Introduction and objectives: A standardised questionnaire may be an excellent tool for epidemiological studies aiming at screening children with suspected food allergies. Thus, the aim of the present study was to develop a screening questionnaire for assessing children with suspected food allergy and to analyse its reproducibility. Materials and methods: A questionnaire of adverse food reactions was developed by literary review of similar questionnaires validated in other countries as well as less well defined, non validated Portuguese questionnaires. Peer review of the questionnaire by a panel of specialists and subsequent exploratory analysis was carried out by applying the questionnaire in children with confirmed food allergy. Test-retest analysis was performed by giving a face-to-face questionnaire to 159 children with suspected adverse food reactions, aged between three and 11 years. Temporal stability using Spearman Rho correlation test and reproducibility was studied using Cohen's Kappa index. Results: 115 children confirmed adverse food reactions that occurred with one or more foods. Retest was given about three weeks after the test, to 50 of these children who were randomly selected. The questionnaire showed good temporal stability (Spearman correlation coefficient of 0.834), and good reproducibility (only two of the 27 items had a Kappa index <0.60). Conclusions: This questionnaire showed good temporal stability and reproducibility. Its validation for screening children with suspected food allergy will allow a standardised approach to diagnosis and comparison of results obtained in different centres. (C) 2018 SEICAP. Published by Elsevier Espana, S.L.U. All rights reserved.

Background and Objective: Vitamin A has been linked to the development of allergic diseases although its role is not fully understood, Retinoic acid (RA), a metabolite of Vitamin A, has been previously associated with the prostaglandin pathway, and PTGDR, a receptor of PGD2, has been proposed as a candidate gene in allergy and asthma. Considering the role of PTGDR in allergy, the goal of this study was to analyze the effect of RA on the activation of the promoter region of the PTGDR gene.Methods: A549 lung epithelial cells were transfected with 4 combinations of genetic variants of the PTGDR promoter and stimulated with all-trans RA (ATRA); luciferase assays were performed using the Dual Luciferase Reporter System, and real-time quantitative polymerase chain reaction was used to measure the expression of PTGDR, CYP26A1, RARA, RARB, RARG, and RXRA in basal A549 cell cultures and after ATRA treatment. We also performed an in silico analysis.Results: After ATRA treatment increased expression of CYP26A1 (12-fold) and RARB (4-fold) was detected. ATRA activated PTGDR promoter activity in transfected cells (P<.001) and RA response element sequences were identified in silico in this promoter region.Conclusions: RA modulated PTGDR promoter activity. Differential response to RA and to new treatments based on PTGDR modulation could depend on genetic background in allergic asthmatic patients.

Background: Allergy and autoimmunity are important immunological entities underlying chronic diseases in children. In some cases both entities develop simultaneously in the same patient. FOXP3 gene codes for a transcription factor involved in regulation of the immune system.Considering that regulatory T cells are involved in controlling immunological disease development, and the relevant role of FOXP3 in this kind of T cells, the objective of this study was to analyse the FOXP3 gene in the most prevalent autoimmune diseases and/or allergies in childhood in a European population.Methods: A total of 255 Caucasian individuals, 95 controls and 160 patients diagnosed with allergic, autoimmune or both diseases were included in this study. The molecular analysis of FOXP3 was performed by DNA sequencing following the recommendations for quality of the European Molecular Genetics Quality Network. Genomic DNA was extracted from peripheral blood of all participants and was amplified using the polymerase chain reaction. After the visualisation of the amplified fragments by agarose gel-electrophoresis, they were sequenced.Results: Thirteen different polymorphisms in FOXP3 gene were found, seven of which had not been previously described. The mutated allele of SNP 7340C>T was observed more frequently in the group of mate children suffering from both allergic and autoimmune diseases simultaneously (p= 0.004, OR= 16.2 [1.34-195.15]).Conclusions: In this study we identified for first time genetic variants of FOXP3 that are significantly more frequent in children who share allergic and autoimmune diseases. These variants mainly affect regulatory sequences that could alter the expression levels of FOXP3 modifying its function including its role in Treg cells. (C) 2014 SEICAP. Published by Elsevier Espana, S.L.U. All rights reserved.

BACKGROUND Asthma is one of the most common chronic inflammatory diseases in developed countries. Susceptibility to asthma is associated with interaction between multiple genes and environmental factors. Several cytokines play a major role in the pathophysiology of the disease. OBJECTIVE We analyzed the distribution of cytokine gene polymorphisms in a group of patients with asthma and a control group in order to determine the effect of these variants, or their combinations, on the development of clinical phenotypes. METHODS We genotyped 22 single-nucleotide polymorphisms (SNPs) corresponding to 13 cytokine genes (IFNG, IL1A, IL1B, IL1R1, IL1RN, IL2, IL4, IL4R, IL6, IL10, IL12B, TGFB1, and TNFA) in 376 individuals (219 asthmatic patients and 157 controls). Genetic association was evaluated using genotype and allele models for different asthma phenotypes. Gene-gene interactions were explored using multifactor dimensionality reduction. RESULTS Genotype AC of IL12B-1188 was associated with the presence of asthma. A significant association was detected between 2 SNPs analyzed in TNFA (-308 and -238) and atopic asthma and severe-persistent asthma. The IL1B TT haplotype (3962T and -511T) was also associated with atopy and moderate-persistent asthma. CONCLUSION Our data show that the presence of SNPs in IL12B, TNFA, and IL1B was significantly associated with asthma, atopy, and severity of asthma.We also highlight the importance of genetic context, haplotype, and gene-gene interaction analysis in genetic association studies.

a Department of Immunoallergy, University Hospital of Salamanca, Salamanca, Spain b Department of Microbiology and Genetics, University of Salamanca, Salamanca, Spain c Department of Clinical Biochemistry, University Hospital of Salamanca, Salamanca, Spain d Department of Obstetrics, Gynecology and Pediatrics, University of Salamanca, Salamanca, Spain e IBSAL Instituto Biosanitario de Salamanca, Salamanca, Spain

Recombinant bacterial colonization plays an indispensable role in disease prevention, alleviation, and treatment. Successful application mainly depends on whether bacteria can efficiently spatiotemporally colonize the host gut. However, a primary limitation of existing methods is the lack of precise spatiotemporal regulation, resulting in uncontrolled methods that are less effective. Herein, we design upconversion microgels (UCMs) to convert near-infrared light (NIR) into blue light to activate recombinant light-responsive bacteria (Lresb) in vivo, where autocrine “functional cellular glues” made of adhesive proteins assist Lresb inefficiently colonizing the gut. The programmable engineering platform is further developed for the controlled and effective colonization of Escherichia coli Nissle 1917 (EcN) in the gut. The colonizing bacteria effectively alleviate DSS-induced colitis in mice. We anticipate that this approach could facilitate the clinical application of engineered microbial therapeutics to accurately and effectively regulate host health.

Tryptase is one of the main proteases located in the secretory granules of the mast cells, and is released through degranulation. It is therefore assumed to play an important role in inflammatory and allergic processes. Four genes are known to encode for these enzymes, with different alleles that give rise to different types of tryptases. The term "tryptase" generally refers to β-tryptase, which in vivo is a heterotetramer, possessing a structure of vital importance for enabling drug and substrate access to the active site of the molecule. Tryptase has been reported to possess antagonistic functions, since it plays an important role both in inflammatory phenomena and as a protector against infection. In allergic processes it is associated to bronchial hyperresponsiveness in asthmatic patients, where PAR-2 is of great importance as an airway receptor. Lastly, the genes that encode for tryptase are highly polymorphic and complex. As a result, it is important to establish a relationship between genotype and phenotype in disorders such as asthma, and to identify mutations that are presumably of pharmacological relevance.

BACKGROUND AND OBJECTIVES Nasal polyposis (NP) is a chronic inflammatory disease of the upper airways with a variable clinical course and unknown pathogenesis that often coexists with other conditions. Considering the possibility of genetic predisposition, we decided to analyze whether polymorphisms in LTC4S, CYSLTR1, PTGDR, and NOS2A were associated with NP. METHODS The study population comprised 486 Caucasian individuals. Polyposis and aspirin intolerance were diagnosed following the recommendations of the European Position Paper on Rhinosinusitis and Nasal Polyps. Genotypes were determined using polymerase chain reaction amplification and direct sequencing. RESULTS The -444A > C LTC4S polymorphism was significantly associated with NP and atopy (P = .033) and with NP and atopic asthma, (P =.012). In addition, a significant association was found when the (CCTTT) repetition of the NOS2A gene was present more than 14 times in patients with NP and asthma (P = .034), in patients with polyposis and intolerance to nonsteroidal anti-inflammatory drugs (P = .009), and in patients with the aspirin triad (P = .005). The PTGDR diplotype CCCT/CCCC (-613CC, -549CC, -441CC and -197TC) was more frequent in patients with NP (P = .043), NP with asthma (P = .013), and the aspirin triad (P = .041). CONCLUSIONS NP was associated with specific polymorphisms only when it occurred with related phenotypes. Our results suggest that this genetic background plays a more relevant role in the development of the associated clinical features of nasal polyposis than in simple polyposis.

RATIONALE: PTGDR gene has been associated with asthma susceptibility by several groups, including ours. In addition we have observed a stronger association of PTGDR with allergic asthma. In the present study we have evaluated the possible association of PTGDR gene with allergen sensitization.METHODS: In this study, 550 individuals (299 atopic patients and 251 non-atopic non-asthmatic controls). Skin prick tests with a battery of common aeroallergens were performed to both patients and controls. After PCR and direct sequencing haplotypic combinations of -613C>T, -549T>C, -441C>T, -197T>C and the non-previously described -95G>T SNPs of PTGDR were analyzed. Assessment of haplotypic interactions were performed with the Shesis software platform. EMQN guidelines were observed in all laboratory procedures.RESULTS: We found that the TCCTG (-613T, -549C, -441C, -197T and -95G) haplotype was more frequent in mite allergic patients (Fisher p=0.05), pollen allergic patients (Fisher p=0.026; OR:1.72[1.06-2.79]) and particularly in patients sensitized to both pollen and mites (Fisher p=0.005; OR:2.39[1.28-4.46] than in controls.CONCLUSIONS: The TCCTG haplotype was significantly associated with pollen and mite allergy. Interestingly, the CCT (-549C, -441C, -197T) haplotype has been previously described to be a high transcriptional efficiency haplotype which could determine a high PTGDR expression. In the present study we described an expanded combination of PTGDR SNPs associated with allergen sensitization. RATIONALE: PTGDR gene has been associated with asthma susceptibility by several groups, including ours. In addition we have observed a stronger association of PTGDR with allergic asthma. In the present study we have evaluated the possible association of PTGDR gene with allergen sensitization. METHODS: In this study, 550 individuals (299 atopic patients and 251 non-atopic non-asthmatic controls). Skin prick tests with a battery of common aeroallergens were performed to both patients and controls. After PCR and direct sequencing haplotypic combinations of -613C>T, -549T>C, -441C>T, -197T>C and the non-previously described -95G>T SNPs of PTGDR were analyzed. Assessment of haplotypic interactions were performed with the Shesis software platform. EMQN guidelines were observed in all laboratory procedures. RESULTS: We found that the TCCTG (-613T, -549C, -441C, -197T and -95G) haplotype was more frequent in mite allergic patients (Fisher p=0.05), pollen allergic patients (Fisher p=0.026; OR:1.72[1.06-2.79]) and particularly in patients sensitized to both pollen and mites (Fisher p=0.005; OR:2.39[1.28-4.46] than in controls. CONCLUSIONS: The TCCTG haplotype was significantly associated with pollen and mite allergy. Interestingly, the CCT (-549C, -441C, -197T) haplotype has been previously described to be a high transcriptional efficiency haplotype which could determine a high PTGDR expression. In the present study we described an expanded combination of PTGDR SNPs associated with allergen sensitization.