Role of HuR in skeletal myogenesis through coordinate regulation of muscle differentiation genes

In this report, we investigate the role of the RNA-binding protein HuR during skeletal myogenesis. At the onset of myogenesis in differentiating C2C12 myocytes and in vivo in regenerating mouse muscle, HuR cytoplasmic abundance increased dramatically, returning to a predominantly nuclear presence upon completion of myogenesis. mRNAs encoding key regulators of myogenesis-specific transcription (myogenin and MyoD) and cell cycle withdrawal (p21), bearing AU-rich regions, were found to be targets of HuR in a differentiation-dependent manner. Accordingly, mRNA half-lives were highest during differentiation, declining when differentiation was completed. Importantly, HuR-overexpressing C2C12 cells displayed increased target mRNA expression and half-life and underwent precocious differentiation. Our findings underscore a critical function for HuR during skeletal myogenesis linked to HuR's coordinate regulation of muscle differentiation genes.

Caracterização molecular de três espécies de Trachycephalus (Anura: Hylidae) : investigando potenciais híbridos interespecíficos

Animais híbridos representam um desafio à taxonomia e sistemática, pois correspondem a unidades evolutivas geralmente sem clara delimitação morfológica, comportamental e genética. Híbridos podem ser morfologicamente intermediários aos parentais ou, devido à introgressão e retrocruzamentos, suas características podem se misturar tornando difícil sua identificação. Uma das formas de identificação de híbridos é por meio de ferramentas de biologia molecular, que ao utilizarem marcadores de DNA mitocondrial (herança exclusiva materna) e DNA nuclear (herança materna e paterna), permitem a comparação entre informações genéticas. Além da hibridização existem outras fontes de conflito entre dados moleculares provenientes do DNA mitocondrial e DNA nuclear, como por exemplo a retenção de polimorfismos ancentrais. Em localidades do Espírito Santo, Brasil, foram coletados indivíduos de morfologia distinta de Trachycephalus mesophaeus e T. nigromaculatus, que são as únicas espécies do gênero conhecidas nesse estado. Porém, estudos piloto usando o gene mitocondrial Citocromo Oxidase subunidade I (COI) agruparam esses espécimes com amostras de T. typhonius. Devido a estas incongruências, foram sequenciados fragmentos de dois genes mitocondriais - COI e Nicotinamida Desidrogenase subunidade 2 (ND2) e um exon nuclear (tirosinase) de 173 indivíduos de Trachycephalus, de forma a esclarecer as identificações taxonômicas e investigar a correspondência entre caracteres morfológicos e genéticos nesta linhagem, na sua área de ocorrência As filogenias moleculares, divergências genéticas, redes de haplótipos e polimorfismos de nucleotídeos únicos (SNPs) confirmaram as três espécies acima mencionadas como linhagens evolutivas distintas e revelaram mais sete indivíduos potencialmente híbridos, mas morfologicamente assinalados a T. mesophaeus, T. nigromaculatus ou T. typhonius.. Devido à taxa de evolução lenta da tirosinase, as espécies mais recentes T. typhonius e T. nigromaculatus parecem não terem sido sorteadas completamente nesse gene. Já T. mesophaeus, que é a espécie mais antiga das três, foi recuperada inequivocamente em todas as análises. De forma inédita, as análises moleculares evidenciaram a ocorrência de introgressão bidirecional entre T. nigromaculatus e T. typhonius e entre T. nigromaculatus e T. mesophaeus, sendo que há indícios de indivíduos F1 (cruzamentos entre espécies parentais puras gerando híbridos). A utilização do gene ND2 mostrou-se mais eficiente do que o gene COI nas filogenias e, apesar da tirosinase ser um gene nuclear de evolução lenta, contribuiu para a identificação de incongruências citonucleares. Nossos resultados mostram que a história filogenética de Trachycephalus é complexa e que o uso de marcadores nucleares de evolução mais rápida e ampliação dessas análises para outras espécies do gênero podem revelar mais eventos de hibridização.

Cidade-paisagem : novas perspectivas sobre a preservação da paisagem urbana no Brasil

The edification of the landscape in the scientific and social field, in speech as in the method, is taken in this study as a complex process, from which were developed relationships of dominance on the perception of space, which persist strongly consolidated, since the genesis of the conceptual practice till its actual praxis. Historically, the landscape studies took place slowly, immersed in many questions, in face of the multiplicity of meanings that the concept offers. In the psyche, the notion of landscape is present since a long time ago, as an unconscious human being practice, even before any ideological hypothesis formulation. However its materialization in the social conscious will come only from painting, and specially with the perspective, through the technicity of the view, at first wandering the infinity, now ordained in a frame‟. Since then, the landscape is perceived according to the order of the view, as the equivalent of nature and beauty, assuming at the same time, an important symbolic value, since it is linked to mnemonic and subjective processes that the being build with the territory. The domain on this space-cognitive experience, characteristic of the contemporary, consolidates in the social imaginary, building consensus on the landscape, whose aesthetic references make a cultural appeal, very pertinent to the actual capitalist dynamics of production the space worldwide, mainly of the spectacle and commodification of cities promoted by the city marketing. In Brazil, this consensual ideology of the landscape surpass the social imaginary and also dominates the political imaginary, whereas the main instrument for preserving the landscape, Decree-law 25, from November 30, 1937, and its limit to those of exceptional value or remarkable feature. The analysis of the processes for putting under governmental trust for inscription of goods by the landscape value, reveals the dominant, if not exclusive, adoption of selection criteria related to aesthetic aspects. Abstain, therefore, from what the nation considers patrimony , other landscapes that, besides not having, at first, remarkable aesthetic value, play a crucial role as an inheritance from ancestor relations between man and space and pre- existing condition for the same present and future relations. From this historical background, the research seeks to transfer into contemporaneity, the ideological analysis of the concept and its relation with the building of the landscape in the collective imaginary, in order to recognize, in current practices to landscape preservation, as much this genesis, rooted in aesthetics, remains strongly consolidated, feeding the current dynamics of consumption and commodification of the city. Therefore, as preliminary conclusion, one can state that the identification of landscapes of different value, especially aesthetic, maintains and intensify the treatment of the city as an object, a standard‟ commodity to be sold / traded on the world market, in detriment of its recognition as a dynamic process that, even though inserted in the global context, develops specificifities and peculiarities, inherent to the production of space, as Lefebvre preconizes, that is, to the production of life, social product , as characteristic, dissent generator

Alianças estratégicas: análise do sector exportador de produtos português

Dissertação de Mestrado em Finanças Empresariais

Cellular polarity in aging: role of redox regulation and nutrition

Cellular polarity concerns the spatial asymmetric organization of cellular components and structures. Such organization is important not only for biological behavior at the individual cell level, but also for the 3D organization of tissues and organs in living organisms. Processes like cell migration and motility, asymmetric inheritance, and spatial organization of daughter cells in tissues are all dependent of cell polarity. Many of these processes are compromised during aging and cellular senescence. For example, permeability epithelium barriers are leakier during aging; elderly people have impaired vascular function and increased frequency of cancer, and asymmetrical inheritance is compromised in senescent cells, including stem cells. Here, we review the cellular regulation of polarity, as well as the signaling mechanisms and respective redox regulation of the pathways involved in defining cellular polarity. Emphasis will be put on the role of cytoskeleton and the AMP-activated protein kinase pathway. We also discuss how nutrients can affect polarity-dependent processes, both by direct exposure of the gastrointestinal epithelium to nutrients and by indirect effects elicited by the metabolism of nutrients, such as activation of antioxidant response and phase-II detoxification enzymes through the transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nrf2). In summary, cellular polarity emerges as a key process whose redox deregulation is hypothesized to have a central role in aging and cellular senescence.

Subunits of the chaperonin CCT are associated with Tetrahymena microtubule structures and are involved in cilia biogenesis

The cytosolic chaperonin CCT is a heterooligomeric complex of about 900 kDa that mediates the folding of cytoskeletal proteins. We observed by indirect immunofluorescence that the Tetrahymena TpCCTalpha, TpCCTdelta, TpCCTepsilon, and TpCCTeta-subunits colocalize with tubulin in cilia, basal bodies, oral apparatus, and contractile vacuole pores. TpCCT-subunits localization was affected during reciliation. These findings combined with atomic force microscopy measurements in reciliating cells indicate that these proteins play a role during cilia biogenesis related to microtubule nucleation, tubulin transport, and/or axoneme assembly. The TpCCT-subunits were also found to be associated with cortex and cytoplasmic microtubules suggesting that they can act as microtubule-associated proteins. The TpCCTdelta being the only subunit found associated with the macronuclear envelope indicates that it has functions outside of the 900 kDa complex. Tetrahymena cytoplasm contains granular/globular-structures of TpCCT-subunits in close association with microtubule arrays. Studies of reciliation and with cycloheximide suggest that these structures may be sites of translation and folding. Combined biochemical techniques revealed that reciliation affects the oligomeric state of TpCCT-subunits being tubulin preferentially associated with smaller CCT oligomeric species in early stages of reciliation. Collectively, these findings indicate that the oligomeric state of CCT-subunits reflects the translation capacity of the cell and microtubules integrity.

Resource sharing in global fixed-priority preemptive multiprocessor scheduling

In this paper we consider global fixed-priority preemptive multiprocessor scheduling of constrained-deadline sporadic tasks that share resources in a non-nested manner. We develop a novel resource-sharing protocol and a corresponding schedulability test for this system. We also develop the first schedulability analysis of priority inheritance protocol for the aforementioned system. Finally, we show that these protocols are efficient (based on the developed schedulability tests) for a class of priority-assignments called reasonable priority-assignments.

A capacity sharing and stealing strategy for open real-time systems

This paper focuses on the scheduling of tasks with hard and soft real-time constraints in open and dynamic real-time systems. It starts by presenting a capacity sharing and stealing (CSS) strategy that supports the coexistence of guaranteed and non-guaranteed bandwidth servers to efficiently handle soft-tasks’ overloads by making additional capacity available from two sources: (i) reclaiming unused reserved capacity when jobs complete in less than their budgeted execution time and (ii) stealing reserved capacity from inactive non-isolated servers used to schedule best-effort jobs. CSS is then combined with the concept of bandwidth inheritance to efficiently exchange reserved bandwidth among sets of inter-dependent tasks which share resources and exhibit precedence constraints, assuming no previous information on critical sections and computation times is available. The proposed Capacity Exchange Protocol (CXP) has a better performance and a lower overhead when compared against other available solutions and introduces a novel approach to integrate precedence constraints among tasks of open real-time systems.

Handling shared resources and precedence constraints in open systems

There is an increasing demand for highly dynamic realtime systems where several independently developed applications with different timing requirements can coexist. This paper proposes a protocol to integrate shared resources and precedence constraints among tasks in such systems assuming no precise information on critical sections and computation times is available. The concept of bandwidth inheritance is combined with a capacity sharing and stealing mechanism to efficiently exchange bandwidth among needed tasks, minimising the cost of blocking.

The capacity exchange protocol

This paper proposes a new strategy to integrate shared resources and precedence constraints among real-time tasks, assuming no precise information on critical sections and computation times is available. The concept of bandwidth inheritance is combined with a capacity sharing and stealing mechanism to efficiently exchange bandwidth among tasks to minimise the degree of deviation from the ideal system’s behaviour caused by inter-application blocking. The proposed Capacity Exchange Protocol (CXP) is simpler than other proposed solutions for sharing resources in open real-time systems since it does not attempt to return the inherited capacity in the same exact amount to blocked servers. This loss of optimality is worth the reduced complexity as the protocol’s behaviour nevertheless tends to be fair and outperforms the previous solutions in highly dynamic scenarios as demonstrated by extensive simulations. A formal analysis of CXP is presented and the conditions under which it is possible to guarantee hard real-time tasks are discussed.

Shared resources and precedence constraints with capacity sharing and stealing

This paper proposes a new strategy to integrate shared resources and precedence constraints among real-time tasks, assuming no precise information on critical sections and computation times is available. The concept of bandwidth inheritance is combined with a greedy capacity sharing and stealing policy to efficiently exchange bandwidth among tasks, minimising the degree of deviation from the ideal system's behaviour caused by inter-application blocking. The proposed capacity exchange protocol (CXP) focus on exchanging extra capacities as early, and not necessarily as fairly, as possible. This loss of optimality is worth the reduced complexity as the protocol's behaviour nevertheless tends to be fair in the long run and outperforms other solutions in highly dynamic scenarios, as demonstrated by extensive simulations.

Comparative studies of Yersinia pestis outer membrane isolation techniques and their potential use in plague epidemiology

In the present study three techniques for obtaining outer membrane enriched fractions from Yersinia pestis were evaluated. The techniques analysed were: differential solubilization of the cytoplasmic membrane with Sarkosyl or Triton X-100, and centrifugation in sucrose density gradients. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of outer membrane isolated by the different methods resulted in similar protein patterns. The measurement of NADH-dehydrogenase and succinate dehydrogenase (inner membrane enzymes) indicated that the outer membrane preparations obtained by the three methods were pure enough for analytical studies. In addition, preliminary evidences on the potential use of outer membrane proteins for the identification of geographic variants of Y. pestis wild isolates are presented.

Laboratório de criação a partir de Macbeth de William Shakespeare: relatório de estágio

Relatório de Estágio submetido à Escola Superior de Teatro e Cinema para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Teatro - especialização em Encenação.

Characterization of the [NiFe] Hydrogenase from the sulfate reducer Desulfovibrio vulgaris Hildenborough

The [NiFe] hydrogenase from Desulfovibrio vulgaris Hildenborough was isolated from the cytoplasmic membranes and characterized by EPR spectroscopy. It has a total molecular mass of 98.7 kDa (subunits of 66.4 and 32.3 kDa), and contains 1 nickel and 12 Fe atoms per heterodimer. The catalytic activities for hydrogen consumption and production were determined to be 174 and 89 umol H2 min-1 mg -1, respectively. As isolated, under aerobic conditions, this hydrogenase exhibits EPR signals characteristic of the nickel centers in [NiFe] hydrogenases (Ni-A signal at gx,y,z=2.32, 2.23 and ~2.0 and Ni-B signal at gx,y,z=2.33, 2.16 and ~2.0) as well as an intense quasi-isotropic signal centered at g=2.02 due to the oxidized [3Fe-4S] center. The redox profile under hydrogen atmosphere is remarkably similar to that of other [NiFe] hydrogenases. The signals observed for the oxidized state disappear, first being substituted by the Ni-C type signal (gx,y,z=2.19, 2.14, ~2.01), which upon long incubation under hydrogen yields the split Ni-C signal due to interaction with the reduced [4Fe-4S] centers.

Mitosis and protein Na-terminal acetylation

Dissertation presented to obtain the Ph.D degree in Developmental Biology