Assessment of single-nucleotide variant discovery protocols in RNA-seq data from human cells exposed to mycotoxins.

Food and feed contamination by nonlegislated mycotoxins beauvericin (BEA) and enniatin B (ENB) is a worldwide health concern in the present. The principal objective of this work is to assess some of the existing protocols to discover the single nucleotide variants (SNVs) in transcriptomic data obtained by RNA-seq from Jurkat cells in vitro samples individually exposed to BEA and ENB at three concentration levels (1.5, 3 and 5 µM). Moreover, previous transcriptomic results will be compared with new findings obtained using a different protocol. SNVs rs201003509 in BEA exposed cells and the rs36045790 in ENB were found in the differentially expressed genes in all doses compared to controls by means of the Genome Analysis Toolkit (GATK) Best Practices workflow. SNV-RNA-seq complementary pipeline did not show any SNV. Concerning gene expression, discrepant results were found for 1.5 µM BEA exposed cells compared with previous findings. However, 354 overlapped differentially expressed genes (DEGs) were identified in the three ENB concentrations used, with 147 matches with respect to the 245 DEGs found in the previous results. In conclusion, the two discovery SNVs protocols based on variant calling from RNA-seq used in this work displayed very different results and there were SNVs found manually not identified by any pipeline. Additionally, the new gene expression analysis reported comparable but non identical DEGs to the previous transcriptomic results obtained from these RNA-seq data.

Effect of culture parameters on the heat tolerance and inorganic polyphosphate accumulation by Lacticaseibacillus rhamnosus CRL1505, a multifunctional bacterium.

Lacticaseibacillus rhamnosus CRL1505 can be used in functional products as a probiotic powder (dried live cells) or as a postbiotic intracellular extract containing inorganic polyphosphate as a functional biopolymer. Thus, the aim of this work was to optimize the production of Lr-CRL1505 depending on the target of the functional product (probiotic or postbiotic). For this purpose, the effect of culture parameters (pH, growth phase) on cell viability, heat tolerance and polyphosphate accumulation by Lacticaseibacillus rhamnosus CRL1505 was evaluated. Fermentations at free pH produced less biomass (0.6 log units) than at controlled pH while the growth phase affected both polyphosphate accumulation and cell heat tolerance. Exponential phase cultures showed 4-15 times greater survival rate against heat shock and 49-62% increased polyphosphate level, compared with the stationary phase. Results obtained allowed setting the appropriate culture conditions for the production of this strain according to its potential application, i.e., as live probiotic cells in powder form or postbiotic. In the first case, running fermentations at pH 5.5 and harvesting the cells at the exponential phase are the best conditions for obtaining a high live biomass yield capable of overcoming heat stress. Whereas the postbiotic formulations production requires fermentations at free pH and harvesting the cells in exponential phase to increase the intracellular polyphosphate level as a first step.

Psychometric validation of the behavioral indicators of pain scale for the assessment of pain in mechanically ventilated and unable to self-report critical care patients.

OBJECTIVE: To assess the psychometric properties of the behavioral indicators of pain scale (ESCID) when applied to a wide range of medical and surgical critical patients. DESIGN: A multicentre, prospective observational study was designed to validate a scale measuring instrument. SETTING: Twenty Intensive Care Units of 14 hospitals belonging to the Spanish National Health System. PARTICIPANTS: A total of 286 mechanically ventilated, unable to self-report critically ill medical and surgical adult patients. PROCEDURE: Pain levels were measured by two independent evaluators simultaneously, using two scales: ESCID and the behavioral pain scale (BPS). Pain was observed before, during, and after two painful procedures (turning, tracheal suctioning) and one non-painful procedure. MAIN VARIABLES: ESCID reliability was measured on the basis of internal consistency using the Cronbach-α coefficient. Inter-rater and intra-rater agreement were measured. The Spearman correlation coefficient was used to assess the correlation between ESCID and BPS. RESULTS: A total of 4386 observations were made in 286 patients (62% medical and 38% surgical). High correlation was found between ESCID and BPS (r=0.94-0.99; p<0.001), together with high intra-rater and inter-rater concordance. ESCID was internally reliable, with a Cronbach-α value of 0.85 (95%CI 0.81-0.88). Cronbach-α coefficients for ESCID domains were high: facial expression 0.87 (95%CI 0.84-0.89), calmness 0.84 (95%CI 0.81-0.87), muscle tone 0.80 (95%CI 0.75-0.84), compliance with mechanical ventilation 0.70 (95%CI 0.63-0.75) and consolability 0.85 (95%CI 0.81-0.88). CONCLUSION: ESCID is valid and reliable for measuring pain in mechanically ventilated unable to self-report medical and surgical critical care patients. CLINICALTRIALS.GOV: NCT01744717.

Appropriateness of behavioural scales in the monitoring of pain in the critically ill patient unable to self-report.

Appropriateness is a dimension of quality that evaluates the effective use of technologies, resources or interventions in specific situations or populations, assessing whether our interventions do more benefit than harm. The evidence regarding pain monitoring in the critically ill patient points to the periodic assessment of pain using appropriate tools, with the aim of improving pain management and more efficient use of analgesics in the intensive care unit. The first step would be to assess the patient's ability to communicate or self-report and, based on this, to select the most appropriate pain assessment tool. In patients who are unable to self-report, behavioural pain assessment tools are recommended. When we talk about the suitability of behavioural scales for pain monitoring in critically ill patients unable to self-report, we refer to their use with a clear clinical benefit, i.e. using the right tool for pain assessment to be effective, efficient and consistent with bioethical principles. To our knowledge, there are no published data on the suitability of pain assessment tools in unable to self-report critically ill patients, so, in the framework of continuous quality improvement in pain care, new research should incorporate this approach by integrating the best scientific evidence with current clinical practice.

BCN Checkpoint, a community-based centre for men who have sex with men in Barcelona, Catalonia, Spain, shows high efficiency in HIV detection and linkage to care.

OBJECTIVES: We assessed the efficiency of BCN Checkpoint in detecting new cases of HIV infection and efficiently linking newly diagnosed individuals to care. METHODS: This study analysed during 2007-2012 the number of tests performed and the number of persons tested in BCN Checkpoint, the HIV prevalence, global and in first visits, the capacity of HIV detection compared to the reported cases in MSM in Catalonia, and the linkage to care rate. RESULTS: During the six years a total of 17.319 tests were performed and 618 HIV-positive cases were detected. Median prevalence of clients who visited the centre for the first time was 5.4% (4.1-5.8). BCN Checkpoint detected 36.3% (35.0-40.4) of all reported cases in MSM during 2009-2011. Linkage to care was achieved directly in 90.5% of the cases and only 2.4% of cases were lost to follow-up. CONCLUSIONS: A community-based centre, addressed to a key population at risk, can be less effort consuming (time and funding) and show high efficiency in HIV detection and linkage to care.

Effect of polyphenols on enniatins-induced cytotoxic effects in mammalian cells.

Enniatins (ENs) are fungal secondary metabolites produced by genus Fusarium. The ENs exert antimicrobial and insecticidal effect, and has also been demonstrated cytotoxic effects on several mammalian cell lines. On the other hands, it has been proved that natural polyphenols have antioxidant effect. In this study, cell effects at low levels of exposure of four ENs (A, A(1), B and B(1)) and five polyphenols (quercetin, quercetin-3-ß-D-glucoside, rutin, myricetin and t-pterostilbene) present in wine; and the cytoprotective effect of these polyphenols exposed simultaneously with ENs in Chinese Hamster Ovary (CHO-K1) cells, were studied. Cell effects were determined by the MTT test after 24 h of exposure. All ENs showed cytotoxic effect. The IC(50) obtained ranged from 4.5 ± 1.2 to 11.0 ± 2.7 µM. The concentration of polyphenols tested ranged from 5 to 50 µM. Polyphenols did not show cytotoxicity and the cytoprotective effect of polyphenols varies depending on the EN tested. The cytoprotective effect of polyphenols in CHO-K1 cells exposed to ENs was as follow: quercetin, from 24 to 84%; quercetin-3-ß-D-glucoside, from 12 to 76%; rutin, from 17 to 83%; myricetin, from 16 to 92% and pterostilbene from 25 to 100%. All polyphenols protected CHO-K1 cells against EN A(1) exposure.

The importance of transcriptomics and proteomics for studying molecular mechanisms of mycotoxin exposure: A review.

This review aims to highlight recent advances where transcriptomics and proteomics have been used as a key tool to understand molecular toxicity of mycotoxins. The most studied mycotoxin by using transcriptomic approach is deoxynivalenol (DON), followed by aflatoxins (AFs) and zearalenone (ZEA). Instead, proteomics mostly focuses on AFs but also in this case, mildly to ZEA and DON. However, in both omics approaches, fewer studies investigated the toxicological effect of emerging mycotoxins, patulin, ochratoxin A, T-2 toxin, alternariol and amino-14,16-dimethyloctadecan-3-ol. The study of changes in the expression of genes involved in immune system are the most common purposes for transcriptomics whereas cellular processes in proteomics field. Concerning the techniques used to perform the experiments, RT-qPCR is the most employed in gene expression analysis whereas liquid chromatography coupled with mass spectrometry is the master technique for proteomics assays. The gathered data have reported that the interest in using these omic approaches has increased in the last five years. However, in vitro models take precedence over the in vivo and ex vivo ones. Therefore, there is a need to enhance the use of in vivo models and alternative methods to better understand mycotoxins mode of action on animal and human health.

In vitro blood brain barrier exposure to mycotoxins and carotenoids pumpkin extract alters mitochondrial gene expression and oxidative stress.

Food and feed are daily exposed to mycotoxin contamination which effects may be counteracted by antioxidants like carotenoids. Some mycotoxins as well as carotenoids penetrate the blood brain barrier (BBB) inducing alterations related to redox balance in the mitochondria. Therefore, the in vitro BBB model ECV304 was subcultured for 7 days and exposed to beauvericine, enniatins, ochratoxin A, zearalenone (100 nM each), individually and combined, and pumpkin extract (500 nM). Reactive oxygen species were measured by fluorescence using the dichlorofluorescein diacetate probe at 0 h, 2 h and 4 h. Intracellular ROS generation reported was condition dependent. RNA extraction was performed and gene expression was analyzed by qPCR after 2 h exposure. The selected genes were related to the Electron Transport Chain (ETC) and mitochondrial activity. Gene expression reported upregulation for exposures including mycotoxins plus pumpkin extract versus individual mycotoxins. Beauvericin and Beauvericin-Enniatins exposure significantly downregulated Complex I and pumpkin addition reverted the effect upregulating Complex I. Complex IV was the most downregulated structure of the ETC. Thioredoxin Interacting Protein was the most upregulated gene. These data confirm that mitochondrial processes in the BBB could be compromised by mycotoxin exposure and damage could be modulated by dietary antioxidants like carotenoids.

Bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey fermented with lactic acid bacteria.

Microbial fermentation with lactic acid bacteria (LAB) is a natural food biopreservation method. Yellow mustard and milk whey are optimum substrates for LAB fermentation. The aim of the present study was to evaluate the bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey both with and without LAB fermentation. All extracts were subjected to a simulated digestion process. Total polyphenols, DL-3-phenyllactic acid (PLA), lactic acid, and the antioxidant activity were determined in the studied matrices before and after simulated digestion. Yellow mustard flour was significantly richer in total polyphenols, whereas significantly higher concentrations of PLA and lactic acid were observed in milk whey. Similar antioxidant activity was determined in both ingredients being in all cases strongly reduced after in vitro digestion. Higher bioaccessibility was found for polyphenols and PLA in milk whey. Transepithelial transport of total polyphenols was higher in yellow mustard flour compared to milk whey, reaching bioavailability values between 3-7% and 1-2%, respectively. PLA transepithelial transport was only significant in both fermented matrices with bioavailability around 4-6%. Transepithelial transport of lactic acid reached values of 31-34% (bioavailability ∼ 22%) and 15-78% (bioavailability ∼ 3%) in milk whey and yellow mustard flour, respectively. LAB fermentation showed beneficial effects on enriching extracts with PLA, lactic acid, and antioxidant activity, as well as increasing bioaccessibility of these acids in yellow mustard flour and total polyphenol bioavailability in milk whey. Results pointed to yellow mustard flour and milk whey as natural preservative ingredients used in the food industry, especially when fermented with LAB.

Proteomics evaluation of enniatins acute toxicity in rat liver.

Enniatins (ENs) are emerging mycotoxins produced by Fusarium fungi which are cytotoxic also at low concentrations due to its ionophoric properties. The aim of this study was to evaluate the hepatic toxicity of ENs exposure at different concentrations in Wistar rats through a proteomic approach. Animals were intoxicated by oral gavage with medium (EN A 256, ENA1 353, ENB 540, ENB1 296 µg/mL) and high concentrations (ENA 513, ENA1 706, ENB 1021, ENB1 593 µg/mL) of an ENs mixture and sacrificed after 8 h. Protein extraction was performed using powdered liver. Peptides were analyzed using a liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer. Proteins were filtered by abundance using Mass Professional Profiler software (Agilent Technologies) and 57 were differentially expressed when compared to the control. In terms of abundance, the liver biomarker Carboamoyl-phosphate synthase showed the highest levels in all conditions employed while actin-1 had the lowest. Bioinformatic analysis using DAVID platform reported acetylation, nucleotide phosphate-binding region:NAD and catalytic activity as the most represented terms. Furthermore, metabolism was the most significant and enriched pathway in Reactome overrepresentation. In conclusion, ENs acute exposure caused protein expression changes related to major cellular processes in rats, hinting its involvement in liver disturbance.

Toxicity of mycotoxins in vivo on vertebrate organisms: A review.

Mycotoxins are considered to be a major risk factor affecting human and animal health as they are one of the most dangerous contaminants of food and feed. This review aims to compile the research developed up to date on the toxicological effects that mycotoxins can induce on human health, through the examination of a selected number of studies in vivo. AFB1 shows to be currently the most studied mycotoxin in vivo, followed by DON, ZEA and OTA. Scarce data was found for FBs, PAT, CIT, AOH and Fusarium emerging mycotoxins. The majority of them concerned the investigation of immunotoxicity, whereas the rest consisted in the study of genotoxicity, oxidative stress, hepatotoxicity, cytotoxicity, teratogenicity and neurotoxicity. In order to assess the risk, a wide range of different techniques have been employed across the reviewed studies: qPCR, ELISA, IHC, WB, LC-MS/MS, microscopy, enzymatic assays, microarray and RNA-Seq. In the last decade, the attention has been drawn to immunologic and transcriptomic aspects of mycotoxins' action, confirming their toxicity at molecular level. Even though, more in vivo studies are needed to further investigate their mechanism of action on human health.

Mitochondrial transcriptional study of the effect of aflatoxins, enniatins and carotenoids in vitro in a blood brain barrier model.

C. maxima (var. Delica), a variety of pumpkin, is well known for its high concentration on carotenoids, possessing dietary benefits and antioxidant properties. Aflatoxins and enniatins are common mycotoxins present in food and feed with an extended toxicity profile in humans and animals. Both types of substances reach a wide range of tissues and organs and have the capability to penetrate the blood brain barrier. Since carotenoids and mycotoxins have been reported to modify diverse mitochondrial processes individually, transcriptional in vitro studies on human epithelial cells ECV 304 were conducted to analyze the relative expression of 13 mitochondria related genes. ECV 304 cells were differentiated for 9 days and treated for 2 h with: a) pumpkin (500 nM); b) aflatoxins (100 nM); c) enniatins (100 nM); d) aflatoxins (100 nM) and pumpkin (500 nM); e) enniatins (100 nM) and pumpkin (500 nM). Even at low concentrations, dietary carotenoids activity on mitochondrial genes expression reported a beneficial effect and, for most of the genes studied across the Electron Transport Chain (ETC), developed a protective effect when mixed with aflatoxins (AFs) or enniatins (ENs).

Pesticide residue determination in surface waters by stir bar sorptive extraction and liquid chromatography/tandem mass spectrometry.

In this stir bar sorptive extraction (SBSE) method, 16 pesticides were extracted from surface water samples by sorption onto 1 mm polydimethylsiloxane layer coated on a 10-mm-length stir bar magnet. After liquid desorption of the analytes with 1 ml of methanol, the detection was performed on a liquid chromatography-tandem mass spectrometry with a triple quadrupole (QqQ) analyzer using selected reaction monitoring mode via electrospray ionization. Parameters affecting SBSE operation, including sample volume, salt addition, extraction time, stirring rate, and desorption conditions, have been evaluated. The optimized SBSE method required two 50 ml aliquots of surface water samples, one aliquot was added of 30% NaCl and stirred at 900 rpm during 1 h for testing five pesticides with log K(o/w) < 3, and the other aliquot was directly extracted following the same procedure for the rest of the pesticides with log K(o/w) > 3. The method was validated in spiked surface water samples at limits of quantifications (LOQs) and ten times the LOQs showing recoveries <62%, and the LOQs reached were from 0.03 microg l(-1) for diazinon to 3 microg l(-1) for simazine. The proposed methodology was applied to the determination of these compounds in samples from Albufera Lake and surrounding channels, showing that SBSE is a powerful tool for routine control analysis of pesticide residues in surface water.

Enniatin B induces expression changes in the electron transport chain pathway related genes in lymphoblastic T-cell line.

Enniatin B is a ionophoric and lipophilic mycotoxin which reaches the bloodstream and has the ability to penetrate into cellular membranes. The purpose of this study was to reveal changes in the gene expression profile caused by enniatin B in human Jurkat lymphoblastic T-cells after 24 h of exposure at 1.5, 3 and 5 µM by next generation sequencing. It was found that up to 27% of human genome expression levels were significantly altered (5750 genes for both down-regulation and up-regulation). In the three enniatin B concentrations studied 245 differentially expressed genes were found to be overlapped, 83 were down and 162 up-regulated. ConsensusPathDB analysis of over-representation of differentially expressed genes provided a list of gene ontology terms in which several biological processes related to nucleoside monophosphate metabolic process, respiratory chain complex, electron transport chain, oxidative phosphorylation and cellular respiration were the most altered. Also, an interesting correlation was found between enniatin B toxicity and the up-regulation of the UCP protein complex. In summary, the transcriptomic analysis revealed that mitochondria are the organelles showing more related differentially expressed genes. Consequently, differentially expressed genes involved in biological processes, molecular functions and pathways related to mitochondrial metabolism and respiration were significantly changed.

Implementation of a food safety management system: a practical guide for micro-companies / Implementación de un sistema de gestión de seguridad alimentaria: una guía práctica para microempresas

Food system is continuously exposed to several safety hazards throughout manufacturing processes. Consequently, the understanding of these mechanisms, the likelihood of their occurrence, and the consequences involves the implementation of an appropriate risk assessment program. Hazard Analysis and Safety Critical Control Points (HACCP) is a powerful tool for self-control system which is fundamental to adapt the requirements of each sector and size of a company but however, is currently limited it at industrial level. In fact, as a result to its flexible application, the recommendations at European level for companies with less than 10 workers focused on a food safety management system (FSMS) based on the principles of good hygiene and manufacturing practices. Therefore, in order to help companies implementing self-monitoring systems, several reference guides have been created under the supervision of health authorities. For this purpose, the aim of these study was to create an abbreviated guide as an introduction to self-control in food safety for spanish micro-enterprises. It is presented as a summary of each prerequisite focused solely on understanding the meaning and documentation necessary to implement it. To carry out the research, European legislation and the main guides at national level, particularly from autonomous communities were took into account. Starting from this information, was provided a classification of the main plans related to prerequisites for hygiene and traceability fulfilling. Moreover, indications concerning the documentation which should be prepared for each one of the prerequisites was detailed. (AU)

El sistema alimentario está continuamente expuesto a varios riesgos de seguridad a lo largo de los procesos de fabricación. En consecuencia, la comprensión de estos mecanismos, la probabilidad de su ocurrencia y las consecuencias implica la implementación de un programa adecuado de evaluación de riesgos. El Análisis de Peligros y Puntos Críticos de Control de Seguridad (APPCC) es una poderosa herramienta para el sistema de autocontrol que es fundamental para adaptar los requisitos de cada sector y tamaño de una empresa pero que, sin embargo, actualmente está limitado a nivel industrial. De hecho, como consecuencia de su aplicación flexible, las recomendaciones a nivel europeo para empresas de menos de 10 trabajadores se centraron en un sistema de gestión de la seguridad alimentaria (SGSA) basado en los principios de buenas prácticas de higiene y fabricación. Por ello, con el fin de ayudar a las empresas a implementar sistemas de autocontrol, se han elaborado varias guías de referencia bajo la supervisión de las autoridades sanitarias. Para ello, el objetivo de este estudio fue crear una guía abreviada como introducción al autocontrol en seguridad alimentaria para microempresas españolas. Se presenta como un resumen de cada requisito previo enfocado únicamente en comprender el significado y la documentación necesaria para implementarlo. Para realizar la investigación se tuvo en cuenta la legislación europea y las principales guías a nivel nacional, en particular de las comunidades autónomas. A partir de esta información, se proporcionó una clasificación de los principales planes relacionados con los requisitos previos para el cumplimiento de la higiene y la trazabilidad. Además, se detallaron indicaciones sobre la documentación que se debe preparar para cada uno de los requisitos previos. (AU)

Mitigation of enniatins in edible fish tissues by thermal processes and identification of degradation products.

Emerging mycotoxins, such as enniatins and beauvericin, are common contaminants in vegetal matrices, but recently, the occurrence of mycotoxins in foodstuffs from animal origin has been also reported as they can be present in edible tissues of animals fed with contaminated feedstuffs. Sea bass, sea bream, Atlantic salmon and rainbow trout from aquaculture analyzed in the present survey showed contamination by emerging Fusarium mycotoxins enniatins (ENs). ENs were extracted from raw and cooked fish with acetonitrile and analyzed by Liquid Chromatography coupled to Mass Spectrometry. In this study, the stability of ENs was evaluated during food processing by the application of different cooking methods (broiling, boiling, microwaving and baking treatments). All treated samples showed a reduction in mycotoxin levels with different percentages depending on the type of EN and the fish species. Thus, the reduction obtained ranged from 30 to 100%. The thermal treatments have shown to be a good strategy to mitigate ENs content in edible fish tissues. On the other hand, some ENs degradation products originated during the application of thermal treatments were identified.

Una actualización sobre las manifestaciones clínicas y el diagnóstico de la enfermedad celiaca / An update on the clinical manifestations and diagnosis of celiac disease

La enfermedad celíaca (EC) es un trastorno sistémico, crónico y de origen autoinmune que afecta a individuos genéticamente susceptibles. Se presenta con una gran variedad de signos, síntomas y enfermedades asociadas, incluso también puede cursar de manera asintomática, lo que hace difícil su diagnóstico. La EC se diagnostica con mayor frecuencia en las mujeres y a edades más tempranas que a los hombres. Debido a la gran heterogeneidad en la forma de presentación de la EC, se ha llevado a cabo una revisión sistemática, donde estudios recientes, muestran una variación del espectro clínico de la enfermedad en función de la edad. En niños pequeños predomina la forma clásica con síntomas como distensión abdominal, pérdida del apetito, diarrea y pérdida de peso; esta forma de presentación típica es más intensa en niños menores de dos años. En niños mayores son frecuentes las manifestaciones atípicas como el dolor abdominal, reflujo, vómitos, fatiga y talla baja, entre otros. En adultos se observa una reducción de la forma de presentación clásica e incremento de la no clásica o atípica. El incremento en la prevalencia en los últimos años y la variación del espectro clínico hacia una forma atípica con síntomas extradigestivos o incluso asintomática, causa retraso en su diagnóstico, por lo que deben reconocerse para que se detecte de manera más precoz. El diagnóstico de la EC ha mejorado mucho en los últimos años debido a la creciente incidencia de esta patología y a la precisión de los nuevos métodos de detección. Se ha realizado una búsqueda bibliográfica de artículos sobre grupos de riesgo y métodos de diagnóstico en bases de datos como: Pubmed, Scopus, Web of Science de los últimos 6 años (2014-2019), se realizaron dos búsquedas en cada una de las bases de datos consultadas, con las siguientes palabras clave: 1) celiac disease AND sign AND symptom AND prevalence; 2) celiac disease AND sign AND symptom AND autoimmune disease. (AU)

Celiac disease (CD) is a systemic, chronic and autoimmune disorder affecting genetically susceptible individuals. It presents a wide variety of signs, symptoms and associated diseases, it can even be asymptomatic, difficulting their diagnosis. CD is diagnosed more often in women and at a younger age than in men. Due to the great hererogeneity in the presentation of CD, a systematic review has been carried out, where recent studies show a variation in the clinical spectrum of the disease as an age function. The classic form predominates in young children with symptoms such as abdominal distension, loss of appetite, diarrhea and weight loss; this typical presentation is more intense in children under two years of age. Atypical manifestations such as abdominal pain, reflux, vomiting, fatigue and short stature, among others, are frequent in older children. In adults, a reduction in the classic presentation and an increase in the non-classic or atypical form was shown. The increase in prevalence in recent years and the variation of the clinical spectrum towards an atypical form with extradigestive or even asymptomatic symptoms, causes a delay in the diagnosis, so they must be recognized to get an early detection. The diagnosis of CD has improved greatly in recent years due to the increasing incidence of this pathology and the precision of new detection methods. A bibliographic search has been carried out for articles on risk groups and diagnostic methods in databases such as Pubmed, Scopus and Web of Science for the last 6 years (2014-2019), two searches were carried out in each consulted database, with the following keywords: 1) celiac disease AND sign AND sym ptom AND prevalence; 2) celiac disease AND sign AND symptom AND autoimmune disease. (AU)