Bifidobacterium longum subsp. infantis CECT 7210 Reduces Inflammatory Cytokine Secretion in Caco-2 Cells Cultured in the Presence of Escherichia coli CECT 515

Previous works have described the activity of Bifidobacterium longum subsp. infantis CECT 7210 (also commercially named B. infantis IM-1®) against rotavirus in mice and intestinal pathogens in piglets, as well as its diarrhea-reducing effect on healthy term infants. In the present work, we focused on the intestinal immunomodulatory effects of B. infantis IM-1® and for this purpose we used the epithelial cell line isolated from colorectal adenocarcinoma Caco-2 and a co-culture system of human dendritic cells (DCs) from peripheral blood together with Caco-2 cells. Single Caco-2 cultures and Caco-2: DC co-cultures were incubated with B. infantis IM-1® or its supernatant either in the presence or absence of Escherichia coli CECT 515. The B. infantis IM-1® supernatant exerted a protective effect against the cytotoxicity caused by Escherichia coli CECT 515 on single cultures of Caco-2 cells as viability reached the values of untreated cells. B. infantis IM-1® and its supernatant also decreased the secretion of pro-inflammatory cytokines by Caco-2 cells and the co-cultures incubated in the presence of E. coli CECT 515, with the response being more modest in the latter, which suggests that DCs modulate the activity of Caco-2 cells. Overall, the results obtained point to the immunomodulatory activity of this probiotic strain, which might underlie its previously reported beneficial effects.Laboratorios Ordesa, S. L.Center for the Development of Industrial Technology (CDTI) of the Spanish Ministry of Science and Innovation (TOLERA project) 2017087

Sudden hearing loss: National survey in Spain

This is the peer reviewed version of the following article: Acta Otorrinolaringológica 67.2 (2016): 59-65, which has been published in final form at http://dx.doi.org/10.1016/j.otorri.2015.03.003The objective of our study was to identify the diagnostic and therapeutic approaches in the different ENT Departments of Spain with respect to sudden deafness. We wanted to establish a basis to help to create a new nation-wide consensus, unifying treatment, diagnostic and follow-up criteria for this disease. Methods: We carried out an anonymous Internet survey, addressing Spanish ENT doctors nation-wide (n = 2,029), gathering in 33 questions different aspects about diagnostic criteria, additional tests, treatment procedures and prognostic factors in sudden deafness, according to the different protocols and experience of the participants in the survey. Results: A total of 293 Spanish ENT doctors (14%) took part anonymously. In relation to diagnostic criteria, is the most noteworthy was the requisite of a confirmed neurosensorial loss (91.1%) followed by "initiated in less than three days" (75%) and 3 consecutive frequencies affected (76.4%). More than half of the participants requested an MRI of the IAC/CPA (68.7%) and 88.2% used gadolinium in this test. The prognostic factor most frequently considered was delay in commencement of treatment onset (84.8%).As far as treatment of primary cases, most of the responders agreed on the use of corticosteroids (99.7%). Oral administration was the most widely used (66%), followed by intravenous (29.6%) and intratympanic (1.4%) administration. Ninety-two percent had not had any major complications with systemic steroids. Intratympanic treatments were used by 70% of responders for rescue in failure. Conclusions: In Spain there is currently a significant disparity of concepts regarding the diagnosis of sudden deafness, and more agreement as to using steroids as their treatment. This highlights the need to implement measures to promote a better approach, which would be homogeneous and consensual, to this condition.Introducción: El objetivo de nuestro estudio es identificar las actitudes diagnósticas y terapéuticas que se llevan a cabo en los diferentes servicios de otorrinolaringología (tanto del ámbito privado como público) en España con respecto a la sordera súbita. Esto permitirá establecer una base que ayude a generar un nuevo consenso a nivel nacional, unificando criterios para el tratamiento, diagnóstico y seguimiento de esta patología. Material y métodos: Se realizó una encuesta anónima por Internet, dirigida a otorrinolaringólogos españoles a nivel nacional (n = 2.029 especialistas afiliados a la SEORL), recopilando en 33 preguntas diferentes aspectos en relación a los criterios diagnósticos, pruebas complementarias, pautas de tratamiento y factores pronósticos en la sordera súbita, según los diferentes protocolos instaurados y experiencia de los participantes en la encuesta. Resultados: Participaron 293 otorrinolaringólogos españoles de forma anónima (14% del total). Respecto a criterios diagnósticos, destaca el requerimiento de confirmar una hipoacusia neurosensorial (91,1%), de inicio en menos de tres días (75%) y afectación de tres frecuencias consecutivas (76,4%). Más de la mitad de los participantes solicitan resonancia magnética de CAI/APC (68,7%), y el 88,2% utiliza contraste con gadolinio en esta prueba. El factor pronóstico que se consideró con mayor frecuencia en la encuesta fue la demora hasta inicio del tratamiento con un 84,8%. Respecto al tratamiento empleado en casos primarios, la gran mayoría de los encuestados (99,7%) coinciden en la administración de corticoides. La vía oral es la más utilizada (66%), seguida de la administración intravenosa (29,6%), e intratimpánica (1,4%). El 92% no han tenido complicaciones mayores con el tratamiento corticoide sistémico. La vía intratimpánica es empleada en un 70% como rescate en fracasos Conclusiones: En España, existe actualmente una importante disparidad en el uso de los medios diagnósticos en la sordera súbita y un mayor acuerdo en el uso de corticoides como su tratamiento. Sería necesario implementar medidas que permitan un mejor abordaje, homogéneo y consensuado de esta patologí

MANEJO DE LOS RECURSOS NATURALES DESDE LA PERSPECTIVA DE GÉNERO: UNA APROXIMACIÓN AL DESARROLLO SUSTENTABLE

En el presente cartel se muestran avances de una investigación realizada en la Comunidad de “El Pirul”, misma que pertenece al municipio de Salvatierra, Gto; sin embargo, antes de comenzar analizaremos los conceptos claves para que sea un poco más comprensible el tema que presentamos, posteriormente se continuara con una breve descripción del lugar donde se realizó la investigación, concluyendo con los resultados que se obtuvieron en la misma. El objetivo principal de esta investigación es: identificar y analizar el uso y preservación de los recursos naturales, desde la perspectiva de género, además de a) identificar el uso de los recursos naturales por hombres y mujeres y b) identificar los recursos naturales que existen en la comunidad desde la perspectiva de género. Metodología: la metodología que se utilizó para la realización de esta investigación fue mixta, incluyendo herramientas como: entrevistas, encuesta, observación participante (la cual se llevó acabo en todas las visitas realizadas a la comunidad con una duración de aproximadamente 4 horas). En lo que concierne a las otras herramientas que se utilizaron, se decidió aplicar un “censo”, ya que la comunidad es muy pequeña, esto nos permitió aplicar 19 encuestas (siendo este el número de casas que existen en la comunidad). Cada encuesta se aplicó a un integrante de la familia. Las entrevistas nos permitieron tener un primer acercamiento a la comunidad, se entrevistó a la delegada y tres habitantes de la comunidad

Monitoring the phenolic ripening of red grapes using a multisensor system based on metal-oxide nanoparticles

Producción CientíficaThe maturity of grapes is usually monitored by means of the sugar concentration. However, the assessment of other parameters such as the phenolic content is also important because the phenolic maturity has an important impact on the organoleptic characteristics of wines. In this work, voltammetric sensors able to detect phenols in red grapes have been developed. They are based on metal oxide nanoparticles (CeO2, NiO, and TiO2,) whose excellent electrocatalytic properties toward phenols allows obtaining sensors with detection limits in the range of 10−8 M and coefficients of variation lower than 7%. An electronic tongue constructed using a combination of the nanoparticle-based sensors is capable to monitor the phenolic maturity of red grapes from véraison to maturity. Principal Component Analysis (PCA) can be successfully used to discriminate samples according to the ripeness. Regression models performed using Partial Least Squares (PLS-1) have established good correlations between voltammetric data obtained with the electrochemical sensors and the Total Polyphenolic Index, the Brix degree and the Total Acidity, with correlation coefficients close to 1 and low number of latent variables. An advantage of this system is that the electronic tongue can be used for the simultaneous assessment of these three parameters which are the main factors used to monitor the maturity of grapes. Thus the electronic tongue based on metal oxide nanoparticles can be a valuable tool to monitor ripeness. These results demonstrate the exciting possible applications of metal oxide nanoparticles in the field of electronic tongues.Ministerio de Economía, Industria y Competitividad - Fondo Europeo de Desarrollo Regional (project AGL2015-67482- R)Junta de Castilla y Leon - Fondo Europeo de Desarrollo Regional (project VA011U16)Junta de Castilla y León (grant BOCYL-D-24112015-9

Altered thymocyte development observed in EphA4-deficient mice courses with changes in both thymic epithelial and extracellular matrix organization

Eph receptors and their ligands, Ephrins, are involved in the thymocyte-thymic epithelial cell (TEC) interactions, key for the functional maturation of both thymocytes and thymic epithelium. Several years ago, we reported that the lack of EphA4, a Eph of the subfamily A, coursed with reduced proportions of double positive (DP) thymocytes apparently due to an altered thymic epithelial stroma [Munoz et al. in J Immunol 177:804–813, 2006]. In the present study, we reevaluate the lymphoid, epithelial, and extracellular matrix (ECM) phenotype of EphA4−/− mice grouped into three categories with respect to their proportions of DP thymocytes. Our results demonstrate a profound hypocellularity, specifc alterations of T cell diferentiation that afected not only DP thymocytes, but also double negative and single positive T cell subsets, as well as the proportions of positively and negatively selected thymocytes. In correlation, thymic histological organization changed markedly, especially in the cortex, as well as the proportions of both Ly51+UEA-1− cortical TECs and Ly51−UEA-1+ medullary TECs. The alterations observed in the expression of ECM components (Fibronectin, Laminin, Collagen IV), integrin receptors (VLA-4, VLA-6), chemokines (CXCL12, CCL25, CCL21) and their receptors (CXCR4, CCR7, CCR9) and in vitro transwell assays on the capacity of migration of WT and mutant thymocytes suggest that the lack of EphA4 alters T-cell diferentiation by presumably afecting cell adhesion between TECs and T-TEC interactions rather than by thymocyte migration

Electronic Noses and Tongues in Wine Industry

Producción CientíficaThe quality of wines is usually evaluated by a sensory panel formed of trained experts or traditional chemical analysis. Over the last few decades, electronic noses (e-noses) and electronic tongues have been developed to determine the quality of foods and beverages. They consist of arrays of sensors with cross-sensitivity, combined with pattern recognition software, which provide a fingerprint of the samples that can be used to discriminate or classify the samples. This holistic approach is inspired by the method used in mammals to recognize food through their senses. They have been widely applied to the analysis of wines, including quality control, aging control, or the detection of fraudulence, among others. In this paper, the current status of research and development in the field of e-noses and tongues applied to the analysis of wines is reviewed. Their potential applications in the wine industry are described. The review ends with a final comment about expected future developments.CM-P agradece a la Universidad de Valladolid por su beca PIF-UVa y CG-H por su contrato pre-doctoral JCYL (BOCYL-D-24112015-9).Ministerio de Economía, Industria y Competitividad – FEDER (Grant AGL2015-67482-R)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13

An Electrochemical Quartz Crystal Microbalance Multisensor System Based on Phthalocyanine Nanostructured Films: Discrimination of Musts

Producción CientíficaAn array of electrochemical quartz crystal electrodes (EQCM) modified with nanostructured films based on phthalocyanines was developed and used to discriminate musts prepared from different varieties of grapes. Nanostructured films of iron, nickel and copper phthalocyanines were deposited on Pt/quartz crystals through the Layer by Layer technique by alternating layers of the corresponding phthalocyanine and poly-allylamine hydrochloride. Simultaneous electrochemical and mass measurements were used to study the mass changes accompanying the oxidation of electroactive species present in must samples obtained from six Spanish varieties of grapes (Juan García, Prieto Picudo, Mencía Regadío, Cabernet Sauvignon, Garnacha and Tempranillo). The mass and voltammetric outputs were processed using three-way models. Parallel Factor Analysis (PARAFAC) was successfully used to discriminate the must samples according to their variety. Multi-way partial least squares (N-PLS) evidenced the correlations existing between the voltammetric data and the polyphenolic content measured by chemical methods. Similarly, N-PLS showed a correlation between mass outputs and parameters related to the sugar content. These results demonstrated that electronic tongues based on arrays of EQCM sensors can offer advantages over arrays of mass or voltammetric sensors used separately.Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13)University of Valladolid (PIF-UVa

Improvement of electrocatalytic effect in voltammetric sensors based on phthalocyanines

Producción CientíficaVoltammetric sensors based on phthalocyanines have been used to detect a variety of compounds. In this paper, the state of the art of sensors prepared using classical techniques will be revised. Then, new strategies to improve the performance of the sensors will be described using as example sensors chemically modified with lutetium bisphthalocyanine (LuPc2) dedicated to the detection of phenols of interest in the food industry. Classical LuPc2 carbon paste electrodes can detect phenols such as catechol, caffeic acid or pyrogallol with limits of detection in the range of 10-4–10-5 M. The performance can be improved by using nanostructured Langmuir–Blodgett (LB) or Layer by Layer (LbL) films. The enhanced surface to volume ratio produce an increase in the sensitivity of the sensors. Limits of detection of 10-5–10-7 M are attained, which are one order of magnitude lower than those obtained using conventional carbon paste electrodes. Moreover, these techniques can be used to coimmobilize two electrocatalytic materials in the same device. The limits of detection obtained in LB sensors combining LuPc2/AuNPs or LuPc2/CNT are further improved. Finally, the LB technique has been used to prepare biosensors where a phenol oxydase (such as tyrosinase or lacasse) is immobilized in a biomimetic environment that preserves the enzymatic activity. Moreover, LuPc2 can be co-immobilized with the enzyme in a lipidc film formed by arachidic acid (AA). LuPc2 can act as an electron mediator facilitating the electron transfer. These biomimetic sensors formed by LuPc2/AA/enzyme show Limits of detection of 10-8 M and an enhanced selectivity.CM-P agradece a la Universidad de Valladolid por su beca PIF-UVa y CG-H por su contrato pre-doctoral JCYL (BOCYL-D-24112015-9).Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13

An allosteric switch between the activation loop and a c-terminal palindromic phosphomotif controls c-Src function.

Autophosphorylation controls the transition between discrete functional and conformational states in protein kinases, yet the structural and molecular determinants underlying this fundamental process remain unclear. Here we show that c-terminal Tyr 530 is a de facto c-Src autophosphorylation site with slow time-resolution kinetics and a strong intermolecular component. On the contrary, activation-loop Tyr 419 undergoes faster kinetics and a cis-to-trans phosphorylation switch that controls c-terminal Tyr 530 autophosphorylation, enzyme specificity, and strikingly, c-Src non-catalytic function as a substrate. In line with this, we visualize by X-ray crystallography a snapshot of Tyr 530 intermolecular autophosphorylation. In an asymmetric arrangement of both catalytic domains, a c-terminal palindromic phospho-motif flanking Tyr 530 on the substrate molecule engages the G-loop of the active kinase adopting a position ready for entry into the catalytic cleft. Perturbation of the phosphomotif accounts for c-Src dysfunction as indicated by viral and colorectal cancer (CRC)-associated c-terminal deleted variants.Weshow that c-terminal residues 531 to 536 are required for c-Src Tyr 530 autophosphorylation, and such a detrimental effect is caused by the substrate molecule inhibiting allosterically the active kinase. Our work reveals a crosstalk between the activation and c-terminal segments that control the allosteric interplay between substrateand enzyme-acting kinases during autophosphorylation.post-print5137 K

The advantages of disposable screen-printed biosensors in a bioelectronic tongue for the analysis of grapes

Producción CientíficaDisposable screen-printed sensors have been modified with enzymes and used to form a bioelectronic tongue dedicated to the discrimination between different grape varieties. The multisensory system combined serigraphied electrodes modified with carbon, platinum, gold, graphene, Prussian blue and nickel oxide nanoparticles (M-SPE) covered with glucose oxidase (M-GOX-SPE) or tyrosinase (M-Tyr-SPE). The M-GOX-SPE and M-Tyr-SPE sensors produced a variety of responses due to the different behavior of the electron mediators of the six screen-printed materials used for the electro-catalysis of the glucose and phenols by means of glucose oxidase and tyrosinase. This variety of responses, together with the capability of the sensors to detect glucose or phenols, allowed the bioelectronic tongue developed here to discriminate between the juices obtained from different varieties of grape. Partial least-squares (PLS-1) multivariate calibration of electrochemical data has been successfully applied to the simultaneous determination of glucose and polyphenols in musts. The discrimination capability shown by this array of cheap and single-use sensors was similar to that found in other complex bioelectronic tongues. The lower price, ease of use and portability of the modified screen-printed electrode system makes the bioelectronic tongue developed here an alternative tool that can be used in situ in the vineyard block.Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13)University of Valladolid (PIF-UVa