Molecular basis of salt tolerance in Physcomitrella Patens model plant: potassium homeostasis and physiological roles of chx transporters

El suelo salino impone un estrés abiótico importante que causa graves problemas en la agricultura ya que la mayoría de los cultivos se ven afectados por la salinidad debido a efectos osmóticos y tóxicos. Por ello, la contaminación y la escasez de agua dulce, la salinización progresiva de tierras y el aumento exponencial de la población humana representan un grave problema que amenaza la seguridad alimentaria mundial para las generaciones futuras. Por lo tanto, aumentar la tolerancia a la salinidad de los cultivos es un objetivo estratégico e ineludible para garantizar el suministro de alimentos en el futuro. Mantener una óptima homeostasis de K+ en plantas que sufren estrés salino es un objetivo importante en el proceso de obtención de plantas tolerantes a la salinidad. Aunque el modelo de la homeostasis de K+ en las plantas está razonablemente bien descrito en términos de entrada de K+, muy poco se sabe acerca de los genes implicados en la salida de K+ o de su liberación desde la vacuola. En este trabajo se pretende aclarar algunos de los mecanismos implicados en la homeostasis de K+ en plantas. Para ello se eligió la briofita Physcomitrella patens, una planta no vascular de estructura simple y de fase haploide dominante que, entre muchas otras cualidades, hacen que sea un modelo ideal. Lo más importante es que no sólo P. patens es muy tolerante a altas concentraciones de Na+, sino que también su posición filogenética en la evolución de las plantas abre la posibilidad de estudiar los cambios claves que, durante el curso de la evolución, se produjeron en las diversas familias de los transportadores de K+. Se han propuesto varios transportadores de cationes como candidatos que podrían tener un papel en la salida de K+ o su liberación desde la vacuola, especialmente miembros de la familia CPA2 que contienen las familias de transportadores KEA y CHX. En este estudio se intenta aumentar nuestra comprensión de las funciones de los transportadores de CHX en las células de las plantas usando P. patens, como ya se ha dicho. En esta especie, se han identificado cuatro genes CHX, PpCHX1-4. Dos de estos genes, PpCHX1 y PpCHX2, se expresan aproximadamente al mismo nivel que el gen PpACT5, y los otros dos genes muestran una expresión muy baja. La expresión de PpCHX1 y PpCHX2 en mutantes de Escherichia coli defectivos en el transporte de K+ restauraron el crecimiento de esta cepa en medios con bajo contenido de K+, lo que viii sugiere que la entrada de K+ es energizada por un mecanismo de simporte con H+. Por otra parte, estos transportadores suprimieron el defecto asociado a la mutación kha1 en Saccharomyces cerevisiae, lo que sugiere que podrían mediar un antiporte en K+/H+. La proteína PpCHX1-GFP expresada transitoriamente en protoplastos de P. patens co-localizó con un marcador de Golgi. En experimentos similares, la proteína PpCHX2-GFP localizó aparentemente en la membrana plasmática y tonoplasto. Se construyeron las líneas mutantes simples de P. patens ΔPpchx1 y ΔPpchx2, y también el mutante doble ΔPpchx2 ΔPphak1. Los mutantes simples crecieron normalmente en todas las condiciones ensayadas y mostraron flujos de entrada normales de K+ y Rb+; la mutación ΔPpchx2 no aumentó el defecto de las plantas ΔPphak1. En experimentos a largo plazo, las plantas ΔPpchx2 mostraron una retención de Rb+ ligeramente superior que las plantas silvestres, lo que sugiere que PpCHX2 promueve la transferencia de Rb+ desde la vacuola al citosol o desde el citosol al medio externo, actuando en paralelo con otros transportadores. Sugerimos que transportadores de K+ de varias familias están involucrados en la homeostasis de pH de orgánulos ya sea mediante antiporte K+/H+ o simporte K+-H+.ix ABSTRACT Soil salinity is a major abiotic stress causing serious problems in agriculture as most crops are affected by it. Moreover, the contamination and shortage of freshwater, progressive land salinization and exponential increase of human population aggravates the problem implying that world food security may not be ensured for the next generations. Thus, a strategic and an unavoidable goal would be increasing salinity tolerance of plant crops to secure future food supply. Maintaining an optimum K+ homeostasis in plants under salinity stress is an important trait to pursue in the process of engineering salt tolerant plants. Although the model of K+ homeostasis in plants is reasonably well described in terms of K+ influx, very little is known about the genes implicated in K+ efflux or release from the vacuole. In this work, we aim to clarify some of the mechanisms involved in K+ homeostasis in plants. For this purpose, we chose the bryophyte plant Physcomitrella patens, a nonvascular plant of simple structure and dominant haploid phase that, among many other characteristics, makes it an ideal model. Most importantly, not only P. patens is very tolerant to high concentrations of Na+, but also its phylogenetic position in land plant evolution opens the possibility to study the key changes that occurred in K+ transporter families during the course of evolution. Several cation transporter candidates have been proposed to have a role in K+ efflux or release from the vacuole especially members of the CPA2 family which contains the KEA and CHX transporter families. We intended in this study to increase our understanding of the functions of CHX transporters in plant cells using P. patens, in which four CHX genes have been identified, PpCHX1-4. Two of these genes, PpCHX1 and PpCHX2, are expressed at approximately the same level as the PpACT5 gene, but the other two genes show an extremely low expression. PpCHX1 and PpCHX2 restored growth of Escherichia coli mutants on low K+-containing media, suggesting they mediated K+ uptake that may be energized by symport with H+. In contrast, these genes suppressed the defect associated to the kha1 mutation in Saccharomyces cerevisiae, which suggest that they might mediate K+/H+ antiport. PpCHX1-GFP protein transiently expressed in P. patens protoplasts co-localized with a Golgi marker. In similar experiments, the PpCHX2-GFP protein appeared to localize to tonoplast and plasma x membrane. We constructed the ΔPpchx1 and ΔPpchx2 single mutant lines, and the ΔPpchx2 ΔPphak1 double mutant. Single mutant plants grew normally under all the conditions tested and exhibited normal K+ and Rb+ influxes; the ΔPpchx2 mutation did not increase the defect of ΔPphak1 plants. In long-term experiments, ΔPpchx2 plants showed a slightly higher Rb+ retention than wild type plants, which suggests that PpCHX2 mediates the transfer of Rb+ from either the vacuole to the cytosol or from the cytosol to the external medium in parallel with other transporters. We suggest that K+ transporters of several families are involved in the pH homeostasis of organelles by mediating either K+/H+ antiport or K+-H+ symport.

Protein isolate and inulin: chemical, rheological and structural basis

Soy protein isolate is typical vegetable protein with health-enhancing activities. Inulin, a prebiotic no digestible carbohydrate, has functional properties. A mashed potato serving of 200 g with added soy protein isolate and inulin concentrations of 15?60 g kg provides from 3 to 12 g of soy protein isolate and/or inulin, respectively. Currently, no information is available about the possible texture-modifying effect of this non-ionizable polar carbohydrate in different soy-based food systems. In this study, the effect of the addition of soy protein isolate and inulin blends at different soy protein isolate: inulin ratios on the degree of inulin polymerization and the rheological and structural properties of fresh mashed and frozen/thawed mashed potatoes were evaluated. The inulin chemical structure remained intact throughout the various treatments, and soy protein isolate did not affect inulin composition being a protein compatible with this fructan. Small-strain rheology showed that both ingredients behaved like soft fillers. In the frozen/thawed mashed potatoes samples,0 addition of 30 : 30 and 15 : 60 blend ratios significantly increased elasticity (G value) compared with 0 : 0 control, consequently reducing the freeze/thaw stability conferred by the cryoprotectants. Inulin crystallites caused a significant strengthening effect on soy protein isolate gel. Micrographs revealed that soy protein isolate supports the inulin structure by building up a second fine-stranded network. Thereby, possibility of using soy protein isolate and inulin in combination with mashed potatoes to provide a highly nutritious and healthy product is promising.

Farm nitrogen balances in six European landscapes as an indicator for nitrogen losses and basis for improved management.

Improved management of nitrogen (N) in agriculture is necessary to achieve a sustainable balance between the production of food and other biomass, and the unwanted effects of N on water pollution, greenhouse gas emissions, biodiversity deterioration and human health. To analyse farm N-losses and the complex interactions within farming systems, efficient methods for identifying emissions hotspots and evaluating mitigation measures are therefore needed. The present paper aims to fill this gap at the farm and landscape scales. Six agricultural landscapes in Poland (PL), the Netherlands (NL), France (FR), Italy (IT), Scotland (UK) and Denmark (DK) were studied, and a common method was developed for undertaking farm inventories and the derivation of farm N balances, N surpluses and for evaluating uncertainty for the 222 farms and 11 440 ha of farmland included in the study. In all landscapes, a large variation in the farm N surplus was found, and thereby a large potential for reductions. The highest average N surpluses were found in the most livestock-intensive landscapes of IT, FR, and NL; on average 202 ± 28, 179 ± 63 and 178 ± 20 kg N ha−1 yr−1, respectively. All landscapes showed hotspots, especially from livestock farms, including a special UK case with large-scale landless poultry farming. Overall, the average N surplus from the land-based UK farms dominated by extensive sheep and cattle grazing was only 31 ± 10 kg N ha−1 yr−1, but was similar to the N surplus of PL and DK (122 ± 20 and 146 ± 55 kg N ha−1 yr−1, respectively) when landless poultry farming was included. We found farm N balances to be a useful indicator for N losses and the potential for improving N management. Significant correlations to N surplus were found, both with ammonia air concentrations and nitrate concentrations in soils and groundwater, measured during the period of N management data collection in the landscapes from 2007–2009. This indicates that farm N surpluses may be used as an independent dataset for validation of measured and modelled N emissions in agricultural landscapes. No significant correlation was found with N measured in surface waters, probably because of spatial and temporal variations in groundwater buffering and biogeochemical reactions affecting N flows from farm to surface waters. A case study of the development in N surplus from the landscape in DK from 1998–2008 showed a 22% reduction related to measures targeted at N emissions from livestock farms. Based on the large differences in N surplus between average N management farms and the most modern and N-efficient farms, it was concluded that additional N-surplus reductions of 25–50%, as compared to the present level, were realistic in all landscapes. The implemented N-surplus method was thus effective for comparing and synthesizing results on farm N emissions and the potentials of mitigation options. It is recommended for use in combination with other methods for the assessment of landscape N emissions and farm N efficiency, including more detailed N source and N sink hotspot mapping, measurements and modelling.

Zero-G Gauging systems. CONTRACT No 17/8/4

Various systems for measuring propellant content in spacecrafts under weightlessness conditions are reviewed. The cavity resonator method is found to be the most suitable measurement; technique. This method is analyzed in detail. A determination of errors intrinsec to the method is carried out.

Assessment of irrigators’ preferences for different water supply risk management tools: option contract and insurance.

Irrigators face the risk of not having enough water to meet their crops’ demand. There are different mechanisms to cope with this risk, including water markets (option contracts) or insurance. A farmer will purchase them when the expected utility change derived from the tool is positive. This paper presents a theoretical assessment of the farmer’s expected utility under two different option contracts, a drought insurance and a combination of an option contract and the insurance. We analyze the conditions that determine farmer’s reference for one instrument or the other and perform a numerical application that is relevant for a Spanish region.

Image analysis by moment invariants using a set of step-like basis functions

Moment invariants have been thoroughly studied and repeatedly proposed as one of the most powerful tools for 2D shape identification. In this paper a set of such descriptors is proposed, being the basis functions discontinuous in a finite number of points. The goal of using discontinuous functions is to avoid the Gibbs phenomenon, and therefore to yield a better approximation capability for discontinuous signals, as images. Moreover, the proposed set of moments allows the definition of rotation invariants, being this the other main design concern. Translation and scale invariance are achieved by means of standard image normalization. Tests are conducted to evaluate the behavior of these descriptors in noisy environments, where images are corrupted with Gaussian noise up to different SNR values. Results are compared to those obtained using Zernike moments, showing that the proposed descriptor has the same performance in image retrieval tasks in noisy environments, but demanding much less computational power for every stage in the query chain.

A Maut aprroach for reusing domain ontologies on the basis of the NeOn Methodlogy

Knowledge resource reuse has become a popular approach within the ontology engineering field, mainly because it can speed up the ontology development process, saving time and money and promoting the application of good practices. The NeOn Methodology provides guidelines for reuse. These guidelines include the selection of the most appropriate knowledge resources for reuse in ontology development. This is a complex decision-making problem where different conflicting objectives, like the reuse cost, understandability, integration workload and reliability, have to be taken into account simultaneously. GMAA is a PC-based decision support system based on an additive multi-attribute utility model that is intended to allay the operational difficulties involved in the Decision Analysis methodology. The paper illustrates how it can be applied to select multimedia ontologies for reuse to develop a new ontology in the multimedia domain. It also demonstrates that the sensitivity analyses provided by GMAA are useful tools for making a final recommendation.

Application of the agency theory for the analysis of performance-based mechanisms in road management

El WCTR es un congreso de reconocido prestigio internacional en el ámbito de la investigación del transporte, y aunque las actas publicadas están en formato digital y sin ISSN ni ISBN, lo consideramos lo suficientemente importante como para que se considere en los indicadores. This paper develops a model based on agency theory to analyze road management systems (under the different contract forms available today) that employ a mechanism of performance indicators to establish the payment of the agent. The base assumption is that of asymmetric information between the principal (Public Authorities) and the agent (contractor) and the risk aversion of this latter. It is assumed that the principal may only measure the agent?s performance indirectly and by means of certain performance indicators that may be verified by the authorities. In this model there is presumed to be a relation between the efforts made by the agent and the performance level measured by the corresponding indicators, though it is also considered that there may be dispersion between both variables that gives rise to a certain degree of randomness in the contract. An analysis of the optimal contract has been made on the basis of this model and in accordance with a series of parameters that characterize the economic environment and the particular conditions of road infrastructure. As a result of the analysis made, it is considered that an optimal contract should generally combine a fixed component and a payment in accordance with the performance level obtained. The higher the risk aversion of the agent and the greater the marginal cost of public funds, the lower the impact of this performance-based payment. By way of conclusion, the system of performance indicators should be as broad as possible but should not overweight those indicators that encompass greater randomness in their results.

Code assessment and modelling for Design Basis Accident Analysis of the European sodium fast reactor design. Part I: System description, modelling and benchmarking

The new reactor concepts proposed in the Generation IV International Forum (GIF) are conceived to improve the use of natural resources, reduce the amount of high-level radioactive waste and excel in their reliability and safe operation. Among these novel designs sodium fast reactors (SFRs) stand out due to their technological feasibility as demonstrated in several countries during the last decades. As part of the contribution of EURATOM to GIF the CP-ESFR is a collaborative project with the objective, among others, to perform extensive analysis on safety issues involving renewed SFR demonstrator designs. The verification of computational tools able to simulate the plant behaviour under postulated accidental conditions by code-to-code comparison was identified as a key point to ensure reactor safety. In this line, several organizations employed coupled neutronic and thermal-hydraulic system codes able to simulate complex and specific phenomena involving multi-physics studies adapted to this particular fast reactor technology. In the “Introduction” of this paper the framework of this study is discussed, the second section describes the envisaged plant design and the commonly agreed upon modelling guidelines. The third section presents a comparative analysis of the calculations performed by each organisation applying their models and codes to a common agreed transient with the objective to harmonize the models as well as validating the implementation of all relevant physical phenomena in the different system codes.

Code assessment and modelling for Design Basis Accident analysis of the European Sodium Fast Reactor design. Part II: Optimised core and representative transients analysis

The new reactor concepts proposed in the Generation IV International Forum require the development and validation of computational tools able to assess their safety performance. In the first part of this paper the models of the ESFR design developed by several organisations in the framework of the CP-ESFR project were presented and their reliability validated via a benchmarking exercise. This second part of the paper includes the application of those tools for the analysis of design basis accident (DBC) scenarios of the reference design. Further, this paper also introduces the main features of the core optimisation process carried out within the project with the objective to enhance the core safety performance through the reduction of the positive coolant density reactivity effect. The influence of this optimised core design on the reactor safety performance during the previously analysed transients is also discussed. The conclusion provides an overview of the work performed by the partners involved in the project towards the development and enhancement of computational tools specifically tailored to the evaluation of the safety performance of the Generation IV innovative nuclear reactor designs.

Micromechanical basis for shear strength of rock discontinuities

The theoretical basis for evaluating shear strength in rock joints is presented and used to derive an equation that governs the relationship between tangential and normal stress on the joint during situations of slippage between the joint faces. The dependent variables include geometric dilatancy, the instantaneous friction angle, and a parameter that considers joint surface roughness. The effect roughness is studied, and the aforementioned formula is used to analyse joints under different conditions. A mathematical expression is deduced that explains Barton's value for the joint roughness coefficient (JRC) according to the roughness geometry. In particular, when the Hoek and Brown failure criterion is used for a rock in the contact with the surface roughness plane, it is possible to determine the shear strength of the joint as a function of the relationship between the uniaxial compressive strength of the wall with the normal stress acting on the wall. Finally, theoretical results obtained for the geometry of a three-dimensional joint are compared with those of the Barton's formulation

Las Opciones Reales en los Expedientes de Regulación de Empleo

Las empresas en situación económica negativa, recurren en España a los Expedientes de Regulación de Empleo que es la figura legal que permite ajustar sus plantillas y cuya utilización se acrecienta en épocas de crisis. El objetivo de esta investigación se focaliza en la integración de la Teoría de Opciones Reales en los proyectos organizativos de recursos humanos, en particular en el estudio y aplicación de la opción de contracción y de la opción de aplazamiento a los proyectos de desarrollo de los Expedientes de Regulación de Empleo. Las opciones reales son una herramienta de gestión, novedosa y contrastada, que posibilita realizar la valoración de los Expedientes de Regulación de Empleo tomando en consideración las incertidumbres futuras, lo que permite cuantificar mejor el resultado de la decisión y supone un avance sobre la valoración realizada de forma estática. Dos aspectos clave ha sido necesario resolver. En primer lugar determinar la fuente de incertidumbre que ponga en valor las opciones reales para este tipo de proyectos y como consecuencia desarrollar los procedimientos para obtener el valor de la opción. La puesta en valor de la opción de contracción permite, a partir de la consideración de la demanda como factor de incertidumbre, desarrollar un modelo que proporciona el valor óptimo del número de contratos de trabajo a extinguir, de modo que sitúa a la organización en la mejor posición para la toma de decisiones. La resolución del modelo se logra con la ayuda de la herramienta “The Newsvendor Model”, que facilita la obtención el valor óptimo. En la aplicación de la opción de aplazamiento se ha concluido asimismo, que la volatilidad de la demanda es la fuente de incertidumbre que hace viable la opción y a partir de dicha incertidumbre se ha desarrollado un procedimiento basado en el árbol binomial que permite obtener su valor. Una vez desarrollados los dos modelos para las opciones de contracción y aplazamiento, se ha elaborado un modelo conjunto con ambas opciones, integrando los resultados obtenidos en cada caso. Para validar los modelos obtenidos se presenta su aplicación al caso real de una empresa en situación económica negativa que requiere la necesidad de llevar a cabo un Expediente de Regulación de Empleo, lo que ha permitido comprobar la utilidad de dichos desarrollos, al obtener resultados que facilitan la toma de decisiones y que presentan soluciones que pueden ser más equilibradas para los trabajadores de la empresa, aspecto clave a tener en cuenta, dado el coste personal, familiar y social de este tipo de proyectos. La integración de las opciones reales en la gestión de recursos humanos tiene un amplio campo de aplicación, dadas las diferentes inversiones que se pueden llevar a cabo sobre el capital humano. Aunque esta investigación se ha focalizado en la opcionalidad de las inversiones para el ajuste de plantilla, existen otro tipo de inversiones en recursos humanos, como son las dedicadas a formación, motivación, adecuación o incorporación de personal, que también estarán sujetas a opcionalidad si existen incertidumbres a futuro. Con el objetivo de avanzar en el estudio y aplicación de las opciones reales a estas inversiones, se presenta un marco conceptual que identifica las bases de partida en lo referente a los retornos de las inversiones y las incertidumbres futuras. Con los resultados obtenidos en esta investigación se entiende que se ha conseguido el objetivo de aplicar la Teoría de Opciones Reales a los Expedientes de Regulación de Empleo y con ello establecer la posibilidad de mejorar las implicaciones sociales de estos proyectos dentro de la empresa y en la sociedad. ABSTRACT During crisis times, like the one Spain has been going through on the last years, the Collective Dismissal Procedure is an important issue to be considered within companies that are having a negative economic situation. This research focuses on the integration of the Real Options Theory for organizational projects in the management of human resources, and particularly the study and inclusion of the option to contract and the deferral option in Collective Dismissal Procedures. The Real Options Theory, an innovative management tool that has been proven as correct, help to improve the assessment of the Collective Dismissal Procedures using the dynamic demand instead of the static demand, giving a better quantification of the decision that is going to be taken. Two main issues which are hoped to be resolve are examined: the search for a source of uncertainty to make the real options viable for this type of projects, and consequently the development of a procedure to obtain the value of the option. The option to contract allows, from the consideration of the demand as a factor of uncertainty, to develop a model obtaining the optimal value in the reduction of the workforce, therefore allowing the organization to place itself in the best position for the decision making. The resolution of this model is achieved with the help of "The Newsvendor Model" tool, which facilitates obtaining the optimal value. The analysis performed in the deferral option also concluded that the volatility of demand is the source of the uncertainty that makes the option viable and the procedure developed by the binomial tree is the tool that enables the value of the option to be found. Once developed two models for contraction and deferment options, a joint model has been developed including both options by integrating the results obtained in each case. To validate the models, the real case of a company in negative economic situation requiring the need to carry out a procedure for collective redundancies is applied. This has demonstrated the usefulness of this kind of development that can deliver results to improve the decision making and provide more balanced solutions for company employees, key consideration given the impact on personal, family life and social environment of these Collective Dismissal Procedures. The integration of real options in the management of human resources has broad scope, given the different investments that can be performed on the human capital. Although this research has focused on the optionality of investments for downsizing, there are other investments in human resources for training, motivation, arrangement or incorporation of staff that are also subject to optionality if there are future uncertainties. With the aim of advancing the study and application of real options to these investments, a conceptual framework is presented that identifies the basis of research in terms of the investment returns and future uncertainties. With the results obtained on this research it is clearly defined the objectives for which this work was started for, the application of the Real Options Theory to the Collective Dismissal Procedure. And thereby achieve the final goal of improving the social impact of these projects within a company and society.