950 resultados para strategy based organization
Resumo:
La sostenibilidad de los sistemas olivareros situados en zonas de pendiente y montaña (SMOPS) en Andalucía se encuentra actualmente amenazada por las elevadas tasas de abandono que afectan a estos sistemas productivos. Así, la consumación de este proceso de abandono, no sólo pondría en peligro a las propias explotaciones, sino a todo el conjunto de bienes y servicios no productivos y al patrimonio cultural generado por este sistema productivo. En este contexto, la búsqueda de alternativas políticas enfocadas a revertir este proceso se erige como una necesidad categórica en aras de garantizar en el largo plazo la sostenibilidad de los olivares de montaña. Esta tesis pretende hacer frente a esta necesidad a través de la construcción de un marco político alternativo para los SMOPS, que permita la integración simultánea de todas las dimensiones que pueden influir en su desarrollo; esto es: el marco político actual, principalmente determinado por la Política Agraria Común (PAC) de la Unión Europea (UE); las preferencias de la sociedad hacia la oferta de bienes y servicios públicos generados por los SMOPS; y las preferencias y voluntad de innovación hacia nuevos manejos y sistemas de gestión de los agricultores y propietarios de las explotaciones. Para ello, se emplea una metodología de investigación mixta, que abarca la realización de cuatro encuestas (personales y online) llevadas a cabo a los agentes o grupos de interés involucrados directa o indirectamente en la gestión de los SMOPS –ciudadanos, agricultores y propietarios y expertos-; una profunda revisión de las herramientas de política agroambiental actuales y posibles alternativas a las mismas; y el desarrollo de nuevas estrategias metodológicas para dotar de mayor precisión y fiabilidad las estimaciones obtenidas a partir del Método del Experimento de Elección (MEE) en el campo de la valoración medioambiental. En general, los resultados muestran que una estrategia de política agroambiental basada en la combinación de los Contratos Territoriales de Zona Rural (CTZR) y el manejo ecológico supondría una mejora en la sostenibilidad de los sistemas olivareros de montaña andaluces, que, al mismo tiempo, propiciaría una mejor consideración de las necesidades y demandas de los agentes implicados en su gestión. Asimismo, los hallazgos obtenidos en esta investigación demandan un cambio de paradigma en los actuales pagos agroambientales, que han de pasar de una estrategia basada en la implementación de acciones, a otra enfocada al logro de objetivos, la cual, en el caso del olivar, se podría centrar en el aumento del secuestro de carbono en el suelo. Desde un punto de vista metodológico, los resultados han contribuido notablemente a mejorar la fiabilidad y precisión de las conclusiones estimadas a partir del MEE, mediante el diseño de un novedoso proceso iterativo para detectar posibles comportamientos inconsistentes por parte de los entrevistados con respecto a su máxima Disposición al Pago (DAP) para lograr la situación considerada como “óptima” en los olivares ecológicos de montaña andaluces. En líneas generales, el actual marco institucional favorece la puesta en práctica de la mayoría de las estrategias propuestas en esta tesis; sin embargo son necesarios mayores esfuerzos para reconducir los actuales Pagos Agroambientales y Climáticos de la PAC, hacia una estrategia de política agroambiental adaptada a las necesidades y requisitos del territorio en el que se aplica, enfocada al logro de objetivos y que sea capaz de integrar y coordinar al conjunto de agentes y grupos de interés involucrados -directa o indirectamente- en la gestión de los olivares de montaña. En este contexto, se espera que la puesta en práctica de nuevas estructuras y acuerdos de gobernanza territorial juegue un importante papel en el desarrollo de una política agroambiental realmente adaptada a las necesidades de los sistemas olivareros de montaña andaluces. ABSTRACT The long-term sustainability of Andalusian sloping and mountainous olive production systems (SMOPS) is currently threatened by the high abandonment rates that affect these production systems. The effective occurrence of this abandonment process is indeed menacing not only farms themselves, but also the wide array of public goods and services provided by SMOPS and the cultural heritage held by this production system. The search of policy alternatives aimed at tackling this process is thus a central necessity. This thesis aims to undertake this necessity by building an alternative policy framework for SMOPS that simultaneously integrates the several dimensions that are susceptible to influence it, namely: the current policy framework, mainly determined by the European Union’s (EU) Common Agricultural Policy (CAP); the social preferences toward the supply of SMOPS’ public goods and services; and farmers’ preferences and willingness to innovate toward new management practices in their farms. For this purpose, we put into practice a mixed-method strategy that combines four face-to-face and online surveys carried out with SMOPS’ stakeholders -including citizens, farmers and experts-; in-depth analysis of current and alternative agrienvironmental policy (AEP) instruments; and the development of novel methodological approaches to advance toward more reliable Discrete Choice Experiment’s (DCE) outcomes in the field of environmental valuation. Overall, results show that a policy strategy based on the combination of Territorial Management Contracts (TMC) and organic management would further enhance Andalusian SMOPS’ sustainability by simultaneously taking into account stakeholders’ demands and needs. Findings also call for paradigm shift of the current action-oriented design of Agri-Environmental-Climate Schemes (AECS), toward a result-based approach, that in the case of olive orchards should particularly be focused on enhancing soil carbon sequestration. From a methodological perspective, results have contributed to improve the accuracy and feasibility of DCE outcomes by designing a novel and iterative procedure focused in ascertaining respondents’ inconsistent behaviour with respect to their stated maximum WTP for the attainment of an ideal situation to be achieved in organic Andalusian SMOPS. Generally, the present institutional framework favours the implementation of the main policy strategies proposed in this thesis, albeit further efforts are required to better conduct current CAP’s agri-environmental instruments toward a territorially targeted result-oriented strategy capable to integrate and coordinate the whole set of stakeholders involved in the management of SMOPS. In this regard, alternative governance structures and arrangements are expected to play a major role on the process of tackling SMOPS’ agri-environmental policy challenge.
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In this work a p-adaptation (modification of the polynomial order) strategy based on the minimization of the truncation error is developed for high order discontinuous Galerkin methods. The truncation error is approximated by means of a truncation error estimation procedure and enables the identification of mesh regions that require adaptation. Three truncation error estimation approaches are developed and termed a posteriori, quasi-a priori and quasi-a priori corrected. Fine solutions, which are obtained by enriching the polynomial order, are required to solve the numerical problem with adequate accuracy. For the three truncation error estimation methods the former needs time converged solutions, while the last two rely on non-converged solutions, which lead to faster computations. Based on these truncation error estimation methods, algorithms for mesh adaptation were designed and tested. Firstly, an isotropic adaptation approach is presented, which leads to equally distributed polynomial orders in different coordinate directions. This first implementation is improved by incorporating a method to extrapolate the truncation error. This results in a significant reduction of computational cost. Secondly, the employed high order method permits the spatial decoupling of the estimated errors and enables anisotropic p-adaptation. The incorporation of anisotropic features leads to meshes with different polynomial orders in the different coordinate directions such that flow-features related to the geometry are resolved in a better manner. These adaptations result in a significant reduction of degrees of freedom and computational cost, while the amount of improvement depends on the test-case. Finally, this anisotropic approach is extended by using error extrapolation which leads to an even higher reduction in computational cost. These strategies are verified and compared in terms of accuracy and computational cost for the Euler and the compressible Navier-Stokes equations. The main result is that the two quasi-a priori methods achieve a significant reduction in computational cost when compared to a uniform polynomial enrichment. Namely, for a viscous boundary layer flow, we obtain a speedup of a factor of 6.6 and 7.6 for the quasi-a priori and quasi-a priori corrected approaches, respectively. RESUMEN En este trabajo se ha desarrollado una estrategia de adaptación-p (modificación del orden polinómico) para métodos Galerkin discontinuo de alto orden basada en la minimización del error de truncación. El error de truncación se estima utilizando el método tau-estimation. El estimador permite la identificación de zonas de la malla que requieren adaptación. Se distinguen tres técnicas de estimación: a posteriori, quasi a priori y quasi a priori con correción. Todas las estrategias requieren una solución obtenida en una malla fina, la cual es obtenida aumentando de manera uniforme el orden polinómico. Sin embargo, mientras que el primero requiere que esta solución esté convergida temporalmente, el resto utiliza soluciones no convergidas, lo que se traduce en un menor coste computacional. En este trabajo se han diseñado y probado algoritmos de adaptación de malla basados en métodos tau-estimation. En primer lugar, se presenta un algoritmo de adaptacin isótropo, que conduce a discretizaciones con el mismo orden polinómico en todas las direcciones espaciales. Esta primera implementación se mejora incluyendo un método para extrapolar el error de truncación. Esto resulta en una reducción significativa del coste computacional. En segundo lugar, el método de alto orden permite el desacoplamiento espacial de los errores estimados, permitiendo la adaptación anisotropica. Las mallas obtenidas mediante esta técnica tienen distintos órdenes polinómicos en cada una de las direcciones espaciales. La malla final tiene una distribución óptima de órdenes polinómicos, los cuales guardan relación con las características del flujo que, a su vez, depenen de la geometría. Estas técnicas de adaptación reducen de manera significativa los grados de libertad y el coste computacional. Por último, esta aproximación anisotropica se extiende usando extrapolación del error de truncación, lo que conlleva un coste computational aún menor. Las estrategias se verifican y se comparan en téminors de precisión y coste computacional utilizando las ecuaciones de Euler y Navier Stokes. Los dos métodos quasi a priori consiguen una reducción significativa del coste computacional en comparación con aumento uniforme del orden polinómico. En concreto, para una capa límite viscosa, obtenemos una mejora en tiempo de computación de 6.6 y 7.6 respectivamente, para las aproximaciones quasi-a priori y quasi-a priori con corrección.
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The partial molecular characterization of multiple sclerosis (MS)-associated retrovirus (MSRV), a novel retrovirus previously called LM7, is reported. MSRV has been isolated repeatedly from leptomeningeal, choroid plexus and from Epstein–Barr virus-immortalized B cells of MS patients. A strategy based on reverse transcriptase PCR with RNA-purified extracellular virions yielded an initial pol fragment from which other regions of the retroviral genome were subsequently obtained by sequence extension. MSRV-specific PCR primers amplified a pol region from RNA present at the peak of reverse transcriptase activity, coinciding with extracellular viral particles in sucrose density gradients. The same sequence was detected in noncellular RNA from MS patient plasma and in cerebrospinal fluid from untreated MS patients. MSRV is related to, but distinct from, the endogenous retroviral sequence ERV9. Whether MSRV represents an exogenous retrovirus with closely related endogenous elements or a replication-competent, virion-producing, endogenous provirus is as yet unknown. Further molecular epidemiological studies are required to determine precisely the apparent association of virions containing MSRV RNA with MS.
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A protein engineering strategy based on efficient and focused mutagenesis implemented by codon-based mutagenesis was developed. Vitaxin, a humanized version of the antiangiogenic antibody LM609 directed against a conformational epitope of the αvβ3 integrin complex, was used as a model system. Specifically, focused mutagenesis was used in a stepwise fashion to rapidly improve the affinity of the antigen binding fragment by greater than 90-fold. In the complete absence of structural information about the Vitaxin-αvβ3 interaction, phage-expressed antibody libraries for all six Ig heavy and light chain complementarity-determining regions were expressed and screened by a quantitative assay to identify variants with improved binding to αvβ3. The Vitaxin variants in these libraries each contained a single mutation, and all 20 amino acids were introduced at each complementarity-determining region residue, resulting in the expression of 2,336 unique clones. Multiple clones displaying 2- to 13-fold improved affinity were identified. Subsequent expression and screening of a library of 256 combinatorial variants of the optimal mutations identified from the primary libraries resulted in the identification of multiple clones displaying greater than 50-fold enhanced affinity. These variants inhibited ligand binding to receptor more potently as demonstrated by inhibition of cell adhesion and ligand competition assays. Because of the limited mutagenesis and combinatorial approach, Vitaxin variants with enhanced affinity were identified rapidly and required the synthesis of only 2,592 unique variants. The use of such small focused libraries obviates the need for phage affinity selection approaches typically used, permitting the use of functional assays and the engineering of proteins expressed in mammalian cell culture.
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Effective chemotherapy remains a key issue for successful cancer treatment in general and neuroblastoma in particular. Here we report a chemotherapeutic strategy based on catalytic antibody-mediated prodrug activation. To study this approach in an animal model of neuroblastoma, we have synthesized prodrugs of etoposide, a drug widely used to treat this cancer in humans. The prodrug incorporates a trigger portion designed to be released by sequential retro-aldol/retro-Michael reactions catalyzed by aldolase antibody 38C2. This unique prodrug was greater than 102-fold less toxic than etoposide itself in in vitro assays against the NXS2 neuroblastoma cell line. Drug activity was restored after activation by antibody 38C2. Proof of principle for local antibody-catalyzed prodrug activation in vivo was established in a syngeneic model of murine neuroblastoma. Mice with established 100-mm3 s.c. tumors who received one intratumoral injection of antibody 38C2 followed by systemic i.p. injections with the etoposide prodrug showed a 75% reduction in s.c. tumor growth. In contrast, injection of either antibody or prodrug alone had no antitumor effect. Systemic injections of etoposide at the maximum tolerated dose were significantly less effective than the intratumoral antibody 38C2 and systemic etoposide prodrug combination. Significantly, mice treated with the prodrug at 30-fold the maximum tolerated dose of etoposide showed no signs of prodrug toxicity, indicating that the prodrug is not activated by endogenous enzymes. These results suggest that this strategy may provide a new and potentially nonimmunogenic approach for targeted cancer chemotherapy.
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The study of the origin and pathogenetic relevance of the oligoclonal antibodies present in the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients has been hampered by a lack of specific ligands. We recently reported a general strategy, based on phage-displayed random peptide libraries, to identify ligands for disease-specific antibodies even in the absence of any information on the nature of the pathologic antigen. With this procedure, we identified several peptides specifically recognized by antibodies present in the CSF of MS patients. Using these peptides as reagents, we demonstrated that they mimic different natural epitopes and react with antibodies enriched in the CSF of MS patients. Antibodies recognizing the selected peptides are commonly found with equal frequency in the sera of MS patients and of normal individuals. In contrast, the repertoire of CSF antibodies appears to be individual-specific and is probably the result of a nonspecific immunodysregulation rather than a stereotyped response to a single antigen/agent.
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Many biological processes rely upon protein-protein interactions. Hence, detailed analysis of these interactions is critical for their understanding. Due to the complexities involved, genetic approaches are often needed. In yeast and phage, genetic characterizations of protein complexes are possible. However, in multicellular organisms, such characterizations are limited by the lack of powerful selection systems. Herein we describe genetic selections that allow single amino acid changes that disrupt protein-protein interactions to be selected from large libraries of randomly generated mutant alleles. The strategy, based on a yeast reverse two-hybrid system, involves a first-step negative selection for mutations that affect interaction, followed by a second-step positive selection for a subset of these mutations that maintain expression of full-length protein (two-step selection). We have selected such mutations in the transcription factor E2F1 that affect its ability to heterodimerize with DP1. The mutations obtained identified a putative helix in the marked box, a region conserved among E2F family members, as an important determinant for interaction. This two-step selection procedure can be used to characterize any interaction domain that can be tested in the two-hybrid system.
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Eph and its homologues form the largest subfamily of receptor tyrosine kinases. Normal expression patterns of this subfamily indicate roles in differentiation and development, whereas their overexpression has been linked to oncogenesis. This study investigated the potential role of Eph-related molecules during very early embryonic development by examining their expression in embryonic stem (ES) cells and embryoid bodies differentiated from ES cells in vitro. By use of a strategy based on reverse transcriptase-mediated PCR, nine clones containing Eph-subfamily sequence were isolated from ES cells. Of these, eight were almost identical to one of four previously identified molecules (Sek, Nuk, Eck, and Mek4). However, one clone contained sequence from a novel Eph-subfamily member, which was termed embryonic stem-cell kinase or Esk. Northern analysis showed expression of Esk in ES cells, embryoid bodies, day 12 mouse embryos, and some tissues of the adult animal. Levels of expression were similar in ES cells and embryoid bodies. By comparison, Mek4 showed no significant transcription in the ES cell cultures by Northern analysis, whereas Eck displayed stronger signals in ES cells than in the embryoid bodies. These results suggest that Eph-subfamily molecules may play roles during the earliest phases of embryogenesis. Furthermore, the relative importance of different members of this subfamily appears to change as development proceeds.
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A strategy based on the gene trap was developed to prescreen mouse embryonic stem cells for insertional mutations in genes encoding secreted and membrane-spanning proteins. The "secretory trap" relies on capturing the N-terminal signal sequence of an endogenous gene to generate an active beta-galactosidase fusion protein. Insertions were found in a cadherin gene, an unc6-related laminin (netrin) gene, the sek receptor tyrosine kinase gene, and genes encoding two receptor-linked protein-tyrosine phosphatases, LAR and PTP kappa. Analysis of homozygous mice carrying insertions in LAR and PTP kappa showed that both genes were effectively disrupted, but neither was essential for normal embryonic development.
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Non-suicidal self-injury (NSSI), such as cutting and burning, is a widespread social problem among lesbian, gay, bisexual, transgender, queer, and questioning (LGBTQ) youth. Extant research indicates that this population is more than twice as likely to engage in NSSI than heterosexual and cisgender (non-transgender) youth. Despite the scope of this social problem, it remains relatively unexamined in the literature. Research on other risk behaviors among LGBTQ youth indicates that experiencing homophobia and transphobia in key social contexts such as families, schools, and peer relationships contributes to health disparities among this group. Consequently, the aims of this study were to examine: (1) the relationship between LGBTQ youth's social environments and their NSSI behavior, and (2) whether/how specific aspects of the social environment contribute to an understanding of NSSI among LGBTQ youth. This study was conducted using an exploratory, sequential mixed methods design with two phases. The first phase of the study involved analysis of transcripts from interviews conducted with 44 LGBTQ youth recruited from a community-based organization. In this phase, five qualitative themes were identified: (1) Violence; (2) Misconceptions, Stigma, and Shame; (3) Negotiating LGBTQ Identity; (4) Invisibility and Isolation; and (5) Peer Relationships. Results from the qualitative phase were used to identify key variables and specify statistical models in the second, quantitative, phase of the study, using secondary data from a survey of 252 LGBTQ youth. The qualitative phase revealed how LGBTQ youth, themselves, described the role of the social environment in their NSSI behavior, while the quantitative phase was used to determine whether the qualitative findings could be used to predict engagement in NSSI among a larger sample of LGBTQ youth. The quantitative analyses found that certain social-environmental factors such as experiencing physical abuse at home, feeling unsafe at school, and greater openness about sexual orientation significantly predicted the likelihood of engaging in NSSI among LGBTQ youth. Furthermore, depression partially mediated the relationships between family physical abuse and NSSI and feeling unsafe at school and NSSI. The qualitative and quantitative results were compared in the interpretation phase to explore areas of convergence and incongruence. Overall, this study's findings indicate that social-environmental factors are salient to understanding NSSI among LGBTQ youth. The particular social contexts in which LGBTQ youth live significantly influence their engagement in this risk behavior. These findings can inform the development of culturally relevant NSSI interventions that address the social realities of LGBTQ youth's lives.
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From a gender perspective, protection and advertising political actions about work-family should promote sharing responsibilities between sexes. Next to political action and specific measures, the project of equal opportunities needs a long-term strategy based on the education on equality. This article proposes the methodologic exposition of a study based on these premises. It facilitates and explains the protocol used for the analysis of the audio-visual advertising campaigns on conciliation emitted by the Woman’s Institute. The evaluation of the actions is focused on the effectiveness from the point of view of mass media. It provides some data that illustrates the proposed study. Finally, it considers the difficulties of the available sources of information.
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This paper analyses the application of the cluster concept to tourist destinations using Benidorm as a case study. A questionnaire was administered to tourism firms based in Benidorm in order to determine whether this destination currently constitutes a tourism cluster or whether it possesses the ideal characteristics to become a cluster with the private agents' collaboration, that is, whether it is a potential cluster. The results obtained from this research indicate that Benidorm's success is not derived from the presence of a cluster due to a series of elements that prevent its existence. In this destination there is a need to strengthen cooperation between public and private agents (especially in those areas that determine the competitive advantage of the destination) and to design a strategy based on shared goals. Both of these elements are fundamental for the characterisation of a cluster.
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We have employed an inverse engineering strategy based on quantitative proteome analysis to identify changes in intracellular protein abundance that correlate with increased specific recombinant monoclonal antibody production (qMab) by engineered murine myeloma (NSO) cells. Four homogeneous NSO cell lines differing in qMab were isolated from a pool of primary transfectants. The proteome of each stably transfected cell line was analyzed at mid-exponential growth phase by two-dimensional gel electrophoresis (2D-PAGE) and individual protein spot volume data derived from digitized gel images were compared statistically. To identify changes in protein abundance associated with qMab clatasets were screened for proteins that exhibited either a linear correlation with cell line qMab or a conserved change in abundance specific only to the cell line with highest qMab. Several proteins with altered abundance were identified by mass spectrometry. Proteins exhibiting a significant increase in abundance with increasing qMab included molecular chaperones known to interact directly with nascent immunoglobulins during their folding and assembly (e.g., BiP, endoplasmin, protein disulfide isomerase). 2D-PAGE analysis showed that in all cell lines Mab light chain was more abundant than heavy chain, indicating that this is a likely prerequisite for efficient Mab production. In summary, these data reveal both the adaptive responses and molecular mechanisms enabling mammalian cells in culture to achieve high-level recombinant monoclonal antibody production. (C) 2004 Wiley Periodicals, Inc.
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We study here the adsorption of hexane on nanoporous MCM-41 silica at 303, 313, and 323 K, for various pore diameters between 2.40 and 4.24 nm. Adsorption equilibria, measured thermogravimetrically, show that all the isotherms, that are somewhat akin to those of type V, exhibit remarkably sharp capillary adsorption phase transition steps and are reversible. The position of the phase transition step gradually shifts from low to high relative pressure with an increase in the temperature as well as the pore sizes. The isosteric heats of adsorption derived from the equilibrium information using the Clapeyron equation reveal a gradual decrease with increasing adsorbed amount because of the surface heterogeneity but approach a constant value near the phase transition. A decrease in the pore size results in an increase in the isosteric heat of adsorption because of the increased dispersion forces. A simple strategy, based on the Broekhoff and De Boer adsorption theory, successfully interprets the hexane adsorption isotherms for the different pore size MCM-41 samples. The parameters of an empirical expression, used to represent the potential of interaction between the adsorbate and adsorbent, are obtained by fitting the monolayer region prior to capillary condensation and the experimental phase transition simultaneously, for some pore sizes. Subsequently, the parameters are used to predict the adsorption isotherm on other pore size samples, which showed good agreement with experimental data.
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In electronic support, receivers must maintain surveillance over the very wide portion of the electromagnetic spectrum in which threat emitters operate. A common approach is to use a receiver with a relatively narrow bandwidth that sweeps its centre frequency over the threat bandwidth to search for emitters. The sequence and timing of changes in the centre frequency constitute a search strategy. The search can be expedited, if there is intelligence about the operational parameters of the emitters that are likely to be found. However, it can happen that the intelligence is deficient, untrustworthy or absent. In this case, what is the best search strategy to use? A random search strategy based on a continuous-time Markov chain (CTMC) is proposed. When the search is conducted for emitters with a periodic scan, it is shown that there is an optimal configuration for the CTMC. It is optimal in the sense that the expected time to intercept an emitter approaches linearity most quickly with respect to the emitter's scan period. A fast and smooth approach to linearity is important, as other strategies can exhibit considerable and abrupt variations in the intercept time as a function of scan period. In theory and numerical examples, the optimum CTMC strategy is compared with other strategies to demonstrate its superior properties.