Association with {beta}-COP regulates the trafficking of the newly synthesized Na,K-ATPase.

Plasma membrane expression of the Na,K-ATPase requires assembly of its α- and β-subunits. Using a novel labeling technique to identify Na,K-ATPase partner proteins, we detected an interaction between the Na,K-ATPase α-subunit and the coat protein, β-COP, a component of the COP-I complex. When expressed in the absence of the Na,K-ATPase β-subunit, the Na,K-ATPase α-subunit interacts with β-COP, is retained in the endoplasmic reticulum, and is targeted for degradation. In the presence of the Na,K-ATPase β-subunit, the α-subunit does not interact with β-COP and traffics to the plasma membrane. Pulse-chase experiments demonstrate that in cells expressing both the Na,K-ATPase α- and β-subunits, newly synthesized α-subunit associates with β-COP immediately after its synthesis but that this interaction does not constitute an obligate intermediate in the assembly of the α- and β-subunits to form the pump holoenzyme. The interaction with β-COP was reduced by mutating a dibasic motif at Lys(54) in the Na,K-ATPase α-subunit. This mutant α-subunit is not retained in the endoplasmic reticulum and reaches the plasma membrane, even in the absence of Na,K-ATPase β-subunit expression. Although the Lys(54) α-subunit reaches the cell surface without need for β-subunit assembly, it is only functional as an ion-transporting ATPase in the presence of the β-subunit.

[Phot. d'une pierre tumulaire à Brignoles (Var), don Octave Teissier en 1892]

Donateur : Teissier, Octave (1825-1904)

Conseqüências do manejo do solo na distribuição de metais pesados em um agrossistema com feijão- de-vagem (Phaseolus vulgaris L.)

Com o objetivo de avaliar a concentração de metais pesados no solo e nas plantas de feijão-de-vagem (Phaseolus vulgaris L.) em diferentes sistemas de preparo do solo, foi realizado um experimento em Paty do Alferes (RJ), em um Latossolo Vermelho-Amarelo com 30 % de declividade. Foram instaladas parcelas do tipo Wischmeier com dimensões de 22,0 x 4,0 m; cada parcela era constituída de um sistema de preparo do solo distinto: plantio convencional, aração com trator no sentido morro abaixo e queima dos restos vegetais (PC); plantio em nível com aração tração animal e cordões de vegetação a cada 7,0 m (PN); cultivo mínimo com apenas abertura de covas para plantio e conservação dos restos vegetais (CM). Foram coletadas amostras de solo, de plantas e de agroquímicos, bem como analisados os teores totais de metais pesados. Verificou-se que as maiores adições de Cu no sistema deveram-se ao uso do fungicida Peprosan. Os sistemas de preparo PN e CM favoreceram acúmulo de Zn, Mn, Cd e Pb no solo, quando comparados ao PC, provavelmente, em virtude das menores perdas por erosão. A concentração de Pb na vagem in natura, nos sistemas de preparo PN e CM, esteve acima dos limites permitidos para alimentos in natura, em níveis impróprios para o consumo humano.

Dramatic influence of V beta gene polymorphism on an antigen-specific CD8+ T cell response in vivo.

According to recent crystallographic studies, the TCR-alpha beta contacts MHC class I-bound antigenic peptides via the polymorphic V gene-encoded complementarity-determining region 1 beta (CDR1 beta) and the hypervariable (D)J-encoded CDR3 beta and CDR3 alpha domains. To evaluate directly the relative importance of CDR1 beta polymorphism on the fine specificity of T cell responses in vivo, we have taken advantage of congenic V beta a and V beta b mouse strains that differ by a CDR1 polymorphism in the V beta 10 gene segment. The V beta 10-restricted CD8+ T cell response to a defined immunodominant epitope was dramatically reduced in V beta a compared with V beta b mice, as measured either by the expansion of V beta 10+ cells or by the binding of MHC-peptide tetramers. These data indicate that V beta polymorphism has an important impact on TCR-ligand binding in vivo, presumably by modifying the affinity of CDR1 beta-peptide interactions.

Clonal deletion of V beta 14-bearing T cells in mice transgenic for mammary tumour virus.

Autoreactive T lymphocytes are clonally deleted during maturation in the thymus. Deletion of T cells expressing particular receptor V beta elements is controlled by poorly defined autosomal dominant genes. A gene has now been identified by expression of transgenes in mice which causes deletion of V beta 14+ T cells. The gene lies in the open reading frame of the long terminal repeat of the mouse mammary tumour virus.

Nodulação e micorrização em Anadenanthera peregrina var. falcata em solo de cerrado autoclavado e não autoclavado

Anadenanthera peregrina var. falcata (angico-do-cerrado), uma leguminosa arbórea, forma associações simbióticas com bactérias fixadoras de nitrogênio (rizóbios) e com fungos micorrízicos arbusculares. Com o objetivo de avaliar a eficiência da inoculação de fungos micorrízicos e rizóbios no crescimento inicial de plantas de angico-do-cerrado, crescidas em solo autoclavado e em solo não autoclavado com e sem inoculação, foi desenvolvido um experimento em casa de vegetação, utilizando raízes micorrizadas de milho e uma mistura de isolados de rizóbios como inoculantes. O crescimento das plantas foi influenciado positivamente pela concomitante inoculação do fungo micorrízico e do rizóbio, tendo as plantas desse tratamento apresentado biomassa cerca de 60 % maior do que o controle no décimo mês. A inoculação de apenas um dos microssimbiontes, entretanto, não provocou diferença na produção de biomassa das plantas. A percentagem de colonização micorrízica foi significativamente mais alta e o número de nódulos maior nas raízes das plantas crescidas no solo não autoclavado, ocasionados pela população de fungos e rizóbios nativos. Nesse tratamento, houve pequeno acúmulo de matéria no xilopódio, provavelmente em virtude do dreno fotossintético por parte dos microssimbiontes, e a concentração de P na parte aérea e xilopódio dessas plantas foi cerca de 1,2 e 8 vezes maior, respectivamente, por causa da colonização micorrízica.

Prospectus dictionarii quinque linguarum, armenicae scilicet litteralis, armenicae vulgaris, latinae, italicae nec non gallicae. [Obsignatus nomine Fr. P. Gabrielis Villa-For-Alexandrini, missionarii cappucini.]

[Prospectus. Livres]

Specific Silencing of the REST Target Genes in Insulin-Secreting Cells Uncovers Their Participation in Beta Cell Survival.

The absence of the transcriptional repressor RE-1 Silencing Transcription Factor (REST) in insulin-secreting beta cells is a major cue for the specific expression of a large number of genes. These REST target genes were largely ascribed to a function of neurotransmission in a neuronal context, whereas their role in pancreatic beta cells has been poorly explored. To identify their functional significance, we have generated transgenic mice expressing REST in beta cells (RIP-REST mice), and previously discovered that REST target genes are essential to insulin exocytosis. Herein we characterized a novel line of RIP-REST mice featuring diabetes. In diabetic RIP-REST mice, high levels of REST were associated with postnatal beta cell apoptosis, which resulted in gradual beta cell loss and sustained hyperglycemia in adults. Moreover, adenoviral REST transduction in INS-1E cells led to increased cell death under control conditions, and sensitized cells to death induced by cytokines. Screening for REST target genes identified several anti-apoptotic genes bearing the binding motif RE-1 that were downregulated upon REST expression in INS-1E cells, including Gjd2, Mapk8ip1, Irs2, Ptprn, and Cdk5r2. Decreased levels of Cdk5r2 in beta cells of RIP-REST mice further confirmed that it is controlled by REST, in vivo. Using siRNA-mediated knock-down in INS-1E cells, we showed that Cdk5r2 protects beta cells against cytokines and palmitate-induced apoptosis. Together, these data document that a set of REST target genes, including Cdk5r2, is important for beta cell survival.

UVBY-Beta Photometry for 67 Stars in the Region of Alpha-Persei

UVBY-Beta photometric data are presented for 67 stars in the region of alpha-Persei and their relationship to the cluster is analyzed. These data allowed us to confirm at least 16 new members of the alpha-Persei open cluster.

A type stars - physical parameters from UVBY and H-beta photometry

uvby H-beta photometry has been obtained for a sample of 93 selected main sequence A stars. The purpose was to determine accurate effective temperatures, surface gravities, and absolute magnitudes for an individual determination of ages and parallaxes, which have to be included in a more extensive work analyzing the kinematic properties of A V stars. Several calibrations and methods to determine the above mentioned parameters have been reviewed, allowing the design of a new algorithm for their determination. The results obtained using this procedure were tested in a previous paper using uvby H-beta data from the Hauck and Mermilliod catalogue, and comparing the rusulting temperatures, surface gravities and absolute magnitudes with empirical determinations of these parameters.

Futile cycling of intermediates of fatty acid biosynthesis toward peroxisomal beta-oxidation in Saccharomyces cerevisiae.

The flux of fatty acids toward beta-oxidation was analyzed in Saccharomyces cerevisiae by monitoring polyhydroxyalkanoate synthesis in the peroxisome from the polymerization, by a bacterial polyhydroxyalkanoate synthase, of the beta-oxidation intermediates 3-hydroxyacyl-CoAs. Synthesis of polyhydroxyalkanoate was dependent on the beta-oxidation enzymes acyl-CoA oxidase and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase multifunctional protein, which are involved in generating 3-hydroxyacyl-CoAs, and on the peroxin PEX5, which is involved in the import of proteins into the peroxisome. In wild type cells grown in media containing fatty acids, the polyhydroxyalkanoate monomer composition was largely influenced by the nature of the external fatty acid, such that even-chain monomers are generated from oleic acid and odd-chain monomers are generated from heptadecenoic acid. In contrast, polyhydroxyalkanoate containing predominantly 3-hydroxyoctanoate, 3-hydroxydecanoate, and 3-hydroxydodecanoate was synthesized in a mutant deficient in the peroxisomal 3-ketothiolase (fox3 Delta 0) growing either on oleic acid or heptadecenoic acid as well as in wild type and fox3 Delta 0 mutants grown on glucose or raffinose, indicating that 3-hydroxyacyl-CoAs used for polyhydroxyalkanoate synthesis were generated from the degradation of intracellular short- and medium-chain fatty acids by the beta-oxidation cycle. Inhibition of fatty acid biosynthesis with cerulenin blocked the synthesis of polyhydroxyalkanoate from intracellular fatty acids but still enabled the use of extracellular fatty acids for polymer production. Mutants affected in the synthesis of lipoic acid showed normal polyhydroxyalkanoate synthesis capacity. Together, these results uncovered the existence of a substantial futile cycle whereby short- and medium-chain intermediates of the cytoplasmic fatty acid biosynthetic pathway are directed toward the peroxisomal beta-oxidation pathway.

Eficiência e diversidade fenotípica de bactérias diazotróficas que nodulam caupi [Vigna unguiculata (L.) Walp] e feijoeiro (Phaseolus vulgaris L.) em solos de mineração de bauxita em reabilitação

A atividade de mineração provoca degradação ambiental em várias partes do mundo, e técnicas de revegetação, empregando diversas espécies vegetais, têm sido indicadas, com vistas em reabilitar essas áreas. A eficiência e a diversidade de grupos-chave de bactérias, como as que nodulam leguminosas e fixam N2 (BNLFN), são de extrema importância, uma vez que estas participam de processos de ciclagem de nutrientes e contribuem para a sustentabilidade dessas áreas. Com o objetivo de avaliar a eficiência e a diversidade de BNLFN, coletaram-se amostras de solo de áreas mineradas, sob diferentes estratégias e cronosseqüências de reabilitação, no verão de 1999, para instalação de ensaios, empregando feijoeiro e caupi como espécies de plantas-isca de BNLFN. Na floração, coletaram-se as plantas e avaliaram-se matéria seca da parte aérea, número, matéria fresca e atividade de nódulos. Não houve influência das diferentes estratégias de reabilitação na eficiência das populações de BNLFN no crescimento do feijoeiro. A diversidade fenotípica dos isolados de BNLFN foi avaliada por meio das características culturais destes em meio de cultivo 79. Após a caracterização fenotípica de 328 isolados de feijoeiro e 420 de caupi, verificou-se que este é mais indicado que o feijoeiro nos estudos de nodulação, eficiência e diversidade de BNLFN. O impacto negativo da mineração é maior na diversidade fenotípica cultural de BNLFN do que na nodulação das plantas-isca utilizadas. A revegetação contribuiu para aumentar a diversidade dessas bactérias em solos de áreas de mineração de bauxita, especialmente quando ocorreu a introdução de leguminosas.

Differential sensitivity of GLUT1- and GLUT2-expressing beta cells to streptozotocin.

Streptozotocin injection in animals destroys pancreatic beta cells, leading to insulinopenic diabetes. Here, we evaluated the toxic effect of streptozotocin (STZ) in GLUT2(-/-) mice reexpressing either GLUT1 or GLUT2 in their beta cells under the rat insulin promoter (RIPG1 x G2(-/-) and RIPG2 x G2(-/-) mice, respectively). We demonstrated that injection of STZ into RIPG2 x G2(-/-) mice induced hyperglycemia (>20 mM) and an approximately 80% reduction in pancreatic insulin content. In vitro, the viability of RIPG2 x G2(-/-) islets was also strongly reduced. In contrast, STZ did not induce hyperglycemia in RIPG1 x G2(-/-) mice and did not reduce pancreatic insulin content. The viability of in vitro cultured RIPG1 x G2(-/-) islets was also unaffected by STZ. As islets from each type of transgenic mice were functionally indistinguishable, these data strongly support the notion that STZ toxicity toward beta cells depends on the expression of GLUT2.