Differential in vitro and in vivo effect of barley cysteine and serine protease inhibitors on phytopathogenic microorganisms

Protease inhibitors from plants have been involved in defence mechanisms against pests and pathogens. Phytocystatins and trypsin/α-amylase inhibitors are two of the best characterized protease inhibitor families in plants. In barley, thirteen cystatins (HvCPI-1 to 13) and the BTI-CMe trypsin inhibitor have been previously studied. Their capacity to inhibit pest digestive proteases, and the negative in vivo effect caused by plants expressing these inhibitors on pests support the defence function of these proteins. Barley cystatins are also able to inhibit in vitro fungal growth. However, the antifungal effect of these inhibitors in vivo had not been previously tested. Moreover, their in vitro and in vivo effect on plant pathogenous bacteria is still unknown. In order to obtain new insights on this feature, in vitro assays were made against different bacterial and fungal pathogens of plants using the trypsin inhibitor BTI-CMe and the thirteen barley cystatins. Most barley cystatins and the BTI-CMe inhibitor were able to inhibit mycelial growth but no bacterial growth. Transgenic Arabidopsis plants independently expressing the BTI-CMe inhibitor and the cystatin HvCPI-6 were tested against the same bacterial and fungal pathogens. Neither the HvCPI-6 expressing transgenic plants nor the BTI-CMe ones were more resistant to plant pathogen fungi and bacteria than control Arabidopsis plants. The differences observed between the in vitro and in planta assays against phytopathogenic fungi are discussed

Aparcamiento subterráneo en el Hospital Clínico San Carlos de Madrid

El presente documento tiene por objetivo la explicacion y justificacion de todos aquellos elementos que conforman el proyecto de construccion del Aparcamiento subterraneo en el Hospital Clinico San Carlos. En primer lugar se ha realizado el Estudio Previo de Soluciones, en el que se han estudiado distintas alternativas posibles para la realizacion del aparcamiento subterraneo. Tras haberlas disenado, descrito, y contrastado, se escogio la que conllevaba el mejor compendio de virtudes segun los parametros o conceptos establecidos. Una vez escogida la solucion, en la fase de estudios que nos ocupa, se procedio a redactar el proyecto de construccion propiamente dicho. Esto implica desarrollar la solucion a fondo, definiendo todos sus elementos. Para ello, se preciso de unos estudios con gran nivel de detalle que sirvieron de base para los el diseno, los calculos, y el desarrollo tecnico que ello conlleva.

Computational study of ligand binding in lipid transfer proteins: Structures, interfaces, and free energies of protein-lipid complexes

Plant nonspecific lipid transfer proteins (nsLTPs) bind a wide variety of lipids, which allows them to perform disparate functions. Recent reports on their multifunctionality in plant growth processes have posed new questions on the versatile binding abilities of these proteins. The lack of binding specificity has been customarily explained in qualitative terms on the basis of a supposed structural flexibility and nonspecificity of hydrophobic protein-ligand interactions. We present here a computational study of protein-ligand complexes formed between five nsLTPs and seven lipids bound in two different ways in every receptor protein. After optimizing geometries inmolecular dynamics calculations, we computed Poisson- Boltzmann electrostatic potentials, solvation energies, properties of the protein-ligand interfaces, and estimates of binding free energies of the resulting complexes. Our results provide the first quantitative information on the ligand abilities of nsLTPs, shed new light into protein-lipid interactions, and reveal new features which supplement commonly held assumptions on their lack of binding specificity.

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.

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.

Estudio para la implantación de Programas en Inglés en los títulos de grado de la ETSI Agrónomos

El proceso actual de convergencia entre instituciones universitarias europeas ha promovido los Programas Académicos en Inglés como un marco fructífero para su internacionalización con el fin de desarrollar la movilidad de estudiantes y la cooperación institucional dentro y fuera del EEES. El objetivo es alcanzar la Internacionalización de la Universidad a través de dos ejes de actuación: la mejora del nivel de inglés de los alumnos egresados y la captación de alumnos extranjeros. En España, la práctica totalidad de las Universidades Públicas ofrecen algún tipo de Programa en Inglés, existiendo una variada tipología de propuestas. En este contexto, se realizó un estudio para la implantación de Programas en Inglés en los títulos de grado de la ETSI Agrónomos de la Universidad Politécnica de Madrid. Para ello se analizaron las diferentes experiencias existentes en Programas en Inglés en las Universidades Españolas, haciendo especial hincapié en los estudios del ámbito agrario. Se evaluó la opinión de alumnos y profesores de la ETSI Agrónomos sobre tres cuestiones: interés en participar en un Programa en Inglés, tipo de programa en el que estaría dispuesto a participar y autoevaluación del nivel de inglés. También se examinó el interés de los alumnos de bachillerato y del mundo laboral, empresas e instituciones del ámbito agroalimentario, en los Programas en Inglés. Por último, se exploraron las implicaciones administrativas y necesidades de recursos que conlleva la implantación de un Programa Académico Universitario en Inglés. Las conclusiones del trabajo destacan la dispar oferta nacional en programas en inglés y en requerimientos lingüísticos a profesores y alumnos. Existe un gran interés tanto por parte de profesores y alumnos de la ETSI Agrónomos como de alumnos de Bachillerato por participar en este tipo de programas.

The Role of N Plant Glycosilation in act d 2 allergenicity.

Plant allergens have hitherto been included in only several protein families that share no common biochemical features. Their physical, biochemical and immunological characteristics have been widely studied, but no definite conclusion has been reached about what makes a protein an allergen. N-glycosylation is characteristic of plant allergen sources but is not present in mammals.

Enriched mannose glycosylation contributes to Act d 2 allergenicity.

Allergens are responsible for the Th2 response in patients as part of complex mixtures of proteins, fatty acids and other molecules. Plant allergens have hitherto been included in several protein families that share no common biochemical features. Their physical, biochemical and immunological characteristics have been widely studied, but no definite conclusion has been reached about what makes a protein an allergen. N-glycosylation is characteristic of plant allergen sources but is not present in mammals.

Alt a 1 from Alternaria interacts with PR5 thaumatin-like proteins

Alt a 1 is a protein found in Alternaria alternata spores related to virulence and pathogenicity and considered to be responsible for chronic asthma in children. We found that spores of Alternaria inoculated on the outer surface of kiwifruits did not develop hyphae. Nevertheless, the expression of Alt a 1 gene was upregulated, and the protein was detected in the pulp where it co-localized with kiwi PR5. Pull-down assays demonstrated experimentally that the two proteins interact in such a way that Alt a 1 inhibits the enzymatic activity of PR5. These results are relevant not only for plant defense, but also for human health as patients with chronic asthma could suffer from an allergic reaction when they eat fruit contaminated with Alternaria.