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The number of foreign owners in football clubs has recently increased in Europe and especially in England. This study aims to analyze whether clubs owned by foreign investors show different parameters of sporting and managerial efficiency. We use data on expected performance (from betting odds), wage costs, and the results of football clubs in the English Premier League (EPL) and English Football League Championship (EFL) over 13 seasons. We estimate different measures of efficiency using three alternative methods: expected vs. actual performance index, nonparametric conditional data envelopment analysis (DEA), and stochastic frontier, while considering foreign ownership as an exogenous factor. Our results suggest that there are not significant differences between domestic-and foreign-owned clubs.

This paper uses a novel semi‐non‐parametric approach, the so‐called stochastic non‐parametric envelopment on Z‐variables data (StoNEZD), to measure the performance of 279 European regions (in 28 EU member states and Norway) in the years 2000, 2007, and 2014. The StoNEZD approach combines the main virtues of the parametric and non‐parametric methods in a unifying framework. The proposed model accounts explicitly for the presence of contextual/exogenous factors that might affect the regional performance and allows for the use of statistical inference methods to explore the effects of such variables (agriculture‐to‐ gross value added (GVA) share, employment rate, and euro area membership). According to our results, a larger agriculture‐to‐GVA ratio has a negative impact on regional growth rates, whereas a higher employment rate has a positive influence on regional economic levels. In overall terms, the euro area membership appears to reduce regional average growth rates but seems to enhance regional average efficiency scores. Este trabajo utiliza un novedoso enfoque semi no parametrico, conocido como analisis envolvente estocastico no parametrico sobre datos de Z variables (StoNEZD, por sus siglas en ingles), para medir el desempeno de 279 regiones europeas (en 28 estados miembros de la UE y Noruega) en los anos 2000, 2007 y 2014. El enfoque StoNEZD combina las principales virtudes de los metodos parametricos y no parametricos en un solo marco. El modelo propuesto tiene en cuenta explicitamente la presencia de factores contextuales o exogenos que podrian afectar el desempeno regional y posibilita el uso de metodos de inferencia estadistica para explorar los efectos de esas variables (contribucion de la agricultura al valor anadido bruto (VAB), la tasa de empleo y la pertenencia a la zona del euro). De acuerdo con los resultados, una mayor contribucion de la agricultura al VAB tiene un efecto negativo en las tasas de crecimiento regional, mientras que una tasa de empleo mas elevada influye positivamente en los niveles economicos regionales. En terminos generales, la pertenencia a la zona del euro parece reducir las tasas promedio de crecimiento regional, pero parece mejorar los valores regionales promedio de eficiencia. 土地データは計画立案と行政に非常に重要であるが、開発途上国においてこのリソースは管理が不十分かつ利用できない場合が多い。最近独立した国では、データが削除されるなど問題が深刻化しており、新たな取り組みが必要となっている。この問題に対処するため、東ティモールにおける土地データを統合する際の法的、行政的、技術的な問題の解決策を検討し、提案する。ユーザーの視点から、データ・アクセス・インターフェースのプロトタイプを作成しテストした。このプロトタイプの実行可能性と、国土を所管する国家機関と公的と民間の両方の利害関係者に対する有用性を測定した。このプロトタイプは実用的であり、東ティモールにおける将来の土地情報システム (LIS)のあるべきかたちを予測するのに役立つことが分かった。

Lignin is a heterogeneous aromatic polymer responsible for cell wall stiffness and protection from pathogen attack. However, lignin represents a bottleneck to biomass degradation due to its recalcitrance related to the natural cell wall resistance to release sugars for fermentation or further processing. A biological approach involving genetics and molecular biology was used to disrupt lignin pathway synthesis and decrease lignin deposition. Here, we imaged three-dimensional fragments of the petioles of wild type and C4H lignin mutant Arabidopsis thaliana plants by synchrotron cryo-ptychography. The three-dimensional images revealed the heterogeneity of vessels, parenchyma, and fibre cell wall morphologies, highlighting the relation between disturbed lignin deposition and vessel implosion (cell collapsing and obstruction of water flow). We introduce a new parameter to accurately define cell implosion conditions in plants, and we demonstrate how cryo-ptychographic X-ray computed tomography (cryo-PXCT) provides new insights for plant imaging in three dimensions to understand physiological processes.

Background In Latin American and Caribbean countries the main concern of public health care managers has been traditionally placed on problems related to funding, payment mechanisms and equity of access. However, more recently, there is a growing interest in improving the levels of efficiency and reducing costs in the provision of health services. In this paper we focus on measuring the efficiency of public hospitals in Panama, where no such studies have been conducted so far. One of the most interesting features of the public hospital system in this country is that there are two different management schemes that coexist, thus we can make a distinction between hospitals operating under the Social Security Fund (SSF) and those belonging to the Ministry of Health (MoH). Methods Our dataset includes data about 22 public hospitals (11 for each model) during the period 2005–2015. We rely on the use of Data Envelopment Analysis and the Malmquist Productivity index to calculate technical efficiency and productivity change of hospitals. In addition, we also apply bootstrapping techniques to calculate confidence intervals for the obtained efficiency and productivity scores in order to obtain more robust results. Results We find that, until the period 2012–2013, the performance of hospitals belonging to different systems experienced a similar trend. However, during the last years of the evaluated period the hospitals operating under the Social Security Fund clearly outperformed Ministry´s hospitals. The main explanation for these divergences seems to be the growth of technological change. Nevertheless, the use of bootstrapping to make statistical inferences reveals that some differences detected may not be significant. Conclusion We demonstrate that the results of traditional DEA and Malmquist index analyses need to be tested for statistical significance. Otherwise, the conclusions reached could be wrong.

La utilización de la técnica del Real Decreto Ley en materia de Seguridad Social: la prolongación de la vida activa de los trabajadores y su conformidad constitucional

The chemical properties of materials are dependent on dynamic changes in their three-dimensional (3D) structure as well as on the reactive environment. We report an in situ 3D imaging study of defect dynamics of a single gold nanocrystal. Our findings offer an insight into its dynamic nanostructure and unravel the formation of a nanotwin network under CO oxidation conditions. In situ/operando defect dynamics imaging paves the way to elucidate chemical processes at the single nano-object level towards defect-engineered nanomaterials.

This paper tests whether competition and collaboration as well as a broad range of other factors lead to improved efficiency in the UK independent school sector. These schools have always operated in a competitive environment, but collaborative groupings are also observed. To answer our main aim, a robust conditional efficiency estimation is employed. Greater efficiency is associated with higher market share, at least at the low levels of market share observed in the sector. There is also some evidence of a positive effect on efficiency of collaboration. Findings regarding the efficiency of schools in the independent sector will be of interest to both independent and state schools as Government policy in the UK has over time encouraged schools in the state sector to become more competitive as an initiative designed to enhance efficiency. The Government is also encouraging greater collaboration between state schools such that they gain benefits from collaboration and sharing of good practice.

Mitochondria are dynamic organelles that change morphology to adapt to cellular energetic demands under both physiological and stress conditions. Cardiomyopathies and neuronal disorders are associated with structure-related dysfunction in mitochondria, but three-dimensional characterizations of the organelles are still lacking. In this study, we combined high-resolution imaging and 3D electron density information provided by cryo-soft X-ray tomography to characterize mitochondria cristae morphology isolated from murine. Using the linear attenuation coefficient, the mitochondria were identified (0.247 ± 0.04 µm−1) presenting average dimensions of 0.90 ± 0.20 µm in length and 0.63 ± 0.12 µm in width. The internal mitochondria structure was successfully identified by reaching up the limit of spatial resolution of 35 nm. The internal mitochondrial membranes invagination (cristae) complexity was calculated by the mitochondrial complexity index (MCI) providing quantitative and morphological information of mitochondria larger than 0.90 mm in length. The segmentation to visualize the cristae invaginations into the mitochondrial matrix was possible in mitochondria with MCI ≥ 7. Altogether, we demonstrated that the MCI is a valuable quantitative morphological parameter to evaluate cristae modelling and can be applied to compare healthy and disease state associated to mitochondria morphology.

BackgroundThe aim of this paper is to analyze the differences in the coordination of chronic illness care between the different public hospital management models coexisting in the Spanish region of Madrid (25 hospitals) during the period 2013–2017.MethodsThe performance of hospitals might be affected by the characteristics of the population they serve and, therefore, this information should be taken into account when estimating efficiency measures. For this purpose, we apply the nonparametric Data Envelopment Analysis (DEA) conditioned to some contextual variables and adapted to a dynamic framework, so that we can assess hospitals during a five-year period. The outputs considered are preventable hospitalizations, readmissions for heart failure and readmissions for chronic obstructive pulmonary disease, whereas the inputs considered are the number of beds, personnel (physicians and other healthcare professionals) and total expenditure on goods and services.ResultsThe results suggest that the level of efficiency demonstrated by the public-private collaboration models of hospital management is higher than traditionally managed hospitals throughout the analyzed period. Nevertheless, we notice that efficiency differences among hospitals are significantly reduced when contextual factors were taken into account.ConclusionsHospitals managed under public-private collaboration models are more efficient than those under traditional management in terms of chronic illness care coordination, being this difference attributable to more agile and flexible management under the collaborative models.

The glycoside hydrolase family 39 (GH39) is a functionally expanding family with limited understanding about the molecular basis for substrate specificity and extremophilicity. In this work, we demonstrate the key role of the positive-subsite region in modulating substrate affinity and how the lack of a C-terminal extension impacts on oligomerization and structural stability of some GH39 members. The crystallographic and SAXS structures of a new GH39 member from the phytopathogen Xanthomonas citri support the importance of an extended C-terminal to promote oligomerization as a molecular strategy to enhance thermal stability. Comparative structural analysis along with site-directed mutagenesis showed that two residues located at the positive-subsite region, Lys166 and Asp167, are critical to substrate affinity and catalytic performance, by inducing local changes in the active site for substrate binding. These findings expand the molecular understanding of the mechanisms involved in substrate recognition and structural stability of the GH39 family, which might be instrumental for biological insights, rational enzyme engineering and utilization in biorefineries.

This paper reports an efficiency analysis of local tax management by provincial tax agencies in Spain based on supramunicipal delegation. To conduct this study, we used the robust order-m conditional model that directly accounts for some socioeconomic environmental variables to estimate the efficiency scores. This is a key issue, as tax agencies do not have control over the context in which they operate, and this may have a severe impact on their performance. Our results suggest that several of the provincial contextual variables accounted for (the net property tax base, population density and inhabitants of the municipalities that have delegated management to the provincial tier of government) have a negative impact on efficiency, especially at higher variable value levels. Considering that the provincial tier of government can opt to set up specific self-governing agencies to perform these tasks, we also applied metafrontier analysis to assess their share in inefficiency. We concluded that the establishment of such self-governing agencies does not lead to higher efficiency levels.

Measuring local government efficiency is a complex task that has to take into account that they usually operate in a heterogeneous context. Therefore, the estimation of relative efficiency measures of their performance needs to account for the effect of contextual and exogenous variables. This should assure that the respective measures adequately reflect the portion of inefficiency that may be attributable to local authorities. In this paper, we apply time-dependent conditional frontier estimators to assess the performance of the 278 Portuguese mainland municipalities for the 2009–2014 period. By applying this nonparametric approach, we can avoid the strong assumptions on the specification of the estimated production function required by traditional two-stage methods. Furthermore, we examine the effect of contextual variables and time on the shape of the frontier and the distribution of inefficiencies. The results reveal that the local reforms do not seem to have achieved the pursued objective of improving the efficiency of local government performance. However, we can observe a small improvement in the last year of the period after the amalgamation of some civil parishes, thus it seems that this specific process might have had a certain impact in the short term. Moreover, we find that the economic and demographic indicators included as contextual variables in our model play an important role as influencing the production set, although those effects do not seem to vary much over time. This evidence is corroborated after conducting a second-stage nonparametric regression of the conditional efficiency measures over those variables.

El envejecimiento de la población mundial es, ante todo, un logro social que se vincula a la mejora en las expectativas de vida de los ciudadanos. El hecho de llegar a cierta edad desemboca, sin embargo, en la salida del mercado laboral de trabajadores que, en principio, se encontrarían en condiciones de proseguir su actividad profesional y de aportar experiencia y conocimientos. Este fenómeno, generalizado en la mayoría de países desarrollados, es aún más relevante en España, donde confluyen simultáneamente dos características que inciden en la toma de decisiones a la hora de fijar una edad ordinaria de jubilación, no siempre tenida en cuenta por las empresas: la baja tasa de natalidad y la segunda mayor esperanza de vida del mundo, que pueden provocar en un futuro más o menos próximo, problemas de financiación en el sistema de Seguridad Social.Este dato se complica aún más si tenemos en cuenta la brecha de género en materia de protección social, que sitúa a las mujeres en una peor posición a causa de su tardía incorporación al mercado laboral y de las frecuentes interrupciones ligadas a la maternidad, que limitan la duración de su carrera profesional y que, en definitiva, culminan en una discriminación que debería ser eliminada.En este estudio se pretende analizar algunas medidas de acción positiva adoptadas en España en los últimos años, tendentes a acabar con estereotipos de género que impiden la plena igualdad entre mujeres y hombres, haciendo también referencia a posibles efectos adversos que de tales actuaciones pudieran derivarse., Le vieillissement de la population mondiale est, tout d´abord, un accomplissement social lié à l´amélioration de l´espérance de vie des citoyens. Le fait d´atteindre un certain âge conduit cependant à la sortie du marché du travail des travailleurs qui, en principe, devraient être en mesure de poursuivre leur activité professionnelle et contribuer à l´expérience et à la connaissance. Ce phénomène, répandu dans la majorité des pays développés, est encore plus pertinent en Espagne, où deux caractéristiques convergent simultanément dans la prise de décision, au moment de fixer un âge ordinaire de retraite, pas toujours pris en compte par les entreprises : le faible taux de natalité et la deuxième plus grande espérance de vie dans lemonde, ce qui peut conduire à des problèmes de financement futurs, dans un avenir plus ou moins proche dans le système de sécurité sociale.C’est encore plus compliqué si l’on tient compte de l´écart entre les sexes, en ce qui concerne la protection sociale, qui désavantage les femmes en raison de leur incorporation tardive sur le marché de travail, et des interruptions fréquentes liées à la maternité, qui limitent la durée de leur carrière professionnelle et aboutissent finalement à une discrimination que devrait être éliminée.Cette étude a pour objectif l’analyse de certaines mesures d´action positive, adoptées en Espagne ces dernières années, qui visent à mettre fin aux stéréotypes sexistes qui entravent l`égalité totale entre les hommes et les femmes, en mentionnant également les éventuels effets néfastes qui pourraient résulter de telles actions., O envelhecimento da população mundial é, antes de tudo, uma conquista social que se vincula à melhoria nas expectativas de vida dos cidadãos. O facto de chegar a uma certa idade desemboca, porém, na saída do mercado laboral de trabalhadores que, em princípio, se encontrariam em condições de seguir com a sua actividade profissional e de contribuir com experiência e conhecimentos. Este fenómeno, generalizado na maioria dos países desenvolvidos, é ainda mais relevante em Espanha, onde confluem simultaneamente duas características que incidem na tomada de decisões na hora de definir uma idade ordinária de aposentadoria, nem sempre tida em conta pelas empresas: a baixa taxa de natalidade e a segunda maior esperança de vida do mundo, que podem provocar num futuro mais ou menos próximo problemas de financiamento no sistema de Segurança Social.Este dado complica-se mais ainda se temos em conta a diferença de género em matéria de protecção social, que situa as mulheres numa pior posição devido à sua tardia incorporação no mercado laboral e das frequentes interrupções ligadas à maternidade, que limitam a duração da sua carreira profissional e que, em definitivo, culminam numa discriminação que deveria ser eliminada.Neste Estudo pretende-se analisar algumas medidas de acção positiva adotadas em Espanha nos últimos anos, tendentes a acabar com estereótipos de género que impedem a igualdade plena entre mulheres e homens, fazendo também referência a possíveis efeitos adversos que de tais actuações pudessem derivar., The ageing of the population worldwide is, above all, a social achievement that links to the improvement in the expectations of citizens. The fact of reaching a certain age ends, however, at the exit from the labour market of workers who, in principle, be able to continue their professional activity and bring experience and knowledge. This phenomenon, widespread in most developed countries, is even more relevant in Spain, where converge simultaneously two features that influence decision-making in an ordinary retirement age not always considered by the companies: the low birth rate and the second highest life expectancy in the world, which can result in the more or less near future funding problems in the social security system. This fact is further complicated if we take into account the gender gap in the field of social protection, which places women in a worse position because of their late incorporation into the labour market and the frequent work interruptions linked to motherhood that limit the length of his professional career and that, ultimately, culminate in a discrimination that should be eliminated. This Study seeks to analyse some positive action measures taken in Spain in recent years, aimed at ending gender stereotypes impeding full equality between men and women, also referring to possible adverse effects arising from such actions, Trayectorias Humanas Trascontinentales, N° 5 | 2019

Halophytic plants can provide an economical and environmentally sustainable source of products for human and animal feeding, in the context of the increase of worldwide emergent semi-arid landscapes. This work reports a comprehensive evaluation of the qualitative and quantitative composition of fatty acids (FA) of nineteen Mediterranean halophytes collected in southern Portugal, with the purpose of establishing their possible uses as food and feed. For FA determination, lipids and free FA were converted to the corresponding fatty acid methyl esters (FAME) and analyzed by GC-MS. Beta maritima had the highest FAME levels (7.3 mg/g DW) while Suaeda vera had the lowest content (1.0 mg/g DW). The most common saturated fatty acid (SFA) across all studied species was palmitic acid. The most prevalent monounsaturated fatty acid (MUFA) was oleic acid. Polyunsaturated fatty acids (PUFA) levels were led by linoleic acid. Less common FAMEs were also detected, namely eicosadienoic and hexadecatrienoic acids. Cotula coronopifolia, Phragmites australis and Suaeda vera displayed the best FA nutritional profiles. These species also showed bioactivities relevant for both human and animal health according to the literature and thus, collectively with this study, they could be further explored as food and feed. Agência financiadora Portuguese Foundation for Science and Technology Portuguese National Budget through the XtremeBio project PTDC/MAR-EST/4346/2012 Portuguese National Budget through GreenVet project ALG-01-0145-FEDER-028876 Portuguese Foundation for Science and Technology SFRH/BD/81425/2011 SFRH/BD/116604/2016 IF/00049/2012 FCT Scientific Employment Stimulus CEECIND/00425/2017 Portuguese National Budget UID/Multi/04326/2019 info:eu-repo/semantics/publishedVersion

This experiment examines the effect of movies portraying stereotypical sex roles on sexual gender stereotypes and rape myths among adolescents and emerging adults (N = 116). After watching a stereotypical sex role movie or a control movie, participants completed a survey on their acceptance of sexual stereotypes and rape myths. Results showed that male and female adolescents who watched the stereotyped movie reported more acceptance of gender stereotypes and rape myths. No effects were found among emerging adults. Social learning theory and media priming are used in discussing these findings. Este experimento investiga el efecto de las películas que representan roles sexuales de género sobre los estereotipos sexuales de género y mitos de la violación de los y las adolescentes, y jóvenes (N = 116). Los y las participantes, tras ver una película representando estereotipos sexuales de género over una película sin dichos estereotipos (grupo de control), completaron un cuestionario sobre suaceptación de estos estereotipos y los mitos de la violación. Los resultados señalaron que, los y las adolescentes que vieron la película estereotípica, mostraron mayor tolerancia hacia estos estereotipos y los mitos de la violación. No se encontraron efectos mediáticos entre los y las jóvenes. Para explicar estos resultados, utilizamos las teorías del aprendizaje social y del efecto de priming.

Adenosine Kinase (ADK) regulates the cellular levels of adenosine (ADO) by fine-tuning its metabolic clearance. The transfer of γ-phosphate from ATP to ADO by ADK involves regulation by the substrates and products, as well as by Mg2+ and inorganic phosphate. Here we present new crystal structures of mouse ADK (mADK) binary (mADK:ADO; 1.2 Å) and ternary (mADK:ADO:ADP; 1.8 Å) complexes. In accordance with the structural demonstration of ADO occupancy of the ATP binding site, kinetic studies confirmed a competitive model of auto-inhibition of ADK by ADO. In the ternary complex, a K+ ion is hexacoordinated between loops adjacent to the ATP binding site, where Asp310 connects the K+ coordination sphere to the ATP binding site through an anion hole structure. Nuclear Magnetic Resonance 2D 15N-1H HSQC experiments revealed that the binding of K+ perturbs Asp310 and residues of adjacent helices 14 and 15, engaging a transition to a catalytically productive structure. Consistent with the structural data, the mutants D310A and D310P are catalytically deficient and loose responsiveness to K+. Saturation Transfer Difference spectra of ATPγS provided evidence for an unfavorable interaction of the mADK D310P mutant for ATP. Reductions in K+ concentration diminish, whereas increases enhance the in vitro activity of mADK (maximum of 2.5-fold; apparent Kd = 10.4 mM). Mechanistically, K+ increases the catalytic turnover (Kcat) but does not affect the affinity of mADK for ADO or ATP. Depletion of intracellular K+ inhibited, while its restoration was accompanied by a full recovery of cellular ADK activity. Together, this novel dataset reveals the molecular basis of the allosteric activation of ADK by K+ and highlights the role of ADK in connecting depletion of intracellular K+ to the regulation of purine metabolism.

Analyzing the efficiency of educational systems is one of the main focuses of the policy debate to promote national competitiveness and future economic growth. In this paper, we assess the performance of secondary schools from 36 countries (26 OECD countries and 10 partners) participating in PISA 2012. For this purpose, we apply a robust conditional nonparametric approach that allows us to incorporate the effect of contextual factors at both school and country level in the estimation of efficiency measures. Our results suggest that there is a greater heterogeneity across countries than across schools. Particularly, we find that differences in efficiency estimates are mainly explained by economic indicators and cultural values. In contrast, some factors previously identified as potential determinants of student achievement, like the existence of tracking or central examinations, do not seem to significantly affect the efficiency of secondary schools.

La presunción de accidente de trabajo en función de la aparición del hecho causante en el lugar y durante la jornada de trabajo prevalece sobre la falta de constatación del nexo causal con el medio de trabajo utilizado

In this study, we investigated the effect of low density lipoprotein receptor (LDLr) deficiency on gap junctional connexin 36 (Cx36) islet content and on the functional and growth response of pancreatic beta-cells in C57BL/6 mice fed a high-fat (HF) diet. After 60 days on regular or HF diet, the metabolic state and morphometric islet parameters of wild-type (WT) and LDLr−/− mice were assessed. HF diet-fed WT animals became obese and hypercholesterolaemic as well as hyperglycaemic, hyperinsulinaemic, glucose intolerant and insulin resistant, characterizing them as prediabetic. Also they showed a significant decrease in beta-cell secretory response to glucose. Overall, LDLr−/− mice displayed greater susceptibility to HF diet as judged by their marked cholesterolaemia, intolerance to glucose and pronounced decrease in glucose-stimulated insulin secretion. HF diet induced similarly in WT and LDLr−/− mice, a significant decrease in Cx36 beta-cell content as revealed by immunoblotting. Prediabetic WT mice displayed marked increase in beta-cell mass mainly due to beta-cell hypertrophy/replication. Nevertheless, HF diet-fed LDLr−/− mice showed no significant changes in beta-cell mass, but lower islet–duct association (neogenesis) and higher beta-cell apoptosis index were seen as compared to controls. The higher metabolic susceptibility to HF diet of LDLr−/− mice may be explained by a deficiency in insulin secretory response to glucose associated with lack of compensatory beta-cell expansion.

Arabinanases (ABNs, EC 3.2.1.99) are promising catalysts for environmentally friendly biomass conversion into energy and chemicals. These enzymes catalyze the hydrolysis of the α-1,5-linked L-arabinofuranoside backbone of plant cell wall arabinans releasing arabino-oligosaccharides and arabinose, the second most abundant pentose in nature. In this work, new findings about the molecular mechanisms governing activation, functional differentiation, and catalysis of GH43 ABNs are presented. Biophysical, mutational, and biochemical studies with the hyperthermostable two-domain endo-acting ABN from Thermotoga petrophila (TpABN) revealed how some GH43 ABNs are activated by calcium ions via hyperpolarization of the catalytically relevant histidine and the importance of the ancillary domain for catalysis and conformational stability. On the other hand, the two GH43 ABNs from rumen metagenome, ARN2 and ARN3, presented a calcium-independent mechanism in which sodium is the most likely substituent for calcium ions. The crystal structure of the two-domain endo-acting ARN2 showed that its ability to efficiently degrade branched substrates is due to a larger catalytic interface with higher accessibility than that observed in other ABNs with preference for linear arabinan. Moreover, crystallographic characterization of the single-domain exo-acting ARN3 indicated that its cleavage pattern producing arabinose is associated with the chemical recognition of the reducing end of the substrate imposed by steric impediments at the aglycone-binding site. By structure-guided rational design, ARN3 was converted into a classical endo enzyme, confirming the role of the extended Arg(203)-Ala(230) loop in determining its action mode. These results reveal novel molecular aspects concerning the functioning of GH43 ABNs and provide new strategies for arabinan degradation.

Orientador: Mário Tyago Murakami Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: A parede celular vegetal é composta por um complexo polissacarídico junto a proteínas estruturais capazes de conter e comprimir todos as organelas citoplasmáticas. Dentre seus componentes estão polímeros tais como a celulose, hemicelulose, pectina e lignina. Para tanto, a natureza usufrui de uma grande diversidade de enzimas especializadas na formação e degradação desses açúcares complexos, as chamadas CAZymes (do inglês, Carbohydrate-Active enZymes) e classificadas em famílias de acordo com sua estrutura primária e enovelamento. As hidrolases glicosídicas (GHs) estão entre as CAZymes mais abundantes, representadas por 135 famílias. Dentre elas, a família GH43, apresenta-se poliespecífica com uma variedade de atividades (ECs), tais como arabinanase (EC 3.2.1.99). Entretanto, são escassos os estudos estruturais a fim de elucidar os mecanismos de degradação e regulação destas enzimas. Seus substratos, as arabinanas, são a segunda pentose mais abundante presente na parede vegetal e tem considerável valor comercial na indústria alimentícia e de degradação da biomassa. Portanto, o presente estudo teve como alvo a caracterização estrutural e bioquímica de duas novas arabinanases, uma endo-?-1,5-L-arabinanase (ARN2) e uma exo-?-1,5-L-arabinanase (ARN3) extraídas da microflora do rúmen bovino e que apresentaram propriedades funcionais únicas em relação as suas homólogas. A resolução das estruturas por cristalografia de raios X revelou o enovelamento canônico da família GH43, 5-fold ?-propeller, com conservação dos resíduos catalíticos que participam de um mecanismo de catálise por inversão do carbono anomérico. Pela caracterização da enzima ARN2, foi possível verificar que sua interface catalítica é extremamente acessível ao solvente e volumosa, explicando assim, sua preferência por substratos ramificados e contrastando com as homólogas de interfaces mais estreitas que permitem principalmente a acomodação de substratos lineares. A enzima ARN3 apresentou, também, características peculiares quanto ao seu mecanismo de ação envolvendo o reconhecimento da extremidade redutora do polímero para liberação de arabinose. Esta região encontra-se parcialmente bloqueada por um longo e divergente loop (R203-A230). Para confirmar o papel desta região no modo de ação da enzima, uma quimera foi desenhada e o longo segmento foi substituto pela sequência SRGEEP, conservada em endo-arabinanases da família. Como predito, a enzima quimérica adquiriu propriedades de uma endo-enzima gerando arabinobiose, majoritariamente. Interessantemente, as análises estruturais permitiram verificar o papel do domínio acessório na estabilização da fenda de ligação ao substrato em enzimas compostas por dois domínios, como ARN2. Em enzimas que compreendem um único domínio, como ARN3, a região é estabilizada por duas cisteínas adjacentes que formam uma ponte dissulfeto. Além disso, foi observado que estas enzimas não possuem um mecanismo regulatório cálcio-dependente, um comportamento inédito na família GH43. Por fim, este trabalho contribuiu coletivamente com novas estratégias para degradação da fração hemicelulósica/péctica da biomassa vegetal e expande o atual conhecimento a respeito da família GH43. Dentro dos elementos pós-textuais desta tese está o artigo I, referente aos resultados obtidos na tese, além dos artigos II e III que consistem de colaborações. No anexo IV consta um breve resumo a respeito dos trabalhos realizados durante o doutorado sanduiche na VTT Technical Research Centre of Finland Abstract: The plant cell wall is composed by a polysaccharide complex together with structural proteins, which are able to hold and compress all the cytosolic organelles. Among these components, there are polymers such as cellulose, hemicellulose, pectin and lignin. To do so, the nature employs a wide range of specialized enzymes in the synthesis and degradation of these complex sugars, known as CAZymes (Carbohydrate-Active enZymes). Glycosyl hydrolases are among the most abundant CAZymes, represented by 135 families. The GH43 family is a polyspecific group of enzymes and contains a variety of ECs activities, for example that ones with arabinanase activity (EC 3.2.1.99), with application in different industrial fields. However, structural studies in order to elucidate the mechanisms underlying their regulation and mode of operation are still scarce. Their substrates, arabinans, are the second most abundant pentose in the plant cell wall, possessing considerable commercial value in food industry and biomass conversion. Therefore, the present study aimed the structural and biochemical characterization of two novel arabinanases, an endo-?-1,5-L-arabinanase (ARN2) and exo-?-1,5-L-arabinanase (ARN3) retrieved from the bovine rumen microflora and, which presented unique functional properties compared to their homologues. Their structures, solved by X-ray crystallography, revealed a canonical fold of the GH43 family, the 5-fold ?-propeller conserving the classical inversion mechanism that involves two conserved glutamic acid residues. Trough ARN2 characterization, it was possible to verify an extremely solvent accessible and large catalytic interface, explaining its preference to branched substrates and, which contrasts with the narrow interface of homologues that favors the binding of linear substrates. ARN3 also presented unusual features in its mode of action involving the recognition of the reducing end of the substrate and the release of arabinose. This region is found partially blocked by a long and divergent loop (R203-A230). To confirm the role of this region in the mode of action, a chimera was designed and produced and the long segment was substituted by the SRGEEP sequence that is conserved in classical GH43 endo-arabinanases. As predicted, the chimera acquired endo-enzyme properties, generating arabinobiose, mainly, as a final product. Interestingly, structural analyses also allowed to verify the role of the accessory domain in the stabilization of the catalytic cleft in two-domain enzymes such as ARN2. In single domain enzymes, such as ARN3, this region is stabilized by a pair of cysteines forming a disulfide bond. Furthermore, the ruminal enzymes showed a calcium-independent regulatory mechanism, an unheard behavior in the GH43 family. Finally, this work contributed collectively to new strategies to the hemicellulose/pectin degradation and expanded the current knowledge about GH43 family. In the post-text elements, there are the manuscript (I) related to the results presented in this thesis and the articles II and III from collaborative works. The final annex (IV) describes the work developed during the sandwich PhD at VTT Technical Research Centre of Finland Doutorado Bioquímica Doutora em Biologia Funcional e Molecular FAPESP 2012/09119-8

As mammals are unable to synthesize essential polyunsaturated fatty acids (PUFA), these compounds need to be taken in through diet. Nowadays, obtaining essential PUFA in diet is becoming increasingly difficult; therefore this work investigated the suitability of using macroalgae as novel dietary sources of PUFA. Hence, 17 macroalgal species from three different phyla (Chlorophyta, Phaeophyta and Rhodophyta) were analyzed and their fatty acid methyl esters (FAME) profile was assessed. Each phylum presented a characteristic fatty acid signature as evidenced by clustering of PUFA profiles of algae belonging to the same phylum in a Principal Components Analysis. The major PUFA detected in all phyla were C-18 and C-20, namely linoleic, arachidonic and eicosapentaenoic acids. The obtained data showed that rhodophytes and phaeophytes have higher concentrations of PUFA, particularly from the n-3 series, thereby being a better source of these compounds. Moreover, rhodophytes and phaeophytes presented. healthier. Sigma(n-6/)Sigma(n-3) and PUFA/saturated fatty acid ratios than chlorophytes. Ulva was an exception within the Chlorophyta, as it presented high concentrations of n-3 PUFA, alpha-linolenic acid in particular. In conclusion, macroalgae can be considered as a potential source for large-scale production of essential PUFA with wide applications in the nutraceutical and pharmacological industries. SEABIOMED [PTDC/MAR/103957/2008]; Foundation for Science and Technology (FCT); Portuguese National Budget; FCT [SFRH/BPD/65116/2009, SFRH/BD/81425/2011]

El articulo trata sobre la futura legislacion de los servicios sociales de Castilla y Leon. Para ello se realiza una exposicion donde se reflexiona sobre la concepcion de necesidad social ligada a la idea de servicio social. Se presenta el informe previo del Consejo Economico y Social de 26 de Junio de 2009 sobre el Anteproyecto de Ley. Para a continuacion llevar a cabo un analisis juridico comparativo a partir del borrador de anteproyecto de ley de servicios sociales y atencion a la dependencia de 7 de enero de 2009 y de la ley 18/1988, de 28 de diciembre, de accion social y servicios sociales.