Planta: un proyecto de aprendizaje basado en juegos en Fisiología Vegetal

[ES] Planta es un juego de cartas pensado como herramienta docente para introducir o repasar conceptos científicotécnicos relacionados con las plantas y su cultivo. A través de este juego se pretende alcanzar el objetivo descrito de manera sencilla, rigurosa, en un ambiente distendido y para distintos niveles de la educación. El argumento del juego es asumir el reto de hacer crecer, mantener y cosechar un cultivo afectado por diversos factores de estrés provocados por los jugadores adversarios. Durante el juego se tratan aspectos básicos del cultivo de las plantas, su fisiología, los aspectos adversos que condicionan su productividad, y los recursos disponibles para remediarlos. El primer paso del proyecto fue crear el juego en sí: mediante un proceso de ensayo y error se alcanzó una versión básica, con diseños de cartas originales, que fue probada entre los propios autores. Ello permitió afinar las reglas del juego y ajustarlas para que el desarrollo de una partida no durara más de treinta minutos. Posteriormente, se encargó una impresión profesional con la cual se realizaron diferentes sesiones con alumnos voluntarios universitarios y de bachillerato. Tras las sesiones, su opinión se recogió en una encuesta. La valoración general del juego fue muy positiva. En la actualidad el juego está en fase de protección de la propiedad intelectual. Por último, está planteada una ampliación del juego (versión avanzada) consistente en la incorporación de cartas con preguntas (de necesaria respuesta para el progreso del jugador en el juego) de conceptos clave en las diferentes asignaturas del área de Fisiología Vegetal en las que se aplique esta herramienta. En las encuestas, los alumnos expresaron de manera clara que les gustaría que el plan de ampliación se llevara a cabo y se aplicara como herramienta docente en las asignaturas del grado

Studying the Impact of Different Field Environmental Conditions on Seed Quality of Quinoa: The Case of Three Different Years Changing Seed Nutritional Traits in Southern Europe

Chenopodium quinoa Willd (quinoa) has acquired an increased agronomical and nutritional relevance due to the capacity of adaptation to different environments and the exceptional nutritional properties of their seeds. These include high mineral and protein contents, a balanced amino acid composition, an elevated antioxidant capacity related to the high phenol content, and the absence of gluten. Although it is known that these properties can be determined by the environment, limited efforts have been made to determine the exact changes occurring at a nutritional level under changing environmental conditions in this crop. To shed light on this, this study aimed at characterizing variations in nutritional-related parameters associated with the year of cultivation and different genotypes. Various nutritional and physiological traits were analyzed in seeds of different quinoa cultivars grown in the field during three consecutive years. We found differences among cultivars for most of the nutritional parameters analyzed. It was observed that the year of cultivation was a determinant factor in every parameter studied, being 2018 the year with lower yields, germination rates, and antioxidant capacity, but higher seed weights and seed protein contents. Overall, this work will greatly contribute to increase our knowledge of the impact of the environment and genotype on the nutritional properties of quinoa seeds, especially in areas that share climatic conditions to Southern Europe.This work was supported by the Ministerio de Ciencia e Innovación (MICINN, Spain) (PID2019-105748RA-I00), the Madrid Government (Comunidad de Madrid-Spain) under the Multiannual Agreement with Universidad Autónoma de Madrid in the line of action encouraging youth research doctors, in the context of the V PRICIT (Regional Programme of Research and Technological Innovation) (SI1/PJI/2019-00124), the CYTED (ValSe-Food 119RT0567), the FPI UAM Fellowship Programme 2019 (to SG-R), and the Ramón y Cajal Programme 2019 (to MR).Peer reviewe

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Computational prediction method to decipher receptor-glycoligand interactions in plant immunity

A lo largo de la evolución, el sistema inmune de las plantas ha seleccionado las paredes celulares de plantas y microorganismos como fuente de Patrones Moleculares Asociados a Daño (DAMPs) o a Microorganismos (MAMPs). La percepción de estos patrones moleculares por los dominios extracelulares (ECD) de Receptores de Reconocimiento de Patrones (PRRs) desencadena respuestas inmunes. Dentro del inmenso número de ligandos que los PRRs vegetales pueden percibir, aquellos compuestos por azúcares están muy poco estudiados, así, un número muy reducido de complejos PRR/glicanos han sido determinados y caracterizados. En esta tesis presentamos un método computacional de cribado in silico que permite predecir nuevas interacciones de ECDs de PRRs con glicanos. Este método se basa en el primer paso de una simulación de dinámica molecular y ha sido desarrollado y optimizado con la familia LysM de PRRs de Arabidopsis thaliana. PRRs de esta familia como CERK1, LYK4 y LYK5 están implicados en la percepción de MAMPs de hongos tales como la quitina o ciertos β-1,3-glucanos. Nuestros resultados in silico predijeron las interacciones directas, ya descritas previamente, de CERK1 y LYK5 con el hexasacárido de quitina [β-1,4-D-(GlcNAc)6], a la vez que descartaban la interacción de este MAMP con LYK4. Por el contrario, no se detectó una unión de CERK1 con laminarihexaosa [β- 1,3-D-(Glc)6] a pesar de que CERK1 es necesario para la activación inmune mediada por este ligando, lo que indicaría que CERK1 podría ser un co-receptor para ciertos glicoligandos. Estos resultados in silico fueron validados in vitro mediante ensayos de calorimetría de los ECD recombinantes de CERK1, LYK4 y LYK5 con β-1,4-D- (GlcNAc)6 y β-1,3-D-(Glc)6. Para aumentar la robustez del método, se testó in silico la interacción de CERK1 con el carbohidrato β-1,4-D-(Glc)6 (celohexaosa), y se comprobó la ausencia de interacción, que fue validada experimentalmente estudiando las respuestas inmunes activadas por celohexaosa, que son similares en plantas silvestres y en el mutante cerk1 de Arabidopsis. Adicionalmente, el método desarrollado se validó analizando interacciones ya descritas de β-1,4-D-(GlcNAc)6 con CfAvr4 y Mg1LysM, así como la desaparición de interacciones entre CERK1 y dicho ligando tras mutaciones en residuos clave del ECD. En una segunda parte de esta tesis, se procedió al testado de PRRs del resto de familias de Arabidopsis con ECDs hipotéticamente capacitados para unir azúcares. Entre los PRRs testados se encuentran AtC-lectina, GmSusD, MmDECTIN-1, PDLP5, LORE, ANX1/2, THE1 y PR5K entre otros. Dichos PRRs fueron testados contra numerosos glicoligandos, entre los que se incluyen: oligosacáridos de quitina, β-glucanos, mananos y xilanos. Los resultados obtenidos permitieron refinar el protocolo de predicción, así como profundizar en el reconocimiento y caracterización de los ECDs de estos receptores. El poder predictivo de este protocolo computacional podría acelerar el descubrimiento y la caracterización de la interacción de glicanos con ECDs de PRRs o de otras proteínas, así como generar información relacionada con las respuestas inmunes activadas por nuevos glicoligandos y los aminoácidos de los dominios ECDs de los PRRs implicados en estas uniones. Estos avances contribuirán al desarrollo futuro de nuevos inmunomoduladores de las repuestas de resistencia de plantas a patógenos. ----------ABSTRACT---------- Throughout evolution, the plant immune system has selected plant and microorganisms cell walls as a source of Damage- or Microbe-Associated Molecular Patterns (MAMPs/DAMPs). The perception of those molecular patterns by the extracellular domains (ECDs) of Pattern Recognition Receptors (PRRs) triggers immune responses. From the vast number of ligands that plant PRRs can bind, those composed of carbohydrate moieties are poorly studied, and only a handful of PRR/glycan complexes have been determined and characterized. In this thesis we present a computational method for in silico predictions of new interactions of PRR-ECDs with glycans. This method is based on the first step of a Molecular Dynamics simulation and has been developed and optimized with the Arabidopsis thaliana LysM-PRR family. PRRs from this family like CERK1, LYK4 and LYK5 are involved in the perception of fungal MAMPs like chitin or some β-1,3-glucans. Our in silico results predicted the direct interactions, previously published, of CERK1 and LYK5 with the chitin hexasaccharide [β-1,4-D-(GlcNAc)6], at the same time that discarded the interaction of this MAMP with LYK4. On the contrary, an interaction of CERK1 with laminarihexaose [β-1,3-D-(Glc)6] was not detected, despite CERK1 is necessary for the immune activation mediated by this ligand, which suggests that CERK1 may act as a co-receptor in its recognition. These in silico results were validated in vitro with isothermal titration calorimetry assays of the recombinant ECDs of CERK1, LYK4 and LYK5 with β-1,4-D-(GlcNAc)6 and β-1,3-D-(Glc)6. To improve the robustness of the method, it was tested in silico the interaction of CERK1 with the carbohydrate β-1,4-D- (GlcNAc)6 (cellohexaose). The absence of interaction predicted was validated experimentally with the study of the immune responses activated by cellohexaose, similar in Arabidopsis wild-type and cerk1 mutant plants. Additionally, the developed method was corroborated by the study of other published interactions like CfAvr4 and Mg1LysM with β-1,4-D-(GlcNAc)6, as well as the disappearance of interactions between CERK1 and β-1,4-D-(GlcNAc)6 upon in silico mutations of CERK1-ECD key residues. In a second part of this thesis, we proceeded to the testing of the rest of PRR families of Arabidopsis putatively able to bind carbohydrates. The PRR tested were AtClectin, GmSusD, MmDECTIN-1, PDLP5, LORE, ANX1/2, THE1 and PR5K amongst others. Those PRRs were tested against several glycoligands like chitin oligosacharides, β-glucans, mannans and xylans. The retrieved results allowed the refinement of the prediction protocol and the study of the recognition and characterization of the receptor- ECDs. The predictive power of this computational protocol may accelerate the discovery and characterization of the interaction of glycans with PRR-ECDs or other proteins, as well as to generate information related to the immune responses activated by both novel glycoligands and the ECD-PRR residues involved in those interactions. These advances could contribute to the development of novel immunomodulators that can provide plant resistance to pathogens

Arabinoxylan-Oligosaccharides Act as Damage Associated Molecular Patterns in Plants Regulating Disease Resistance

Immune responses in plants can be triggered by damage/microbe-associated molecular patterns (DAMPs/MAMPs) upon recognition by plant pattern recognition receptors (PRRs). DAMPs are signaling molecules synthesized by plants or released from host cellular structures (e.g., plant cell walls) upon pathogen infection or wounding. Despite the hypothesized important role of plant cell wall-derived DAMPs in plant-pathogen interactions, a very limited number of these DAMPs are well characterized. Recent work demonstrated that pectin-enriched cell wall fractions extracted from the cell wall mutant impaired in Arabidopsis Response Regulator 6 (arr6), that showed altered disease resistance to several pathogens, triggered more intense immune responses than those activated by similar cell wall fractions from wild-type plants. It was hypothesized that arr6 cell wall fractions could be differentially enriched in DAMPs. In this work, we describe the characterization of the previous immune-active fractions of arr6 showing the highest triggering capacities upon further fractionation by chromatographic means. These analyses pointed to a role of pentose-based oligosaccharides triggering plant immune responses. The characterization of several pentose-based oligosaccharide structures revealed that β-1,4-xylooligosaccharides of specific degrees of polymerization and carrying arabinose decorations are sensed as DAMPs by plants. Moreover, the pentasaccharide 33-α-L-arabinofuranosyl-xylotetraose (XA3XX) was found as a highly active DAMP structure triggering strong immune responses in Arabidopsis thaliana and enhancing crop disease resistance

Cell wall-derived mixed-linked β-1,3/1,4-glucans trigger immune responses and disease resistance in plants

Pattern-triggered immunity (PTI) is activated in plants upon recognition by pattern recognition receptors (PRRs) of damage- and microbe-associated molecular patterns (DAMPs and MAMPs) derived from plants or microorganisms, respectively. To understand better the plant mechanisms involved in the perception of carbohydrate-based structures recognized as DAMPs/MAMPs, we have studied the ability of mixed-linked β-1,3/1,4-glucans (MLGs), present in some plant and microbial cell walls, to trigger immune responses and disease resistance in plants. A range of MLG structures were tested for their capacity to induce PTI hallmarks, such as cytoplasmic Ca2+ elevations, reactive oxygen species production, phosphorylation of mitogen-activated protein kinases and gene transcriptional reprogramming. These analyses revealed that MLG oligosaccharides are perceived by Arabidopsis thaliana and identified a trisaccharide, β-d-cellobiosyl-(1,3)-β-d-glucose (MLG43), as the smallest MLG structure triggering strong PTI responses. These MLG43-mediated PTI responses are partially dependent on LysM PRRs CERK1, LYK4 and LYK5, as they were weaker in cerk1 and lyk4 lyk5 mutants than in wild-type plants. Cross-elicitation experiments between MLG43 and the carbohydrate MAMP chitohexaose [β-1,4-d-(GlcNAc)6 ], which is also perceived by these LysM PRRs, indicated that the mechanism of MLG43 recognition could differ from that of chitohexaose, which is fully impaired in cerk1 and lyk4 lyk5 plants. MLG43 treatment confers enhanced disease resistance in A. thaliana to the oomycete Hyaloperonospora arabidopsidis and in tomato and pepper to different bacterial and fungal pathogens. Our data support the classification of MLGs as a group of carbohydrate-based molecular patterns that are perceived by plants and trigger immune responses and disease resistance

Unidades didácticas para la educación secundaria obligatoria

Esta obra se compone de seis unidades didácticas, destinadas a alumnos de 3õ y 4õ de la E.S.O, e incluso de Bachillerato y C.O.U. La primera de ellas va destinada al conocimiento geográfico y del medio ambiente; la segunda al estudio de cuestiones relacionadas con la España Antigua, mediante un ejemplo de disputa territorial en el yacimiento de Tiermes; la tercera está dedicada al expresionismo abstracto en la colección Thyssen-Bornemisza; la cuarta unidad hace referencia a las técnicas de estampación dentro de actividades relacionadas con las artes plásticas. Las dos últimas unidades presentan experiencias para ser desarrolladas en el campo de los idiomas inglés y francés respectivamente..MadridBiblioteca de Educación del Ministerio de Educación, Cultura y Deporte; Calle San Agustín, 5; 28014 Madrid; Tel. +34917748000; [email protected]

Copper-functionalized nanostructured silica-based systems: Study of the antimicrobial applications and ROS generation against gram positive and gram negative bacteria

A series of copper-functionalized SBA-15 (Santa Barbara Amorphous) materials containing the ligands triethoxysilylpropylmaleamic acid (maleamic) or triethoxy-3-(2-imidazolin-1-yl)propylsilane (imidazoline) have been prepared. The nanostructured silica-based systems SBA-maleamic, SBA-imidazoline, SBA-maleamic-Cu and SBA-imidazoline-Cu were characterized by several methods observing that the functionalization took place mainly inside the pores of the mesoporous system. The antimicrobial behaviour of the synthesized materials against Staphylococcus aureus and Escherichia coli was tested observing a very potent activity of the copper-functionalized systems (minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values for SBA-maleamic-Cu of ca. 31.25 μg/mL, which correspond with ca. 1.13 μg/mL of Cu). A study of the oxidative stress promoted by the synthesized materials showed that the SBA-maleamic-Cu and the SBA-imidazoline-Cu were able to increase the reactive oxygen species (ROS) production in S. aureus by 427% and 373%, respectively, while this increase was slightly lower in E. coli (387 and 324%, respectively). Furthermore, an electrochemical study was carried out in order to determine if these materials interact with lysine or alanine to validate a potential antimicrobial mechanism based on the inhibition of the synthesis of the peptidoglycan of the bacterial wall. Finally, these studies were also performed to determine the potential interaction of the copper-containing materials with glutathione in order to assess if they are able to perturb the metabolism of this tripeptide.Fil: Díaz-García, Diana. Universidad Rey Juan Carlos; EspañaFil: Ardiles, Perla R.. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Farmacia; ArgentinaFil: Díaz-Sánchez, Miguel. Universidad Rey Juan Carlos; EspañaFil: Mena-Palomo, Irene. Universidad Rey Juan Carlos; EspañaFil: del Hierro, Isabel. Universidad Rey Juan Carlos; EspañaFil: Prashar, Sanjiv. Universidad Rey Juan Carlos; EspañaFil: Rodríguez-Diéguez, Antonio. Universidad Rey Juan Carlos; EspañaFil: Páez, Paulina Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Gómez Ruiz, Santiago. Universidad Rey Juan Carlos; Españ

Whole-Exome Sequencing of 24 Spanish Families: Candidate Genes for Non-Syndromic Pediatric Keratoconus

Keratoconus is a corneal dystrophy that is one of the main causes of corneal transplantation and for which there is currently no effective treatment for all patients. The presentation of this disease in pediatric age is associated with rapid progression, a worse prognosis and, in 15–20% of cases, the need for corneal transplantation. It is a multifactorial disease with genetic variability, which makes its genetic study difficult. Discovering new therapeutic targets is necessary to improve the quality of life of patients. In this manuscript, we present the results of whole-exome sequencing (WES) of 24 pediatric families diagnosed at the University Hospital La Paz (HULP) in Madrid. The results show an oligogenic inheritance of the disease. Genes involved in the structure, function, cell adhesion, development and repair pathways of the cornea are proposed as candidate genes for the disease. Further studies are needed to confirm the involvement of the candidate genes described in this article in the development of pediatric keratoconus