Effect of Aspergillus niger prolyl endopeptidase in patients with celiac disease on a long-term gluten-free diet.

BACKGROUND: The gluten-free diet (GFD) has limitations, and there is intense research in the development of adjuvant therapies. AIM: To examine the effects of orally administered Aspergillus niger prolyl endopeptidase protease (AN-PEP) on inadvertent gluten exposure and symptom prevention in adult celiac disease (CeD) patients following their usual GFD. METHODS: This was an exploratory, double-blind, randomized, placebo-controlled trial that enrolled CeD patients on a long-term GFD. After a 4-wk run-in period, patients were randomized to 4 wk of two AN-PEP capsules (GliadinX; AVI Research, LLC, United States) at each of three meals per day or placebo. Outcome endpoints were: (1) Average weekly stool gluten immunogenic peptides (GIP) between the run-in and end of treatments and between AN-PEP and placebo; (2) celiac symptom index (CSI); (3) CeD-specific serology; and (4) quality of life. Stool samples were collected for GIP testing by ELISA every Tuesday and Friday during run-ins and treatments. RESULTS: Forty patients were randomized for the intention-to-treat analysis, and three were excluded from the per-protocol assessment. Overall, 628/640 (98.1%) stool samples were collected. GIP was undetectable (< 0.08 µg/g) in 65.6% of samples, and no differences between treatment arms were detected. Only 0.5% of samples had GIP concentrations sufficiently high (> 0.32 µg/g) to potentially cause mucosal damage. Median GIP concentration in the AN-PEP arm was 44.7% lower than in the run-in period. One-third of patients exhibiting GIP > 0.08 µg/g during run-in had lower or undetectable GIP after AN-PEP treatment. Compared with the run- in period, the proportion of symptomatic patients (CSI > 38) in the AN-PEP arm was significantly lower (P < 0.03). AN-PEP did not result in changes in specific serologies. CONCLUSION: This exploratory study conducted in a real-life setting revealed high adherence to the GFD. The AN-PEP treatment did not significantly reduce the overall GIP stool concentration. However, given the observation of a significantly lower prevalence of patients with severe symptoms in the AN-PEP arm, further clinical research is warranted.

Who are the sexual and gender minorities who frequently interact with children and their association with healthcare. / Quem são as minorias sexuais e de gênero que convivem frequentemente com filhos(as) e sua associação a cuidados em saúde.

This study aimed to examine the sociodemographic profile of sexual and gender minorities who regularly interact with children and investigate whether such frequent interactions are associated with healthcare factors. This cross-sectional study utilized data from the LGBT+ Health Survey in Brazil, conducted online and anonymously from August to November 2020 with 958 participants. Regular interaction with children was defined as living with children or engaging in bi-weekly face-to-face meetings with children residing in different households. Healthcare factors encompass having a professional or reference service, feeling comfortable in discussing personal issues, and receiving worse quality medical or hospital care. The statistical analysis used the Poisson regression with robust variance. The prevalence of interaction with children was 5.3%. We observed a statistically higher prevalence among cisgender women (13.4%) and Black/brown and other non-white people (7.9%) after adjusting for age. The results showed a positive association only between regular interaction with children and worse-quality medical or hospital care received (PR=6.00; 95%CI 1.22-29.67). These findings highlight a persistent stigma and prejudice within healthcare services.

Objetivou-se analisar as características sociodemográficas das minorias sexuais e de gênero que convivem frequentemente com filhos(as) e verificar se existe associação entre convívio frequente com filhos(as) e os cuidados em saúde. Trata-se de um estudo transversal com dados do inquérito de saúde LGBT+, realizado no Brasil em 2020 (agosto-novembro) de forma on-line e anônima, totalizado 958 participantes. O convívio frequente com filhos(as) foi avaliado pela moradia com filhos(as) ou encontros presenciais quinzenais com filhos(as) que moram em outro domicílio. Os cuidados em saúde incluíram ter um profissional ou serviço de referência, sentir-se à vontade para contar seus problemas e receber tratamento médico ou hospitalar de pior qualidade. A regressão de Poisson com variância robusta foi usada na análise estatística. A prevalência de convívio com filhos(as) foi de 5,3%. Após o ajuste por idade, verificou-se uma prevalência estatisticamente maior em mulheres cisgênero (13,4%) e entre pretos/pardos e outras raças/cores não brancas (7,9%). Observou-se que o convívio frequente com filhos(as) foi positivamente associado apenas a receber tratamento médico ou hospitalar de pior qualidade (RP=6,00; IC95% 1,22-29,67). Esses achados destacam que ainda há estigma/preconceito nos serviços de saúde.

Case report: Primary CDK4/6 inhibitor and endocrine therapy in locally advanced breast cancer and its effect on gut and intratumoral microbiota.

Locally advanced breast cancer poses significant challenges to the multidisciplinary team, in particular with hormone receptor (HR) positive, HER2-negative tumors that classically yield lower pathological complete responses with chemotherapy. The increasingly significant use of CDK 4/6 inhibitors (CDK4/6i) plus endocrine therapy (ET) in different breast cancer settings has led to clinical trials focusing on this strategy as a primary treatment, with promising results. The impact of the microbiota on cancer, and vice-versa, is an emerging topic in oncology. The authors report a clinical case of a postmenopausal female patient with an invasive breast carcinoma of the right breast, Luminal B-like, staged as cT4cN3M0 (IIIB). Since the lesion was considered primarily inoperable, the patient started letrozole and ribociclib. Following 6 months of systemic therapy, the clinical response was significant, and surgery with curative intent was performed. The final staging was ypT3ypN2aM0, R1, and the patient started adjuvant letrozole and radiotherapy. This case provides important insights on primary CDK4/6i plus ET in locally advanced unresectable HR+/HER2- breast cancer and its potential implications in disease management further ahead. The patient's gut microbiota was analyzed throughout the disease course and therapeutic approach, evidencing a shift in gut microbial dominance from Firmicutes to Bacteroidetes and a loss of microbial diversity following 6 months of systemic therapy. The analysis of the intratumoral microbiota from the surgical specimen revealed high microbial dissimilarity between the residual tumor and respective margins.

Thiazolyl-isatin derivatives: Synthesis, in silico studies, in vitro biological profile against breast cancer cells, mRNA expression, P-gp modulation, and interactions of Akt2 and VIM proteins.

The literature reports that thiazole and isatin nuclei present a range of biological activities, with an emphasis on anticancer activity. Therefore, our proposal was to make a series of compounds using the molecular hybridization strategy, which has been used by our research group, producing hybrid molecules containing the thiazole and isatin nuclei. After structural planning and synthesis, the compounds were characterized and evaluated in vitro against breast cancer cell lines (T-47D, MCF-7 and MDA-MB-231) and against normal cells (PBMC). The activity profile on membrane proteins involved in chemoresistance and tumorigenic signaling proteins was also evaluated. Among the compounds tested, the compounds 4c and 4a stood out with IC50 values of 1.23 and 1.39 µM, respectively, against the MDA-MB-231 cell line. Both compounds exhibited IC50 values of 0.45 µM for the MCF-7 cell line. Compounds 4a and 4c significantly decreased P-gp mRNA expression levels in MCF-7, 4 and 2 folds respectively. Regarding the impact on tumorigenic signaling proteins, compound 4a inhibited Akt2 in MDA-MB-231 and compound 4c inhibited the mRNA expression of VIM in MCF-7.

Rapid Anti-tTG-IgA Screening Test for Early Diagnosis of Celiac Disease in Pediatric Populations.

A large number of patients with celiac disease (CD) remain undiagnosed because they do not fulfill the criteria for entry into the conventional diagnostic workflow. This study evaluated the clinical utility of anti-tissue transglutaminase IgA antibody lateral flow immunoassays (anti-tTG-IgA LFIA) in the undiagnosed-CD-based pediatric population and the impact of a gluten-free diet (GFD) on screening-detected CD. A total of 576 volunteers were tested for anti-tTG-IgA. Gluten consumption habits, CD related symptoms, and risk factors for CD development were evaluated. Volunteers testing positive for anti-tTG-IgA were referred to the conventional CD diagnostic workflow, and the impact of the GFD on health-related quality of life (HR-QoL) was measured. Among them, 13 had a positive anti-tTG-IgA LFIA test result: 11 had confirmed CD (1.91%), one refused confirmatory tests, and another is undergoing diagnosis. Regarding the CD prevalence, no significant differences were observed among risk (1.89%) and symptomatic (2.65%) groups and the entire tested population (1.55%). Rapid anti-tTG-IgA LFIAs could be of clinical utility in primary care for the early identification of children with CD unidentified by the conventional diagnostic workflow. It could potentially reduce the costs of undiagnosed CD, avoiding unnecessary referrals to gastroenterologists, reducing diagnosis delays and long-term problems, and improving patients' HR-QoL.

Evaluation of the Usefulness of an Automatable Immunoassay for Monitoring Celiac Disease by Quantification of Immunogenic Gluten Peptides in Urine.

A gluten-free diet (GFD) is currently the only treatment available for patients with celiac disease (CD). However, adherence to a GFD can be challenging because gluten is present in many foods. A lifelong follow-up of patients with CD must be performed to promote adherence to a GFD and to identify the appearance of symptoms and the associated diseases. Therefore, the development of tools to analyze gluten exposure in these patients is important. This study proposes the development of the first automatable ELISA to monitor adherence to a GFD through the quantification of urine gluten immunogenic peptides (u-GIP). Seven healthy volunteers without suspicion of CD and 23 patients with CD were monitored as part of this study to optimize, validate, and apply this assay. Non-interference was found in the urine matrix, and the recovery percentage for spiked samples was 81-101%. The u-GIP was stable for up to 16 days when the samples were stored at different temperatures. Overall, 100% of the patients had detectable u-GIP at diagnosis (range of 0.39-2.14 ng GIP/mL), which reduced to 27% after 12 months on a GFD. Therefore, this highly sensitive immunoassay would allow the analysis of u-GIP from a large battery of samples in clinical laboratories of specialized healthcare centers.

Clinical utility of urinary gluten immunogenic peptides in the follow-up of patients with coeliac disease.

BACKGROUND: Gluten-free diet (GFD) is the only treatment for patients with coeliac disease (CD) and its compliance should be monitored to avoid cumulative damage. AIMS: To analyse gluten exposures of coeliac patients on GFD for at least 24 months using different monitoring tools and its impact on duodenal histology at 12-month follow-up and evaluate the interval of determination of urinary gluten immunogenic peptides (u-GIP) for the monitoring of GFD adherence. METHODS: Ninety-four patients with CD on a GFD for at least 24 months were prospectively included. Symptoms, serology, CDAT questionnaire, and u-GIP (three samples/visit) were analysed at inclusion, 3, 6, and 12 months. Duodenal biopsy was performed at inclusion and 12 months. RESULTS: At inclusion, 25.8% presented duodenal mucosal damage; at 12 months, this percentage reduced by half. This histological improvement was indicated by a reduction in u-GIP but did not correlate with the remaining tools. The determination of u-GIP detected a higher number of transgressions than serology, regardless of histological evolution type. The presence of >4 u-GIP-positive samples out of 12 collected during 12 months predicted histological lesion with a specificity of 93%. Most patients (94%) with negative u-GIP in ≥2 follow-up visits showed the absence of histological lesions (p < 0.05). CONCLUSION: This study suggests that the frequency of recurrent gluten exposures, according to serial determination of u-GIP, could be related to the persistence of villous atrophy and that a more regular follow-up every 6 months, instead of annually, provides more useful data about the adequate adherence to GFD and mucosal healing.

Enhanced bioprocess control to advance the manufacture of mesenchymal stromal cell-derived extracellular vesicles in stirred-tank bioreactors.

Extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) act as signaling mediators of cellular responses. However, despite representing a promising alternative to cell-based therapies, clinical translation of EVs is currently limited by their lack of scalability and standardized bioprocessing. Herein, we integrated scalable downstream processing protocols with standardized expansion of large numbers of viable cells in stirred-tank bioreactors to improve EV production. Higher EV yields were linked to EV isolation by tangential flow filtration followed by size exclusion chromatography, rendering 5 times higher number of EVs comparatively to density gradient ultracentrifugation protocols. Additionally, when compared to static culture, EV manufacture in bioreactors resulted in 2.2 higher yields. Highlighting the role of operating under optimal cell culture conditions to maximize the number of EVs secreted per cell, MSCs cultured at lower glucose concentration favored EV secretion. While offline measurements of metabolites concentration can be performed, in this work, Raman spectroscopy was also applied to continuously track glucose levels in stirred-tank bioreactors, contributing to streamline the selection of optimal EV collection timepoints. Importantly, MSC-derived EVs retained their quality attributes and were able to stimulate angiogenesis in vitro, therefore highlighting their promising therapeutic potential.

Molecular profile and peripheral markers of neurodegeneration in patients with Niemann-Pick type C: Decrease in Plasminogen Activator Inhibitor type 1 and Platelet-Derived Growth Factor type AA.

Niemann-Pick type C1 (NPC1) is a fatal inherited disease, caused by pathogenic variants in NPC1 gene, which leads to intracellular accumulation of non-esterified cholesterol and glycosphingolipids. This accumulation leads to a wide range of clinical manifestations, including neurological and cognitive impairment as well as psychiatric disorders. The pathophysiology of cerebral damage involves loss of Purkinje cells, synaptic disturbance, and demyelination. Miglustat, a reversible inhibitor of glucosylceramide synthase, is an approved treatment for NPC1 and can slow neurological damage. The aim of this study was to assess the levels of peripheric neurodegeneration biomarkers of NPC1 patients, namely brain-derived neurotrophic factor (BDNF), platelet-derived growth factors (PDGF-AA and PDGF-AB/BB), neural cell adhesion molecule (NCAM), PAI-1 Total and Cathepsin-D, as well as the levels of cholestane-3ß,5α,6ß-triol (3ß,5α,6ß-triol), a biomarker for NPC1. Molecular analysis of the NPC1 patients under study was performed by next generation sequencing (NGS) in cultured fibroblasts. We observed that NPC1 patients treated with miglustat have a significant decrease in PAI-1 total and PDGF-AA concentrations, and no alteration in BDNF, NCAM, PDGF-AB/BB and Cathepsin D. We also found that NPC1 patients treated with miglustat have normalized levels of 3ß,5α,6ß-triol. The molecular analysis showed four described mutations, and for two patients was not possible to identify the second mutated allele. Our results indicate that the decrease of PAI-1 and PDGF-AA in NPC1 patients could be involved in the pathophysiology of this disease. This is the first work to analyze those plasmatic markers of neurodegenerative processes in NPC1 patients.

Adherence to glucose monitoring with intermittently scanned continuous glucose monitoring in patients with type 1 diabetes.

PURPOSE: This study aims to predict the Intermittently scanned continuous glucose monitoring (isCGM) adherence behavior of patients with Type 1 Diabetes. METHODS: Patients with Type 1 Diabetes mellitus using FreeStyle Libre™ System (FL), a isCGM device, that attended the "Insulin Infusion Pump clinic" at Centro Hospitalar de São João were enrolled and evaluated for sociodemographic and clinical characterization, beliefs and concerns about Diabetes Mellitus, as well as isCGM's perceptions. Intermittently scanned continuous glucose monitoring data were collected to characterize monitoring patterns and to measure isCGM's adherence-FL average of scans/day. RESULTS: Seventy-two patients with a mean of 30.36 years (sd=11.35) participate in this study. A median of 7 scans/day was performed. The adherence predictors found was Age (ß = 0.191, p = 0.006), Time in target (ß = 0.530, p = 0.002), isCGM Necessity (ß = 2.631, p = 0.048), Body Mass Index (ß = -0.549, p = 0.017) and Sex (ß = -3.996; p = 0.011). CONCLUSIONS: This study emphasizes the relevance of glucose monitoring adherence in disease control and shows that males of younger ages, presenting with higher body mass index levels, lower time in target, and reporting lower isCGM necessity are less adherent to isCGM. Therefore, these patients should be closely followed and object of personalized strategies to promote treatment adherence.

Complicações cardiovasculares em gestante com COVID-19: revisão integrativa / Cardiovascular complications in pregnant womem with COVID-19: integrative review

OBJETIVO: Identificar as complicações cardiovasculares na gestante com COVID-19 e seus desfechos. MÉTODO: A revisão integrativa foi norteada pela pergunta sobre complicações cardiovasculares da gestante com Covid-19. Foram incluídos estudos primários cujos desfechos eram complicações cardiovasculares em gestantes com covid-19, publicados em inglês, português ou espanhol nas fontes do PubMed ou BVS entre 2016 e 2021. RESULTADOS: Observou-se como principais complicações cardiovasculares da gestação com Covid-19, a trombose arterial, venosa, coagulopatias, cardiomiopatias, insuficiência cardíaca e injúria miocárdica, e o principal desfecho encontrado foi a necessidade de parto cesárea relacionado ao sofrimento fetal, medo de transmissão vertical de COVID-19 pelos profissionais de saúde e complicações de saúde da própria gestante. CONCLUSÕES: As complicações cardiovasculares foram identificadas por exames laboratoriais, de imagem e eletrocardiograma e os resultados possibilitaram ampliar o conhecimento dos profissionais da área da saúde, das gestantes nas internações e dos riscos das complicações cardiovasculares para a gestante com Covid-19.

Nurses' Involvement in the Development and Usability Assessment of an Innovative Peripheral Intravenous Catheterisation Pack: A Mix-Method Study.

Guaranteeing peripheral venous access is one of the cornerstones of modern healthcare. Recent evidence shows that the lack of adequate clinical devices can result in the provision of substandard care to patients who require peripheral intravenous catheterization (PIVC). To address this challenge, we aimed to develop a PIVC pack for adult patients and assess the usability of this new device. METHODS: Following a mix-method design, the PIVC pack development and usability assessment were performed in two phases with the involvement of its potential end-users (nurses). In phase one (concept and semi-functional prototype assessment), focus group rounds were conducted, and a usability assessment questionnaire was applied at each stage. In phase two (pre-clinical usability assessment), a two-arm crossover randomised controlled trial (PIVC pack versus traditional material) was conducted with nurses in a simulated setting. Final interviews were conducted to further explore the PIVC pack applicability in a real-life clinical setting. RESULTS: High average usability scores were identified in each study phase. During the pre-clinical usability assessment, the PIVC pack significantly reduced procedural time (Z = -2.482, p = 0.013) and avoided omissions while preparing the required material (Z = -1.977, p = 0.048). The participating nurses emphasised the pack's potential to standardise practices among professionals, improve adherence to infection control recommendations, and enhance stock management. CONCLUSIONS: The developed pack appears to be a promising device that can assist healthcare professionals in providing efficient and safe care to patients requiring a PIVC. Future studies in real clinical settings are warranted to test its cost-effectiveness.

Análisis bifactorial de la Escala de Autoestima de Rosenberg y relación entre actividad física y autoestima en adolescentes / Bifactorial analysis of the Rosenberg Self-esteem Scale and relationship between physical activity and self-esteem in adolescents

La autoestima es un constructo de gran importancia en la adolescencia que favorece la salud y el bienestar psicológico de los escolares. En este trabajo se analiza la bidimensionalidad de la Escala de Autoestima de Rosenberg (EAR) mediante análisis factorial confirmatorio, así como las relaciones existentes entre la práctica de actividad física, Autoestima Global y las dimensión positiva y negativa de la autoestima. Se llevó a cabo en una muestra de 429 estudiantes de Educación Secundaria Obligatoria y Bachillerato de la ciudad de Sevilla (Andalucía). Los instrumentos utilizados fueron Cuestionario de Actividad Física en adolescentes (PaqA de Martínez-Gómez et al., 2009) y Escala de Autoestima de Rosenberg (EAR, 1965) versión de Atienza et al. (2000). Los resultados obtenidos permiten aceptar la bidimensionalidad de la escala, así mismo se constató buenos niveles de autoestima global y una tendencia a una mejor autoestima en los adolescentes activos. La práctica de actividad física se asocia con la dimensión Autoestima Positiva, pero no con la Autoestima Negativa y la Autoestima Global. (AU)

Self-esteem is a construct of great importance in adolescence that favors the health and psychological well-being of schoolchildren. In this paper, the two-dimensionality of the Rosenberg Self-Esteem Scale (RAS) is analyzed through confirmatory factor analysis, as well as the relationships between the practice of physical activity (PA), Global self-esteem, and the positive and negative dimensions of self-esteem. It was carried out in a sample of 429 students in the high school of the city of Seville (Andalusia). The instruments used were the Physical Activity Questionnaire in adolescents (PaqA, Martínez-Gómez et al., 2009) and the Rosenberg Self-Esteem Scale (EAR, 1965) version of Atienza et al. (2000). The results obtained allow us to accept the two-dimensionality of the scale, as well as good levels of global self-esteem and a tendency to better self-esteem in active adolescents. The practice of PA is associated with the Positive Self-esteem dimension, but not with Negative Self-esteem or Global Self-esteem. (AU)

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 CD.

Consumption of Tritordeum Bread Reduces Immunogenic Gluten Intake without Altering the Gut Microbiota.

Gluten proteins are responsible for the wheat breadmaking quality. However, gluten is also related to human pathologies for which the only treatment is a gluten-free diet (GFD). GFD has gained popularity among individuals who want to reduce their gluten intake. Tritordeum is a cereal species that originated after crossing durum wheat with wild barley and differs from bread wheat in its gluten composition. In this work, we have characterized the immunogenic epitopes of tritordeum bread and results from a four-phase study with healthy adults for preferences of bread and alterations in the gut microbiota after consuming wheat bread, gluten-free bread, and tritordeum bread are reported. Tritordeum presented fewer peptides related to gluten proteins, CD-epitopes, and IgE binding sites than bread wheat. Participants rated tritordeum bread higher than gluten-free bread. Gut microbiota analysis revealed that the adherence to a strict GFD involves some minor changes, especially altering the species producing short-chain fatty acids. However, the short-term consumption of tritordeum bread does not induce significant changes in the diversity or community composition of the intestinal microbiota in healthy individuals. Therefore, tritordeum bread could be an alternative for healthy individuals without wheat-related pathologies who want to reduce their gluten consumption without harming their gut health.

Advances in Celiac Disease and Gluten-Free Diet.

Celiac disease (CD) is a systemic disease that causes chronic enteropathy of the small intestine and develops through an inadequate immune response to gluten in genetically predisposed individuals [...].

An efficient urine peptidomics workflow identifies chemically defined dietary gluten peptides from patients with celiac disease.

Celiac disease (CeD) is an autoimmune disorder induced by consuming gluten proteins from wheat, barley, and rye. Glutens resist gastrointestinal proteolysis, resulting in peptides that elicit inflammation in patients with CeD. Despite well-established connections between glutens and CeD, chemically defined, bioavailable peptides produced from dietary proteins have never been identified from humans in an unbiased manner. This is largely attributable to technical challenges, impeding our knowledge of potentially diverse peptide species that encounter the immune system. Here, we develop a liquid chromatographic-mass spectrometric workflow for untargeted sequence analysis of the urinary peptidome. We detect over 600 distinct dietary peptides, of which ~35% have a CeD-relevant T cell epitope and ~5% are known to stimulate innate immune responses. Remarkably, gluten peptides from patients with CeD qualitatively and quantitatively differ from controls. Our results provide a new foundation for understanding gluten immunogenicity, improving CeD management, and characterizing the dietary and urinary peptidomes.

Individual variability in patterns and dynamics of fecal gluten immunogenic peptides excretion after low gluten intake.

PURPOSE: Determination of Gluten Immunogenic Peptides (GIP) in feces is a direct tool for gluten exposure detection. The sensitivity of GIP detection methods for cases of unintentional low gluten intakes is unknown. We studied the interindividual variability in the kinetic of excretion under homogeneously controlled dietary conditions, and the sensitivity of fecal GIP tests after low amounts of punctual gluten ingestions. METHODS: Participants (n = 20) followed the same gluten-free menu for 12 days in which two separated doses of gluten (50 mg and 2 g) were ingested and all the depositions were collected. GIP from stool samples were analyzed by ELISA and lateral flow immunoassay (LFIA) tests. RESULTS: Most participants had detectable GIP after 50 mg and 2 g gluten ingestions using ELISA test (72.2% and 95%, respectively), whereas the LFIA test showed less sensitivity (22.2% and 80%, respectively). GIP were detected at higher either frequency or concentration in the range of 12-36 h after 50 mg intake, and 12-84 h after 2 g consumption. Considering this period, diagnostic sensitivity of GIP detection after a single 50 mg ingestion may be significatively increased analyzing three stool samples per individual. High variability among participants was found in the time and amount of GIP excretion; however, some individuals showed common patterns for both gluten intakes. CONCLUSION: Sporadic gluten exposure detection may require several fecal samples to achieve level of sensitivity above 90%. Interindividual variability in the dynamic of GIP excretion may suggest patterns of gluten metabolism.