Group 2 innate lymphoid cells are key in lipid transfer protein allergy pathogenesis.

Background: Immunopathology in food allergy is characterized by an uncontrolled type 2 immune response and specific-IgE production. Recent studies have determined that group 2 innate lymphoid cells (ILC2) participate in the food allergy pathogenic mechanism and their severity. Our objective was to investigate the role of ILC2 in peach-allergic patients due to non-specific lipid transfer protein (Pru p 3) sensitization. Methods: The immune response in peripheral blood mononuclear cells was characterized in lipid transfer protein-allergic patients and healthy controls. We have analyzed the Pru p 3 uptake on ILC2, the expression of costimulatory molecules, and their involvement on the T-cell proliferative response and cytokine production under different experimental conditions: cytokines involved in group 2 innate lymphoid cell activation (IL-33 and IL-25), Pru p 3 as main food allergen, and the combination of both components (IL-33/IL-25+Pru p 3) using cell sorting, EliSpot, flow cytometry, and confocal microscopy. Results: Our results show that Pru p 3 allergen is taken up by group 2 innate lymphoid cells, regulating their costimulatory molecule expression (CD83 and HLA-DR) depending on the presence of Pru p 3 and its combination with IL-33/IL-25. The Pru p 3-stimulated ILC2 induced specific GATA3+Th2 proliferation and cytokine (IL-4, IL-5, and IL-13) production in lipid transfer protein-allergic patients in a cell contact-dependent manner with no changes in Tbet+Th1- and FOXP3+Treg cell differentiation. Conclusions: The results indicate that in lipid transfer protein-allergic patients, the responsible allergen, Pru p 3, interacts with group 2 innate lymphoid cells, promoting a Th2 cell response. Our results might be of interest in vivo, as they show a role of group 2 innate lymphoid cells as antigen-presenting cells, contributing to the development of food allergy. Consequently, group 2 innate lymphoid cells may be considered as potential therapeutic targets.

Evolutionary analysis reveals the role of a non-catalytic domain of peptidyl arginine deiminase 2 in transcriptional regulation.

Peptidyl arginine deiminases (PADIs) catalyze protein citrullination, a post-translational conversion of arginine to citrulline. The most widely expressed member of this family, PADI2, regulates cellular processes that impact several diseases. We hypothesized that we could gain new insights into PADI2 function through a systematic evolutionary and structural analysis. Here, we identify 20 positively selected PADI2 residues, 16 of which are structurally exposed and maintain PADI2 interactions with cognate proteins. Many of these selected residues reside in non-catalytic regions of PADI2. We validate the importance of a prominent loop in the middle domain that encompasses PADI2 L162, a residue under positive selection. This site is essential for interaction with the transcription elongation factor (P-TEFb) and mediates the active transcription of the oncogenes c-MYC, and CCNB1, as well as impacting cellular proliferation. These insights could be key to understanding and addressing the role of the PADI2 c-MYC axis in cancer progression.

A synthetic glycodendropeptide induces methylation changes on regulatory T cells linked to tolerant responses in anaphylactic-mice.

Introduction: Lipid transfer proteins (LTPs) are allergens found in a wide range of plant-foods. Specifically, Pru p 3, the major allergen of peach, is commonly responsible for severe allergic reactions. The need for new alternatives to conventional food allergy treatments, like restrictive diets, suggests allergen immunotherapy as a promising option. It has been demonstrated that sublingual immunotherapy (SLIT) with synthetic glycodendropeptides, such as D1ManPrup3, containing mannose and Pru p 3 peptides induced tolerance in mice and that the persistence of this effect depends on treatment dose (2nM or 5nM). Moreover, it produces changes associated with differential gene expression and methylation profile of dendritic cells, as well as phenotypical changes in regulatory T cells (Treg). However, there are no works addressing the study of epigenetic changes in terms of methylation in the cell subsets that sustain tolerant responses, Treg. Therefore, in this work, DNA methylation changes in splenic-Treg from Pru p 3 anaphylactic mice were evaluated. Methods: It was performed by whole genome bisulphite sequencing comparing SLIT-D1ManPrup3 treated mice: tolerant (2nM D1ManPrup3), desensitized (5nM D1ManPrup3), and sensitized but not treated (antigen-only), with anaphylactic mice. Results: Most of the methylation changes were found in the gene promoters from both SLIT-treated groups, desensitized (1,580) and tolerant (1,576), followed by the antigen-only (1,151) group. Although tolerant and desensitized mice showed a similar number of methylation changes, only 445 genes were shared in both. Remarkably, interesting methylation changes were observed on the promoter regions of critical transcription factors for Treg function like Stat4, Stat5a, Stat5b, Foxp3, and Gata3. In fact, Foxp3 was observed exclusively as hypomethylated in tolerant group, whereas Gata3 was only hypomethylated in the desensitized mice. Discussion: In conclusion, diverse D1ManPrup3 doses induce different responses (tolerance or desensitization) in mice, which are reflected by differential methylation changes in Tregs.

Implementation of Exome Sequencing in Clinical Practice for Neurological Disorders.

Neurological disorders (ND) are diseases that affect the brain and the central and autonomic nervous systems, such as neurodevelopmental disorders, cerebellar ataxias, Parkinson's disease, or epilepsies. Nowadays, recommendations of the American College of Medical Genetics and Genomics strongly recommend applying next generation sequencing (NGS) as a first-line test in patients with these disorders. Whole exome sequencing (WES) is widely regarded as the current technology of choice for diagnosing monogenic ND. The introduction of NGS allows for rapid and inexpensive large-scale genomic analysis and has led to enormous progress in deciphering monogenic forms of various genetic diseases. The simultaneous analysis of several potentially mutated genes improves the diagnostic process, making it faster and more efficient. The main aim of this report is to discuss the impact and advantages of the implementation of WES into the clinical diagnosis and management of ND. Therefore, we have performed a retrospective evaluation of WES application in 209 cases referred to the Department of Biochemistry and Molecular Genetics of the Hospital Clinic of Barcelona for WES sequencing derived from neurologists or clinical geneticists. In addition, we have further discussed some important facts regarding classification criteria for pathogenicity of rare variants, variants of unknown significance, deleterious variants, different clinical phenotypes, or frequency of actionable secondary findings. Different studies have shown that WES implementation establish diagnostic rate around 32% in ND and the continuous molecular diagnosis is essential to solve the remaining cases.

Identification of Asthma Phenotypes in the Spanish MEGA Cohort Study Using Cluster Analysis

Introduction: The definition of asthma phenotypes has not been fully established, neither there are cluster studies showing homogeneous results to solidly establish clear phenotypes. The purpose of this study was to develop a classification algorithm based on unsupervised cluster analysis, identifying clusters that represent clinically relevant asthma phenotypes that may share asthma-related outcomes. Methods: We performed a multicentre prospective cohort study, including adult patients with asthma (N=512) from the MEGA study (Mechanisms underlying the Genesis and evolution of Asthma). A standardised clinical history was completed for each patient. Cluster analysis was performed using the kernel k-groups algorithm. Results: Four clusters were identified. Cluster 1 (31.5% of subjects) includes adult-onset atopic patients with better lung function, lower BMI, good asthma control, low ICS dose, and few exacerbations. Cluster 2 (23.6%) is made of adolescent-onset atopic asthma patients with normal lung function, but low adherence to treatment (59% well-controlled) and smokers (48%). Cluster 3 (17.1%) includes adult-onset patients, mostly severe non-atopic, with overweight, the worse lung function and asthma control, and receiving combination of treatments. Cluster 4 (26.7%) consists of the elderly-onset patients, mostly female, atopic (64%), with high BMI and normal lung function, prevalence of smokers and comorbidities. Conclusion: We defined four phenotypes of asthma using unsupervised cluster analysis. These clusters are clinically relevant and differ from each other as regards FEV1, age of onset, age, BMI, atopy, asthma severity, exacerbations, control, social class, smoking and nasal polyps. (AU)

Identification of Asthma Phenotypes in the Spanish MEGA Cohort Study Using Cluster Analysis.

INTRODUCTION: The definition of asthma phenotypes has not been fully established, neither there are cluster studies showing homogeneous results to solidly establish clear phenotypes. The purpose of this study was to develop a classification algorithm based on unsupervised cluster analysis, identifying clusters that represent clinically relevant asthma phenotypes that may share asthma-related outcomes. METHODS: We performed a multicentre prospective cohort study, including adult patients with asthma (N=512) from the MEGA study (Mechanisms underlying the Genesis and evolution of Asthma). A standardised clinical history was completed for each patient. Cluster analysis was performed using the kernel k-groups algorithm. RESULTS: Four clusters were identified. Cluster 1 (31.5% of subjects) includes adult-onset atopic patients with better lung function, lower BMI, good asthma control, low ICS dose, and few exacerbations. Cluster 2 (23.6%) is made of adolescent-onset atopic asthma patients with normal lung function, but low adherence to treatment (59% well-controlled) and smokers (48%). Cluster 3 (17.1%) includes adult-onset patients, mostly severe non-atopic, with overweight, the worse lung function and asthma control, and receiving combination of treatments. Cluster 4 (26.7%) consists of the elderly-onset patients, mostly female, atopic (64%), with high BMI and normal lung function, prevalence of smokers and comorbidities. CONCLUSION: We defined four phenotypes of asthma using unsupervised cluster analysis. These clusters are clinically relevant and differ from each other as regards FEV1, age of onset, age, BMI, atopy, asthma severity, exacerbations, control, social class, smoking and nasal polyps.

Analysis of dermoscopic changes of blue nevi on digital follow-up: A 21-year retrospective cohort study.

BACKGROUND: Blue nevi are benign dermal melanocytic proliferations that are often easy to recognize clinically. Rarely, these lesions can display atypical features, suggesting the presence of a malignant blue nevus or mimicking cutaneous metastases of melanoma. OBJECTIVE: To describe the clinical evolution of blue nevi over time and to assess the need for monitoring these lesions. METHODS: We conducted a retrospective cohort study of 103 patients who were followed between December 1998 and November 2019. An artificial intelligence algorithm was used to identify blue nevi from the databases of two digital epiluminescence devices. Changes in the area of each lesion were calculated with a segmentation neural network. RESULTS: We included 123 blue nevi from 103 patients. Most of the lesions segmented, 99 (91.7%), were considered stable. Of the 9 (8.3%) growing blue nevi identified, 2 (1.85%) showed significant growth. The studied growing blue nevi turned out to be cellular blue nevi, presented with a low tumour mutation burden and GNAQ c.626A>T alteration was identified in both lesions. LIMITATIONS: Some clinical variants of blue nevi might not be included. CONCLUSIONS: Most blue nevi remain stable during their evolution. Rarely, they can show progressive growth, although histopathological or molecular signs of malignancy have not been identified.

[External Quality Control in Immunohematology of National Medical Center's Blood Bank. History and present]. / Control de Calidad Externo en Inmunohematología del Banco de Sangre del Centro Médico Nacional. Historia y presente.

For the proper functioning of the Immunohematology Area, an External Quality Control was established since 1973 through a program that evaluates the performance of the laboratories of the Blood Bank and transfusion services that carry out immunohematology tests. This program consists of sending panel cells to participating blood banks or services, which are phenotyped erythrocyte samples previously studied as problem cases but whose results are unknown by the participating laboratories. The processes in which the program is of most importance are determination of the ABO group, determination of Rh, performance of the direct and indirect Coombs test, and pre-transfusion compatibility tests. It was carried out an observational and retrospective study of the results obtained in the 2020 period from 104 units participating in the Immunohematology Quality Control Program of the National Medical Center's Blood Bank. A panel of cells was sent for external quality control of immunohematology every 45 days, resulting in 9 panels for each unit in the studied period. Compliance with the program was observed in the general result (79.6%), i.e., there was a decrease in the participation of the registered units. Of a maximum score of 100% to be obtained, it was observed a general result of 95.3% compliance of the participating units. The results obtained confirm the good general training of the immunohematology laboratories of the participating units. Yet, as in any external control program, it becomes clear that obtaining an erroneous result is a risk that can occur in any laboratory.

Para el buen funcionamiento del Área de Inmunohematología, desde 1973 se instauró el Control de Calidad Externo con un programa que evalúa los laboratorios del Banco de Sangre y los servicios de transfusión que hacen pruebas de inmunohematología. El programa consiste en enviar a los bancos de sangre, o servicios participantes, células panel, que son muestras de eritrocitos fenotipados y previamente estudiados como casos problema, pero cuyos resultados son desconocidos por los laboratorios participantes. Los procesos en los que el programa es de suma importancia son determinación del grupo ABO, determinación del Rh, realización de la prueba de Coombs directa e indirecta y las pruebas de compatibilidad pretransfusionales. Se hizo un estudio observacional y retrospectivo de los resultados de 2020 de 104 unidades participantes en el Programa de Control de Calidad de Inmunohematología del Banco de Sangre del Centro Médico Nacional Siglo XXI. Se envió un panel de células para el control de calidad externo de inmunohematología cada 45 días y dio como resultado nueve paneles para cada unidad. El cumplimiento del programa se observó en el resultado general (79.6%), es decir, disminuyó la participación de las unidades inscritas. De una calificación máxima de 100%, hubo un resultado general de las unidades participantes del 95.3% de cumplimiento. Los resultados confirman la buena capacitación general de los laboratorios de inmunohematología de las unidades participantes. Aun así, como en cualquier programa de control externo, obtener un resultado erróneo es un riesgo que puede presentarse en cualquier laboratorio.

Corrigendum: MicroRNAs as potential regulators of immune response networks in asthma and chronic obstructive pulmonary disease.

[This corrects the article DOI: 10.3389/fimmu.2020.608666.].

Lethal Congenital Contracture Syndrome 11: A Case Report and Literature Review.

Lethal congenital contracture syndrome 11 (LCCS11) is caused by homozygous or compound heterozygous variants in the GLDN gene on chromosome 15q21. GLDN encodes gliomedin, a protein required for the formation of the nodes of Ranvier and development of the human peripheral nervous system. We report a fetus with ultrasound alterations detected at 28 weeks of gestation. The fetus exhibited hydrops, short long bones, fixed limb joints, absent fetal movements, and polyhydramnios. The pregnancy was terminated and postmortem studies confirmed the prenatal findings: distal arthrogryposis, fetal growth restriction, pulmonary hypoplasia, and retrognathia. The fetus had a normal chromosomal microarray analysis. Exome sequencing revealed two novel compound heterozygous variants in the GLDN associated with LCCS11. This manuscript reports this case and performs a literature review of all published LCCS11 cases.

Open Source Assessment of Deep Learning Visual Object Detection.

This paper introduces Detection Metrics, an open-source scientific software for the assessment of deep learning neural network models for visual object detection. This software provides objective performance metrics such as mean average precision and mean inference time. The most relevant international object detection datasets are supported along with the most widely used deep learning frameworks. Different network models, even those built from different frameworks, can be fairly compared in this way. This is very useful when developing deep learning applications or research. A set of tools is provided to manage and work with different datasets and models, including visualization and conversion into several common formats. Detection Metrics may also be used in automatic batch processing for large experimental tests, saving researchers time, and new domain-specific datasets can be easily created from videos or webcams. It is open-source, can be audited, extended, and adapted to particular requirements. It has been experimentally validated. The performance of the most relevant state-of-the-art neural models for object detection has been experimentally compared. In addition, it has been used in several research projects, guiding in selecting the most suitable network model architectures and training procedures. The performance of the different models and training alternatives can be easily measured, even on large datasets.

[Anti-SARS-CoV-2 seroprevalence in administrative staff of the Mexican Institute for Social Security]. / Seroprevalencia anti-SARS-CoV-2 en personal administrativo del Instituto Mexicano del Seguro Social.

Background: Asymptomatic subjects, the lack of diagnostic tests and, in countries like Mexico, the epidemiological surveillance method does not allow to establish the real number of infections in the COVID-19 pandemic. Frontline health personnel, as well as other groups related to priority activities are considered of high risk. We included administrative workers in contact with health personnel in the hospital units of the Mexican Institute for Social Security (IMSS, according to its initials in Spanish). Objective: To identify the seroprevalence of antibodies to SARS-CoV-2 in IMSS' administrative staff who does not treat patients. Material and methods: 76 volunteer participating individuals were incluided; IgG antibodies against the SARS-CoV-2 nucleoprotein were measured. A questionnaire was administered to the participants in order to identify possible risk factors. Results: 76 participants were included (39 men, 51.7%), with a median age of 42 years. 29 out of 76 subjects (38.2%), whose median age was 38 years (range 18-69 years); 15 men (51.7%), and 14 women (48.3%). A higher percentage of positive subjects under 45 years of age (n = 20, 84.2%) was observed than those aged 45 or over (n = 9, 25%), with an OR of 3 (95% CI 1.13-7.96, p = 0.03). No statistically significant difference was found regarding the type of comorbidity. Conclusions: The prevalence identified shows an important circulation of the virus in the administrative staff.

Introducción: los sujetos asintomáticos, la falta de pruebas diagnósticas y, en países como México, el método de vigilancia epidemiológica no permiten establecer el número real de contagios en la pandemia de COVID-19. El personal de salud de primera línea y otros grupos con actividades prioritarias son de alto riesgo. Se incluyeron trabajadores administrativos en contacto con personal de salud en las unidades hospitalarias del Instituto Mexicano del Seguro Social (IMSS). Objetivo: identificar la seroprevalencia de anticuerpos contra SARS-CoV-2 en personal administrativo del IMSS que no atiende a enfermos. Material y métodos: se incluyeron 76 individuos a los cuales se les midieron los anticuerpos IgG contra la nucleoproteína del SARS-CoV-2. También se les aplicó un cuestionario para identificar factores de riesgo. Resultados: se incluyeron 76 participantes (39 hombres, 51.7%), con una mediana de 42 años de edad. Fueron positivos 29 de 76 sujetos (38.2%), cuya mediana de edad fue de 38 años (rango 18-69 años); 15 hombres y 14 mujeres. Hubo mayor porcentaje de sujetos positivos menores de 45 años (n = 20, 84.2%) que aquellos de edad ≥ 45 años (n = 9, 25%), con una RM de 3 (IC 95% 1.13-7.96, p = 0.03). No hubo diferencia estadísticamente significativa respecto al tipo de comorbilidad. Conclusiones: la prevalencia identificada muestra una circulación importante del virus en el personal administrativo.