Structural basis of the allergenicity to strawberries due to Fra a 1.02

Strawberry fruits are highly valued due to their flavor, aroma, and benefits for human health. Despite this, 30% of the population with food hypersensitivity also shows adverse reactions to strawberry (Franz-Oberdorf et al, 2016). The FaFra a 1 protein family, homologs of the major birch pollen allergen Bet v 1, is involved in this allergenicity to strawberry. By RNAseq we have identified transcripts for 18 members of the FaFra a 1 family (from 1.01 to 1.18) in strawberry fruits. Although expressed in all tissues analyzed, each family member presents a unique pattern of expression, which suggests functional specialization for each FaFra a 1 protein. FaFra a 1.02 (Fra2 from now on) is the most expressed one in red fruits and is also the most allergenic among the family members tested (Muñoz et al. 2010; Franz- Oberdorf et al, 2016). In order to understand the molecular bases of this allergenicity we crystalized Fra2 and obtained its structure by X-ray diffraction. Fra2 showed a very high structural homology to Bet v 1, and we asked whether the two proteins were recognized by the immune system in a similar way. For this, we generated five different mutant versions of Fra2 in sites described as important for allergenicity in Bet v 1 (Fernandes et al, 2016), and studied their potential allergenicity as well as their crystal structures. Three of the mutants had substitutions in loop 4 (E46R, D48R, E46/48A) and the other two facing the cavity (A141F and Q64W). Compared to Fra2, all the mutants showed a significant reduction in their capacity to be recognized by the serum of patients with allergies to Bet v 1, and their crystal structures revealed conformational changes in the Bet v 1- IgG interaction sites. Together, these results support that Fra2 and Bet v 1 have similar allergenic determinants We hope this research will aid in understanding how human IgGs interact with Fra2 and might help in the development of new cultivars with a lesser allergenic potential.Grants BIO2013-44199R and BES-2014-068723 (MINECO). The authors also acknowledge the support by the Plan Propio from University of Malaga, Campus de Excelencia Internacional de Andalucía

Estrategias para mejorar la salud mental frente al confinamiento por el COVID-19 en estudiantes universitarios

Introduction: Due to the COVID-19 pandemic, containment measures were taken, including confinement that negatively affected the mental health, a situation that forced the establishment of strategies. Objective: to determine the strategies to improve mental health of confinement, to know all the strategies that are most prevalent in use and to establish which are the most used. Methodology: review of the literature in the databases PUBMED, SCIELO, GOOGLE SCHOLAR, obtaining 40 articles: 34 cross-sectional observational, 4 longitudinal observational and 2 opinion articles. Results: 14 coping strategies were determined, grouped into 3 categories: cognitive, behavioural and emotional. Conclusions: The most prevalent strategies were active, positive rethinking at a cognitive level, problem-focused coping: level, and affective support: emotional.Introducción: Por la pandemia de COVID-19 se tomaron medidas de contención, entre ellas el confinamiento que afectó de forma negativa la salud mental de los universitarios, situación que obligó a establecer estrategias para mitigar el impacto psicológico. Objetivo: determinar las estrategias para mejorar la salud mental que utilizan los universitarios frente al confinamiento, conocer todas las estrategias que tienen mayor preponderancia en uso y establecer cuáles son las más utilizadas. Metodología: revisión de la literatura en las bases de datos PUBMED, SCIELO, GOOGLE SCHOLAR, obteniendo 40 artículos: 34 observacionales transversales, 4 observacionales longitudinales y 2 artículos de opinión. Resultados: se determinaron 14 estrategias de afrontamiento, agrupadas en 3 categorías: cognitivo, conductual y emocional. Conclusiones: Las estrategias con mayor preponderancia fueron las activas, a nivel cognitivo: replanteamiento positivo, conductual: afrontamiento centrado en el problema, y emocional: apoyo afectivo. Las más utilizada de las 3 categorías fue el apoyo emocional

Effect of binary raw materials replacement (quartz and feldspar) for porcelain chamotte on the electro-technical siliceous porcelain properties

The hurry for ecological practices and waste control has emerged as an obligation in modern times, demanding precise strategies to restrain waste accumulation and to stimulate recycling and reuse actions to lower the climate effect. The replacement of binary raw materials for porcelain chamotte waste in siliceous porcelain was studied to obtain eco-friendly high-voltage porcelain. Quartz and feldspar were progressively replaced by 5, 10, and 15 wt.% of porcelain chamotte in a conventional siliceous electro-technical porcelain composition. The replacement effect on sintered samples at 1250°C under industrial heat treatment was evaluated by measuring the linear shrinkage, bulk density, porosity, flexural strength, and microhardness technological properties. Phase analysis was carried out by X-ray diffraction. Microstructural characteristics were studied using a scanning electron microscope. The results showed that chamotte-containing samples reached bulk densities of about 2.36 g/cm3 and a porosity percentage near zero. The maximum flexural strength value at glazed states was 87.8 MPa, for 15 wt.% scrap-containing samples. X-ray diffraction studies revealed a higher mullite phase content in chamotte-containing samples. Scanning electronic microscopy images of the polished and etched specimens show the presence of quartz grains and secondary mullite needles embedded in a feldspathic vitreous matrix. The properties reached by the chamotte-containing samples are attractive since the values obtained in terms of flexural strength, density, and porosity are compared to those reported for conventional siliceous porcelain were obtained. The most noticeable result was observed in flexural resistance. The glazed porcelain bodies showed a flexural strength improvement of about 15%. Then, these porcelain compositions suggest an alternative to produce a more sustainable, affordable, and environmentally-friendly porcelain insulator product

Graph based study of allergen cross-reactivity of plant lipid transfer proteins (LTPs) using microarray in a multicenter study.

The study of cross-reactivity in allergy is key to both understanding. the allergic response of many patients and providing them with a rational treatment In the present study, protein microarrays and a co-sensitization graph approach were used in conjunction with an allergen microarray immunoassay. This enabled us to include a wide number of proteins and a large number of patients, and to study sensitization profiles among members of the LTP family. Fourteen LTPs from the most frequent plant food-induced allergies in the geographical area studied were printed into a microarray specifically designed for this research. 212 patients with fruit allergy and 117 food-tolerant pollen allergic subjects were recruited from seven regions of Spain with different pollen profiles, and their sera were tested with allergen microarray. This approach has proven itself to be a good tool to study cross-reactivity between members of LTP family, and could become a useful strategy to analyze other families of allergens

El registro fósil de vertebrados y patrimonio paleontológico más representativo de los pirineos occidentales

Los Pirineos occidentales son un territorio con una notable diversidad geológica y un importante registro fósil, que constituye un gran archivo de paleobiodiversidad. En lo que respecta al registro fósil de vertebrados, los Pirineos occidentales (Región Vasco-Cantábrica, Cuenca de Pamplona y sector noroccidental de la Cuenca del Ebro) son relevantes tanto desde el punto de vista de los yacimientos paleontológicos como de las colecciones de fósiles procedentes de ellos. Varios yacimientos de vertebrados forman parte del Inventario de Lugares de Interés Geológico (LIG) con alto o muy alto interés paleontológico en la Comunidad Autónoma del País Vasco. Destacan los yacimientos de peces cretácicos de Zeanuri en Bizkaia, mamíferos eocenos de Zambrana e icnitas miocenas de Salinas de Añana en Álava/Araba y la asociación de vertebrados pleistocenos de Kiputz cerca de Mutriku, en Gipuzkoa. Otros yacimientos notables en el área de estudio deberían ser objeto de iniciativas de geoconservación: es el caso de la cantera de Laño (Condado de Treviño), con asociaciones fi nicretácicas de vertebrados continentales y marinos, los afloramientos con icnitas de aves y mamíferos del Eoceno al Mioceno inferior de Navarra/Nafarroa y la cueva vizcaína de Santa Isabel de Ranero (Pleistoceno), por citar únicamente los ejemplos más sobresalientes. Asimismo, existen colecciones de fósiles de vertebrados en los Pirineos occidentales que destacan por su elevado interés paleontológico, aunque algunos no provengan de LIG reconocidos. Estas colecciones están depositadas en diversos centros e instituciones de Bilbao, Irún, Leioa, Pamplona-Iruñea, y Vitoria-Gasteiz. La valoración de la paleobiodiversidad y el registro fósil es un paso importante para el conocimiento del patrimonio paleontológico de un territorio y la creación de fi guras de geoconservación (e.g., LIG, geosites). Sin embargo, la mera inclusión de los yacimientos paleontológicos en estas fi guras no garantiza su protección y conservación, ni la adecuada gestión de las colecciones de fósiles procedentes de ellos. Por lo tanto, es necesario diseñar estrategias de conservación transversales teniendo en cuenta las particularidades de cada yacimiento y/o colección. La divulgación científi ca y cultural del patrimonio paleontológico de un territorio, como el aquí estudiado, debe contar con una mayor colaboración entre los paleontólogos que han valorado los bienes paleontológicos y los técnicos que los gestionan. Para ello, es necesaria la disposición de partidas presupuestarias específicas que posibiliten el desarrollo de diferentes acciones encaminadas a la investigación, conservación y difusión social de este patrimonio.The western Pyrenees is a territory with a remarkable geodiversity and an important fossil record, which constitutes a large archive of palaeobiodiversity. With respect to the vertebrate fossil record, the western Pyrenees (Basque-Cantabrian Region, Pamplona Basin and the north-western sector of the Ebro Basin) are relevant both from the point of view of palaeontological sites and fossil collections. Several vertebrate fossil sites are integrated into the Inventory of Geological Interest Sites (LIG) with high to very high palaeontological interest of the Autonomous Community of the Basque Country. It is worth highlighting the Cretaceous fish sites of Zeanuri in Biscay, Eocene mammals of Zambrana and Miocene ichnites of Salinas de Añana in Araba, and the Pleistocene vertebrate fossil assemblage of Kiputz near Mutriku, in Gipuzkoa. Other highly significant sites in the study area should be subject to geoconservation initiatives. This is the case of the Laño quarry in the Condado de Treviño, which has provided continental and marine vertebrate fossil assemblages from the latest Cretaceous, Eocene to Early Miocene sites with bird and mammal tracks in Navarre, and the Pleistocene cave deposits of Santa Isabel de Ranero in Biscay, to mention only the most outstanding examples. There are also vertebrate fossil collections in the western Pyrenees that stand out for their high palaeontological interest, although some do not come from recognized LIG. These collections are deposited in several centres and institutions of Bilbao, Irun, Leioa, Pamplona-Iruñea and Vitoria-Gasteiz. The valuation of the palaeobiodiversity and fossil record is an important step for the study of palaeontological heritage of a territory and the set-up of geoconservation designations (e.g., LIG, geosites). However, the mere inclusion of the palaeontological assents in these geoconservation designations does not guarantee either their protection and conservation or the suitable management of the fossil collection. Therefore, it is necessary to design conservation strategies taking into account the particularities of each palaeontological site and/or fossil collection. The scientific and cultural dissemination of the palaeontological heritage of a territory like one studied here should have greater collaboration between the palaeontologists that have valued the palaeontological assets and the technicians and institutions that manage them. This requires the provision of specific budgetary items that enable the development of the different activities of research, conservation and social dissemination of this heritage.Fil: Badiola, Ainara. Universidad del País Vasco; EspañaFil: Arlegi, Mikel. Universidad del País Vasco; España. Université de Bordeaux; FranciaFil: Astibia, Humberto. Universidad del País Vasco; EspañaFil: Bardet, Nathalie. Centre National de la Recherche Scientifique; Francia. Muséum National d'Histoire Naturelle; Francia. Université Pierre et Marie Curie; FranciaFil: Berreteaga, Ana. Universidad del País Vasco; EspañaFil: Corral, J. Carmelo. Museo de Ciencias Naturales de Álava/; EspañaFil: Díaz Martínez, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigación en Paleobiología y Geología; Argentina. Universidad Nacional de Río Negro. Sede Alto Valle. Instituto de Investigaciones en Paleobiología y Geología; ArgentinaFil: Gómez Olivencia, Asier. Universidad del País Vasco; España. IKERBASQUE. Basque Foundation for Science; España. Universidad Carlos III de Madrid. Instituto de Salud; EspañaFil: Lopez Horgue, Mikel A.. Universidad del País Vasco; EspañaFil: Perales Gogenola, Leire. Universidad del País Vasco; EspañaFil: Pereda Suberbiola, Xabier. Universidad del País Vasco; Españ

Robustness of dead Cas9 activators in human pluripotent and mesenchymal stem cells

Human pluripotent stem cells (hPSCs) and mesenchymal stromal/stem cells (hMSCs) are clinically relevant sources for cellular therapies and for modeling human development and disease. Many stem cell-based applications rely on the ability to activate several endogenous genes simultaneously to modify cell fate. However, genetic intervention of these cells remains challenging. Several catalytically dead Cas9 (dCas9) proteins fused to distinct activation domains can modulate gene expression when directed to their regulatory regions by a specific single-guide RNA (sgRNA). In this study, we have compared the ability of the first-generation dCas9-VP64 activator and the second-generation systems, dCas9-SAM and dCas9-SunTag, to induce gene expression in hPSCs and hMSCs. Several stem cell lines were tested for single and multiplexed gene activation. When the activation of several genes was compared, all three systems induced specific and potent gene expression in both single and multiplexed settings, but the dCas9-SAM and dCas9-SunTag systems resulted in the highest and most consistent level of gene expression. Simultaneous targeting of the same gene with multiple sgRNAs did not result in additive levels of gene expression in hPSCs nor hMSCs. We demonstrate the robustness and specificity of second-generation dCas9 activators as tools to simultaneously activate several endogenous genes in clinically relevant human stem cells.We thank CERCA/Generalitat de Catalunya and Fundació Josep Carreras-Obra Social la Caixa for their institutional support. We thank Jose Luis Sardina (IJC, Barcelona) for technical assistance with the teratoma assays. Financial support for this work was obtained from the Catalunya Goverment (SGR330 and PERIS 2017-2019), the Spanish Ministry of Economy and Competitiveness (SAF2016-80481-R), the European Research Council (CoG-2014-646903), and the Fundación Leo Messi to P.M.; the Spanish Association against Cancer (AECC-CI-2015) and the Health Institute Carlos III (ISCIII/FEDER, PI17/01028) to C.B.; the Biotechnology and Biological Sciences Research Council (BBRSC) to L.M.F. and A.F.; and the Spanish National Research and Development Plan (ISCIII/FEDER, PI17/02303) and the AEI/MICIU EXPLORA Project (BIO2017-91272-EXP) to S.R.-P. P.M. is an investigator of the Spanish Cell Therapy Cooperative Network (TERCEL). R.T.-R. is supported by a postdoctoral fellowship from the Asociación Española Contra el Cáncer (AECC).S

Antigenic Proteins Involved in Occupational Rhinitis 1 and Asthma Caused by Obeche Wood (Triplochiton Scleroxylon)

Background Obeche wood dust is a known cause of occupational asthma where an IgE-mediated mechanism has been demonstrated. Objective To characterize the allergenic profile of obeche wood dust and evaluate the reactivity of the proteins by in vitro, ex vivo and in vivo assays in carpenters with confirmed rhinitis and/or asthma Materials and methods An in-house obeche extract was obtained, and two IgE binding bands were purified (24 and 12 kDa) and sequenced by N-terminal identity. Specific IgE and IgG, basophil activation tests and skin prick tests (SPTs) were performed with whole extract and purified proteins. CCD binding was analyzed by ELISA inhibition studies. Results Sixty-two subjects participated: 12 with confirmed occupational asthma/rhinitis (ORA+), 40 asymptomatic exposed (ORA−), and 10 controls. Of the confirmed subjects, 83% had a positive SPT to obeche. There was a 100% recognition by ELISA in symptomatic subjects vs. 30% and 10% in asymptomatic exposed subjects and controls respectively (p<0.05). Two new proteins were purified, a 24 kDa protein identified as a putative thaumatin-like protein and a 12 kDa gamma-expansin. Both showed allergenic activity in vitro, with the putative thaumatin being the most active, with 92% recognition by ELISA and 100% by basophil activation test in ORA+ subjects. Cross-reactivity due to CCD was ruled out in 82% of cases. Conclusions Two proteins of obeche wood were identified and were recognized by a high percentage of symptomatic subjects and by a small proportion of asymptomatic exposed subjects. Further studies are required to evaluate cross reactivity with other plant allergens

The involvement of thaumatin-like proteins in plant food cross-reactivity: a multicenter study using a specific protein microarray.

Cross-reactivity of plant foods is an important phenomenon in allergy, with geographical variations with respect to the number and prevalence of the allergens involved in this process, whose complexity requires detailed studies. We have addressed the role of thaumatin-like proteins (TLPs) in cross-reactivity between fruit and pollen allergies. A representative panel of 16 purified TLPs was printed onto an allergen microarray. The proteins selected belonged to the sources most frequently associated with peach allergy in representative regions of Spain. Sera from two groups of well characterized patients, one with allergy to Rosaceae fruit (FAG) and another against pollens but tolerant to food-plant allergens (PAG), were obtained from seven geographical areas with different environmental pollen profiles. Cross-reactivity between members of this family was demonstrated by inhibition assays. Only 6 out of 16 purified TLPs showed noticeable allergenic activity in the studied populations. Pru p 2.0201, the peach TLP (41%), chestnut TLP (24%) and plane pollen TLP (22%) proved to be allergens of probable relevance to fruit allergy, being mainly associated with pollen sensitization, and strongly linked to specific geographical areas such as Barcelona, Bilbao, the Canary Islands and Madrid. The patients exhibited mayor que50% positive response to Pru p 2.0201 and to chestnut TLP in these specific areas. Therefore, their recognition patterns were associated with the geographical area, suggesting a role for pollen in the sensitization of these allergens. Finally, the co-sensitizations of patients considering pairs of TLP allergens were analyzed by using the co-sensitization graph associated with an allergen microarray immunoassay. Our data indicate that TLPs are significant allergens in plant food allergy and should be considered when diagnosing and treating pollen-food allergy

Transport of Pru p 3 across gastrointestinal epithelium - an essential step towards the induction of food allergy?

Background Since intestinal absorption of food protein can trigger an allergic reaction, the effect of plant food allergen on intestinal epithelial cell permeability and its ability to cross the epithelial monolayer was evaluated. Objective To study the interaction of Pru p 3 with intestinal epithelium, its natural entrance, analyzing transport kinetics and cellular responses that trigger. Methods This was achieved using Pru p 3, the peach LTP, as a model. Enterocytic monolayers were established by culturing Caco 2 cells, as a model of enterocytes, on permeable supports that separate the apical and basal compartments. Pru p 3 was added to the apical compartment, the transepithelial resistance (TEER) was measured, and the transport was quantified. Results The peach allergen that crossed the cell monolayer was detected in the cell fraction and in the basal medium by immunodetection with specific antibodies and the quantity was measured by ELISA assay. Pru p 3 was able to cross the monolayer without disturbing the integrity of the tight junctions. This transport was significantly higher than that of a non-allergenic peach LTP, LTP1, and occurred via lipid raft pathway. The incubation of Caco 2 cells with Pru p 3 and LTP1 produced the expression of epithelial-specific cytokines TSLP, IL33 and IL25. Conclusion These results suggest that Pru p 3 was able to cross the cell monolayer by the transcellular route and then induce the production of Th2 cytokines. The results of the present study represent a step towards clarifying the importance of Pru p 3 as a sensitizer. Clinical relevance The capacity of food allergens to cross the intestinal monolayer could explain their high allergenic capacity and its fast diffusion through the body associating to severe symptoms