Growth response of broiler chickens to inclusion of hydrolyzed porcine mucosa (Palbio) in diets varying in total lysine content.

Growth response of broiler chickens to inclusion of hydrolyzed porcine mucosa (Palbio) in diets varying in total lysine content

Caracterización molecular del mutante riso 1508 de cebada : modificaciones transcripcionales y posttranscripcionales en el desarrollo de la semilla

La semilla es el principal órgano reproductivo de las plantas espermatofitas, permitiendo la dispersión de las poblaciones y asegurando su supervivencia gracias a su tolerancia a la desecación y a su capacidad para germinar bajo condiciones ambientales óptimas. El rendimiento y valor económico de los cereales, que constituyen la primera cosecha mundial, depende, en buena medida, de la eficacia con que se acumulan en la semilla sustancias de reserva: proteínas, carbohidratos y lípidos. El principal carbohidrato acumulado en la semilla de cebada es el almidón y la fracción mayoritaria de proteínas es la de las prolaminas (solubles en etanol al 70%); estas proteínas tienen muy bajo contenido en lisina, un aminoácido esencial en la dieta de animales monogástricos. Con el fin de mejorar el valor nutricional de la semilla de cebada, se han obtenido diferentes mutantes con un mayor contenido en este aminoácido. Riso 1508 es un mutante de cebada rico en lisina cuya mutación lys3a, de efectos pleiotrópicos, segrega como un único gen mendeliano. Entre otros, presenta una reducción drástica de la expresión de algunos genes que codifican proteínas de reserva de tipo prolamina, en concreto, presenta reducida la expresión de los genes que codifican B-, C- y ϒ-Hordeínas y del inhibidor de tripsina CMe, pero no tiene alterada la expresión del gen que codifica las D-Hordeínas. Este último gen carece en su promotor del motivo GLM (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), que es reconocido por factores transcripcionales bZIP. En este trabajo, el mutante de cebada Riso 1508 se ha utilizado como herramienta para profundizar en el conocimiento de la regulación génica en semillas durante las fases de la maduración y la germinación. Para ello, en una primera aproximación, se llevó a cabo un análisis transcriptómico comparando el genotipo mutante con el silvestre durante la maduración de la semilla. Además de confirmar variaciones en los genes que codifican proteínas de reserva, este análisis indicó que también estaban afectados los genes relacionados con metabolismo de carbohidratos. Por ello se decidió caracterizar la familia multigénica de sacarosas sintasa (SUSy) en cebada. Se anotaron dos nuevos genes, HvSs3 y HvSs4, cuya expresión se comparó con la de los genes HvSs1 y HvSs2, previamente descritos en el laboratorio. La expresión de los cuatro genes en tejidos diferentes y su respuesta a estreses abióticos se analizó mediante RT-qPCR. HvSs1 y HvSs2 se expresaron preferencialmente durante el desarrollo del endospermo, y HvSs1 también fue un tránscrito abundante durante la germinación. HvSs1 se indujo en hojas en condiciones de anoxia y HvSs3 por estrés hídrico, y ambos genes se indujeron por tratamientos de frío. La localización subcelular de las cuatro isoformas no fue sólo citoplásmica, sino que también se localizaron en zonas próximas a retículo endoplásmico y en la cara interna de la membrana plasmática; además, se observó una co-localización de HvSS1 con el marcador de mitocondrias. Estos datos sugieren un papel distinto aunque parcialmente solapante de las cuatro Sacarosa Sintasas de cebada, descritas hasta la fecha. Las cinéticas de expresión de los genes que codifican los TFs más importantes implicados en la regulación génica durante el desarrollo del endospermo de cebada, se analizaron por RT-qPCR en ambos genotipos, demostrando que los TFs de la clase DOF aparecieron desregulados durante todo el proceso en Riso 1508 comparado con el cv. Bomi, aunque también se observaron diferencias significativas en algunos de los que codifican bZIPs. Estudios previos indicaban que el ortólogo de BLZ2 en maíz, O2, se regula post-traduccionalmente mediante un mecanismo de fosforilación/defosforilación reversible, y que la forma defosforilada es la fisiológicamente activa. En este trabajo se demostró que BLZ2 está sujeto a este tipo de regulación y que la proteín-fosfatasa HvPP2C2 está implicada en el proceso. La interacción de HvPP2C2 y BLZ2 tiene lugar en el núcleo celular únicamente en presencia de 100 μM ABA. En el mutante Riso 1508, BLZ2 se encuentra en un estado hiperfosforilado tanto durante la maduración como durante la germinación de la semilla, lo que dificultaría la unión de BLZ2 a las secuencias GLM en los promotores de los genes que codifican B-, C-,y ϒ- Hordeínas y CMe. Summary The seed is the main reproductive organ of spermatophyte plants allowing the spread of populations and ensuring their survival through its desiccation tolerance and because of their ability to germinate under optimum environmental conditions. Yield and economic value of cereal crops, that constitute the first world crop, depend largely on the efficiency with which they accumulate in the seed reserve substances: proteins, carbohydrates and lipids. The main carbohydrate accumulated in the barley seed is starch and the major protein fraction is that of prolamins (soluble in 70% ethanol); these proteins have a very low lysine content, an essential amino-acid for the diet of monogastric animals. In order to improve the nutritional value of the barley seed, different mutants have been obtained with a higher content of this amino-acid. Riso 1508 is one lysine-rich mutant whose mutation (lys3a) segregates as a single Mendelian gene with pleiotropic effects, such as a drastic reduction of genes encoding the trypsin inhibitor CMe and the B-, C-and ϒ-hordeins, but has not altered the expression of the gene encoding the D-hordeins. This latter gene lacks in its promotor the GLM motif (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), that is recognised by bZIP transcription factors In this work we have used the barley mutant Riso 1508 as a tool for better understanding gene regulation in seeds during the maturation and germination phases. To this aim, a transcriptomic analysis was performed comparing wild and mutant genotypes during seed maturation. Besides confirming variations in the expression of genes encoding reserve proteins, this analysis indicated that some genes related with carbohydrate metabolism were also affected. It was therefore decided to characterize the multigene family of sucrose synthases (SUSy) in barley. Two new genes were annotated, HvSs3 and HvSs4, and its expression was compared with that of genes HvSs1 and HvSs2, previously described in our laboratory. The expression of the four genes in different tissues and in response to abiotic stresses was analyzed by RTqPCR. HvSs1 and HvSs2 were preferentially expressed during the development of the endosperm, and the HvSs1 transcript was also abundant upon germination. HvSs1 was induced in leaves by anoxic conditions, HvSs3 by water stress, and both genes were induced by cold treatments. The subcellular localization of all four isoforms was not only cytoplasmic, but they could be found along the endoplasmic reticulum and at the inner side of the cell membrane; HvSS1, was also associated with the mitochondrial marker. These data suggest a distinct but partially overlapping roles for the barley sucrose synthases, described so far. The expression kinetics of the genes encoding the most important TFs involved in gene regulation during barley endosperm development was analyzed by RT-qPCR in both genotypes. These data show that the genes encoding DOF TFs were mis-regulated throughout the process in Riso 1508, although significant differences were also found among some of those encoding bZIPs. Previous studies indicated that the BLZ2 orthologue in maize, O2, was post-translationally regulated by reversible phosphorylation/dephosphorylation and that the dephosphorylated protein is the physiologically active form. In this work we demostrate that BLZ2 is under a similar regulation and that the proteinphosphatase HvPP2C2 is implicated in the process. The interaction between HvPP2C2 and BLZ2 takes place in the cell nucleus only in the presence of 100 μM ABA. In the Riso 1508 mutant, BLZ2 is found in a hyperphosphorylated state in the maturation phase and upon seed germination; because of this, the BLZ2 binding to the GLM promoter sequences of genes encoding B-, C- y ϒ- Hordeins and CMe would be decreased in the mutant.

Automatic Inference of Determinacy and Mutual Exclusion for Logic Programs Using Mode and Type Analyses

We propose an analysis for detecting procedures and goals that are deterministic (i.e., that produce at most one solution at most once),or predicates whose clause tests are mutually exclusive (which implies that at most one of their clauses will succeed) even if they are not deterministic. The analysis takes advantage of the pruning operator in order to improve the detection of mutual exclusion and determinacy. It also supports arithmetic equations and disequations, as well as equations and disequations on terms,for which we give a complete satisfiability testing algorithm, w.r.t. available type information. Information about determinacy can be used for program debugging and optimization, resource consumption and granularity control, abstraction carrying code, etc. We have implemented the analysis and integrated it in the CiaoPP system, which also infers automatically the mode and type information that our analysis takes as input. Experiments performed on this implementation show that the analysis is fairly accurate and efficient.

Architectures for Video-on-Demand content distribution in managed networks

La demanda de contenidos de vídeo ha aumentado rápidamente en los últimos años como resultado del gran despliegue de la TV sobre IP (IPTV) y la variedad de servicios ofrecidos por los operadores de red. Uno de los servicios que se ha vuelto especialmente atractivo para los clientes es el vídeo bajo demanda (VoD) en tiempo real, ya que ofrece una transmisión (streaming) inmediata de gran variedad de contenidos de vídeo. El precio que los operadores tienen que pagar por este servicio es el aumento del tráfico en las redes, que están cada vez más congestionadas debido a la mayor demanda de contenidos de VoD y al aumento de la calidad de los propios contenidos de vídeo. Así, uno de los principales objetivos de esta tesis es encontrar soluciones que reduzcan el tráfico en el núcleo de la red, manteniendo la calidad del servicio en el nivel adecuado y reduciendo el coste del tráfico. La tesis propone un sistema jerárquico de servidores de streaming en el que se ejecuta un algoritmo para la ubicación óptima de los contenidos de acuerdo con el comportamiento de los usuarios y el estado de la red. Debido a que cualquier algoritmo óptimo de distribución de contenidos alcanza un límite en el que no se puede llegar a nuevas mejoras, la inclusión de los propios clientes del servicio (los peers) en el proceso de streaming puede reducir aún más el tráfico de red. Este proceso se logra aprovechando el control que el operador tiene en las redes de gestión privada sobre los equipos receptores (Set-Top Box) ubicados en las instalaciones de los clientes. El operador se reserva cierta capacidad de almacenamiento y streaming de los peers para almacenar los contenidos de vídeo y para transmitirlos a otros clientes con el fin de aliviar a los servidores de streaming. Debido a la incapacidad de los peers para sustituir completamente a los servidores de streaming, la tesis propone un sistema de streaming asistido por peers. Algunas de las cuestiones importantes que se abordan en la tesis son saber cómo los parámetros del sistema y las distintas distribuciones de los contenidos de vídeo en los peers afectan al rendimiento general del sistema. Para dar respuesta a estas preguntas, la tesis propone un modelo estocástico preciso y flexible que tiene en cuenta parámetros como las capacidades de enlace de subida y de almacenamiento de los peers, el número de peers, el tamaño de la biblioteca de contenidos de vídeo, el tamaño de los contenidos y el esquema de distribución de contenidos para estimar los beneficios del streaming asistido por los peers. El trabajo también propone una versión extendida del modelo matemático mediante la inclusión de la probabilidad de fallo de los peers y su tiempo de recuperación en el conjunto de parámetros del modelo. Estos modelos se utilizan como una herramienta para la realización de exhaustivos análisis del sistema de streaming de VoD asistido por los peers para la amplia gama de parámetros definidos en los modelos. Abstract The demand of video contents has rapidly increased in the past years as a result of the wide deployment of IPTV and the variety of services offered by the network operators. One of the services that has especially become attractive to the customers is real-time Video on Demand (VoD) because it offers an immediate streaming of a large variety of video contents. The price that the operators have to pay for this convenience is the increased traffic in the networks, which are becoming more congested due to the higher demand for VoD contents and the increased quality of the videos. Therefore, one of the main objectives of this thesis is finding solutions that would reduce the traffic in the core of the network, keeping the quality of service on satisfactory level and reducing the traffic cost. The thesis proposes a system of hierarchical structure of streaming servers that runs an algorithm for optimal placement of the contents according to the users’ behavior and the state of the network. Since any algorithm for optimal content distribution reaches a limit upon which no further improvements can be made, including service customers themselves (the peers) in the streaming process can further reduce the network traffic. This process is achieved by taking advantage of the control that the operator has in the privately managed networks over the Set-Top Boxes placed at the clients’ premises. The operator reserves certain storage and streaming capacity on the peers to store the video contents and to stream them to the other clients in order to alleviate the streaming servers. Because of the inability of the peers to completely substitute the streaming servers, the thesis proposes a system for peer-assisted streaming. Some of the important questions addressed in the thesis are how the system parameters and the various distributions of the video contents on the peers would impact the overall system performance. In order to give answers to these questions, the thesis proposes a precise and flexible stochastic model that takes into consideration parameters like uplink and storage capacity of the peers, number of peers, size of the video content library, size of contents and content distribution scheme to estimate the benefits of the peer-assisted streaming. The work also proposes an extended version of the mathematical model by including the failure probability of the peers and their recovery time in the set of parameters. These models are used as tools for conducting thorough analyses of the peer-assisted system for VoD streaming for the wide range of defined parameters.

A holistic experience in the integrated learning of specialized English and content in Engineering degrees

This paper aims to outline a theory-based Content and Language Integrated Learning course and to establish the rationale for adopting a holistic approach to the teaching of languages in tertiary education. Our work focuses on the interdependence between Content and Language Integrated Learning (CLIL), and the use of Information and Communication Technologies (ICT), in particular regarding the learning of English within the framework of Telecommunications Engineering. The study first analyses the diverse components of the instructional approach and the extent to which this approach interrelates with technologies within the context of what we have defined as a holistic experience, since it also aims to develop a set of generic competences or transferable skills. Second, an example of a course project framed in this holistic approach is described in order to exemplify the specific actions suggested for learner autonomy and CLIL. The approach provides both an adequate framework as well as the conditions needed to carry out a lifelong learning experience within our context, a Spanish School of Engineering. In addition to specialized language and content, the approach integrates the learning of skills and capacities required by the new plans that have been established following the Bologna Declaration in 1999.

Study of the influence of actin-binding proteins using linear analyses of cell deformability

The actin cytoskeleton plays a key role in the deformability of the cell and in mechanosensing. Here we analyze the contributions of three major actin cross-linking proteins, myosin II, a-actinin and filamin, to cell deformability, by using micropipette aspiration of Dictyostelium cells. We examine the applicability of three simple mechanical models: for small deformation, linear viscoelasticity and drop of liquid with a tense cortex; and for large deformation, a Newtonian viscous fluid. For these models, we have derived linearized equations and we provide a novel, straightforward methodology to analyze the experiments. This methodology allowed us to differentiate the effects of the cross-linking proteins in the different regimes of deformation. Our results confirm some previous observations and suggest important relations between the molecular characteristics of the actin-binding proteins and the cell behavior: the effect of myosin is explained in terms of the relation between the lifetime of the bond to actin and the resistive force; the presence of a-actinin obstructs the deformation of the cytoskeleton, presumably mainly due to the higher molecular stiffness and to the lower dissociation rate constants; and filamin contributes critically to the global connectivity of the network, possibly by rapidly turning over crosslinks during the remodeling of the cytoskeletal network, thanks to the higher rate constants, flexibility and larger size. The results suggest a sophisticated relationship between the expression levels of actinbinding proteins, deformability and mechanosensing.