EoE CONNECT, the European registry of clinical, environmental, and genetic determinants in eosinophilic esophagitis: rationale, design, and study protocol of a large-scale epidemiological study in Europe

Artículo escrito por un elevado número de autores, solo se referencian el que aparece en primer lugar, el nombre del grupo de colaboración, si lo hubiere, y los autores pertenecientes a la UAMThe growing prevalence of eosinophilic esophagitis (EoE) represents a considerable burden to patients and health care systems. Optimizing cost-effective management and identifying mechanisms for disease onset and progression are required. However, the paucity of large patient cohorts and heterogeneity of practice hinder the defining of optimal management of EoE. Methods: EoE CONNECT is an ongoing, prospective registry study initiated in 2016 and currently managed by EUREOS, the European Consortium for Eosinophilic Diseases of the Gastrointestinal Tract. Patients are managed and treated by their responsible specialists independently. Data recorded using a web-based system include demographic and clinical variables; patient allergies; environmental, intrapartum, and early life exposures; and family background. Symptoms are structurally assessed at every visit; endoscopic features and histological findings are recorded for each examination. Prospective treatment data are registered sequentially, with new sequences created each time a different treatment (active principle, formulation, or dose) is administered to a patient. EoE CONNECT database is actively monitored to ensure the highest data accuracy and the highest scientific and ethical standards. Results: EoE CONNECT is currently being conducted at 39 centers in Europe and enrolls patients of all ages with EoE. In its aim to increase knowledge, to date EoE CONNECT has provided evidence on the effectiveness of first- and second-line therapies for EoE in clinical practice, the ability of proton pump inhibitors to induce disease remission, and factors associated with improved response. Drug effects to reverse fibrous remodeling and endoscopic features of fibrosis in EoE have also been assessed. Conclusion: This prospective registry study will provide important information on the epidemiological and clinical aspects of EoE and evidence as to the real-world and long-term effectiveness and safety of therapy. These data will potentially be a vital benchmark for planning future EoE health care services in EuropeThe authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: The establishment and design of the EoE CONNECT registry was developed with a grant from the United European Gastroenterology through the National Societies Link Award program. The maintenance of the database is financed by EUREOS (European Society of Eosinophilic Oesophagitis). Funding agencies had no role in the study design, in the writing of this manuscript, or the decision to submit for publicatio

Piphillin predicts metagenomic composition and dynamics from DADA2-corrected 16S rDNA sequences

Shotgun metagenomic sequencing reveals the potential in microbial communities. However, lower-cost 16S ribosomal RNA (rRNA) gene sequencing provides taxonomic, not functional, observations. To remedy this, we previously introduced Piphillin, a software package that predicts functional metagenomic content based on the frequency of detected 16S rRNA gene sequences corresponding to genomes in regularly updated, functionally annotated genome databases. Piphillin (and similar tools) have previously been evaluated on 16S rRNA data processed by the clustering of sequences into operational taxonomic units (OTUs). New techniques such as amplicon sequence variant error correction are in increased use, but it is unknown if these techniques perform better in metagenomic content prediction pipelines, or if they should be treated the same as OTU data in respect to optimal pipeline parameters

Poor sensitivity of fecal gluten immunogenic peptides and serum antibodies to detect duodenal mucosal damage in celiac disease monitoring

A lifelong gluten-free diet (GFD) is the only current treatment for celiac disease (CD), but strict compliance is complicated. Duodenal biopsies are the “gold standard” method for diagnosing CD, but they are not generally recommended for disease monitoring. We evaluated the sensitivity and specificity of fecal gluten immunogenic peptides (GIPs) to detect duodenal lesions in CD patients on a GFD and compared them with serum anti-tissue transglutaminase (tTG) IgA antibodies. A prospective study was conducted at two tertiary centers in Spain on a consecutive series of adolescents and adults with CD who maintained a long-lasting GFD. Adherence to a GFD and health-related quality of life were scored with validated questionnaires. Mucosal damage graded according to the Marsh–Oberhüber classification (Marsh 1/2/3) was used as the reference standard. Of the 97 patients included, 27 presented duodenal mucosal damage and 70 had normal biopsies (Marsh 0). The sensitivity (33%) and specificity (81%) of GIPs were similar to those provided by the two assays used to measure anti-tTG antibodies. Scores in questionnaires showed no association with GIP, but an association between GIPs and patients’ self-reported gluten consumption was found (p = 0.003). GIP displayed low sensitivity but acceptable specificity for the detection of mucosal damage in CD.This research was funded by a grant from Asociación Castellana de Aparato Digestivo (year 2018) to A.J.L

Evaluation of a New Monoclonal Chemiluminescent Immunoassay Stool Antigen Test for the Diagnosis of Helicobacter pylori Infection: A Spanish Multicentre Study

The stool antigen test (SAT) represents an attractive alternative for detection of Helicobacter pylori. The aim of this study was to assess the accuracy of a new SAT, the automated LIAISON(R) Meridian H. pylori SA based on monoclonal antibodies, compared to the defined gold standard C-13-urea breath test (UBT). This prospective multicentre study (nine Spanish centres) enrolled patients >= 18 years of age with clinical indication to perform UBT for the initial diagnosis and for confirmation of bacterial eradication. Two UBT methods were used: mass spectrometry (MS) including citric acid (CA) or infrared spectrophotometry (IRS) without CA. Overall, 307 patients (145 naive, 162 with confirmation of eradication) were analysed. Using recommended cut-off values (negative SAT = 1.10) the sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 67%, 97%, 86%, 92% and 91%, respectively, obtaining an area under the receiver operating characteristic (ROC) curve (AUC) of 0.85. Twenty-eight patients, including seven false positives and 21 false negatives, presented a discordant result between SAT and UBT. Among the 21 false negatives, four of six tested with MS and 11 of 15 tested with IRS presented a borderline UBT delta value. In 25 discordant samples, PCR targeting H. pylori DNA was performed to re-assess positivity and SAT accuracy was re-analysed: sensitivity, specificity, positive predictive value, negative predictive value, accuracy and AUC were 94%, 97%, 86%, 99%, 97% and 0.96, respectively. The new LIAISON(R) Meridian H. pylori SA SAT showed a good accuracy for diagnosis of H. pylori infection

EoE CONNECT, the European Registry of Clinical, Environmental, and Genetic Determinants in Eosinophilic Esophagitis:rationale, design, and study protocol of a large-scale epidemiological study in Europe

BACKGROUND: The growing prevalence of eosinophilic esophagitis (EoE) represents a considerable burden to patients and health care systems. Optimizing cost-effective management and identifying mechanisms for disease onset and progression are required. However, the paucity of large patient cohorts and heterogeneity of practice hinder the defining of optimal management of EoE. METHODS: EoE CONNECT is an ongoing, prospective registry study initiated in 2016 and currently managed by EUREOS, the European Consortium for Eosinophilic Diseases of the Gastrointestinal Tract. Patients are managed and treated by their responsible specialists independently. Data recorded using a web-based system include demographic and clinical variables; patient allergies; environmental, intrapartum, and early life exposures; and family background. Symptoms are structurally assessed at every visit; endoscopic features and histological findings are recorded for each examination. Prospective treatment data are registered sequentially, with new sequences created each time a different treatment (active principle, formulation, or dose) is administered to a patient. EoE CONNECT database is actively monitored to ensure the highest data accuracy and the highest scientific and ethical standards. RESULTS: EoE CONNECT is currently being conducted at 39 centers in Europe and enrolls patients of all ages with EoE. In its aim to increase knowledge, to date EoE CONNECT has provided evidence on the effectiveness of first- and second-line therapies for EoE in clinical practice, the ability of proton pump inhibitors to induce disease remission, and factors associated with improved response. Drug effects to reverse fibrous remodeling and endoscopic features of fibrosis in EoE have also been assessed. CONCLUSION: This prospective registry study will provide important information on the epidemiological and clinical aspects of EoE and evidence as to the real-world and long-term effectiveness and safety of therapy. These data will potentially be a vital benchmark for planning future EoE health care services in Europe

EoE CONNECT, the European Registry of Clinical, Environmental, and Genetic Determinants in Eosinophilic Esophagitis: rationale, design, and study protocol of a large-scale epidemiological study in Europe

Background: The growing prevalence of eosinophilic esophagitis (EoE) represents a considerable burden to patients and health care systems. Optimizing cost-effective management and identifying mechanisms for disease onset and progression are required. However, the paucity of large patient cohorts and heterogeneity of practice hinder the defining of optimal management of EoE. Methods: EoE CONNECT is an ongoing, prospective registry study initiated in 2016 and currently managed by EUREOS, the European Consortium for Eosinophilic Diseases of the Gastrointestinal Tract. Patients are managed and treated by their responsible specialists independently. Data recorded using a web-based system include demographic and clinical variables; patient allergies; environmental, intrapartum, and early life exposures; and family background. Symptoms are structurally assessed at every visit; endoscopic features and histological findings are recorded for each examination. Prospective treatment data are registered sequentially, with new sequences created each time a different treatment (active principle, formulation, or dose) is administered to a patient. EoE CONNECT database is actively monitored to ensure the highest data accuracy and the highest scientific and ethical standards. Results: EoE CONNECT is currently being conducted at 39 centers in Europe and enrolls patients of all ages with EoE. In its aim to increase knowledge, to date EoE CONNECT has provided evidence on the effectiveness of first- and second-line therapies for EoE in clinical practice, the ability of proton pump inhibitors to induce disease remission, and factors associated with improved response. Drug effects to reverse fibrous remodeling and endoscopic features of fibrosis in EoE have also been assessed. Conclusion: This prospective registry study will provide important information on the epidemiological and clinical aspects of EoE and evidence as to the real-world and long-term effectiveness and safety of therapy. These data will potentially be a vital benchmark for planning future EoE health care services in Europe

Accurate and timely diagnosis of Eosinophilic Esophagitis improves over time in Europe. An analysis of the EoE CONNECT Registry

BACKGROUND: Poor adherence to clinical practice guidelines for eosinophilic esophagitis (EoE) has been described and the diagnostic delay of the disease continues to be unacceptable in many settings. OBJECTIVE: To analyze the impact of improved knowledge provided by the successive international clinical practice guidelines on reducing diagnostic delay and improving the diagnostic process for European patients with EoE. METHODS: Cross‐sectional analysis of the EoE CONNECT registry based on clinical practice. Time periods defined by the publication dates of four major sets of guidelines over 10 years were considered. Patients were grouped per time period according to date of symptom onset. RESULTS: Data from 1,132 patients was analyzed and median (IQR) diagnostic delay in the whole series was 2.1 (0.7‐6.2) years. This gradually decreased over time with subsequent release of new guidelines (p < 0.001), from 12.7 years up to 2007 to 0.7 years after 2017. The proportion of patients with stricturing of mixed phenotypes at the point of EoE diagnosis also decreased over time (41.3% vs. 16%; p < 0.001), as did EREFS scores. The fibrotic sub‐score decreased from a median (IQR) of 2 (1‐2) to 0 (0‐1) when patients whose symptoms started up to 2007 and after 2017 were compared (p < 0.001). In parallel, symptoms measured with the Dysphagia Symptoms Score reduced significantly when patients with symptoms starting before 2007 and after 2012 were compared. A reduction in the number of endoscopies patients underwent before the one that achieved an EoE diagnosis, and the use of allergy testing as part of the diagnostic workout of EoE, also reduced significantly over time (p = 0.010 and p < 0.001, respectively). CONCLUSION: The diagnostic work‐up of EoE patients improved substantially over time at the European sites contributing to EoE CONNECT, with a dramatic reduction in diagnostic delay

Análisis de los cambios en la fosforilación de proteínas inducidos por Acido Retinoico en células de Neuroblastoma

Póster original presentado XXVIII Congreso Nacional de la Sociedad Española de Bioquímica y Biología Molecular, celebrado en Zaragoza, septiembre, 2005El Acido Retinoico (RA), la forma activa de la vitamina A, induce en células de neuroblastoma SH-SY5Y la diferenciación neural. Los efectos del RA están mediados por su Receptor RAR, que pertenece a la superfamilia de los Receptores Nucleares de Hormonas. Además de sus acciones transcripcionales clásicas regulando la expresión de genes concretos, el RA actúa también de un modo extra-genómico, modulando la actividad de rutas de transducción de señal. El tratamiento con RA activa, entre otras la vía de señalización, PI3K/Akt, cuya activación por RA es un requisito para la diferenciación neural (López-Carballo G. et al., J. Biol. Chem. 277, 25297-25304, 2002). Estamos interesados en conocer cuáles son las dianas de estas acciones extra-genómicas del RA en células de neuroblastoma, y que influencia pueden tener sobre la respuesta transcripcional a RA. Para ello hemos iniciado un abordaje proteómico, basado en el hecho de que el resultado final de estas acciones extra-genómicas debe ser un cambio en los patrones de fosforilación de proteínas específicas. Para ello realizamos un enriquecimiento en proteínas fosforiladas a partir de extractos nucleares o celulares totales de células control y tratadas con RA, mediante cromatografía de afinidad. Estos extractos enriquecidos en proteínas fosforiladas son separados mediante electroforesis bidimensional (IEF/SDS-PAGE), y los geles son analizados y comparados informáticamente para identificar las bandas expresadas diferencialmente en ambas condiciones. Finalmente, las manchas identificadas son aisladas del gel, digeridas con tripsina y analizadas mediante espectrometría de masas (MALDI-TOF y LC/MS/MS).E. J. Laserna es becario FPI de la Generalitat Valenciana Financiado por el PNI+D+I (SAF2003-00311).Peer reviewe

Rapid, non-genomic actions of Retinoic Acid on Phosphatidyl-inositol-3-Kinase signalling pathway mediated by the Retinoic Acid Receptor

Póster original presentado EMBO Conference on Nuclear Receptors: Structure and Function in Health and Disease, celebrado en Italia en 2007Retinoic Acid (RA) treatment of SH-SY5Y neuroblastoma cells results in activation of phosphatidyl-inositol-3-kinase (PI3K) signalling pathway, and this activation is required for RA-induced differentiation. RA activates PI3K and ERK1/2 MAP Kinase signalling pathways through a rapid, non-genomic mechanism that does not require new gene transcription or newly synthesized proteins. Activation of PI3K by RA appears to involve the classical nuclear receptor RAR, on the basis of the pharmacological profile of the activation, loss and gain of function experiments with MEF-RAR(αβγ)L-/L- null cells, and the physical association between liganded RAR and PI3K kinase activity. Ligand binding differentially regulated the association of RAR with the two subunits of PI3K. Immunoprecipitation experiments performed in SH-SY5Y cells showed stable association between RARα and p85, the regulatory subunit of PI3K, independently of the presence of RA. In contrast, ligand administration increased the association of p110, the catalytic subunit of PI3K, to this complex. The intracellular localization of RAR resulted to be relevant for PI3K activation. A chimerical RAR receptor fusing c-Src myristylation domain to the N-terminal of RARα (Myr-RARα) was targeted to plasma membrane. Transfection of Myr-RARα to MEF-RAR(αβγ)L-/L- null cells and COS-7 cells results in strong activation of PI3K signalling pathway, although both in the absence as well in the presence of RA. Our results suggest a mechanism in which ligand binding to RAR would play a major role in the assembly and intracellular location of a signalling complex involving RAR and the subunits of PI3K. Next we have investigated the consequences of the activation of signalling pathways by RA in neuroblastoma cells. Downstream targets of the PI3K/Akt signalling pathway, like mTOR or p70S6K, are activated upon RA addition. In addition we show that RA addition rapidly increased phosphorylation of several Akt kinase target proteins in the nucleus, identified by an antibody recognizing the phosphorylated Akt motif. We have analyzed possible nuclear protein targets for RA-induced phosphorylation, with the aim of finding functional links between non-genomic actions and classical transcriptional actions mediated by RAR. We have found that RA treatment rapidly results in phosphorylation of chromatin proteins (core histone H3), as well as proteins involved in transcriptional activation (transcription factor CREB). In a second approach, nuclear phospho-proteins from control and RA-treated neuroblastoma cells were purified by affinity chromatography and analysed by 2D-electrophoresis. The differentially expressed spots were identified by Mass Spectrometry. We have found that RA treatment induces phosphorylation of chromatin proteins (HMGB1 and Histone H1.5) and proteins involved in the processing and transport of mRNA (PABP, hnRNP-K, hnRNP-C1/C2, Nucleophosmin).E. J. Laserna was supported by a FPI fellowship from Generalitat Valenciana.Peer reviewe

Non-genomic actions of retinoic acid induce pi3k signaling pathway and phosphorylation of nuclear proteins

Póster original presentado Bregenz Summer School on Endocrinology: Nuclear Receptors in Health and Disease, celebrado en Bregenz (Austria), 2007Retinoic acid (RA), the active form of vitamin A, induces neuroblastoma cells SH-SY5Y to differentiate. In addition to its classical transcriptional actions regulating the expression of specific genes, RA acts in an extra-genomic way, modulating the activity of relevant signalling cascades. In particular, RA treatment of SH-SY5Y neuroblastoma cells results in activation of phosphatidyl-inositol-3-kinase (PI3K) signaling pathway, and this activation is required for RA-induced differentiation (Lopez-Carballo G. et al., J. Biol. Chem. 277, 25297-25304, 2002). RA activates PI3K and also ERK1/2 MAP Kinase signaling pathways through a rapid, non-genomic mechanism that does not require new gene transcription or newly synthesized proteins. Activation of PI3K by RA appears to involve the classical nuclear receptor RAR, on the basis of the pharmacological profile of the activation, loss and gain of function experiments with MEF-RAR(αβγ)L-/L- null cells, and the physical association between liganded RAR and PI3K kinase activity. Immunoprecipitation experiments performed in SH-SY5Y cells showed stable association between RARα and p85, the regulatory subunit of PI3K, independently of the presence of RA. In contrast, ligand administration increased the association of p110, the catalytic subunit of PI3K, to this complex. The intracellular localization of RAR resulted to be relevant for PI3K activation. A chimerical RAR receptor fusing c-Src myristylation domain to the N-terminal of RARα (Myr-RARα) was targeted to plasma membrane. Transfection of Myr-RARα to MEF-RAR(αβγ)L-/L- null cells and COS-7 cells results in strong activation of PI3K signalling pathway. Our results suggest a mechanism in which ligand binding to RAR would play a major role in the assembly and intracellular location of a signalling complex involving RAR and the subunits of PI3K( Masiá S. et al., Mol. Endrocinol., in press, 2007). Next, we have investigated the consequences of the activation of signalling pathways by RA in neuroblastoma cells. Western blots using an antibody which specifically recognizes the phosphorylation motif of Akt have demonstrated that Akt activation in response to RA treatment produced a rapid phosphorylation of downstream substrates. Moreover, RA treatment resulted in rapid phosphorylation of well-known targets of Akt, such as mTOR and the ribosomal kinase p70S6, and other proteins, as CREB and histone H3, which are likely targets of RA non-genomic actions according to the scientific literature. In a second approach, nuclear phospho-proteins from control and RA-treated neuroblastoma cells were purified by affinity chromatography and analysed by 2D-electrophoresis. The differentially expressed spots were identified by Mass Spectrometry. We have found that RA treatment induces phosphorylation of chromatin proteins (HMGB1 and Histone H1.5) and proteins involved in the processing and transport of mRNA (PABP, hnRNP-K, hnRNP-C1/C2, Nucleophosmin). Finally, the observed RA-induced phosphorylation changes in these proteins were confirmed in 2-D western blots and immunoprecipitation experiments.Supported by grants SAF2003-00311 and SAF2006-00647 from D. G. Investigación (MEC, Spain). E. J. Laserna was supported by a FPI fellowship from Generalitat Valenciana. S. Masiá was supported by a FPI fellowship (MEC) and a training contract from Programa de Personal Técnico de Apoyo (MEC/Fondo Social Europeo).Peer reviewe