Structural and functional conservation of key domains in InsP3 and ryanodine receptors.

Inositol-1,4,5-trisphosphate receptors (InsP(3)Rs) and ryanodine receptors (RyRs) are tetrameric intracellular Ca(2+) channels. In each of these receptor families, the pore, which is formed by carboxy-terminal transmembrane domains, is regulated by signals that are detected by large cytosolic structures. InsP(3)R gating is initiated by InsP(3) binding to the InsP(3)-binding core (IBC, residues 224-604 of InsP(3)R1) and it requires the suppressor domain (SD, residues 1-223 of InsP(3)R1). Here we present structures of the amino-terminal region (NT, residues 1-604) of rat InsP(3)R1 with (3.6 Å) and without (3.0 Å) InsP(3) bound. The arrangement of the three NT domains, SD, IBC-β and IBC-α, identifies two discrete interfaces (α and β) between the IBC and SD. Similar interfaces occur between equivalent domains (A, B and C) in RyR1 (ref. 9). The orientations of the three domains when docked into a tetrameric structure of InsP(3)R and of the ABC domains docked into RyR are remarkably similar. The importance of the α-interface for activation of InsP(3)R and RyR is confirmed by mutagenesis and, for RyR, by disease-causing mutations. Binding of InsP(3) causes partial closure of the clam-like IBC, disrupting the β-interface and pulling the SD towards the IBC. This reorients an exposed SD loop ('hotspot' (HS) loop) that is essential for InsP(3)R activation. The loop is conserved in RyR and includes mutations that are associated with malignant hyperthermia and central core disease. The HS loop interacts with an adjacent NT, suggesting that activation re-arranges inter-subunit interactions. The A domain of RyR functionally replaced the SD in full-length InsP(3)R, and an InsP(3)R in which its C-terminal transmembrane region was replaced by that from RyR1 was gated by InsP(3) and blocked by ryanodine. Activation mechanisms are conserved between InsP(3)R and RyR. Allosteric modulation of two similar domain interfaces within an N-terminal subunit reorients the first domain (SD or A domain), allowing it, through interactions of the second domain of an adjacent subunit (IBC-β or B domain), to gate the pore

The OBAA Standard for Developing Repositories of Learning Objects: the Case of Ocean Literacy in Azores

This paper describes the existing web resources of learning objects to promote ocean literacy. The several projects and sites are explored, and the shortcomings revealed. The limitations identified include insufficient metadata about registered learning objects and lack of support for intelligent applications. As solution, we promote the seaThings project that relies on a multi-disciplinary approach to promote literacy in the marine environment by implementing a specific Learning Objects repositories (LOR) and a federation of repositories (FED), supported by a OBAA, a versatile and innovative standard that will provide the necessary support for intelligent applications for education purposes, to be used in schools and other educational institutions.info:eu-repo/semantics/publishedVersio

Osteoblasts and Bone Marrow Mesenchymal Stromal Cells Control Hematopoietic Stem Cell Migration and Proliferation in 3D In Vitro Model

BACKGROUND: Migration, proliferation, and differentiation of hematopoietic stem cells (HSCs) are dependent upon a complex three-dimensional (3D) bone marrow microenvironment. Although osteoblasts control the HSC pool, the subendosteal niche is complex and its cellular composition and the role of each cell population in HSC fate have not been established. In vivo models are complex and involve subtle species-specific differences, while bidimensional cultures do not reflect the 3D tissue organization. The aim of this study was to investigate in vitro the role of human bone marrow-derived mesenchymal stromal cells (BMSC) and active osteoblasts in control of migration, lodgment, and proliferation of HSCs. METHODOLOGY/PRINCIPAL FINDINGS: A complex mixed multicellular spheroid in vitro model was developed with human BMSC, undifferentiated or induced for one week into osteoblasts. A clear limit between the two stromal cells was established, and deposition of extracellular matrix proteins fibronectin, collagens I and IV, laminin, and osteopontin was similar to the observed in vivo. Noninduced BMSC cultured as spheroid expressed higher levels of mRNA for the chemokine CXCL12, and the growth factors Wnt5a and Kit ligand. Cord blood and bone marrow CD34(+) cells moved in and out the spheroids, and some lodged at the interface of the two stromal cells. Myeloid colony-forming cells were maintained after seven days of coculture with mixed spheroids, and the frequency of cycling CD34(+) cells was decreased. CONCLUSIONS/SIGNIFICANCE: Undifferentiated and one-week osteo-induced BMSC self-assembled in a 3D spheroid and formed a microenvironment that is informative for hematopoietic progenitor cells, allowing their lodgment and controlling their proliferation

Modelagem para prognose precoce do número de árvores e área basal por classe diamétrica para Tectona grandis

O presente estudo teve como objetivo modelar os atributos da floresta: sítio, sobrevivência, variância dos diâmetros, área basal e média aritmética dos diâmetros e; ajustar a função probabilística de Weibull pelo método de momentos (recuperação de parâmetros) para prognose precoce da área basal e número de árvores por classe diamétrica. O estudo foi desenvolvido com dados de Tectona grandis provenientes da região norte do Estado do Mato Grosso. Para o desenvolvimento do estudo, foi feita a classificação de sítio, o ajuste dos modelos de sobrevivência e área basal, bem como o ajuste da distribuição diamétrica pela função de Weibull. Utilizaram-se as informações de cada parcela (18 ao todo) e definiu-se a idade de 7 anos para ter-se a projeção do número de árvores e área basal. Os principais resultados obtidos para os atributos do povoamento, de acordo com as estatísticas encontradas, propiciaram estimativas precisas. A metodologia empregada utilizando-se o método de recuperação de parâmetros da função Weibull representou uma aproximação realística à estimação da estrutura do povoamento

Déficit de volume de líquidos: perfil de características definidoras no paciente portador de queimadura

The present study aimed at describing the profile of defining characteristics in patients with the nursing diagnosis "Fluid volume deficit" related to active loss of fluid secondary to burns. Data were collected by means of a tool, containing 29 possible defining characteristics of this diagnosis. Seven nurses, that worked at the Burnt Unit for at least five years ago, provided opinions about the degree to which each defining characteristic is indicative of this diagnosis. Nurses rated each defining characteristic of diagnosis being tested on a scale of zero to one. The results confirmed all, except one (increased body temperature) defining characteristics presented by NANDA for this diagnosis and indicated 10 new defining characteristics.El objetivo de este estudio fue describir el perfil de las características que la definen en paciente con diagnóstico de enfermería "Déficit de volumen de líquidos" relacionado a la pérdida activa de líquidos, secundaria a quemadura. La recolección de datos fue hecha a través de un instrumento que conttene 29 características posibiles que definen ese diagnóstico. El instrumento fue desarrollado con siete enfermeros que trabajaban en una Unidad de Quemados, los quales poseían una experiencia mínima de cinco años. Los enfermeros evaluaron las características en cuanto al grado que cada una de estas características eran indicativas de ese diagnóstico, atribuyendo valores en una escala de zero a uno. Los resultados confirmaron todas las características presentadas por NANDA para ese diagnóstico, excepto una (temperatura corporal elevada) y indicaron 10 nuevas características.O objetivo deste estudo foi descrever o perfil das características definidoras do paciente com diagnóstico de enfermagem "Déficit de volume de líquidos" relacionado a perda ativa de líquidos, secundária à queimadura. Os dados foram coletados através de um instrumento, contendo 29 características definidoras possíveis para esse diagnóstico, aplicado a sete enfermeiros, que trabalhavam há pelo menos cinco anos em serviços especializados em tratamento de queimaduras. Os enfermeiros avaliaram as características definidoras, quanto ao grau que cada uma é indicativa desse diagnóstico, atribuindo valores numa escala de zero a um. Os resultados confirmaram todas as características definidoras apresentadas pela North American Nursing Diagnosis Association (NANDA), exceto uma (temperatura corporal elevada), e apontaram 10 novas características

A construção das personagens em The Zoo Story: uma abordagem sistêmico-funcional

Este trabalho busca mostrar como se dá a interação das personagens Peter e Jerry na peça The Zoo Story, de Edward Albee, tomando como aporte teórico a linguística sistêmico-funcional. Foram analisadas e quantificadas as trocas de informação e de bens e serviços e do recurso à modalidade nas falas das personagens, bem como dos tipos de processo associados a cada uma nas rubricas do texto, de modo a observar como se deu a construção dessas personagens e a dinâmica de sua interação ao longo da peça. A contabilização das funções atribuídas ao Finito mostrou que as personagens deram primazia a localizar aquilo que expressavam no tempo e no espaço; no entanto, os casos de modalidade e de modalização e modulação elucidaram um forte contraste na personalidade de ambas, reflexo de suas diferentes condições socioeconômicas. Finalmente, as manifestações das personagens no plano físico, através da quantificação dos tipos de processos associados a elas, serviram para mostrar as atitudes de cada uma com relação a sua contraparte, atitudes essas nem sempre manifestadas verbalmente, daí a utilidade de observar os processos que guiam as rubricas do texto. Este trabalho busca elucidar algumas propriedades das metafunções experiencial e interpessoal da LSF e mostrar como ela pode ser aplicada à análise do texto literário, revelando aspectos não somente formais do texto, mas também a maneira como as personagens são construídas em relação umas às outras.This paper seeks to show how the interaction between the characters Peter and Jerry takes place in the play The Zoo Story, by Edward Albee, taking systemic-functional linguistics as a theoretical contribution. The exchanges of information and goods and services and the use of modality in the characters’ dialogues were analyzed and quantified, as well as the types of process associated with each one in the performance rubrics, in order to observe the construction of these characters and the dynamics of their interaction throughout the play. The accounting of the functions attributed to Finite showed that the characters gave priority to locating what they expressed in time and space; however, modality and modulation cases elucidated a strong contrast in the personality of both, reflecting their different socioeconomic conditions. Finally, the physical manifestations of the, through the quantification of the types of processes associated with them, served to show the attitudes of each one in relation to their counterpart, attitudes not always manifested verbally, hence the utility of observing the processes that guide the text rubrics. This paper seeks to elucidate some properties of the experiential and interpersonal metafunctions and to show how they can be applied to the analysis of the literary text, revealing not only formal aspects of the text but also the way the characters are constructed in relation to one another.info:eu-repo/semantics/publishedVersio

Métodos de avaliação ambiental e experiências construídas: uma revisão da teoria e prática sobre sustentabilidade urbana / Environmental assessment methods and built experiences: a review of the theory and practice on urban sustainability

Ao mesmo tempo em que os estudos sobre desenvolvimento sustentável se tornam cada vez mais presentes no âmbito da construção civil e da arquitetura brasileiras, presencia-se o surgimento de empreendimentos habitacionais de grandes dimensões que apresentam características muito distantes do que se conceitua por sustentabilidade no meio urbano. Essa realidade ocorre principalmente em empreendimentos destinados à população de baixa renda que, apesar de contribuírem para a diminuição do déficit habitacional brasileiro, pouco têm ajudado no surgimento de uma cidade mais bela e eficiente, socialmente mais inclusiva, menos consumidora de recursos naturais e com menor impacto no ambiente natural. Este artigo tem por objetivo apresentar uma revisão bibliográfica dos conceitos de sustentabilidade urbana e dos critérios presentes nos métodos de avaliação ambiental mais utilizados atualmente na escala urbana no Brasil e no exterior, com destaque na literatura e na prática profissional na França, por seu avanço em termos de certificação para edificações sustentáveis e por apresentar referências mais acessíveis à área de pesquisa dos autores deste artigo. Espera-se, a partir dos dados levantados no âmbito desta pesquisa, obter um entendimento mais claro a respeito do conceito e dos critérios de sustentabilidade urbana, na teoria e na prática, e identificar elementos que serão utilizados para a futura elaboração de um modelo de referência para a análise da sustentabilidade urbana em escala de bairro. 

Regulation of cellular contractile force, shape and migration of fibroblasts by oncogenes and Histone deacetylase 6

The capacity of cells to adhere to, exert forces upon and migrate through their surrounding environment governs tissue regeneration and cancer metastasis. The role of the physical contractile forces that cells exert in this process, and the underlying molecular mechanisms are not fully understood. We, therefore, aimed to clarify if the extracellular forces that cells exert on their environment and/or the intracellular forces that deform the cell nucleus, and the link between these forces, are defective in transformed and invasive fibroblasts, and to indicate the underlying molecular mechanism of control. Confocal, Epifluorescence and Traction force microscopy, followed by computational analysis, showed an increased maximum contractile force that cells apply on their environment and a decreased intracellular force on the cell nucleus in the invasive fibroblasts, as compared to normal control cells. Loss of HDAC6 activity by tubacin-treatment and siRNA-mediated HDAC6 knockdown also reversed the reduced size and more circular shape and defective migration of the transformed and invasive cells to normal. However, only tubacin-mediated, and not siRNA knockdown reversed the increased force of the invasive cells on their surrounding environment to normal, with no effects on nuclear forces. We observed that the forces on the environment and the nucleus were weakly positively correlated, with the exception of HDAC6 siRNA-treated cells, in which the correlation was weakly negative. The transformed and invasive fibroblasts showed an increased number and smaller cell-matrix adhesions than control, and neither tubacin-treatment, nor HDAC6 knockdown reversed this phenotype to normal, but instead increased it further. This highlights the possibility that the control of contractile force requires separate functions of HDAC6, than the control of cell adhesions, spreading and shape. These data are consistent with the possibility that defective force-transduction from the extracellular environment to the nucleus contributes to metastasis, via a mechanism that depends upon HDAC6. To our knowledge, our findings present the first correlation between the cellular forces that deforms the surrounding environment and the nucleus in fibroblasts, and it expands our understanding of how cells generate contractile forces that contribute to cell invasion and metastasisis

The Fourteenth Data Release of the Sloan Digital Sky Survey: First Spectroscopic Data from the extended Baryon Oscillation Spectroscopic Survey and from the second phase of the Apache Point Observatory Galactic Evolution Experiment

The fourth generation of the Sloan Digital Sky Survey (SDSS-IV) has been in operation since July 2014. This paper describes the second data release from this phase, and the fourteenth from SDSS overall (making this, Data Release Fourteen or DR14). This release makes public data taken by SDSS-IV in its first two years of operation (July 2014-2016). Like all previous SDSS releases, DR14 is cumulative, including the most recent reductions and calibrations of all data taken by SDSS since the first phase began operations in 2000. New in DR14 is the first public release of data from the extended Baryon Oscillation Spectroscopic Survey (eBOSS); the first data from the second phase of the Apache Point Observatory (APO) Galactic Evolution Experiment (APOGEE-2), including stellar parameter estimates from an innovative data driven machine learning algorithm known as "The Cannon"; and almost twice as many data cubes from the Mapping Nearby Galaxies at APO (MaNGA) survey as were in the previous release (N = 2812 in total). This paper describes the location and format of the publicly available data from SDSS-IV surveys. We provide references to the important technical papers describing how these data have been taken (both targeting and observation details) and processed for scientific use. The SDSS website (www.sdss.org) has been updated for this release, and provides links to data downloads, as well as tutorials and examples of data use. SDSS-IV is planning to continue to collect astronomical data until 2020, and will be followed by SDSS-V.Comment: SDSS-IV collaboration alphabetical author data release paper. DR14 happened on 31st July 2017. 19 pages, 5 figures. Accepted by ApJS on 28th Nov 2017 (this is the "post-print" and "post-proofs" version; minor corrections only from v1, and most of errors found in proofs corrected

Induction of Eosinophil Apoptosis by the Cyclin-Dependent Kinase Inhibitor AT7519 Promotes the Resolution of Eosinophil-Dominant Allergic Inflammation

Eosinophils not only defend the body against parasitic infection but are also involved in pathological inflammatory allergic diseases such as asthma, allergic rhinitis and contact dermatitis. Clearance of apoptotic eosinophils by macrophages is a key process responsible for driving the resolution of eosinophilic inflammation and can be defective in allergic diseases. However, enhanced resolution of eosinophilic inflammation by deliberate induction of eosinophil apoptosis using pharmacological agents has not been previously demonstrated. Here we investigated the effect of a novel cyclin-dependent kinase inhibitor drug, AT7519, on human and mouse eosinophil apoptosis and examined whether it could enhance the resolution of a murine model of eosinophil-dominant inflammation in vivo.Eosinophils from blood of healthy donors were treated with AT7519 and apoptosis assessed morphologically and by flow-cytometric detection of annexin-V/propidium iodide staining. AT7519 induced eosinophil apoptosis in a concentration dependent manner. Therapeutic administration of AT7519 in eosinophil-dominant allergic inflammation was investigated using an established ovalbumin-sensitised mouse model of allergic pleurisy. Following ovalbumin challenge AT7519 was administered systemically at the peak of pleural inflammation and inflammatory cell infiltrate, apoptosis and evidence of macrophage phagocytosis of apoptotic eosinophils assessed at appropriate time points. Administration of AT7519 dramatically enhanced the resolution of allergic pleurisy via direct induction of eosinophil apoptosis without detriment to macrophage clearance of these cells. This enhanced resolution of inflammation was shown to be caspase-dependent as the effects of AT7519 were reduced by treatment with a broad spectrum caspase inhibitor (z-vad-fmk).Our data show that AT7519 induces human eosinophil apoptosis and enhances the resolution of a murine model of allergic pleurisy by inducing caspase-dependent eosinophil apoptosis and enhancing macrophage ingestion of apoptotic eosinophils. These findings demonstrate the utility of cyclin-dependent kinase inhibitors such as AT7519 as potential therapeutic agents for the treatment of eosinophil dominant allergic disorders