Purificação e caracterização parcial de uma Fucana C de Dictyota mentrualis e estudo do seu efeito antiinflamatório e nociceptivo

In recent years, sulfated polysaccharides from marine algae have emerged as an important class of natural biopolymers with potential application in human and veterinary health care, while taking advantage of the absence of potential risk of contamination by animal viruses. Among these, fucans isolated from the cell walls of marine brown alga have been study due to their anticoagulant, antithrombotic, anti-inflammatory and antiviral activities. These biological effects of fucans have been found to depend on the degree of sulfation and molecular size of the polysaccharide chains. In the present study, we examined structural features of a fucan extracted from brown alga Dictyota menstrualis and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolytic digestion, followed by sequential acetone precipitation producing 5 fractions. Gel lectrophoresis using 0.05 M 1,3-diaminopropane-acetate buffer, pH 9.0, stained with 0.1% toluidine blue, showed the presence of sulfated polysaccharides in all fractions. The chemical analyses demonstrated that all fractions are composed mainly of fucose, xylose, galactose, uronic acid, and sulfate. Electrophoresis in agarose gel in three different buffers demonstrated that the fraction 2.0v have only one population of fucan. This compound was purify by exclusion molecular. It has shown composition of fucose, xilose, sulfate and uronic acid in molar ration of 1.0: 1.7: 1.1: 0.5 respectively. The effect of this heterofucan on the leukocyte migration was observed 6h after zymozan (mg/g) administration into the peritoneum. The heterofucan showed higher antimigratory activity, it decrease the migration of leukocyte in 83.77% to peritoneum. The results suggest that this fucan is a new antimigratory compound with potential pharmacological appications

Cogestion des ressources naturelles : une approche structurale pour quantifier la contribution des réseaux d'acteurs à la résilience des systèmes socio-écologiques

Alors que les activités anthropiques font basculer de nombreux écosystèmes vers des régimes fonctionnels différents, la résilience des systèmes socio-écologiques devient un problème pressant. Des acteurs locaux, impliqués dans une grande diversité de groupes — allant d’initiatives locales et indépendantes à de grandes institutions formelles — peuvent agir sur ces questions en collaborant au développement, à la promotion ou à l’implantation de pratiques plus en accord avec ce que l’environnement peut fournir. De ces collaborations répétées émergent des réseaux complexes, et il a été montré que la topologie de ces réseaux peut améliorer la résilience des systèmes socio-écologiques (SSÉ) auxquels ils participent. La topologie des réseaux d’acteurs favorisant la résilience de leur SSÉ est caractérisée par une combinaison de plusieurs facteurs : la structure doit être modulaire afin d’aider les différents groupes à développer et proposer des solutions à la fois plus innovantes (en réduisant l’homogénéisation du réseau), et plus proches de leurs intérêts propres ; elle doit être bien connectée et facilement synchronisable afin de faciliter les consensus, d’augmenter le capital social, ainsi que la capacité d’apprentissage ; enfin, elle doit être robuste, afin d’éviter que les deux premières caractéristiques ne souffrent du retrait volontaire ou de la mise à l’écart de certains acteurs. Ces caractéristiques, qui sont relativement intuitives à la fois conceptuellement et dans leur application mathématique, sont souvent employées séparément pour analyser les qualités structurales de réseaux d’acteurs empiriques. Cependant, certaines sont, par nature, incompatibles entre elles. Par exemple, le degré de modularité d’un réseau ne peut pas augmenter au même rythme que sa connectivité, et cette dernière ne peut pas être améliorée tout en améliorant sa robustesse. Cet obstacle rend difficile la création d’une mesure globale, car le niveau auquel le réseau des acteurs contribue à améliorer la résilience du SSÉ ne peut pas être la simple addition des caractéristiques citées, mais plutôt le résultat d’un compromis subtil entre celles-ci. Le travail présenté ici a pour objectifs (1), d’explorer les compromis entre ces caractéristiques ; (2) de proposer une mesure du degré auquel un réseau empirique d’acteurs contribue à la résilience de son SSÉ ; et (3) d’analyser un réseau empirique à la lumière, entre autres, de ces qualités structurales. Cette thèse s’articule autour d’une introduction et de quatre chapitres numérotés de 2 à 5. Le chapitre 2 est une revue de la littérature sur la résilience des SSÉ. Il identifie une série de caractéristiques structurales (ainsi que les mesures de réseaux qui leur correspondent) liées à l’amélioration de la résilience dans les SSÉ. Le chapitre 3 est une étude de cas sur la péninsule d’Eyre, une région rurale d’Australie-Méridionale où l’occupation du sol, ainsi que les changements climatiques, contribuent à l’érosion de la biodiversité. Pour cette étude de cas, des travaux de terrain ont été effectués en 2010 et 2011 durant lesquels une série d’entrevues a permis de créer une liste des acteurs de la cogestion de la biodiversité sur la péninsule. Les données collectées ont été utilisées pour le développement d’un questionnaire en ligne permettant de documenter les interactions entre ces acteurs. Ces deux étapes ont permis la reconstitution d’un réseau pondéré et dirigé de 129 acteurs individuels et 1180 relations. Le chapitre 4 décrit une méthodologie pour mesurer le degré auquel un réseau d’acteurs participe à la résilience du SSÉ dans lequel il est inclus. La méthode s’articule en deux étapes : premièrement, un algorithme d’optimisation (recuit simulé) est utilisé pour fabriquer un archétype semi-aléatoire correspondant à un compromis entre des niveaux élevés de modularité, de connectivité et de robustesse. Deuxièmement, un réseau empirique (comme celui de la péninsule d’Eyre) est comparé au réseau archétypique par le biais d’une mesure de distance structurelle. Plus la distance est courte, et plus le réseau empirique est proche de sa configuration optimale. La cinquième et dernier chapitre est une amélioration de l’algorithme de recuit simulé utilisé dans le chapitre 4. Comme il est d’usage pour ce genre d’algorithmes, le recuit simulé utilisé projetait les dimensions du problème multiobjectif dans une seule dimension (sous la forme d’une moyenne pondérée). Si cette technique donne de très bons résultats ponctuellement, elle n’autorise la production que d’une seule solution parmi la multitude de compromis possibles entre les différents objectifs. Afin de mieux explorer ces compromis, nous proposons un algorithme de recuit simulé multiobjectifs qui, plutôt que d’optimiser une seule solution, optimise une surface multidimensionnelle de solutions. Cette étude, qui se concentre sur la partie sociale des systèmes socio-écologiques, améliore notre compréhension des structures actorielles qui contribuent à la résilience des SSÉ. Elle montre que si certaines caractéristiques profitables à la résilience sont incompatibles (modularité et connectivité, ou — dans une moindre mesure — connectivité et robustesse), d’autres sont plus facilement conciliables (connectivité et synchronisabilité, ou — dans une moindre mesure — modularité et robustesse). Elle fournit également une méthode intuitive pour mesurer quantitativement des réseaux d’acteurs empiriques, et ouvre ainsi la voie vers, par exemple, des comparaisons d’études de cas, ou des suivis — dans le temps — de réseaux d’acteurs. De plus, cette thèse inclut une étude de cas qui fait la lumière sur l’importance de certains groupes institutionnels pour la coordination des collaborations et des échanges de connaissances entre des acteurs aux intérêts potentiellement divergents.

As rodovias e as aves de rapina nocturnas relação entre a densidade, o habitat e a mortalidade

As estradas e tráfego inerente surgem como a criação antrópica mais conspícua e penetrante na paisagem natural, sendo considerados os principais agentes causadores de fragmentação e destruição de habitats, assim como representam um obstáculo físico sem precedentes, limitando as relações directas entre os indivíduos, por diminuição da frequência de dispersão e aumento da mortalidade por atropelamento, impedindo o fluxo natural de genes e suscitando o aumento de fenómenos de inbreeding e perda de heterozigotia. Todos os impactes deletérios associados às rodovias são claramente perceptíveis em vertebrados, onde as aves de rapinas nocturnas não são excepção. Uma vez que estas rapaces beneficiam das suas bermas e orlas, como locais de poiso, nidificação ou como corredores de dispersão através da paisagem, são frequentemente vítimas de mortalidade por atropelamento em estradas, sendo esta problemática considerada actualmente uma das mais recentes e importantes formas de mortalidade não natural em rapinas nocturnas e vinculada como um dos maiores problemas de conservação que afecta este grupo. Não obstante, esse mútuo efeito de atracção/repulsa das estradas a estas rapaces, as rodovias criam uma barreira específica que limita a dinâmica, comportamento e densidade populacional das espécies residentes, reconhecendo-se que o isolamento daí resultante, pode comprometer a viabilidade populacional a longo prazo, podendo mesmo conduzir a altos riscos de extinção das populações locais devido a efeitos estocásticos. Mediante esta problemática, este trabalho debruçou-se sobre um único objectivo principal: a avaliação do impacte das rodovias e do tráfego, na densidade das aves de rapina nocturnas. Este estudo foi efectuado na região Alentejana, abrangendo uma área de cariz tipicamente mediterrânico, delimitada pelas localidades de Montemor-o-Novo, Arraiolos e Évora, sendo seccionada por 143 quilómetros de estradas, divididas em autoestrada, rodovias com elevada e reduzida densidade de tráfego. A monitorização das rapinas nocturnas foi conduzida em dois anos amostrais (2005 e 2007), tendo sido focalizada sobretudo em duas espécies de Strigiformes, a Coruja­ do-mato Strix aluco e o Mocho-galego Athene noctua, recorrendo ao uso de playbacks com reprodução de vocalizações de indivíduos conspecíficos. Foram usadas 32 variáveis explicativas integradas em três grandes grupos: variáveis de estrada, métricas da paisagem, uso do solo, tendo sido analiticamente testadas, recorrendo à aplicação de Modelos Lineares Generalizados. Os principais resultados obtidos demonstram que as variáveis de estrada, aliadas à densidade de tráfego e ruído inerente à sua circulação, são provavelmente, responsáveis por um comportamento de repulsa das espécies de aves de rapina nocturnas em estudo, apresentando estas densidades mais elevadas longe de áreas antropicamente perturbadas e, portanto, de menor qualidade que se encontram adjacentes às rodovias. Todavia a presença de habitat favorável a estas rapaces é provavelmente o descritor com maior poder estatístico no que concerne à sua distribuição e densidade, sendo os montados densos e a presença de zonas agrícolas de sequeiro, positivamente correlacionadas, respectivamente com a densidade de casais reprodutores de Coruja-do-mato e Mocho-galego. Mediante a observação dos resultados será veemente a aplicação de medidas de mitigação específicas, que fundamentalmente considerem o afastamento dos efectivos populacionais longe das estradas e tráfego, conservando e assegurando as características estruturais, requisitos e qualidade dos habitats, de modo a incrementar e garantir a viabilidade e densidade das populações, fidelizando a territorialidade e permanência destas aves nestas áreas. Adicionalmente deverá investir-se na conectividade das manchas de paisagem fragmentada pelas rodovias, criando opções de conservação estratégicas, em zonas ecologicamente mais sensíveis, que não somente minimizem o efeito de repulsa reconhecido nestas aves, mas também os níveis de mortalidade por atropelamento, tornando os ecossistemas mais funcionais para a sobrevivência destes rapaces. ABSTRACT; Roads and traffic are the most conspicuous and pervasive human creation, being the great responsible for fragmentation and habitats destruction, reducing animal movement through landscape, which implies decrease of gene flow and loss of variability that can fragmented populations, thus reducing their sizes and densities. All deleterious impacts associated with roads are clearly visible in vertebrates, where owls aren't exception, being frequent victims of road mortality, since they can use roadside habitats and edges for hunting, nesting or dispersal corridors through the landscape, being nowadays one of the most recent and important causes of non­natural mortality in owls and has been recognized as one of the largest conservation problems affecting this group. However, the attractive and avoidance effect of roads and his edges on owls creates a barrier effect that limits dynamics, behaviour and breeding density of resident species, recognizing that possible isolation, could compromise populations survival, make them more vulnerable to high risks of local extinction due to stochastic effects. Despite that, several authors suggest that owls use roads to hunt, as marginal habitats, or for navigation corridors through the landscape ln this context, the major aims of this study was to verify if there are negative effects of roads on the density of owls, considering traffic as an influencing factor. This study was conducted in Alentejo, covering a typical Mediterranean area, including three main localities Montemor-o-Novo, Évora and Arraiolos, being sectioned by 143 km of roads, including highway, roads with high traffic density, and the remaining with low traffic density. The owl census was conducted in two sampling years (2005 and 2007) and was focused mainly on Little owl Athene noctua and Tawny Owl Strix aluco species, using the playback technique, with conspecific calls. We used 32 explanatory variables, mainly included in three groups: road variables, landscape metrics and land use, having been analytically tested, with application of Generalised Linear Models. The main results show that noisy roads with high traffic density are probably the most responsible for the avoidance behaviour of owls, under the study area, showing density depression near high anthropogenic disturbed areas adjacent to roads. However, the presence of habitat quality to these birds is probably the descriptor with greater statistical power, considering its distribution and density, with the dense oak woodland and croplands and arable lands, positively correlated, respectively with Tawny owl and Little owl density. ln consequence, the great conservation effort should be done in order to keep breeding populations away from roads and traffic, ensuring the structural features, requirements and quality of its habitats in order to enhance and ensure the viability and density of owl's populations in these areas. ln addition, it is important to invest in connectivity between roadside fragmented patches, creating strategic conservation options, in sensitive areas, which minimize the avoidance effect, recognized in owls, but also road-kill levels, making ecosystems more functional to survival of these top predators.

Evaluation of the conductor-like screening model for real solvents for the prediction of the water activity coefficient at infinite dilution in ionic liquids

Ionic liquids (ILs) have attracted great attention, from both industry and academia, as alternative fluids for very different types of applications. The large number of cations and anions allow a wide range of physical and chemical characteristics to be designed. However, the exhaustive measurement of all these systems is impractical, thus requiring the use of a predictive model for their study. In this work, the predictive capability of the conductor-like screening model for real solvents (COSMO-RS), a model based on unimolecular quantum chemistry calculations, was evaluated for the prediction water activity coefficient at infinite dilution, gamma(infinity)(w), in several classes of ILs. A critical evaluation of the experimental and predicted data using COSMO-RS was carried out. The global average relative deviation was found to be 27.2%, indicating that the model presents a satisfactory prediction ability to estimate gamma(infinity)(w) in a broad range of ILs. The results also showed that the basicity of the ILs anions plays an important role in their interaction with water, and it considerably determines the enthalpic behavior of the binary mixtures composed by Its and water. Concerning the cation effect, it is possible to state that generally gamma(infinity)(w) increases with the cation size, but it is shown that the cation-anion interaction strength is also important and is strongly correlated to the anion ability to interact with water. The results here reported are relevant in the understanding of ILs-water interactions and the impact of the various structural features of its on the gamma(infinity)(w) as these allow the development of guidelines for the choice of the most suitable lLs with enhanced interaction with water.

L’alerte éthique : vers une gouvernance omnicanal ?

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Síntese química e avaliação das propriedades antibacterianas e antiparasitárias de análogos de peptídeos antimicrobianos de anuros

Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Molecular, 2015.

CONDICIONES INICIALES PARA UNA POLÍTICA DE DESARROLLO Y DEL MEDIO AMBIENTE. Un enfoque a partir de los derechos concretos a la vida

Resumen En este ensayo se pretende analizar, críticamente, un aspecto que consideramos crucial en toda propuesta de desarrollo, ya sea que éste se entienda como “desarrollo económico”, como “desarrollo humano sostenible”, o como desarrollo a secas: los criterios formales de decisión y sus correspondientes marcos categoriales. Estos criterios se insertan necesariamente en un sistema de coordinación del trabajo social, y condicionan los fines y las metas de la acción. Además, lo hacemos teniendo especialmente en cuenta aquellos rasgos estructurales del subdesarrollo capitalista que creemos necesario enfrentar y superar en toda propuesta de desarrollo: el desempleo, las desigualdades sociales y regionales, la exclusión social y la destrucción del medio ambiente. El análisis presupone una imagen del ser humano que concibe a éste como un sujeto de derechos concretos a la vida, imagen que parte del trabajo humano en el conjunto de la división social del trabajo, y por tanto, de un sujeto en comunidad. Adjudica al ser humano determinados derechos a la vida que tienen que impregnar a la sociedad entera para que pueda ser realmente una sociedad libre.   Abstract This paper attempts analyze, in a critical way, a crucial issue concerning every development proposal: the formal criteria of decision and their respective theoretical frameworks, either we talk about economic development, human development or just development. These criteria are necessarily inserted in a social labor coordination system and they determine those ends and goals of the human action. Moreover, we emphasize those structural features from capitalist underdeveloped countries that are urgent to face and overcome: unemployment, social and regional inequalities, social exclusion and environment destruction. Analysis we do presupposes the human being conceived as a person with concrete rights of live, as from the human labor in the whole of social labor division, and therefore, as a person in a community. Every society, in order to be a free society, must to assign these concrete rights of live to every human being.

Crystal structure of Bn IV in complex with myristic acid: A Lys49 myotoxic phospholipase A(2) from Bothrops neuwiedi venom

The LY549-PLA(2)s myotoxins have attracted attention as models for the induction of myonecrosis by a catalytically independent mechanism of action. Structural studies and biological activities have demonstrated that the myotoxic activity of LYS49-PLA(2) is independent of the catalytic activity site. The myotoxic effect is conventionally thought to be to due to the C-terminal region 111-121, which plays an effective role in membrane damage. In the present study, Bn IV LYS49-PLA(2) was isolated from Bothrops neuwiedi snake venom in complex with myristic acid (CH3(CH2)(12)COOH) and its overall structure was refined at 2.2 angstrom resolution. The Bn IV crystals belong to monoclinic space group P2(1) and contain a dimer in the asymmetric unit. The unit cell parameters are a = 38.8, b = 70.4, c = 44.0 angstrom. The biological assembly is a "conventional dimer" and the results confirm that dimer formation is not relevant to the myotoxic activity. Electron density map analysis of the Bn IV structure shows clearly the presence of myristic acid in catalytic site. The relevant structural features for myotoxic activity are located in the C-terminal region and the Bn IV C-terminal residues NKKYRY are a probable heparin binding domain. These findings indicate that the mechanism of interaction between Bn IV and muscle cell membranes is through some kind of cell signal transduction mediated by heparin complexes. (C) 2010 Elsevier Masson SAS. All rights reserved.

Design, Fabrication, and Mechanical Property Analysis of 3D Nanoarchitected Materials

Recent developments in micro- and nanoscale 3D fabrication techniques have enabled the creation of materials with a controllable nanoarchitecture that can have structural features spanning 5 orders of magnitude from tens of nanometers to millimeters. These fabrication methods in conjunction with nanomaterial processing techniques permit a nearly unbounded design space through which new combinations of nanomaterials and architecture can be realized. In the course of this work, we designed, fabricated, and mechanically analyzed a wide range of nanoarchitected materials in the form of nanolattices made from polymer, composite, and hollow ceramic beams. Using a combination of two-photon lithography and atomic layer deposition, we fabricated samples with periodic and hierarchical architectures spanning densities over 4 orders of magnitude from ρ=0.3-300kg/m³ and with features as small as 5nm. Uniaxial compression and cyclic loading tests performed on different nanolattice topologies revealed a range of novel mechanical properties: the constituent nanoceramics used here have size-enhanced strengths that approach the theoretical limit of materials strength; hollow aluminum oxide (Al₂O₃) nanolattices exhibited ductile-like deformation and recovered nearly completely after compression to 50% strain when their wall thicknesses were reduced below 20nm due to the activation of shell buckling; hierarchical nanolattices exhibited enhanced recoverability and a near linear scaling of strength and stiffness with relative density, with E∝ρ^1.04 and σy∝ρ^1.17 for hollow Al₂O₃ samples; periodic rigid and non-rigid nanolattice topologies were tested and showed a nearly uniform scaling of strength and stiffness with relative density, marking a significant deviation from traditional theories on “bending” and “stretching” dominated cellular solids; and the mechanical behavior across all topologies was highly tunable and was observed to strongly correlate with the slenderness λ and the wall thickness-to-radius ratio t/a of the beams. These results demonstrate the potential of nanoarchitected materials to create new highly tunable mechanical metamaterials with previously unattainable properties.

Explanation, Dialogue and Default Logic

La distinción entre argumentación y explicación es una tarea complicada pero necesaria por diversas razones. Una de ellas es la necesidad de incorporar la explicación en un movimiento del diálogo como resultado de una obligación dialéctica. Se propusieron distintos sistemas de diálogo que exploran la distinción enfatizando aspectos pragmáticos. En el presente trabajo me ocupo de aspectos estructurales de la explicación analizados en el marco de la lógica por defecto que permite caracterizar ciertas objeciones en el diálogo. Asimismo, considero que la versión operacional de la lógica por defecto constituye una aproximaciónadecuada en la construcción de la explicación y en la representación de la instancia de diálogo en el intercambio dialéctico

Explanation, Dialogue and Default Logic

La distinción entre argumentación y explicación es una tarea complicada pero necesaria por diversas razones. Una de ellas es la necesidad de incorporar la explicación en un movimiento del diálogo como resultado de una obligación dialéctica. Se propusieron distintos sistemas de diálogo que exploran la distinción enfatizando aspectos pragmáticos. En el presente trabajo me ocupo de aspectos estructurales de la explicación analizados en el marco de la lógica por defecto que permite caracterizar ciertas objeciones en el diálogo. Asimismo, considero que la versión operacional de la lógica por defecto constituye una aproximaciónadecuada en la construcción de la explicación y en la representación de la instancia de diálogo en el intercambio dialéctico

Integrated Omics for the global mapping of biomolecules in LDL

Circulating low density lipoproteins (LDL) are thought to play a crucial role in the onset and development of atherosclerosis, though the detailed molecular mechanisms responsible for their biological effects remain controversial. The complexity of biomolecules (lipids, glycans and protein) and structural features (isoforms and chemical modifications) found in LDL particles hampers the complete understanding of the mechanism underlying its atherogenicity. For this reason the screening of LDL for features discriminative of a particular pathology in search of biomarkers is of high importance. Three major biomolecule classes (lipids, protein and glycans) in LDL particles were screened using mass spectrometry coupled to liquid chromatography. Dual-polarity screening resulted in good lipidome coverage, identifying over 300 lipid species from 12 lipid sub-classes. Multivariate analysis was used to investigate potential discriminators in the individual lipid sub-classes for different study groups (age, gender, pathology). Additionally, the high protein sequence coverage of ApoB-100 routinely achieved (≥70%) assisted in the search for protein modifications correlating to aging and pathology. The large size and complexity of the datasets required the use of chemometric methods (Partial Least Square-Discriminant Analysis, PLS-DA) for their analysis and for the identification of ions that discriminate between study groups. The peptide profile from enzymatically digested ApoB-100 can be correlated with the high structural complexity of lipids associated with ApoB-100 using exploratory data analysis. In addition, using targeted scanning modes, glycosylation sites within neutral and acidic sugar residues in ApoB-100 are also being explored. Together or individually, knowledge of the profiles and modifications of the major biomolecules in LDL particles will contribute towards an in-depth understanding, will help to map the structural features that contribute to the atherogenicity of LDL, and may allow identification of reliable, pathology-specific biomarkers. This research was supported by a Marie Curie Intra-European Fellowship within the 7th European Community Framework Program (IEF 255076). Work of A. Rudnitskaya was supported by Portuguese Science and Technology Foundation, through the European Social Fund (ESF) and "Programa Operacional Potencial Humano - POPH".

El teatro cómico de Enrique García Álvarez

Enrique García Álvarez (Madrid, 1883-1931) belonged to the group of writers who stood out in the late nineteenth century and early twentieth in what was known as “género chico”, label used to oppose this kind of theater to the dramaturgy considered serious. It was composed of brief and comic works whose intention was to generate a fun time for the spectators. Our author was one of the leading representatives and his numerous creations appeared profusely in the posters of the theaters per hour. In the twentieth century, he adapted to the new forms imposed on the scene. Despite his enormous fame at the time, nowadays Enrique García Álvarez remains completely unknown, not only in the eyes of the general public but also to the more knowledgeable circles of our literature. Studies of his dramatic work are almost nonexistent and basically limited to their collaborations with other authors who had more luck as Arniches or Muñoz Seca. The purpose of this thesis is, therefore, to complete a thorough study of the figure and production of Enrique García Álvarez, as well as its impact on the literature. The opening chapter is a reconstruction of the life and work of the author. In the following chapters, we proceeded to the analysis of structural features such as theme, time, space, and characters of his dramatic world. At the end of these pages, we have included a photographic appendix that illustrates some moments of his personal and professional career. We have also developed appendices to classify his librettos chronologically, by collaborator and by genre...

Isolation and characterization of plastid terminal oxidase gene from carrot and its relation to carotenoid accumulation

Carrot (Daucus carota L.) is a biennial plant that accumulates considerable amounts of carotenoid pigments in the storage root. To better understand the molecular mechanisms for carotenoid accumulation in developing storage roots, plastid terminal oxidase (PTOX) cDNA was isolated and selected for reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Present in photosynthetic species, PTOX is a plastid-located, nucleus encoded plastoquinone (PQ)-O2 oxidoreductase (plastioquinol oxidase). The enzyme is known to play a role as a cofactor for phytoene desaturase, and consequently plays a key role in the carotenoid biosynthesis pathway. A single PTOX gene was identified (DcPTOX) in carrot. DcPTOX encodes a putative protein with 366 amino acids that contains the typical structural features of PTOXs from higher plants. The expression of DcPTOX was analysed during the development of white, yellow, orange, red, and purple carrot roots, along with five genes known to be involved in the carotenoid biosynthesis pathway, PSY2, PDS, ZDS1, LCYB1, and LCYE. Expression analysis revealed the presence of DcPTOX transcripts in all cultivars, and an increase of transcripts during the time course of the experiment, with differential expression among cultivars in early stages of root growth. Our results demonstrated that DcPTOX showed a similar profile to that of other carotenoid biosynthetic genes with high correlation to all of them. The preponderant role of PSY in the biosynthesis of carotenoid pigments was also confirmed.

Structural and cellular features in metaphyseal and diaphyseal periosteum of osteoporotic rats

Despite the important physiological role of periosteum in the pathogenesis and treatment of osteoporosis, little is known about the structural and cellular characteristics of periosteum in osteoporosis. To study the structural and cellular differences in both diaphyseal and metaphyseal periosteum of osteoporotic rats, samples from the right femur of osteoporotic and normal female Lewis rats were collected and tissue sections were stained with hematoxylin and eosin, antibodies or staining kit against tartrate resistant acid phosphatase (TRAP), alkaline phosphatase (ALP), vascular endothelial growth factor (VEGF), von Willebrand (vWF), tyrosine hydroxylase (TH) and calcitonin gene-related peptide (CGRP). The results showed that the osteoporotic rats had much thicker and more cellular cambial layer of metaphyseal periosteum compared with other periosteal areas and normal rats (P\0.001). The number of TRAP? osteoclasts in bone resorption pits, VEGF? cells and the degree of vascularization were found to be greater in the cambial layer of metaphyseal periosteum of osteoporotic rats (P\0.05), while no significant difference was detected in the number of ALP? cells between the two groups. Sympathetic nerve fibers identified by TH staining were predominantly located in the cambial layer of metaphyseal periosteum of osteoporotic rats. No obvious difference in the expression of CGRP between the two groups was found. In conclusion, periosteum may play an important role in the cortical bone resorption in osteoporotic rats and this pathological process may be regulated by the sympathetic nervous system.