopes presented in the α-gliadin 33-mer, which showed to be highly proteolytic resistant and is considered to be the most immunodominant peptide within gluten in celiac disease (CD). GIP were detectable and quantifiable in very different kind of difficult to analyze food, revealing the potential immunogenicity by detecting T-cell activity of celiac patients. But GIP were also found in stool and urine of celiac patients on a supposedly gluten-free diet (GFD), showing the capacity to resist and be absorbed and excreted from the body, providing the first simple and objective means to assess adherence to the GFD. Methods to specifically and sensitively detect the most active GIP in food and biological fluids are rational candidates may use similar analytical standard references for determination of the immunopathological risk of gluten exposure in gluten-related diseases.España, MINECO AGL2013-48946-CEspaña, Ministerio de Ciencia, Innovación y Universidades y Corporación Tecnológica de Andalucía (CTA

Update on nutritional aspects of gluten-free diet in celiac patients

A strict gluten - free diet is the only currently available therapeutic treatment for patients with celiac disease, an autoimmune disorder of the small intestine associated with a permanent intolerance to gluten proteins. In recent years, the dramatically prompted changes in the dietary habit of an increasingly large population (celiac disease, non - celiac gluten sensitivit y and gluten allergy) has resulted in rising demands for gluten - free products. Before starting gluten - free diet, alteration in intestinal absorption capacity of celiac patients involves deficiencies of nutrients, vitamins and dietary minerals. The habitual poor gluten - free food choices in addition to inherent deficiencies in the gluten - free diet of diagnosed celiac patients may relate with dietary inadequacies. Therefore dietary assessment and counseling at the time of celiac disease diagnosis and ongoing c are are crucial as well as fortification of gluten - free foods also need to be considered. This article reviews the nutritional aspects of gluten - free diet in celiac patients and provides an up - date of dietetic recommendations to correct these deficiencies and to ensure optimum gluten - free diet compliance

Alternative grains as potential raw material for gluten– free food development in the diet of celiac and gluten– sensitive patients

Celiac disease is an autoimmune disorder resulting from gluten intolerance and is based on a genetic predisposition. Gluten is a protein composite found in the cereals wheat, rye, barley and certain oat varieties. A strict gluten-free diet is the only currently available therapeutic treatment for patients with celiac disease. Rising demands for gluten-free products parallels the apparent or real increase in celiac disease, non-celiac gluten sensitivity and gluten allergy. However, gluten removal results in major problems for bakers, and currently, many gluten-free products available on the market are of low quality exhibiting poor mouthfeel and flvor. Thus, an increasing trend in research is focusing on the application of alternative grains potentially healthy to elaborate gluten-free products. A promising area is the use of cereals (rice, corn and sorghum), minor cereals (fonio, teff, millet and job’s tears) or pseudocereals such as amaranth, buckwheat, quinoa. Nevertheless, commercialization of these products is still quite limited. The aim of this work is to review recent advances in research about the nutritional quality and potential health benefis of alternative grains tolerated by patients with gluten related pathologies.Junta de Andalucía P09AGR-478

Accessibility evaluation of Moodle centred in visual impairements

Proceedings of: 7th International Conference on Web Information Systems and Technologies (WEBIST 2011), Noordwijkerhout, The Netherlands, May 6-9, 2011The use of new technologies has been increasing during last years in education. More specifically, the use of e-learning systems provides for students more freedom to learn through Internet when and where they prefer in each moment. However, e-learning tools are not perfect tools. Most of these e-learning tools present accessibility barriers, so not all students are able to use them completely in their studies. The goal of this paper is to evaluate if Moodle e-learning web tool is accessible for visually impaired people using assistive technologies like screen readers. The evaluation has been divided in two main objectives: first, to evaluate Moodle from a user perspective, simulating the interaction of a blind student with the system and using a screen reader; second, to evaluate Moodle from an accessibility expert perspective, analysing if Moodle is in accordance to ATAG and WCAG W3C guidelines.The work presented in this paper has been partially founded by GEMMA (TSI-020302-2010-141) and SAGAS (TSI-020100-2010-184) research projects

Accessibility barriers for users of screen readers in the Moodle learning content management system

In recent decades, the use of the Internet has spread rapidly into diverse social spheres including that of education. Currently, most educational centers make use of e-learning environments created through authoring tool applications like learning content management systems (LCMSs). However, most of these applications currently present accessibility barriers that make the creation of accessible e-learning environments difficult for teachers and administrators. In this paper, the accessibility of the Moodle authoring tool, one of the most frequently used LCMSs worldwide, is evaluated. More specifically, the evaluation is carried out from the perspective of two visually impaired users accessing content through screen readers, as well as a heuristic evaluation considering the World Wide Web Consortium's Authoring Tool Accessibility Guidelines. The evaluation results demonstrate that Moodle presents barriers for screen reader users, limiting their ability to access the tool. One example of accessibility problems for visually impaired users is the frequent inability to publish learning contents without assistance. In light of these results, the paper offers recommendations that can be followed to reduce or eliminate these accessibility barriers.This study has been partially funded by the MA2VICMR (S2009/TIC-1542) research project.Publicad

Role of oats in celiac disease

A gluten-free diet is currently the only effective means of treating individuals with celiac disease. Such a diet enables celiac patients to control their symptoms and avoid various complications associated with this condition. However, while the quality of gluten-free foods has significantly improved during recent decades, maintenance of a gluten-free diet does not necessarily ensure adequate nutritional intake. Because oats are an important source of proteins, lipids, vitamins, minerals, and fibre, their inclusion in a gluten-free diet might improve the nutritional status of a celiac patient. Although oats are included in the list of gluten-free ingredients specified in European regulations, their safety when consumed by celiac patients remains debatable. Some studies claim that pure oats are safe for most celiac people, and contamination with other cereal sources is the main problem facing people with this disease. However, it is necessary to consider that oats include many varieties, containing various amino acid sequences and showing different immunoreactivities associated with toxic prolamins. As a result, several studies have shown that the immunogenicity of oats varies depending on the cultivar consumed. Thus, it is essential to thoroughly study the variety of oats used in a food ingredient before including it in a gluten-free diet

Quality of Life in Teenagers and Adults With Coeliac Disease: From Newly Spanish Coeliac Disease Questionnaire Validation to Assessment in a Population-Based Study

Background: Coeliac disease (CD) is an immune-mediated systemic disorder elicited by the ingestion of gluten in genetically predisposed individuals. Gluten restriction in CD sufferers leads to numerous limitations in various aspects of daily life and can significantly impact the quality-of-life (QoL). The specific and widely used Coeliac Disease Questionnaire (CDQ) is an excellent tool to evaluate QoL in patients with CD, assessing physical, psychological, and social domains. This questionnaire is unavailable in Spain. Therefore, our study is the first to translate, culturally adapt, validate, and apply the Spanish version of CDQ to a representative sample of Spanish teenagers and adults with CD. Methods: A total of 153 CD participants with biopsy-proven and self-reported gluten-free adherence were included in the cross-sectional study, which included four stages: (1) translation and retranslation of the French CDQ version into Spanish; (2) cultural adaptation and semantic evaluation; (3) CDQ validation through the internal consistency determination and reproducibility of the QoL; and (4) application of the questionnaire to Spanish teenagers and adults with CD and estimation of QoL using EQ-5D. Results: The internal consistency and test–retest reliability of the Spanish CDQ were satisfactory and no ceiling or floor effects were detected. Significant correlations were identified between the CDQ scales, and the instrument for validation covering similar dimensions of the QoL was identified. The mean CDQ total score was 131.03 ± 24.1, and the social domain had the highest rating. There was no correlation between the time spent on a gluten-free diet and QoL. A significantly higher QoL score was reported among males and adolescents in the 15–17 age groups. Conclusion: The newly Spanish CDQ is an appropriate tool to assess the QoL of the teenager and adult patients with CD. This study highlights the importance of identifying the affected scales to address actions to reduce the impact of the gluten-free diet burden of the coeliac patients and maintain public health regulations that support patients with chronic diseases such as CDCentro para el Desarrollo Tecnologico Industrial 20/0112Corporación Tecnológica de Andalucía 20/011

Biomarkers to monitor gluten-free diet compliance in celiac patients

Gluten-free diet (GFD) is the only treatment for celiac disease (CD). There is a general consensus that strict GFD adherence in CD patients leads to full clinical and histological remission accompanied by improvement in quality of life and reduced long-term complications. Despite the importance of monitoring the GFD, there are no clear guidelines for assessing the outcome or for exploring its adherence. Available methods are insufficiently accurate to identify occasional gluten exposure that may cause intestinal mucosal damage. Serological tests are highly sensitive and specific for diagnosis, but do not predict recovery and are not useful for follow-up. The use of serial endoscopies, it is invasive and impractical for frequent monitoring, and dietary interview can be subjective. Therefore, the detection of gluten immunogenic peptides (GIP) in feces and urine have been proposed as new non-invasive biomarkers to detect gluten intake and verify GFD compliance in CD patients. These simple immunoassays in human samples could overcome some key unresolved scientific and clinical problems in CD management. It is a significant advance that opens up new possibilities for the clinicians to evaluate the CD treatment, GFD compliance, and improvement in the quality of life of CD patient

A new microbial gluten-degrading prolyl endopeptidase: Potential application in celiac disease to reduce gluten immunogenic peptides

Gluten is a complex of proteins present in barley, wheat, rye and several varieties of oats that triggers celiac disease in genetically predisposed subjects. Gluten is notoriously difficult to digest by mammalian proteolytic enzymes and therefore, proline-rich digestion-resistant peptides contain multiple immunogenic epitopes. Prolyl endopeptidases (PEP) hydrolyse internal proline residues on the carboxyl side of peptides and have been proposed for food gluten detoxification and as oral enzyme supplementation for celiacs. The aim of this study was to identify new gluten-degrading microbial enzymes with the potential to reduce gluten immunogenicity by neutralizing its antigenic epitopes. Using a gluten-degrading colony screening approach, a bacterial isolate (2RA3) displaying the highest glutenase activity was selected, characterized and its genome completely sequenced. The identification through 16S rDNA gene sequencing showed a 99,1% similarity to Chryseobacterium taeanense. Hydrolysis of gluten immunogenic peptides (GIP) was further monitored, over a 48-hour period, by colony encapsulation in gliadin-containing microspheres, followed by detection with the G12 anti-GIP monoclonal antibody. Glutenase activity was detected in the extracellular medium of 2RA3 cultures, where gel electrophoresis and gliadin zymography revealed the presence of a ~50 kDa gluten-degrading enzyme. Nano-ESI-Q-TOF of the excised active band identified 7 peptides contained in the protein product predicted for an open reading frame (ORF) in the 2RA3 genome. Based on sequence similarity to the PEP family, the new enzyme was named PEP 2RA3. The PEP 2RA3 coding sequence was PCR-amplified from C. taeanense 2RA3, cloned and expressed in Escherichia coli as a C-terminally His-tagged recombinant protein and purified by Ni-NTA affinity chromatography. The recombinant protein, with predicted molecular mass and isoelectric point of 78.95 kDa and 6.8, respectively, shows PEP activity with standard chromogenic substrates, works optimally at pH 8.0 and 30°C and remains stable at pH 6.0 and 50°C, indicating a potential use in gluten-containing food process applications. The ability of the recombinant enzyme to degrade GIP in beer into smaller peptides was confirmed.Ministry of Science and Innovation AGL2009-0906