Amiloride-sensitive epithelial Na+ channel is made of three homologous subunits.

The amiloride-sensitive epithelial sodium channel constitutes the rate-limiting step for sodium reabsorption in epithelial cells that line the distal part of the renal tubule, the distal colon, the duct of several exocrine glands, and the lung. The activity of this channel is upregulated by vasopressin and aldosterone, hormones involved in the maintenance of sodium balance, blood volume and blood pressure. We have identified the primary structure of the alpha-subunit of the rat epithelial sodium channel by expression cloning in Xenopus laevis oocytes. An identical subunit has recently been reported. Here we identify two other subunits (beta and gamma) by functional complementation of the alpha-subunit of the rat epithelial Na+ channel. The ion-selective permeability, the gating properties and the pharmacological profile of the channel formed by coexpressing the three subunits in oocytes are similar to that of the native channel.

Comparative phylogeography of mutualists and the effect of the host on the genetic structure of its partners.

Whether or not species participating in specialized and obligate interactions display similar and simultaneous demographic variations at the intraspecific level remains an open question in phylogeography. In the present study, we used the mutualistic nursery pollination occurring between the European globeflower Trollius europaeus and its specialized pollinators in the genus Chiastocheta as a case study. Explicitly, we investigated if the phylogeographies of the pollinating flies are significantly different from the expectation under a scenario of plant-insect congruence. Based on a large-scale sampling, we first used mitochondrial data to infer the phylogeographical histories of each fly species. Then, we defined phylogeographical scenarios of congruence with the plant history, and used maximum likelihood and Bayesian approaches to test for plant-insect phylogeographical congruence for the three Chiastocheta species. We show that the phylogeographical histories of the three fly species differ. Only Chiastocheta lophota and Chiastocheta dentifera display strong spatial genetic structures, which do not appear to be statistically different from those expected under scenarios of phylogeographical congruence with the plant. The results of the present study indicate that the fly species responded in independent and different ways to shared evolutionary forces, displaying varying levels of congruence with the plant genetic structure

cAMP-dependent chloride secretion mediates tubule enlargement and cyst formation by cultured mammalian collecting duct cells.

Polycystic kidney diseases result from disruption of the genetically defined program that controls the size and geometry of renal tubules. Cysts which frequently arise from the collecting duct (CD) result from cell proliferation and fluid secretion. From mCCD(cl1) cells, a differentiated mouse CD cell line, we isolated a clonal subpopulation (mCCD-N21) that retains morphogenetic capacity. When grown in three-dimensional gels, mCCD-N21 cells formed highly organized tubular structures consisting of a palisade of polarized epithelial cells surrounding a cylindrical lumen. Subsequent addition of cAMP-elevating agents (forskolin or cholera toxin) or of membrane-permeable cAMP analogs (CPT-cAMP) resulted in rapid and progressive dilatation of existing tubules, leading to the formation of cystlike structures. When grown on filters, mCCD-N21 cells exhibited a high transepithelial resistance as well as aldosterone- and/or vasopressin-induced amiloride-sensitive and -insensitive current. The latter was in part inhibited by Na(+)-K(+)-2Cl(-) cotransporter (bumetanide) and chloride channel (NPPB) inhibitors. Real-time PCR analysis confirmed the expression of NKCC1, the ubiquitous Na(+)-K(+)-2Cl(-) cotransporter and cystic fibrosis transmembrane regulator (CFTR) in mCCD-N21 cells. Tubule enlargement and cyst formation were prevented by inhibitors of Na(+)-K(+)-2Cl(-) cotransporters (bumetanide or ethacrynic acid) or CFTR (NPPB or CFTR inhibitor-172). These results further support the notion that cAMP signaling plays a key role in renal cyst formation, at least in part by promoting chloride-driven fluid secretion. This new in vitro model of tubule-to-cyst conversion affords a unique opportunity for investigating the molecular mechanisms that govern the architecture of epithelial tubes, as well as for dissecting the pathophysiological processes underlying cystic kidney diseases.

Caracterización especiológica bentodemersal durante el crucero de investigación del recurso merluza BIC Jose Olaya Balandra 9901

Los lances de pesca se efectuaron entre 96 y 560 m de profundidad, del 7 al 26 de enero de 1999, frente al área entre Huarmey (10°5 'S) y Puerto Pizarro (3°29 'S). Se registraron 168 especies: 97 peces y 71 invertebrados (35 crustáceos, 19 moluscos, 8 equinodermos, 3 poliquetos y 6 cnidarios). La captura total fue de 19.962,13 kg, de los cuales 18.041,53 kg (90,38%) correspondieron a peces; 93,88 kg (0.47%) a crustáceos; 11,65 kg (0,06%) a moluscos; 7,69 kg (0,04%) a equinodermos; 0,64 kg (0,03%) a poliquetos y 1.806,74 kg (9,05%) a cnidarios. El falso volador Prionotus stephanophrys destacó por su abundancia y frecuencia, habiéndose capturado 6.755,26 kg (33,8% del volumen total), seguido por la merluza Merluccius gayi peruanus con 4.823,99 kg (24,2%) y la cabrilla Paralabrax humeralis con 4.581,23 kg (23,0%). Se analiza también la distribución latitudinal y batimétrica de la fauna registrada, y se presentan las tallas de tres especies de peces (anguila común Ophichthus pacifici, congrio negro Cherublemma emmelas y el pez guadaña Hoplostethus pacificus) capturadas durante cinco operaciones de pesca. La diversidad específica y equitabilidad referidas a la merluza son objeto de análisis, y se determina que son significativamente bajas en el estrato I de la mayoría de subáreas en estudio, donde alcanzan valores hasta de cero, a excepción de la subárea F, mientras que en los estratos II y III de las subáreas A, B, C, D y F alcanzaron valores significativos.

Respective roles of calcitonin receptor-like receptor (CRLR) and receptor activity-modifying proteins (RAMP) in cell surface expression of CRLR/RAMP heterodimeric receptors.

Receptor activity modifying proteins RAMP1, RAMP2, and RAMP3 are responsible for defining affinity to ligands of the calcitonin receptor-like receptor (CRLR). It has also been proposed that receptor activity-modifying proteins (RAMP) are molecular chaperones required for CRLR transport to the cell surface. Here, we have studied the respective roles of CRLR and RAMP in transporting CRLR/RAMP heterodimers to the plasma membrane by using a highly specific binding assay that allows quantitative detection of cell surface-expressed CRLR or RAMP in the Xenopus oocytes expression system. We show that: (i) heterodimer assembly is not a prerequisite for efficient cell surface expression of CRLR, (ii) N-glycosylated RAMP2 and RAMP3 are expressed at the cell surface and their transport to the plasma membrane requires N-glycans, (iii) RAMP1 is not N-glycosylated and is transported to the plasma membrane only upon formation of heterodimers with CRLR, and (iv) introduction of N-glycosylation sites in the RAMP1 sequence (D58N/G60S, Y71N, and K103N/P105S) allows cell surface expression of these mutants at levels similar to that of wild-type RAMP1 co-expressed with CRLR. Our data argue against a chaperone function for RAMP and identify the role of N-glycosylation in targeting these molecules to the cell surface.

Recurrent deletions and reciprocal duplications of 10q11.21q11.23 including CHAT and SLC18A3 are likely mediated by complex low-copy repeats.

We report 24 unrelated individuals with deletions and 17 additional cases with duplications at 10q11.21q21.1 identified by chromosomal microarray analysis. The rearrangements range in size from 0.3 to 12 Mb. Nineteen of the deletions and eight duplications are flanked by large, directly oriented segmental duplications of >98% sequence identity, suggesting that nonallelic homologous recombination (NAHR) caused these genomic rearrangements. Nine individuals with deletions and five with duplications have additional copy number changes. Detailed clinical evaluation of 20 patients with deletions revealed variable clinical features, with developmental delay (DD) and/or intellectual disability (ID) as the only features common to a majority of individuals. We suggest that some of the other features present in more than one patient with deletion, including hypotonia, sleep apnea, chronic constipation, gastroesophageal and vesicoureteral refluxes, epilepsy, ataxia, dysphagia, nystagmus, and ptosis may result from deletion of the CHAT gene, encoding choline acetyltransferase, and the SLC18A3 gene, mapping in the first intron of CHAT and encoding vesicular acetylcholine transporter. The phenotypic diversity and presence of the deletion in apparently normal carrier parents suggest that subjects carrying 10q11.21q11.23 deletions may exhibit variable phenotypic expressivity and incomplete penetrance influenced by additional genetic and nongenetic modifiers.

Selectividad de la red Granton 400x120 mm PA en la evaluación de merluza peruana. Otoño 2002

En el crucero BIC Olaya 0205 (Callao-Puerto Pizarro), se monitoreó la selectividad de la red de arrastre de fondo Granton tipo 400x120 mm PA, empleando copo (90 mm) y sobrecopo (13 mm). La probabilidad de captura al 50%, en la subárea A fue de 4,52cm; en la B, de 25,46 cm y en la C de 26,14 cm. El L50% total de las subáreas A, B, C fue 25,15 cm, con un factor de selección 0,28 y un rango de selección de 8,55 cm.

Dinâmica da agregação induzida pelo uso de plantas de inverno para cobertura do solo

O impacto de plantas de inverno sobre a estabilidade estrutural do solo, antecedendo a cultura do milho sob plantio direto, foi avaliado em Podzólico Vermelho-Amarelo. O experimento foi realizado no campus da Universidade Federal de Santa Maria, de maio de 1991 a maio de 1992. Os tratamentos utilizados constaram de: chícharo (Lathyrus sativus L.), tremoço azul (Lupinus angustifolius L.), ervilhaca (Vicia sativa L.), aveia preta (Avena strigosa Schieb) e pousio invernal. Em cada parcela, foram coletadas mensalmente, desde a implantação das plantas de inverno até a colheita do milho, totalizando treze coletas, amostras de solo parcialmente deformadas e indeformadas para as determinações. Foram usadas duas subamostras na profundidade de 0-5 cm, com pá de corte, compondo uma amostra para análise de agregados e carbono orgânico e cinco subamostras, na mesma profundidade, com uso de cilindro volumétrico, compondo amostra para análise da atividade microbiana e umidade do solo. As plantas de cobertura induziram variação temporal da estabilidade dos agregados no período de estudo. A aveia preta atingiu maiores valores de estabilidade estrutural durante o ciclo das culturas de inverno, enquanto o tremoço azul maiores valores durante o ciclo do milho. Isso pode ser atribuído ao sistema radicular da gramínea e à taxa de decomposição da leguminosa, criando ambiente favorável à agregação, pela ação de raízes, cobertura do solo, fornecimento de material orgânico e conservação da umidade favoráveis à ação de microrganismos. Tais fatores, provavelmente, favoreceram a formação e conservação dos agregados do solo.

Expertises croisées dans la dégénérescence maculaire liée à l'âge. Focus sur la physiopathologie, l'angiogenèse, les données pharmacologiques et cliniques [Review and expert opinion in age related macular degeneration. Focus on the pathophysiology, angiogenesis and pharmacological and clinical data].

Age related macular degeneration (AMD) is a pathological aging of the macula, brought about by the interaction of genetic and environmental factors. It induces geographic atrophy of the retina and/or choroidal neovascularization. In the latter, abnormal vessels develop from the choriocapillaris, with the involvement of VEGF (vascular endothelial growth factor). The VEGF family includes several factors, including VEGF-A, B, C, D, F and PlGF (placental growth factor). Their biological properties and their affinities to the VEGFR1, VEGFR2 and VEGFR3 receptors found on endothelial cells differ. Exudative AMD involves mainly VEGF-A and VEGF-R2. Anti-VEGF agents used in ophthalmology (ranibizumab, bevacizumab and aflibercept) are designed to primarily target this pathway. In vitro, all have sufficient affinity to their ligands. Their therapeutic efficacy must therefore be judged based on clinical criteria. In clinical practice, the minimum number of injections required for a satisfactory result appears to be comparable with all the three. The few available studies on therapeutic substitutions of anti-VEGF compounds suggest that some patients may benefit from substituting the anti-VEGF in cases of an unsatisfactory response to an initial molecule. Although local side effects, including increased risk of geographic atrophy, and systemic effects, including vascular accidents, have been suggested, these risks remain low, specially compared to the benefits of the treatment. Differences in safety between anti-VEGF are theoretically possible but unproven.

Influência de diferentes sistemas de uso e manejo do solo sobre a flutuação populacional do Oligochaeta edáfico Amynthas spp.

Avaliou-se, mensalmente, a influência dos sistemas plantio direto (PD) e convencional (PC), mata nativa (MN) e campo nativo (CN) sobre a densidade populacional e biomassa do Oligochaeta gênero Amynthas spp., na região dos Campos Gerais (PR), no período de fevereiro de 1993 a janeiro de 1994. No PC e PD, as áreas foram cultivadas com milho, triticale e soja, enquanto, no campo nativo, a vegetação era composta de estepes de gramíneas baixas e de matas de galerias subtropicais para a mata nativa. A densidade populacional foi determinada pelo método do formol a 0,22% e a biomassa pela pesagem direta em balança analítica. A densidade populacional e a biomassa foram superiores no plantio direto e mata nativa, comparadas às do plantio convencional, enquanto, no campo nativo, não ocorreu a presença do gênero estudado. Os valores mais expressivos de densidade populacional e biomassa nos ecossistemas plantio direto e mata nativa foram observados no verão e primavera.

Nitric oxide reduces Cl- absorption in the mouse cortical collecting duct through an ENaC-dependent mechanism.

Since nitric oxide (NO) participates in the renal regulation of blood pressure, in part, by modulating transport of Na(+) and Cl(-) in the kidney, we asked whether NO regulates net Cl(-) flux (JCl) in the cortical collecting duct (CCD) and determined the transporter(s) that mediate NO-sensitive Cl(-) absorption. Cl(-) absorption was measured in CCDs perfused in vitro that were taken from aldosterone-treated mice. Administration of an NO donor (10 μM MAHMA NONOate) reduced JCl and transepithelial voltage (VT) both in the presence or absence of angiotensin II. However, reducing endogenous NO production by inhibiting NO synthase (100 μM N(G)-nitro-l-arginine methyl ester) increased JCl only in the presence of angiotensin II, suggesting that angiotensin II stimulates NO synthase activity. To determine the transport process that mediates NO-sensitive changes in JCl, we examined the effect of NO on JCl following either genetic ablation or chemical inhibition of transporters in the CCD. Since the application of hydrochlorothiazide (100 μM) or bafilomycin (5 nM) to the perfusate or ablation of the gene encoding pendrin did not alter NO-sensitive JCl, NO modulates JCl independent of the Na(+)-dependent Cl(-)/HCO3(-) exchanger (NDCBE, Slc4a8), the A cell apical plasma membrane H(+)-ATPase and pendrin. In contrast, both total and NO-sensitive JCl and VT were abolished with application of an epithelial Na(+) channel (ENaC) inhibitor (3 μM benzamil) to the perfusate. We conclude that NO reduces Cl(-) absorption in the CCD through a mechanism that is ENaC-dependent.

Global radiolarian zonation for the Pliensbachian, Toarcian and Aalenian

Jurassic radiolarians from 220 samples in Queen Charlotte Islands, B.C., Williston Lake, B.C., east-central Oregon, Baja California Sur, southern Spain, Austria, Slovenia, Turkey, Oman, Japan and Argentina were studied in order to construct global zonation for the Pliensbachian, Toarcian and Aalenian stages. Well-preserved faunas from continuous stratigraphic sections in Queen Charlotte Islands provide the most detailed record for this time interval, and all collections are tied to North American ammonite zones or assemblages. Collections from nearly all other areas lack independent dating except for early Toarcian carbon-isotope dating in Slovenia and late Aalenian ammonites in Spain. A database of 197 widely distributed updated taxonomic species was used to construct a Unitary Association (UA) zonation for the interval. A global sequence of 41 UAs was obtained for the top of the Sinemurian to the base of the Bajocian. The first and the last UAs represent the Late Sinemurian and the Early Bajocian respectively. The remaining 39 UAs were merged into nine zones (four Early Pliensbachian, one Late Pliensbachian, one Early Toarcian, one Middle-Late Toarcian, and two Aalenian) according to prominent radiolarian faunal breaks and ammonite data. The new zones are the Canutus tip pen - Katroma clara Zone (latest Sinemurian/earliest Pliensbachian); Zartus mostleri - Pseudoristola megaglobosa, Hsuum mulleri - Trillus elkhornensis and Gigi fustis - Lantus sixi zones (Early Pliensbachian); Eucyrtidiellum nagaiae - Praeparvicingula tlellensis Zone (Late Pliensbachian); Napora relica - Eucyrtidiellum disparile Zone (Early Toarcian); Elodium pessagnoi - Hexasaturnalis hexagonus Zone (Middle and Late Toarcian); Higumastra transversa - Napora nipponica Zone (early Aalenian); and Mirifusus proavus - Transhsuum hisuikyoense Zone (late Aalenian). These zones can be correlated worldwide and link previously established UA zonations for the Hettangian-Sinemurian and the Middle to Upper Jurassic. The new zonation allows high-resolution dating in the studied interval and provides a solid basis for analyzing faunal turnovers and the paleobiogeography of Jurassic radiolarians. (C) 2010 Elsevier B.V. All rights reserved.

Teores de nutrientes e de matéria orgânica afetados pela rotação de culturas e sistema de preparo do solo

Este trabalho foi realizado na Embrapa Arroz e Feijão, em Santo Antônio de Goiás (GO), em Latossolo Vermelho perférrico, sob pivô central, por seis anos consecutivos, durante os quais se efetuaram 12 cultivos. Estudaram-se os efeitos de quatro sistemas de preparo do solo e seis rotações de culturas sobre o pH em água, Al e Ca + Mg trocáveis, teor de matéria orgânica, P extraível e K trocável . Foi utilizado o delineamento inteiramente casualizado, em parcelas subdivididas, com três repetições. As parcelas foram constituídas pelos sistemas de preparo: arado/grade, arado, grade e plantio direto; as subparcelas, pelas rotações: (a) arroz-feijão, (b) milho-feijão, (c) soja-trigo, (d) soja-trigo-soja-feijão-arroz-feijão, (e) arroz consorciado com calopogônio-feijão e (f) milho-feijão-milho-feijão-arroz-feijão. As rotações a, b, c e e foram anuais e as d e f, trienais. As diferenças observadas entre os sistemas de preparo do solo, quanto aos valores de pH, Al e Ca + Mg trocáveis, deveram-se à profundidade de mobilização do solo e à incorporação do calcário. A distribuição do K trocável e do P extraível no perfil do solo variou com a profundidade do solo mobilizada pelos diferentes sistemas de preparo, especialmente no caso do P, em que a menor mobilização causou maior concentração na camada superficial. Os sistemas de rotação que incluíram soja propiciaram maiores valores de pH e Ca + Mg trocáveis e menores de Al trocável. Os sistemas de preparo do solo e de rotação mantiveram, após 12 cultivos, teores de matéria orgânica do solo semelhantes aos iniciais.

Efeitos do sistema de preparo e da rotação de culturas na porosidade e densidade do solo

O trabalho foi realizado na Embrapa Arroz e Feijão, em Santo Antônio de Goiás (GO), em Latossolo Vermelho perférrico, sob pivô central, por seis anos consecutivos, durante os quais se efetuaram 12 cultivos. Estudaram-se os efeitos de quatro sistemas de preparo do solo e seis rotações de culturas sobre a densidade e porosidade do solo. Foi utilizado o delineamento inteiramente casualizado, em parcelas subdivididas, com três repetições. As parcelas foram constituídas pelos sistemas de preparo: arado/grade, arado, grade e plantio direto e as subparcelas pelas rotações: (a) arroz-feijão, (b) milho-feijão, (c) soja-trigo, (d) soja-trigo-soja-feijão-arroz-feijão, (e) arroz consorciado com calopogônio-feijão e (f) milho-feijão-milho-feijão-arroz-feijão. As rotações a, b, c e e foram anuais e as d e f, trienais. O plantio direto ocasionou maior valor de densidade do solo e menores de porosidade total e macroporosidade na camada superficial, enquanto o preparo do solo com grade aradora propiciou o menor valor de densidade e maior de porosidade total. Nas camadas mais profundas, o preparo do solo com arado de aiveca propiciou os menores valores de densidade do solo e maiores de porosidade total e macroporosidade. Na camada superficial do solo, os sistemas de rotação que incluíram soja e trigo levaram a maiores valores de densidade do solo e microporosidade e menores de macroporosidade, enquanto o sistema arroz consorciado com calopogônio-feijão propiciou maior valor de macroporosidade e menor de microporosidade.

Eficiência da fixação biológica de N2 por estirpes de Bradyrhizobium na soja em plantio direto

O presente trabalho foi realizado nas safras de 1994/95, 1996/97, 1997/98 e 1998/99, objetivando avaliar a eficiência das estirpes de Bradyrhizobium recomendadas pela pesquisa, pela determinação da capacidade competitiva e da eficiência na fixação do N2 na cultura da soja, sob o sistema plantio direto. Os experimentos foram instalados em áreas experimentais da FUNDACEP, Cruz Alta (RS), manejadas desde cinco até dez anos em plantio direto. Na safra de 1994/95, foram usados os seguintes tratamentos: testemunha sem inoculação; 200 kg ha-1 de N mineral, parcelados e sem inoculação; e as combinações das quatro estirpes recomendadas comercialmente, SEMIA 587 + SEMIA 5019; SEMIA 587 + SEMIA 5079; SEMIA 587 + SEMIA 5080; SEMIA 5019 + SEMIA 5079; SEMIA 5019 + SEMIA 5080; SEMIA 5079 + SEMIA 5080 e dois isolados do sorogrupo SEMIA 586. Em 1996/97, foi retirado o último tratamento e acrescidos os tratamentos CPAC 40 + CPAC 44 e CPAC 42 + CPAC 45. Na safra de 1997/98, foi acrescido o tratamento 20 kg ha-1 de N mineral na semeadura + inoculação com SEMIA 5079 + SEMIA 5080. A adubação nitrogenada reduziu a nodulação e não resultou em acréscimo na produção da soja. Na avaliação do rendimento de grãos, em cada uma das quatro safras e na análise conjunta, a testemunha sem inoculação não apresentou diferença significativa em relação aos demais tratamentos. A não-resposta da soja à prática da inoculação pode ser atribuída à população naturalizada de Bradyrhizobium, em número adequado e eficiente, e às condições favoráveis à fixação biológica do nitrogênio, como temperatura e umidade adequadas do solo, proporcionadas pelo sistema plantio direto.