La ironía en el Edipo Rey, (Prólogo y Episodio I). Hacia una redefinición y una propuesta tipológica de esta estrategia discursiva

En las últimas décadas los lingüistas dedicaron especial atención a la ironía, proponiendo diversas definiciones, mutuamente excluyentes que, quizás por prejuicios de escuela, priorizaban un solo aspecto de csta estrategia discursiva. Basándonos en la lectura de los primeros tramos del Edipo Rey, paradigmáticamente irónicos, intentaremos llegar a una definición comprensiva del recurso y propondremos una categorización de las ironías presentes en el texto, organizada sobre un doble eje enunciativo y retórico

Seawater carbonate chemistry and length, weight, survival rate, metabolic rate of coral reef fish Amphiprion melanopus in a laboratory experiment

Carbon dioxide concentrations in the surface ocean are increasing owing to rising CO2 concentrations in the atmosphere. Higher CO2 levels are predicted to affect essential physiological processes of many aquatic organisms, leading to widespread impacts on marine diversity and ecosystem function, especially when combined with the effects of global warming. Yet the ability for marine species to adjust to increasing CO2 levels over many generations is an unresolved issue. Here we show that ocean conditions projected for the end of the century (approximately 1,000 µatm CO2 and a temperature rise of 1.5-3.0 °C) cause an increase in metabolic rate and decreases in length, weight, condition and survival of juvenile fish. However, these effects are absent or reversed when parents also experience high CO2 concentrations. Our results show that non-genetic parental effects can dramatically alter the response of marine organisms to increasing CO2 and demonstrate that some species have more capacity to acclimate to ocean acidification than previously thought.

Habitat traits and food availability determine the response of marine invertebrates to ocean acidification

Energy availability and local adaptation are major components in mediating the effects of ocean acidification (OA) on marine species. In a long-term study, we investigated the effects of food availability and elevated pCO2 (ca 400, 1000 and 3000 µatm) on growth of newly settled Amphibalanus (Balanus) improvisus to reproduction, and on their offspring. We also compared two different populations, which were presumed to differ in their sensitivity to pCO2 due to differing habitat conditions: Kiel Fjord, Germany (Western Baltic Sea) with naturally strong pCO2 fluctuations, and the Tjärnö Archipelago, Sweden (Skagerrak) with far lower fluctuations. Over 20 weeks, survival, growth, reproduction and shell strength of Kiel barnacles were all unaffected by elevated pCO2, regardless of food availability. Moulting frequency and shell corrosion increased with increasing pCO2 in adults. Larval development and juvenile growth of the F1 generation were tolerant to increased pCO2, irrespective of parental treatment. In contrast, elevated pCO2 had a strong negative impact on survival of Tjärnö barnacles. Specimens from this population were able to withstand moderate levels of elevated pCO2 over 5 weeks when food was plentiful but showed reduced growth under food limitation. Severe levels of elevated pCO2 negatively impacted growth of Tjärnö barnacles in both food treatments. We demonstrate a conspicuously higher tolerance to elevated pCO2 in Kiel barnacles than in Tjärnö barnacles. This tolerance was carried-over from adults to their offspring. Our findings indicate that populations from fluctuating pCO2 environments are more tolerant to elevated pCO2 than populations from more stable pCO2 habitats. We furthermore provide evidence that energy availability can mediate the ability of barnacles to withstand moderate CO2 stress. Considering the high tolerance of Kiel specimens and the possibility to adapt over many generations, near future OA alone does not seem to present a major threat for A. improvisus

Simulation of the deformation of polycrystalline nanostructured Ti by computational homogenization

Computational homogenization by means of the finite element analysis of a representative volume element of the microstructure is used to simulate the deformation of nanostructured Ti. The behavior of each grain is taken into account using a single crystal elasto-viscoplastic model which includes the microscopic mechanisms of plastic deformation by slip along basal, prismatic and pyramidal systems. Two different representations of the polycrystal were used. Each grain was modeled with one cubic finite element in the first one while many cubic elements were used to represent each grain in the second one, leading to a model which includes the effect of grain shape and size in a limited number of grains due to the computational cost. Both representations were used to simulate the tensile deformation of nanostructured Ti processed by ECAP-C as well as the drawing process of nanostructured Ti billets. It was found that the first representation based in one finite element per grain led to a stiffer response in tension and was not able to predict the texture evolution during drawing because the strain gradient within each grain could not be captured. On the contrary, the second representation of the polycrystal microstructure with many finite elements per grain was able to predict accurately the deformation of nanostructured Ti.

NFC based remote control of services for interactive spaces

Ubiquitous computing (one person, many computers) is the third era in the history of computing. It follows the mainframe era (many people, one computer) and the PC era (one person, one computer). Ubiquitous computing empowers people to communicate with services by interacting with their surroundings. Most of these so called smart environments contain sensors sensing users’ actions and try to predict the users’ intentions and necessities based on sensor data. The main drawback of this approach is that the system might perform unexpected or unwanted actions, making the user feel out of control. In this master thesis we propose a different procedure based on Interactive Spaces: instead of predicting users’ intentions based on sensor data, the system reacts to users’ explicit predefined actions. To that end, we present REACHeS, a server platform which enables communication among services, resources and users located in the same environment. With REACHeS, a user controls services and resources by interacting with everyday life objects and using a mobile phone as a mediator between himself/herself, the system and the environment. REACHeS’ interfaces with a user are built upon NFC (Near Field Communication) technology. NFC tags are attached to objects in the environment. A tag stores commands that are sent to services when a user touches the tag with his/her NFC enabled device. The prototypes and usability tests presented in this thesis show the great potential of NFC to build such user interfaces.

Contribuciones al análisis de problemas supercomplejos de toma de decisiones

Los fundamentos de la Teoría de la Decisión Bayesiana proporcionan un marco coherente en el que se pueden resolver los problemas de toma de decisiones. La creciente disponibilidad de ordenadores potentes está llevando a tratar problemas cada vez más complejos con numerosas fuentes de incertidumbre multidimensionales; varios objetivos conflictivos; preferencias, metas y creencias cambiantes en el tiempo y distintos grupos afectados por las decisiones. Estos factores, a su vez, exigen mejores herramientas de representación de problemas; imponen fuertes restricciones cognitivas sobre los decisores y conllevan difíciles problemas computacionales. Esta tesis tratará estos tres aspectos. En el Capítulo 1, proporcionamos una revisión crítica de los principales métodos gráficos de representación y resolución de problemas, concluyendo con algunas recomendaciones fundamentales y generalizaciones. Nuestro segundo comentario nos lleva a estudiar tales métodos cuando sólo disponemos de información parcial sobre las preferencias y creencias del decisor. En el Capítulo 2, estudiamos este problema cuando empleamos diagramas de influencia (DI). Damos un algoritmo para calcular las soluciones no dominadas en un DI y analizamos varios conceptos de solución ad hoc. El último aspecto se estudia en los Capítulos 3 y 4. Motivado por una aplicación de gestión de embalses, introducimos un método heurístico para resolver problemas de decisión secuenciales. Como muestra resultados muy buenos, extendemos la idea a problemas secuenciales generales y cuantificamos su bondad. Exploramos después en varias direcciones la aplicación de métodos de simulación al Análisis de Decisiones. Introducimos primero métodos de Monte Cario para aproximar el conjunto no dominado en problemas continuos. Después, proporcionamos un método de Monte Cario basado en cadenas de Markov para problemas con información completa con estructura general: las decisiones y las variables aleatorias pueden ser continuas, y la función de utilidad puede ser arbitraria. Nuestro esquema es aplicable a muchos problemas modelizados como DI. Finalizamos con un capítulo de conclusiones y problemas abiertos.---ABSTRACT---The foundations of Bayesian Decisión Theory provide a coherent framework in which decisión making problems may be solved. With the advent of powerful computers and given the many challenging problems we face, we are gradually attempting to solve more and more complex decisión making problems with high and multidimensional uncertainty, múltiple objectives, influence of time over decisión tasks and influence over many groups. These complexity factors demand better representation tools for decisión making problems; place strong cognitive demands on the decison maker judgements; and lead to involved computational problems. This thesis will deal with these three topics. In recent years, many representation tools have been developed for decisión making problems. In Chapter 1, we provide a critical review of most of them and conclude with recommendations and generalisations. Given our second query, we could wonder how may we deal with those representation tools when there is only partial information. In Chapter 2, we find out how to deal with such a problem when it is structured as an influence diagram (ID). We give an algorithm to compute nondominated solutions in ID's and analyse several ad hoc solution concepts.- The last issue is studied in Chapters 3 and 4. In a reservoir management case study, we have introduced a heuristic method for solving sequential decisión making problems. Since it shows very good performance, we extend the idea to general problems and quantify its goodness. We explore then in several directions the application of simulation based methods to Decisión Analysis. We first introduce Monte Cario methods to approximate the nondominated set in continuous problems. Then, we provide a Monte Cario Markov Chain method for problems under total information with general structure: decisions and random variables may be continuous, and the utility function may be arbitrary. Our scheme is applicable to many problems modeled as IDs. We conclude with discussions and several open problems.

The impact of large-scale circulation patterns on summer crop in Iberian Peninsula

Large-scale circulations patterns (ENSO, NAO) have been shown to have a significant impact on seasonal weather, and therefore on crop yield over many parts of the world(Garnett and Khandekar, 1992; Aasa et al., 2004; Rozas and Garcia-Gonzalez, 2012). In this study, we analyze the influence of large-scale circulation patterns and regional climate on the principal components of maize yield variability in Iberian Peninsula (IP) using reanalysis datasets. Additionally, we investigate the modulation of these relationships by multidecadal patterns. This study is performed analyzing long time series of maize yield, only climate dependent, computed with the crop model CERES-maize (Jones and Kiniry, 1986) included in Decision Support System for Agrotechnology Transfer (DSSAT v.4.5).

The universal ancestor

A genetic annealing model for the universal ancestor of all extant life is presented; the name of the model derives from its resemblance to physical annealing. The scenario pictured starts when “genetic temperatures” were very high, cellular entities (progenotes) were very simple, and information processing systems were inaccurate. Initially, both mutation rate and lateral gene transfer levels were elevated. The latter was pandemic and pervasive to the extent that it, not vertical inheritance, defined the evolutionary dynamic. As increasingly complex and precise biological structures and processes evolved, both the mutation rate and the scope and level of lateral gene transfer, i.e., evolutionary temperature, dropped, and the evolutionary dynamic gradually became that characteristic of modern cells. The various subsystems of the cell “crystallized,” i.e., became refractory to lateral gene transfer, at different stages of “cooling,” with the translation apparatus probably crystallizing first. Organismal lineages, and so organisms as we know them, did not exist at these early stages. The universal phylogenetic tree, therefore, is not an organismal tree at its base but gradually becomes one as its peripheral branchings emerge. The universal ancestor is not a discrete entity. It is, rather, a diverse community of cells that survives and evolves as a biological unit. This communal ancestor has a physical history but not a genealogical one. Over time, this ancestor refined into a smaller number of increasingly complex cell types with the ancestors of the three primary groupings of organisms arising as a result.

Growth and viability of macrophages continuously stimulated to produce nitric oxide

Deregulated production of nitric oxide (NO) has been implicated in the development of certain human diseases, including cancer. We sought to assess the damaging potential of NO produced under long-term conditions through the development of a suitable model cell culture system. In this study, we report that when murine macrophage-like RAW264.7 cells were exposed continuously to bacterial lipopolysaccharide (LPS) or mouse recombinant interferon-γ (IFN-γ) over periods of 21–23 days, they continued to grow, but with doubling times 2 to 4 times, respectively, longer than the doubling time of unstimulated cells. Stimulated cells produced NO at rates of 30 to 70 nmol per million cells per day throughout the stimulation period. Within 24 hr after removal of stimulant, cells resumed exponential growth. Simultaneous exposure to LPS and IFN-γ resulted in decreased cell number, which persisted for 2 days after removal of the stimulants. Exponential growth was attained only after an additional 4 days. Addition of N-methyl-l-arginine (NMA), an NO synthase inhibitor, to the medium inhibited NO production by 90% of all stimulated cells, partially reduced doubling time of cells stimulated with LPS or IFN-γ, and partially increased viability and growth rates in those exposed to both LPS and IFN-γ. However, when incubated with LPS and IFN-γ at low densities both in the presence and in the absence of NMA, cells grew at a rate slower than that of unstimulated cells, with no cell death, and they resumed exponential growth 24 hr after removal of stimulants. Results from cell density experiments suggest that macrophages are protected from intracellularly generated NO; much of the NO damaging activity occurred outside of the producer cells. Collectively, results presented in this study suggest that the type of cellular toxicity observed in macrophages is markedly influenced by rate of exposure to NO: at low rates of exposure, cells exhibit slower growth; at higher rates, cells begin to die; at even higher rates, cells undergo growth arrest or die. The ability of RAW264.7 cells to produce NO over many cell generations makes the cell line a useful system for the study of other aspects of cellular damage, including genotoxicity, resulting from exposure to NO under long-term conditions.

A human myeloma cell line suitable for the generation of human monoclonal antibodies

Ever since monoclonal antibodies were produced in 1975 with mouse myeloma cells there has been interest in developing human myeloma cultures for the production of monoclonal antibodies. However, despite multiple attempts, no human myeloma line suitable for hybridoma production has been described. Here we report the derivation of a hypoxanthine–aminopterin–thymidine-sensitive and ouabain-resistant human myeloma cell line (Karpas 707H) that contains unique genetic markers. We show that this line is useful for the generation of stable human hybridomas. It can easily be fused with ouabain-sensitive Epstein–Barr virus-transformed cells as well as with fresh tonsil and blood lymphocytes, giving rise to stable hybrids that continuously secrete very large quantities of human immunoglobulins. The derived hybrids do not lose immunoglobulin secretion over many months of continuous growth. The availability of this cell line should enable the in vitro immortalization of human antibody-producing B cells that are formed in vivo. The monoclonal antibodies produced may have advantages in immunotherapy.

Mutagenesis of the peptidyltransferase center of 23S rRNA: the invariant U2449 is dispensable

U2449 is one of many invariant residues in the central loop of domain V of 23S rRNA, a region that constitutes part of the peptidyltransferase center of the ribosome. In Escherichia coli, this U is post-transcriptionally modified to dihydrouridine (D) and is the only D modification found in E.coli rRNAs. To analyze the role of this base and its modification in ribosomal function, all three base substitutions were constructed on a plasmid copy of the rrnB operon and assayed for their ability to support cell growth in a strain of E.coli lacking chromosomal rrn operons. Both purine substitution mutations were not viable. However, growth and antibiotic sensitivity of cells expressing only the mutant D2449C rRNA was indistinguishable from wild type. We conclude that while a pyrimidine is required at position 2449 for proper ribosomal function, the D modification is dispensable.

Implications of immune-to-brain communication for sickness and pain

This review presents a view of hyperalgesia and allodynia not typical of the field as a whole. That is, exaggerated pain is presented as one of many natural consequences of peripheral infection and injury. The constellation of changes that results from such immune challenges is called the sickness response. This sickness response results from immune-to-brain communication initiated by proinflammatory cytokines released by activated immune cells. In response to signals it receives from the immune system, the brain orchestrates the broad array of physiological, behavioral, and hormonal changes that comprise the sickness response. The neurocircuitry and neurochemistry of sickness-induced hyperalgesia are described. One focus of this discussion is on the evidence that spinal cord microglia and astrocytes are key mediators of sickness-induced hyperalgesia. Last, evidence is presented that hyperalgesia and allodynia also result from direct immune activation, rather than neural activation, of these same spinal cord glia. Such glial activation is induced by viruses such as HIV-1 that are known to invade the central nervous system. Implications of exaggerated pain states created by peripheral and central immune activation are discussed.

α-Catenin-independent Recruitment of ZO-1 to Nectin-based Cell-Cell Adhesion Sites through Afadin

ZO-1 is an actin filament (F-actin)–binding protein that localizes to tight junctions and connects claudin to the actin cytoskeleton in epithelial cells. In nonepithelial cells that have no tight junctions, ZO-1 localizes to adherens junctions (AJs) and may connect cadherin to the actin cytoskeleton indirectly through β- and α-catenins as one of many F-actin–binding proteins. Nectin is an immunoglobulin-like adhesion molecule that localizes to AJs and is associated with the actin cytoskeleton through afadin, an F-actin–binding protein. Ponsin is an afadin- and vinculin-binding protein that also localizes to AJs. The nectin-afadin complex has a potency to recruit the E-cadherin–β-catenin complex through α-catenin in a manner independent of ponsin. By the use of cadherin-deficient L cell lines stably expressing various components of the cadherin-catenin and nectin-afadin systems, and α-catenin–deficient F9 cell lines, we examined here whether nectin recruits ZO-1 to nectin-based cell-cell adhesion sites. Nectin showed a potency to recruit not only α-catenin but also ZO-1 to nectin-based cell-cell adhesion sites. This recruitment of ZO-1 was dependent on afadin but independent of α-catenin and ponsin. These results indicate that ZO-1 localizes to cadherin-based AJs through interactions not only with α-catenin but also with the nectin-afadin system.

Nuclear protein import: Ran-GTP dissociates the karyopherin alphabeta heterodimer by displacing alpha from an overlapping binding site on beta.

The alpha subunit of the karyopherin heterodimer functions in recognition of the protein import substrate and the beta subunit serves to dock the trimeric complex to one of many sites on nuclear pore complex fibers. The small GTPase Ran and the Ran interactive protein, p10, function in the release of the docked complex. Repeated cycles of docking and release are thought to concentrate the transport substrate for subsequent diffusion into the nucleus. Ran-GTP dissociates the karyopherin heterodimer and forms a stoichiometric complex with Ran-GTP. Here we report the mapping of karyopherin beta's binding sites both for Ran-GTP and for karyopherin alpha. We discovered that karyopherin beta's binding site for Ran-GTP shows a striking sequence similarity to the cytoplasmic Ran-GTP binding protein, RanBP1. Moreover, we found that Ran-GTP and karyopherin alpha bind to overlapping sites on karyopherin beta. Having a higher affinity to the overlapping site, Ran-GTP displaces karyopherin alpha and binds to karyopherin beta. Competition for overlapping binding sites may be the mechanism by which GTP bound forms of other small GTPases function in corresponding dissociation-association reactions. We also mapped Ran's binding site for karyopherin beta to a cluster of basic residues analogous to those previously shown to constitute karyopherin alpha's binding site to karyopherin beta.

Reconstitution of p53-ubiquitinylation reactions from purified components: the role of human ubiquitin-conjugating enzyme UBC4 and E6-associated protein (E6AP).

The E6 protein of the high-risk human papillomaviruses inactivates the tumor suppressor protein p53 by stimulating its ubiquitinylation and subsequent degradation. Ubiquitinylation is a multistep process involving a ubiquitin-activating enzyme, one of many distinct ubiquitin-conjugating enzymes, and in certain cases, a ubiquitin ligase. In human papillomavirus-infected cells, E6 and the E6-associated protein are thought to act as a ubiquitin-protein ligase in the ubiquitinylation of p53. Here we describe the cloning of a human ubiquitin-conjugating enzyme that specifically ubiquitinylates E6-associated protein. Furthermore, we define the biochemical pathway of p53 ubiquitinylation and demonstrate that in vivo inhibition of various components in the pathway leads to an inhibition of E6-stimulated p53 degradation.