Proteómica de la madurez del tomate : identificación de dos estados de madurez del fruto por perfiles proteicos totales del pericarpio en RILs de tomate

Durante la madurez del fruto se producen cambios morfológicos, fisiológicos y bioquímicos provocados por la expresión regulada de diferentes genes. El objetivo de este trabajo fue verificar si la presencia de polipéptidos totales del pericarpio en los estados verde maduro (VM) y rojo maduro (RM) permite caracterizar la madurez del tomate. Se analizaron 18 líneas endocriadas recombinantes obtenidas por selección antagónica-divergente de un cruzamiento entre la cv. Caimanta (Solanum lycopersicum) y la entrada LA722 (S. pimpinellifolium), que fueron incluidas junto a la F1 como testigos experimentales. Los extractos proteicos se obtuvieron de dos muestras independientes de cada estado según el protocolo estándar y se resolvieron en SDS-PAGE. Se analizó la presencia/ausencia de bandas por genotipos y por estado, detectándose 26 en VM y 29 en RM. Algunas bandas fueron comunes entre estados, mientras que otras resultaron propias de VM o RM, respectivamente. Se calcularon las distancias de Jaccard y se realizó un análisis de conglomerados según el método UPGMA. En el dendrograma (correlación cofenética = 0,43) se distinguieron dos grandes grupos definidos por el estado de madurez. Se concluye que los perfiles proteicos del pericarpio son una herramienta postgenómica apropiada para identificar dos estados de madurez del fruto de tomate.

Detection histories for eight species of Amazona parrots in Venezuela during the NeoMaps bird surveys in 2010

Documenting changes in distribution is necessary for understanding species' response to environmental changes, but data on species distributions are heterogeneous in accuracy and resolution. Combining different data sources and methodological approaches can fill gaps in knowledge about the dynamic processes driving changes in species-rich, but data-poor regions. We combined recent bird survey data from the Neotropical Biodiversity Mapping Initiative (NeoMaps) with historical distribution records to estimate potential changes in the distribution of eight species of Amazon parrots in Venezuela. Using environmental covariates and presence-only data from museum collections and the literature, we first used maximum likelihood to fit a species distribution model (SDM) estimating a historical maximum probability of occurrence for each species. We then used recent, NeoMaps survey data to build single-season occupancy models (OM) with the same environmental covariates, as well as with time- and effort-dependent detectability, resulting in estimates of the current probability of occurrence. We finally calculated the disagreement between predictions as a matrix of probability of change in the state of occurrence. Our results suggested negative changes for the only restricted, threatened species, Amazona barbadensis, which has been independently confirmed with field studies. Two of the three remaining widespread species that were detected, Amazona amazonica, Amazona ochrocephala, also had a high probability of negative changes in northern Venezuela, but results were not conclusive for Amazona farinosa. The four remaining species were undetected in recent field surveys; three of these were most probably absent from the survey locations (Amazona autumnalis, Amazona mercenaria and Amazona festiva), while a fourth (Amazona dufresniana) requires more intensive targeted sampling to estimate its current status. Our approach is unique in taking full advantage of available, but limited data, and in detecting a high probability of change even for rare and patchily-distributed species. However, it is presently limited to species meeting the strong assumptions required for maximum-likelihood estimation with presence-only data, including very high detectability and representative sampling of its historical distribution.

Three species distribution models for the marine diatom Fragilariopsis kerguelensis of the Southern Ocean, links to NetCDF files

Fragilariopsis kerguelensis, a dominant diatom species throughout the Antarctic Circumpolar Current, is coined to be one of the main drivers of the biological silicate pump. Here, we study the distribution of this important species and expected consequences of climate change upon it, using correlative species distribution modeling and publicly available presence-only data. As experience with SDM is scarce for marine phytoplankton, this also serves as a pilot study for this organism group. We used the maximum entropy method to calculate distribution models for the diatom F. kerguelensis based on yearly and monthly environmental data (sea surface temperature, salinity, nitrate and silicate concentrations). Observation data were harvested from GBIF and the Global Diatom Database, and for further analyses also from the Hustedt Diatom Collection (BRM). The models were projected on current yearly and seasonal environmental data to study current distribution and its seasonality. Furthermore, we projected the seasonal model on future environmental data obtained from climate models for the year 2100. Projected on current yearly averaged environmental data, all models showed similar distribution patterns for F. kerguelensis. The monthly model showed seasonality, for example, a shift of the southern distribution boundary toward the north in the winter. Projections on future scenarios resulted in a moderately to negligibly shrinking distribution area and a change in seasonality. We found a substantial bias in the publicly available observation datasets, which could be reduced by additional observation records we obtained from the Hustedt Diatom Collection. Present-day distribution patterns inferred from the models coincided well with background knowledge and previous reports about F. kerguelensis distribution, showing that maximum entropy-based distribution models are suitable to map distribution patterns for oceanic planktonic organisms. Our scenario projections indicate moderate effects of climate change upon the biogeography of F. kerguelensis.

Organic matter and trace element concentrations in sapropels from ODP Leg 160 sites

A distinct Pliocene eastern Mediterranean sapropel (i-282), recovered from three Ocean Drilling Program (ODP) Leg 160 Sites, has been investigated for its organic and inorganic composition. This sapropel is characterized by high organic carbon (Corg) and trace element contents, and the presence of isorenieratene derivatives. The latter suggests that the base of the photic zone was sulphidic during formation of the sapropel. Combined with evidence of bottom water anoxia (preservation of laminae, high redox-sensitive trace element contents, and the abundance and isotopic composition of pyrite) this leads to the tentative conclusion that almost the entire water column may have been anoxic. This anoxia resulted from high productivity and not from stagnation, because an approximation of the trace element budget during sapropel formation shows that water exchange with the western Mediterranean is needed. Entire water column anoxia has been suggested earlier for several black shales. With regard to the depositional environment and the Corg content, however, only the Cenomanian=Turonian Boundary Event (CTBE) black shales appear to be comparable to this sapropel. The proposed trace element removal mechanism of scavenging and (co-)precipitation in an anoxic water column, is thought to be similar for both types of deposits. The ultimate trace element source for the sapropel, however, is seawater, whereas it is hydrothermal and fluvial input for CTBE black shales (because they have a larger temporal and spatial distribution). Nonetheless, the Corg-rich eastern Mediterranean Pliocene sapropel discussed here may be considered to be a younger analogue of CTBE black shales.

Intelligent context-aware services based on internet of things architectures

La Internet de las Cosas (IoT), como parte de la Futura Internet, se ha convertido en la actualidad en uno de los principales temas de investigación; en parte gracias a la atención que la sociedad está poniendo en el desarrollo de determinado tipo de servicios (telemetría, generación inteligente de energía, telesanidad, etc.) y por las recientes previsiones económicas que sitúan a algunos actores, como los operadores de telecomunicaciones (que se encuentran desesperadamente buscando nuevas oportunidades), al frente empujando algunas tecnologías interrelacionadas como las comunicaciones Máquina a Máquina (M2M). En este contexto, un importante número de actividades de investigación a nivel mundial se están realizando en distintas facetas: comunicaciones de redes de sensores, procesado de información, almacenamiento de grandes cantidades de datos (big--‐data), semántica, arquitecturas de servicio, etc. Todas ellas, de forma independiente, están llegando a un nivel de madurez que permiten vislumbrar la realización de la Internet de las Cosas más que como un sueño, como una realidad tangible. Sin embargo, los servicios anteriormente mencionados no pueden esperar a desarrollarse hasta que las actividades de investigación obtengan soluciones holísticas completas. Es importante proporcionar resultados intermedios que eviten soluciones verticales realizadas para desarrollos particulares. En este trabajo, nos hemos focalizado en la creación de una plataforma de servicios que pretende facilitar, por una parte la integración de redes de sensores y actuadores heterogéneas y geográficamente distribuidas, y por otra lado el desarrollo de servicios horizontales utilizando dichas redes y la información que proporcionan. Este habilitador se utilizará para el desarrollo de servicios y para la experimentación en la Internet de las Cosas. Previo a la definición de la plataforma, se ha realizado un importante estudio focalizando no sólo trabajos y proyectos de investigación, sino también actividades de estandarización. Los resultados se pueden resumir en las siguientes aseveraciones: a) Los modelos de datos definidos por el grupo “Sensor Web Enablement” (SWE™) del “Open Geospatial Consortium (OGC®)” representan hoy en día la solución más completa para describir las redes de sensores y actuadores así como las observaciones. b) Las interfaces OGC, a pesar de las limitaciones que requieren cambios y extensiones, podrían ser utilizadas como las bases para acceder a sensores y datos. c) Las redes de nueva generación (NGN) ofrecen un buen sustrato que facilita la integración de redes de sensores y el desarrollo de servicios. En consecuencia, una nueva plataforma de Servicios, llamada Ubiquitous Sensor Networks (USN), se ha definido en esta Tesis tratando de contribuir a rellenar los huecos previamente mencionados. Los puntos más destacados de la plataforma USN son: a) Desde un punto de vista arquitectónico, sigue una aproximación de dos niveles (Habilitador y Gateway) similar a otros habilitadores que utilizan las NGN (como el OMA Presence). b) Los modelos de datos están basado en los estándares del OGC SWE. iv c) Está integrado en las NGN pero puede ser utilizado sin ellas utilizando infraestructuras IP abiertas. d) Las principales funciones son: Descubrimiento de sensores, Almacenamiento de observaciones, Publicacion--‐subscripcion--‐notificación, ejecución remota homogénea, seguridad, gestión de diccionarios de datos, facilidades de monitorización, utilidades de conversión de protocolos, interacciones síncronas y asíncronas, soporte para el “streaming” y arbitrado básico de recursos. Para demostrar las funcionalidades que la Plataforma USN propuesta pueden ofrecer a los futuros escenarios de la Internet de las Cosas, se presentan resultados experimentales de tres pruebas de concepto (telemetría, “Smart Places” y monitorización medioambiental) reales a pequeña escala y un estudio sobre semántica (sistema de información vehicular). Además, se está utilizando actualmente como Habilitador para desarrollar tanto experimentación como servicios reales en el proyecto Europeo SmartSantander (que aspira a integrar alrededor de 20.000 dispositivos IoT). v Abstract Internet of Things, as part of the Future Internet, has become one of the main research topics nowadays; in part thanks to the pressure the society is putting on the development of a particular kind of services (Smart metering, Smart Grids, eHealth, etc.), and by the recent business forecasts that situate some players, like Telecom Operators (which are desperately seeking for new opportunities), at the forefront pushing for some interrelated technologies like Machine--‐to--‐Machine (M2M) communications. Under this context, an important number of research activities are currently taking place worldwide at different levels: sensor network communications, information processing, big--‐ data storage, semantics, service level architectures, etc. All of them, isolated, are arriving to a level of maturity that envision the achievement of Internet of Things (IoT) more than a dream, a tangible goal. However, the aforementioned services cannot wait to be developed until the holistic research actions bring complete solutions. It is important to come out with intermediate results that avoid vertical solutions tailored for particular deployments. In the present work, we focus on the creation of a Service--‐level platform intended to facilitate, from one side the integration of heterogeneous and geographically disperse Sensors and Actuator Networks (SANs), and from the other the development of horizontal services using them and the information they provide. This enabler will be used for horizontal service development and for IoT experimentation. Prior to the definition of the platform, we have realized an important study targeting not just research works and projects, but also standardization topics. The results can be summarized in the following assertions: a) Open Geospatial Consortium (OGC®) Sensor Web Enablement (SWE™) data models today represent the most complete solution to describe SANs and observations. b) OGC interfaces, despite the limitations that require changes and extensions, could be used as the bases for accessing sensors and data. c) Next Generation Networks (NGN) offer a good substrate that facilitates the integration of SANs and the development of services. Consequently a new Service Layer platform, called Ubiquitous Sensor Networks (USN), has been defined in this Thesis trying to contribute to fill in the previous gaps. The main highlights of the proposed USN Platform are: a) From an architectural point of view, it follows a two--‐layer approach (Enabler and Gateway) similar to other enablers that run on top of NGN (like the OMA Presence). b) Data models and interfaces are based on the OGC SWE standards. c) It is integrated in NGN but it can be used without it over open IP infrastructures. d) Main functions are: Sensor Discovery, Observation Storage, Publish--‐Subscribe--‐Notify, homogeneous remote execution, security, data dictionaries handling, monitoring facilities, authorization support, protocol conversion utilities, synchronous and asynchronous interactions, streaming support and basic resource arbitration. vi In order to demonstrate the functionalities that the proposed USN Platform can offer to future IoT scenarios, some experimental results have been addressed in three real--‐life small--‐scale proofs--‐of concepts (Smart Metering, Smart Places and Environmental monitoring) and a study for semantics (in--‐vehicle information system). Furthermore we also present the current use of the proposed USN Platform as an Enabler to develop experimentation and real services in the SmartSantander EU project (that aims at integrating around 20.000 IoT devices).

Allelic variation of a dehydrin gene cosegregates with chilling tolerance during seedling emergence

Dehydrins (DHNs, LEA D-11) are plant proteins present during environmental stresses associated with dehydration or low temperatures and during seed maturation. Functions of DHNs have not yet been defined. Earlier, we hypothesized that a ≈35-kDa DHN and membrane properties that reduce electrolyte leakage from seeds confer chilling tolerance during seedling emergence of cowpea (Vigna unguiculata L. Walp.) in an additive and independent manner. Evidence for this hypothesis was not rigorous because it was based on correlations of presence/absence of the DHN and slow electrolyte leakage with chilling tolerance in closely related cowpea lines that have some other genetic differences. Here, we provide more compelling genetic evidence for involvement of the DHN in chilling tolerance of cowpea. We developed near-isogenic lines by backcrossing. We isolated and determined the sequence of a cDNA corresponding to the ≈35-kDa DHN and used gene-specific oligonucleotides derived from it to test the genetic linkage between the DHN presence/absence trait and the DHN structural gene. We tested for association between the DHN presence/absence trait and both low-temperature seed emergence and electrolyte leakage. We show that allelic differences in the Dhn structural gene map to the same position as the DHN protein presence/absence trait and that the presence of the ≈35-kDa DHN is indeed associated with chilling tolerance during seedling emergence, independent of electrolyte leakage effects. Two types of allelic variation in the Dhn gene were identified in the protein-coding region, deletion of one Φ-segment from the DHN-negative lines and two single amino acid substitutions.

A natural classification of the basic helix–loop–helix class of transcription factors

A natural (evolutionary) classification is provided for 242 basic helix–loop–helix (bHLH) motif-containing proteins. Phylogenetic analyses of amino acid sequences describe the patterns of evolutionary change within the motif and delimit evolutionary lineages. These evolutionary lineages represent well known functional groups of proteins and can be further arranged into five groups based on binding to DNA at the hexanucleotide E-box, the amino acid patterns in other components of the motif, and the presence/absence of a leucine zipper. The hypothesized ancestral amino acid sequence for the bHLH transcription factor family is given together with the ancestral sequences of the subgroups. It is suggested that bHLH proteins containing a leucine zipper are not a natural, monophyletic group.

Consumo de oxigénio em bivalves

Os organismos vivos obtêm energia degradando moléculas orgânicas através da respiração celular. Esse processo pode ocorrer com ou sem consumo de oxigénio. Em muitos seres vivos com células complexas, a respiração celular ocorre com consumo de oxigénio pelo que se designa de respiração aeróbia. O consumo de oxigénio pode ser observado utilizando uma substância indicadora de presença/ausência de O2. O indicador utilizado neste experiencia foi o azul-de-metileno que muda de cor conforme está na forma reduzida ou na forma oxidada. Os resultados mostram que ocorreu a redução do azul-de-metileno, o que significa que o oxigénio foi consumido. O objetivo desta atividade é observar a mobilização de energia em bivalves (Scrobicularria plana) vulgarmente chamada de Lambujinha através do consumo de O2.

Modelos de regressão para identificação de marcadores preditivos de asma na descendência de mulheres com atopia

Trabalho de projecto de mestrado, Bioestatística, Universidade de Lisboa, Faculdade de Ciências, 2016

Alcoholic neurobiology: Changes in dependence and recovery

This article presents the proceedings of a symposium held at the meeting of the International Society for Biomedical Research on Alcoholism (ISBRA) in Mannheim, Germany, in October, 2004. Chronic alcoholism follows a fluctuating course, which provides a naturalistic experiment in vulnerability, resilience, and recovery of human neural systems in response to presence, absence, and history of the neurotoxic effects of alcoholism. Alcohol dependence is a progressive chronic disease that is associated with changes in neuroanatomy, neurophysiology, neural gene expression, psychology, and behavior. Specifically, alcohol dependence is characterized by a neuropsychological profile of mild to moderate impairment in executive functions, visuospatial abilities, and postural stability, together with relative sparing of declarative memory, language skills, and primary motor and perceptual abilities. Recovery from alcoholism is associated with a partial reversal of CNS deficits that occur in alcoholism. The reversal of deficits during recovery from alcoholism indicates that brain structure is capable of repair and restructuring in response to insult in adulthood. Indirect support of this repair model derives from studies of selective neuropsychological processes, structural and functional neuroimaging studies, and preclinical studies on degeneration and regeneration during the development of alcohol dependence and recovery from dependence. Genetics and brain regional specificity contribute to unique changes in neuropsychology and neuroanatomy in alcoholism and recovery. This symposium includes state-of-the-art presentations on changes that occur during active alcoholism as well as those that may occur during recovery-abstinence from alcohol dependence. Included are human neuroimaging and neuropsychological assessments, changes in human brain gene expression, allelic combinations of genes associated with alcohol dependence and preclinical studies investigating mechanisms of alcohol induced neurotoxicity, and neuroprogenetor cell expansion during recovery from alcohol dependence.

Assessing the taxonomic status of dingoes Canis familiaris dingo for conservation

1. The conservation status of the dingo Canis familiaris dingo is threatened by hybridization with the domestic dog C. familiaris familiaris. A practical method that can estimate the different levels of hybridization in the field is urgently required so that animals below a specific threshold of dingo ancestry (e.g. 1/4 or 1/2 dingoes) can reliably be identified and removed from dingo populations. 2. Skull morphology has been traditionally used to assess dingo purity, but this method does not discriminate between the different levels of dingo ancestry in hybrids. Furthermore, measurements can only be reliably taken from the skulls of dead animals. 3. Methods based on the analysis of variation in DNA are able to discriminate between the different levels of hybridization, but the validity of this method has been questioned because the materials currently used as a reference for dingoes are from captive animals of unproven genetic purity. The use of pre-European materials would improve the accuracy of this method, but suitable material has not been found in sufficient quantity to develop a reliable reference population. Furthermore, current methods based on DNA are impractical for the field-based discrimination of hybrids because samples require laboratory analysis. 4. Coat colour has also been used to estimate the extent of hybridization and is possibly the most practical method to apply in the field. However, this method may not be as powerful as genetic or morphological analyses because some hybrids (e.g. Australian cattle dog x dingo) are similar to dingoes in coat colour and body form. This problem may be alleviated by using additional visual characteristics such as the presence/absence of ticking and white markings.

Optimizing Presence–Absence Surveys For Detecting Population Trends

Presence-absence surveys are a commonly used method for monitoring broad-scale changes in wildlife distributions. However, the lack of power of these surveys for detecting population trends is problematic for their application in wildlife management. Options for improving power include increasing the sampling effort or arbitrarily relaxing the type I error rate. We present an alternative, whereby targeted sampling of particular habitats in the landscape using information from a habitat model increases power. The advantage of this approach is that it does not require a trade-off with either cost or the Pr(type I error) to achieve greater power. We use a demographic model of koala (Phascolarctos cinereus) population dynamics and simulations of the monitoring process to estimate the power to detect a trend in occupancy for a range of strategies, thereby demonstrating that targeting particular habitat qualities can improve power substantially. If the objective is to detect a decline in occupancy, the optimal strategy is to sample high-quality habitats. Alternatively, if the objective is to detect an increase in occupancy, the optimal strategy is to sample intermediate-quality habitats. The strategies with the highest power remained the same under a range of parameter assumptions, although observation error had a strong influence on the optimal strategy. Our approach specifically applies to monitoring for detecting long-term trends in occupancy or abundance. This is a common and important monitoring objective for wildlife managers, and we provide guidelines for more effectively achieving it.

'Taking a sickie':Job satisfaction and job involvement as interactive predictors of absenteeism in a public organization

Correlations between absenteeism and work attitudes such as job satisfaction have often been found to be disappointingly weak. As prior work reveals, this might be due to ignoring interactive effects of attitudes with different attitude targets (e.g. job involvement and organizational commitment). Drawing on basic principles in personality research and insights about the situational variability of job satisfaction judgments, we proposed that similar interactions should be present also for attitudes with the same target. More specifically, it was predicted that job involvement affects absenteeism more if job satisfaction is low as this indicates a situation with weak constraints. Both attitudes were assessed in a sample of 436 employees working in a large civil service organization, and two indexes of absence data (frequency and time lost) were drawn from personnel records covering a 12-month period following the survey. Whereas simple correlations were not significant, a moderated regression documented that the hypothesized interaction was significant for both indicators of absence behaviour. As a range of controls (e.g. age, gender, job level) were accounted for, these findings lend strong support to the importance of this new, specific form of attitude interaction. Thus, we encourage researchers not only to consider interactions of attitudes with a different focus (e.g. job vs. organization) but also interactions between job involvement and job satisfaction as this will yield new insights into the complex function of attitudes in influencing absenteeism. © 2007 The British Psychological Society.

Gaussian process approximations of stochastic differential equations

Stochastic differential equations arise naturally in a range of contexts, from financial to environmental modeling. Current solution methods are limited in their representation of the posterior process in the presence of data. In this work, we present a novel Gaussian process approximation to the posterior measure over paths for a general class of stochastic differential equations in the presence of observations. The method is applied to two simple problems: the Ornstein-Uhlenbeck process, of which the exact solution is known and can be compared to, and the double-well system, for which standard approaches such as the ensemble Kalman smoother fail to provide a satisfactory result. Experiments show that our variational approximation is viable and that the results are very promising as the variational approximate solution outperforms standard Gaussian process regression for non-Gaussian Markov processes.

Resveratrol stimulates hydrogen sulfide (H2S) formation to relax murine corpus cavernosum

Introduction: Resveratrol (RVT) found in red wine protects against erectile dysfunction and relaxes penile tissue (corpus cavernosum) via a nitric oxide (NO) independent pathway. However, the mechanism remains to be elucidated. Hydrogen sulfide (H2S) is a potent vasodilator and neuromodulator generated in corpus cavernosum. Aims: We investigated whether RVT caused the relaxation of mice corpus cavernosum (MCC) through H2S. Methods: H2S formation is measured by methylene blue assay and vascular reactivity experiments have been performed by DMT strip myograph in CD1 MCC strips. Main Outcome Measures: Endothelial NO synthase (eNOS) inhibitor Nω-Nitro-L-arginine (L-NNA, 0.1mM) or H2S inhibitor aminooxyacetic acid (AOAA, 2mM) which inhibits both cystathionine-β-synthase (CBS) and cystathionine-gamma-lyase (CSE) enzyme or combination of AOAA with PAG (CSE inhibitor) has been used in the presence/absence of RVT (0.1mM, 30min) to elucidate the role of NO or H2S pathways on the effects of RVT in MCC. Concentration-dependent relaxations to RVT, L-cysteine, sodium hydrogen sulfide (NaHS) and acetylcholine (ACh) were studied. Results: Exposure of murine corpus cavernosum to RVT increased both basal and L-cysteine-stimulated H2S formation. Both of these effects were reversed by AOAA but not by L-NNA. RVT caused concentration-dependent relaxation of MCC and that RVT-induced relaxation was significantly inhibited by AOAA or AOAA+PAG but not by L-NNA. L-cysteine caused concentration-dependent relaxations, which are inhibited by AOAA or AOAA+PAG significantly. Incubation of MCC with RVT significantly increased L-cysteine-induced relaxation, and this effect was inhibited by AOAA+PAG. However, RVT did not alter the effect of exogenous H2S (NaHS) or ACh-induced relaxations. Conclusions: These results demonstrate that RVT-induced relaxation is at least partly dependent on H2S formation and acts independent of eNOS pathway. In phosphodiesterase 5 inhibitor (PDE-5i) nonresponder population, combination therapy with RVT may reverse erectile dysfunction via stimulating endogenous H2S formation. Yetik-Anacak G, Dereli MV, Sevin G, Ozzayim O, Erac Y, and Ahmed A. Resveratrol stimulates hydrogen sulfide (H2S) formation to relax murine corpus cavernosum.