Determinación del umbral de detección de variaciones del genoma del tipo SNVs y pequeñas indels en base a los valores de la relación Var/Depth en muestras clínicas analizadas mediante tecnología NGS por exoma

[EN] The use of NGS technologies for the genetic diagnosis of hereditary diseases is becoming increasingly common. During the variant calling process, the values of Depth and Var/Depth for each sequenced position are obtained. These values indicate the number of reads obtained for that position, and the ratio between the number of reads for the alternative allele and the total number of reads obtained for that position. In theory, a heterozygous variant (1 copy of the reference allele and 1 copy of the alternative allele) should have a V/D close to 0.5. However, factors such as the Depth or the complexity of the sequence analyzed (homopolymer, GC rich regions, homology regions, etc.) can alter the V/D value (moving it away from the theoretical ratio of 0.5) and can raise doubts about the presence of these variations in the sample. Currently, the presence of a variant with probable clinical relevance must be confirmed by Sanger sequencing, the Gold Standard for sequencing in most laboratories. This confirmation represents an extra cost, and can represent a significant expense depending on the number of variants detected by NGS. This expense increases exponentially when a large number of genes are analyzed as in exome analysis. In this context, the aim of this study is to empirically determine a V/D threshold value that will allow us to discriminate a priori between a false positive and a false negative, avoiding performing the additional Sanger sequencing confirmation. To do this, we will select and confirm by Sanger sequencing a large number of variants identified by means of exome analysis and that have a V/D value far from the theoretical value of a heterozygous variant (0.5). The data obtained will be subjected to a statistical analysis in order to establish this V/D threshold value. The determination of the cut-off point is helpful since it allows increasing the benefit-efficiency ratio of the genetic diagnosis using NGS.[ES] El uso de las tecnologías de secuenciación masiva (NGS) para el diagnóstico de enfermedades de origen genético es cada vez más frecuente. En el proceso de identificación de variantes mediante NGS, para cada posición secuenciada se obtienen unos valores de Depth (D) y Var/Depht (V/D), que indican el número de lecturas obtenidas para esa posición, y el ratio entre el número de veces que se detecta el alelo alternativo con respecto a las lecturas totales para esa posición. Teóricamente, una variante en heterocigosis (1 copia del alelo de referencia y 1 copia del alelo alternativo) debería tener un V/D en torno al 0.5. No obstante, factores como el Depth o la complejidad de la secuencia analizada (zonas de homopolímeros, zonas ricas en GC, zonas de homología etc.) pueden alterar el ratio V/D (alejándolo del ratio teórico de 0.5) y creando, por tanto, dudas acerca de la verdadera presencia de la variante en la muestra analizada. Actualmente la presencia de una variante con posible relevancia clínica ha de ser confirmada mediante secuenciación Sanger, ya que se trata de la metodología Gold Standard de secuenciación en la mayoría de laboratorios. Esta confirmación representa un coste adicional para estos estudios, pudiendo llegar a suponer un gasto relevante en función del número de variantes detectadas mediante NGS. Este gasto se incrementa de forma exponencial con un mayor número de genes analizados, como es el caso del exoma diagnóstico. En este contexto, el objetivo del presente trabajo consiste en determinar el valor umbral de V/D, que permita esclarecer a priori la presencia o ausencia de una variante detectada por NGS, sin necesidad de realizar su confirmación por secuenciación Sanger. Para ello se llevará a cabo la selección y confirmación mediante secuenciación Sanger, de un elevado número de variantes con un V/D discrepante del teórico 0.5, identificadas en muestras de exoma. Estos datos serán sometidos a un análisis estadístico para el establecimiento del valor umbral de V/D que nos permita discriminar entre la presencia de un falso positivo frente a un verdadero positivo. La determinación de dicho punto de corte es de gran ayuda ya que permite aumentar el beneficio coste-eficiencia del diagnóstico genético mediante NGS.Ozáez Martínez, J. (2016). Determinación del umbral de detección de variaciones del genoma del tipo SNVs y pequeñas indels en base a los valores de la relación Var/Depth en muestras clínicas analizadas mediante tecnología NGS por exoma. http://hdl.handle.net/10251/68718TFG

INTELIGENCIA EMOCIONAL EN EDUCACIÓN PRIMARIA.

The central topic of this paper is the emotional intelligence. The emotional intelligence helps to the development of the self-esteem, the empathy, the emotional control and so on, which provides very beneficial properties to be worked out and stimulated in the classroom. First, we will carry out an exhaustive search on the principal components of the emotional intelligence and we will investigate thoroughly this term. After the review of the present programs on the market, we will carry out the explanation of two programs for the development of the emotional intelligenc

Inteligencia emocional en Educación Primaria

El tema central de este documento es la inteligencia emocional. La inteligencia emocional ayuda al desarrollo de la autoestima, la empatía, el control emocional…lo que la dota de propiedades muy beneficiosas para ser trabajada e impulsada en el aula. En primer lugar, realizaremos una búsqueda exhaustiva acerca de los componentes principales de la inteligencia emocional e investigaremos a fondo este término. Tras la revisión de los programas presentes en el mercado, llevaremos a cabo la explicación de dos programas para el desarrollo de la inteligencia emocional

Ecdysone-related biomarkers of toxicity in the model organism Chironomus riparius: stage and sex-dependent variations in gene expression profiles

Despite being considered a model organism in toxicity studies, particularly in assessing the environmental impact of endocrine disrupting compounds (EDCs) and other chemicals, the molecular basis of development is largely unknown in Chironomus riparius. We have characterized the expression patterns of important genes involved in the ecdysone pathway from embryos to pupa, but specially during the different phases of C. riparius fourth larval instar, according to the development of genital and thoracic imaginal discs. Real-Time PCR was used to analyze: EcR and usp, two genes encoding the two dimerizing partners of the functional ecdysone receptor; E74, an early response gene induced by ecdysteroids; vg (vitellogenin), an effector gene; hsp70 and hsc70, two heat-shock genes involved in the correct folding of the ecdysone receptor; and rpL13, as a part of the ribosomal machinery. Our results show for the first time stage and sex-dependent variations in ecdysone-responsive genes, specially during the late larval stage of C. riparius. The induction in the expression of EcR and usp during the VII-VIII phase of the fourth instar is concomitant with a coordinated response in the activity of the other genes analyzed, suggesting the moment where larvae prepare for pupation. This work is particularly relevant given that most of the analyzed genes have been proposed previously in this species as sensitive biomarkers for the toxicological evaluation of aquatic ecosystems. Identifying the natural regulation of these molecular endpoints throughout the Chironomus development will contribute to a more in-depth and accurate evaluation of the disrupting effects of EDCs in ecotoxicological studies.IO's research was funded by an FPI grant from Ministerio de Ciencia e Innovación (http://www.idi.mineco.gob.es; CTM2009-07189). Research funding was provided by Ministerio de Economía y Competitividad (grant number CTM-2012-37547) and Ministerio de Ciencia e Innovación (grant number CGL2009-10868), SpainS

Arabidopsis subtilases promote defense-related pectin methylesterase activity and robust immune responses to botrytis infection

Plants involve a fine modulation of pectin methylesterase (PME) activity against microbes. PME activity can promote the cell wall stiffening and the production of damage signals able to induce defense responses and plant resistance to pathogens. However, the molecular mechanisms underlying PME activation during disease remain largely unknown. In this study, we explored the role of subtilases (SBTs) as PME activators in Arabidopsis immunity. By using biochemical and reverse genetic approaches, we found that the expression of SBT3.3 and SBT3.5 influences the induction of defense-related PME activity and resistance to the fungus Botrytis cinerea. Arabidopsis sbt3.3 and sbt3.5 knockout mutants showed decreased induction of PME activity and increased susceptibility to the fungus. SBT3.3 expression was stimulated by oligogalacturonides. Overexpression of SBT3.3 overactivated PME activity during fungal infection and enhanced resistance to B. cinerea. A negative correlation was observed between SBT3.3 expression and cell wall methyl ester content in the genotypes analyzed after B. cinerea infection. Increased expression of defense-related genes, including PAD3, CYP81F2 and WAK2, was also revealed in SBT3.3 overexpressing lines. We also demonstrated that SBT3.3 and pro-PME17 are both secreted into the cell wall using distinct protein secretion pathways and different kinetics. Our results propose SBT3.3 and SBT3.5 as modulators of PME activity in Arabidopsis against Botrytis to promptly boost immunity limiting the growth-defense trade-off

Comparative Study of Inkjet-Printed Silver Conductive Traces With Thermal and Electrical Sintering

Thermal sintering has traditionally been the most popular sintering method to enhance conductivity after the printing process in the manufacturing of printed electronics. Nevertheless, in recent years, there has been a growing interest in electrical sintering as an alternative method to overcome some of the limitations of thermal curing. This paper makes a comparative study of both sintering methods in terms of surface morphology, electrical dc conductance, and radiofrequency performance for different applied voltage waveforms. To this end, microstrip transmission lines have been inkjet-printed using nanoparticle-based silver ink on flexible polyimide substrate. The traces have been tested under different sintering conditions, achieving electrical sintering resistivity values only 2.3 times higher than that of bulk silver. This implies a 62% reduction in comparison with the best resistivity value achieved using thermal sintering in our samples. The main novelty of this contribution lies in the analysis of RF behavior as a function of electrical sintering conditions. Lower resistivities have been achieved with slower voltage ramps or allowing higher density current during sintering. It has also been proved that electrically sintered lines have similar RF performance than high-temperature thermally sintered lines in terms of insertion losses, regardless of their very different surface topology. Therefore, we can take advantage of the benefits that electrical sintering offers over thermal sintering regarding significant shorter sintering times maintaining suitable RF performance.This work was supported in part by the Spanish Ministry of Economics and Competitiveness under Grant CTQ2016-78754-C2-1-R

A new role for green leaf volatile esters in tomato stomatal defense against Pseudomonas syringe pv. tomato

[EN] The volatile esters of (Z)-3-hexenol with acetic, propionic, isobutyric, or butyric acids are synthesized by alcohol acyltransferases (AAT) in plants. These compounds are differentially emitted when tomato plants are efficiently resisting an infection with Pseudomonas syringae pv. tomato. We have studied the defensive role of these green leaf volatile (GLV) esters in the tomato response to bacterial infection, by analyzing the induction of resistance mediated by these GLVs and the phenotype upon bacterial infection of tomato plants impaired in their biosynthesis. We observed that treatments of plants with (Z)-3-hexenyl propionate (HP) and, to a greater extent with (Z)-3-hexenyl butyrate (HB), resulted in stomatal closure, PR gene induction and enhanced resistance to the bacteria. HB-mediated stomatal closure was also effective in several plant species belonging to Nicotiana, Arabidopsis, Medicago, Zea and Citrus genus, and both stomatal closure and resistance were induced in HB-treated NahG tomato plants, which are deficient in salicylic acid (SA) accumulation. Transgenic antisense AAT1 tomato plants, which displayed a reduction of ester emissions upon bacterial infection in leaves, exhibited a lower ratio of stomatal closure and were hyper-susceptible to bacterial infection. Our results confirm the role of GLV esters in plant immunity, uncovering a SA-independent effect of HB in stomatal defense. Moreover, we identified HB as a natural stomatal closure compound with potential agricultural applications.This work was funded by Grant AICO/2017/048 from the Generalitat Valenciana and by Grant INNVAL10/18/005 from the Agencia Valenciana de la Innovacio (Spain). We would like to thank the Metabolomics Service of the IBMCP (Valencia, Spain), especially to Teresa Caballero for her excellent technical support in the VOCs quantification. We also thank Eduardo Moya for technical assistance.López-Gresa, MP.; Payá, C.; Ozáez-Martínez, M.; Rodrigo Bravo, I.; Conejero Tomás, V.; Klee, H.; Belles Albert, JM.... (2018). A new role for green leaf volatile esters in tomato stomatal defense against Pseudomonas syringe pv. tomato. Frontiers in Plant Science. 9:1-12. https://doi.org/10.3389/fpls.2018.01855S112

Universidad popular, Cabañal - Valencia

Centro de formación continuada, también denominado universidad para adultos o universidad popular, con el condicionante de ofrecer un uso flexible dentro de este ámbito general docente. En periodo estival se utilizará como universidad de verano, con residencia. Cuenta con aulas teóricas y prácticas, así como sala polivalente, de audiovisuales y de exposiciones.Rus Ozáez, M. (2011). Universidad popular, Cabañal - Valencia. http://hdl.handle.net/10251/36777.Archivo delegad