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.

Identification and removal of low-complexity sites in allele-specific analysis of ChIP-seq data.

MOTIVATION: High-throughput sequencing technologies enable the genome-wide analysis of the impact of genetic variation on molecular phenotypes at unprecedented resolution. However, although powerful, these technologies can also introduce unexpected artifacts. Results: We investigated the impact of library amplification bias on the identification of allele-specific (AS) molecular events from high-throughput sequencing data derived from chromatin immunoprecipitation assays (ChIP-seq). Putative AS DNA binding activity for RNA polymerase II was determined using ChIP-seq data derived from lymphoblastoid cell lines of two parent-daughter trios. We found that, at high-sequencing depth, many significant AS binding sites suffered from an amplification bias, as evidenced by a larger number of clonal reads representing one of the two alleles. To alleviate this bias, we devised an amplification bias detection strategy, which filters out sites with low read complexity and sites featuring a significant excess of clonal reads. This method will be useful for AS analyses involving ChIP-seq and other functional sequencing assays.

Effluent particle removal by microirrigation system filters

The aim of this work was to determine whether the filters used in microirrigation systems can remove potentially emitter-clogging particles. The particle size and volume distributions of different effluents and their filtrates were established, and the efficiency of the removal of these particles and total suspended solids by screen, disc and sand filters determined. In most of the effluents and filtrates, the number of particles with a diameter > 20 μm was minimal. By analysing the particle volume distribution it was found that particles larger than the disc and screen filter pores appeared in the filtrates. However, the sand filter was able to retain particles larger than the pore size. The filtration efficiency depended more on the type of effluent than on the filter. It was also found that the particle size distribution followed a potential law. Analysis of the β exponents showed that the filters did not significantly modify the particle size distribution of the effluents

Application of K-feldspar-jadeite-quartz barometry to eclogite facies metagranites and metapelites in the Sesia Lanzo Zone (Western Alps, Italy)

The eclogite facies assemblage K-feldspar-jadeite-quartz in metagranites and metapelites from the Sesia-Lanzo Zone (Western Alps, Italy) records the equilibration pressure by dilution of the reaction jadeite + quartz = albite. The metapelites show partial transformation from a pre-Alpine assemblage of garnet (Alm(63)Prp(26)Grs(10))-K-feldspar-plagioclase-biotite +/- sillimanite to the Eo-Alpine high-pressure assemblage garnet (Alm(50)Prp(14)Grs(35))-jadeite (Jd(80-97)Di(0-4)Hd(0-8)Acm(0-7))=zoisite-phengite. Plagioclase is replaced by jadeite-zoisite-kyanite-K-feldspar-quartz and biotite is replaced by garnet-phengite or omphacite-kyanite-phengite. Equilibrium was attained only in local domains in the metapelites and therefore the K-feldspar-jadeite-quartz (KJQ) barometer was applied only to the plagioclase pseudomorphs and K-feldspar domains. The albite content of K-feldspar ranges from 4 to 11 mol% in less equilibrated assemblages from Val Savenca and from 4 to 7 mol% in the partially equilibrated samples from Monte Mucrone and the equilibrated samples from Montestrutto and Tavagnasco. Thermodynamic calculations on the stability of the assemblage K-feldspar-jadeite-quartz using available mixing data for K-feldspar and pyroxene indicate pressures of 15-21 kbar (+/- 1.6-1.9 kbar) at 550 +/- 50 degrees C. This barometer yields direct pressure estimates in high-pressure rocks where pressures are seldom otherwise fixed, although it is sensitive to analytical precision and the choice of thermodynamic mixing model for K-feldspar. Moreover, the KJQ barometer is independent of the ratio P-H2O/P-T. The inferred limiting a(H2O) for the assemblage jadeite-kyanite in the metapelites from Val Savenca is low and varies from 0.2 to 0.6.

UBVRI photometry of G, K, M Hipparcos stars

UBVRI data are presented for a set of 229 late-type stars, most of them being high proper motion stars. All these data are part of the Input Catalog planned observations for the Hipparcos mission.

UBVRI photometry of G, K, M Hipparcos stars. II.

As indicated by Grenon (1989), the data of the present series of reports on the UBVRI photometry of late-type stars in the Hipparcos Input Catalog are to be employed in computations of Hipparcos observing time, as well as in evaluating the observability of faint stars by the satellite. Attention is here given to late type stars in the V = 8-12 range, including distant red giants in the Galactic plane (Hipparcos proposal 189), as well as high proper motion stars included in the G, LTT, LP, and MCC catalogs.

Role of Na+-K+-ATPase in insulin-induced lactate release by skeletal muscle.

Hyperinsulinemia increases lactate release by various organs and tissues. Whereas it has been shown that aerobic glycolysis is linked to Na+-K+-ATPase activity, we hypothesized that stimulation by insulin of skeletal muscle Na+-K+-ATPase is responsible for increased muscle lactate production. To test this hypothesis, we assessed muscle lactate release in healthy volunteers from the [13C]lactate concentration in the effluent dialysates of microdialysis probes inserted into the tibialis anterior muscles on both sides and infused with solutions containing 5 mmol/l [U-13C]glucose. On one side, the microdialysis probe was intermittently infused with the same solution additioned with 2.10(-5) M ouabain. In the basal state, [13C]lactate concentration in the dialysate was not affected by ouabain. During a euglycemic-hyperinsulinemic clamp, [13C]lactate concentration increased by 135% in the dialysate without ouabain, and this stimulation was nearly entirely reversed by ouabain (56% inhibition compared with values in the dialysate collected from the contralateral probe). These data indicate that insulin stimulates muscle lactate release by activating Na+-K+-ATPase in healthy humans.

K-T transition in Deccan Traps of central India marks major marine Seaway across India

Deccan intertrappean sediments in central India are generally considered as terrestrial deposits of Maastrichtian age, but the Cretaceous-Tertiary (K-T) position is still unknown. Here we report the discovery of the K-T transition, a marine incursion and environmental changes preserved within the intertrappean sediments at Jhilmili, Chhindwara District, Madhya Pradesh. Integrative biostratigraphic, sedimentologic, mineralogic and chemostratigraphic analyses reveal the basal Danian in the intertrappean sediments between lower and upper trap basalts that regionally correspond to C29r and the C29R/C29N transition, respectively. Intertrappean deposition occurred in predominantly terrestrial semi-humid to and environments. But a short aquatic interval of fresh water ponds and lakes followed by shallow coastal marine conditions with brackish marine ostracods and early Danian zone P1a planktic foraminifera mark this interval very close to the K-T boundary. This marine incursion marks the existence of a nearby seaway, probably extending inland from the west through the Narmada and Tapti rift valleys. The Jhilmili results thus identify the K-T boundary near the end of the main phase of Deccan eruptions and indicate that a major seaway extended at least 800 km across India. (C) 2009 Elsevier B.V. All rights reserved.

Cinética de absorção e eficiência nutricional de K+, Ca2+ e Mg2+ em plantas jovens de quatro clones de eucalipto

Modelos mecanísticos baseados em princípios de transporte de solutos pode ser de grande utilidade para prever os impactos do cultivo de florestas plantadas, por exemplo, com eucalipto sobre o capital e fluxo de nutrientes no solo. Dentre as variáveis de entrada demandadas por tais modelos, tem-se os valores das constantes Vmax, Km e Cmin da cinética de absorção iônica. Assim, os objetivos do presente trabalho foram determinar os valores de Vmax, Km e Cmin para K, Ca e Mg, bem como avaliar as respectivas eficiências nutricionais de clones de eucalipto. O estudo consistiu de três ensaios em solução nutritiva (um para cada cátion), sendo utilizadas mudas propagadas vegetativamente de um híbrido de E. grandis x E. urophylla (clone 1213) e de três híbridos de E. grandis (clones 7074, 57 e 129). Com base nos teores de cada um desses nutrientes nas soluções de depleção, em cada tempo de amostragem, no volume inicial e final de solução nos vasos e no peso de matéria fresca de raízes, foram obtidos os valores das constantes cinéticas. Para K, o clone 7074 apresentou o menor valor de Vmax em relação aos demais clones, os quais não diferiram entre si, e em relação ao Km e Cmin, os clones não diferiram estatisticamente. Para Ca, os clones estudados diferiram quanto ao valor de Vmax e Km, não diferindo, entretanto, para o Cmin. Os menores valores de Km para Mg foram verificados para os clones 57 e 7074, ou seja, as proteínas transportadoras de Mg na membrana plasmática das células radiculares apresentaram maior afinidade para esse nutriente. Contudo, os valores de Vmax e Cmin não diferiram entre os clones estudados. Diferenças na eficiência nutricional dos clones estudados quanto a K e a Ca foram devidas às diferenças na eficiência de absorção, e para Mg às diferenças na eficiência de absorção e de utilização.

Inducing effect of skeletal muscle extracts on the appearance of calbindin-immunoreactive dorsal root ganglion cells in culture.

Calbindin D-28k is a calcium-binding protein which is not expressed by dorsal root ganglion cells cultured from 6-day-old (E6) chick embryos. When soluble muscle extracts from embryos at E11, E18 or chickens 2 weeks after hatching were added immediately after seeding, dorsal root ganglia cells grown at E6 displayed neuronal subpopulations expressing calbindin immunoreactivity with time; the effect of muscle extract on the percentage of calbindin-immunoreactive dorsal root ganglia cells followed a dose-response curve. When muscle extract was added to cultures after a 3 day delay, the percentage of calbindin-expressing neurons was unchanged. The effect produced by muscle extract and, to a lesser degree, skin extract on the appearance of calbindin-positive neurons was not reproduced by brain or liver extracts while all four exerted a trophic action on cultured neurons. Hence it is assumed that muscle extract contains a factor which produces an inductive effect on the initiation of calbindin-expression by uncommitted subpopulations of sensory neurons rather than a trophic influence on the selective survival of covertly committed neuronal subpopulations. The fact that muscle extract promoted calbindin expression by dorsal root ganglia cells in neuron-enriched as well as in mixed dorsal root ganglion cell cultures indicates that the factor would act directly on sensory neurons rather than indirectly through mediation of non-neuronal cells. Since the active muscular factor was non-dialysable, heat-inactivated, trypsin-sensitive and retained by molecular filters with a cut-off of 30 K, this factor is probably a protein.

Activity of phosphorus of synthetic Fe-K-P compounds in superphosphate fertilizers as affected by pH and ionic strength

The concentration of orthophosphate ions released from Fe-K-P compounds (Fe3KH8(PO4)6 .6H2O and Fe3KH14(PO4)8 .4H2O) present in superphosphates increases with pH, which initially suggests that the agronomic effectiveness of P fertilizers containing high amounts of these compounds would also increase with soil pH but studies considering activity, instead of concentration, are necessary. With this purpose, both compounds were synthesized under laboratory conditions, characterized by elemental chemical analysis, optical microscopy, X ray diffractometry, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), and used in a solubility study. Solutions of 0.01, 0.05 and 0.1 mol L-1 NaCl with pH adjusted to 3.0, 4.0, 5.0, 5.5, 6.0, 6.5, 7.0 and 7.5 were prepared for the solubility study of H8-syn, H14-syn and a phosphate rock (PR) from Brazil. The orthophosphate activity as H2PO4- and HPO4(2-) was calculated in each situation as related to pH and ionic strength using software MINTEQ. The remaining precipitates after equilibrium were chemically analyzed and subjected to X ray, SEM and EDS. Results of chemical analysis and instrumental techniques confirmed the preparation method. The activity of orthophosphate ions of both compounds tended to decrease under increasing pH and/or ionic strength of the solution, which in turn suggests that an increase in the solution pH does not necessarily promote an increase in the P bioavailability for plant uptake. This can be important when evaluating agronomic data of P fertilizers with high contents of these two Fe-K-P compounds.

Adubação orgânica da batata com esterco e, ou, Crotalaria juncea: II - disponibilidade de N, P e K no solo ao longo do ciclo de cultivo

Os solos do Agreste paraibano têm baixa fertilidade e a prática usual de adubação é a incorporação de esterco na época do plantio. Entretanto, dependendo da qualidade do esterco, essa prática pode causar a imobilização de nutrientes do solo durante os primeiros meses de cultivo. É possível que o cultivo e incorporação de Crotalaria juncea, combinado com o esterco, possa promover mineralização mais sincronizada com a demanda de nutrientes pelas plantas. No presente estudo, realizado em 2003, foram conduzidos experimentos de campo e em casa de vegetação para testar essa hipótese. As parcelas de campo foram previamente cultivadas com batata no período de 1996 a 2002 e submetidas anualmente aos seguintes tratamentos: plantio e incorporação da crotalária na época da floração (C); adição de 15 t ha-1 de esterco (E); plantio e incorporação de crotalária + 7,5 t ha-1 de esterco (CE); e testemunha, sem esterco ou crotalária (T). Em 2003, foram aplicados os mesmos tratamentos de adubação orgânica e foi avaliada a dinâmica da decomposição e liberação de nutrientes pelo material vegetal e esterco contidos em sacolas de náilon incorporadas ao solo das parcelas de campo. Também se avaliou a dinâmica da disponibilidade de N, P e K no solo das parcelas de campo, por meio de coletas periódicas de solo. No ensaio em casa de vegetação, foram realizados cultivos sucessivos de capim-buffel (Cenchrus ciliaris L.) por um período de 300 dias, utilizando-se amostras do solo das parcelas de campo. As perdas de massa e nutrientes do material incorporado ao solo foram maiores nos primeiros 30 dias da incubação, em todos os tratamentos. No final do ensaio, as proporções de matéria seca e nutrientes remanescentes foram maiores nos tratamentos E e CE que nos tratamentos C e T. No solo das parcelas de campo, o tratamento E aumentou os teores de P e K extraíveis (Mehlich-1) do solo ao longo do período do estudo, porém provocou a imobilização de N do solo nas primeiras semanas. O tratamento C aumentou o teor de N mineral do solo no período imediatamente após a incorporação, mas não aumentou o teor de P e K extraíveis. No ensaio em casa de vegetação, o capim-buffel acumulou mais biomassa aérea e nutrientes no primeiro corte, 35 dias após o transplantio do capim, no solo do tratamento CE. Nos cortes subseqüentes, o tratamento E levou à maior produção de biomassa e acumulação de nutrientes, indicando que a limitação de N no período inicial após a incorporação de esterco ao solo prejudicou o crescimento do buffel. Os resultados corroboram a hipótese de que o plantio e incorporação da crotalária, combinado com a aplicação de apenas a metade da dose usual de esterco caprino, promoveu mineralização de nutrientes mais sincronizada com as necessidades das culturas agrícolas, pois foi capaz de evitar a imobilização de N do solo no período inicial de cultivo e elevou os teores de P e K disponíveis ao longo de todo o período.