958 resultados para Ca(2 ) uniporter
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The main Precambrian tectonic units of Uruguay include the Piedra Alta tectonostratigraphic terrane (PATT) and Nico Perez tectonostratigraphic terrane (NPTT), separated by the Sarandi del Yi high-strain zone. Both terranes are well exposed in the Rio de La Plata craton (RPC). Although these tectonic units are geographically small, they record a wide span of geologic time. Therefore improved geological knowledge of this area provides a fuller understanding of the evolution of the core of South America. The PATT is constituted by low-to medium-grade metamorphic belts (ca. 2.1 Ga); its petrotectonic associations such as metavolcanic units, conglomerates, banded iron formations, and turbiditic deposits suggest a back-arc or a trench-basin setting. Also in the PATT, a late to post-orogenic, arc-related layered mafic complex (2.3-1.9 Ga), followed by A-type granites (2.08 Ga), and finally a taphrogenic mafic dike swarm (1.78 Ga) occur. The less thoroughly studied NPTT consists of Palaeoproterozoic high-grade metamorphic sequences (ca. 2.2 Ga), mylonites and postorogenic and rapakivi granites (1.75 Ga). The Brasiliano-Pan African orogeny affected this terrane. Neoproterozoic cover occurs in both tectonostratigraphic terranes, but is more developed in the NPTT. Over the past 15 years, new isotopic studies have improved our recognition of different tectonic events and associated processes, such as reactivation of shear zones and fluids circulation. Transamazonian and Statherian tectonic events were recognized in the RPC. Based on magmatism, deformation, basin development and metamorphism, we propose a scheme for the Precambrian tectonic evolution of Uruguay, which is summarized in the first Palaeoproterozoic tectonic map of the Rio de La Plata craton.
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Leptospixosis, a spirochaetal zoonotic disease caused by Leptospira, has been recognized as an important emerging infectious disease. LipL32 is the major exposed outer membrane protein found exclusively in pathogenic leptospires, where it accounts for up to 75% of the total outer membrane proteins. It is highly immunogenic, and recent studies have implicated LipL32 as an extracellular matrix binding protein, interacting with collagens, fibronectin, and laminin. In order to better understand the biological role and the structural requirements for the function of this important lipoprotein, we have determined the 2.25-angstrom-resolution structure of recombinant LipL32 protein corresponding to residues 21-272 of the wild-type protein (LipL32(21-272)). The LipL32(21-272) monomer is made of a jelly-roll fold core from which several peripheral secondary structures protrude. LipL32(21-272) is structurally similar to several other jelly-roll proteins, some of which bind calcium ions and extracellular matrix proteins. Indeed, spectroscopic data (circular dichroism, intrinsic tryptophan fluorescence, and extrinsic 1-amino-2-naphthol-4-sulfonic acid fluorescence) confirmed the calcium-binding properties of LipL32(21-272). Ca(2+) binding resulted in a significant increase in the thermal stability of the protein, and binding was specific for Ca(2+) as no structural or stability perturbations were observed for Mg(2+), Zn(2+), or Cu(2+). Careful examination of the crystal lographic structure suggests the locations of putative regions that could mediate Ca(2+) binding as well as binding to other interacting host proteins, such as collagens, fibronectin, and lamixidn. (C) 2009 Elsevier Ltd. All rights reserved.
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Nicotinic acetylcholine receptors (nAChR) exert pivotal roles in synaptic transmission, neuroprotection and differentiation. Particularly, homomeric alpha 7 receptors participate in neurite outgrowth, presynaptic control of neurotransmitter release and Ca(2+) influx. However, the study of recombinant alpha 7 nAChRs in transfected cell lines is difficult due to low expression of functional receptor channels. We show that PC12 pheochromocytoma cells induced to differentiation into neurons are an adequate model for studying differential nAChR gene expression and receptor activity. Whole-cell current recording indicated that receptor responses increased during the course of differentiation. Transcription of mRNAs coding for alpha 3, alpha 5, alpha 7, beta 2 and beta 4 subunits was present during the course of differentiation, while mRNAs coding for alpha 2, alpha 4 and beta 3 subunits were not expressed in PC12 cells. alpha 7 subunit expression was highest following 1 day of induction to differentiation. Activity of alpha 7 nAChRs, however, was most elevated on day 2 as revealed by inhibition experiments in the presence of 10 nM methyllycaconitine, rapid current decay and receptor responsiveness to the alpha 7 agonist choline. Increased alpha 7 receptor activity was noted when PC12 were induced to differentiation in the presence of choline, confirming that chronic agonist treatment augments nAChR activity. In summary, PC12 cells are an adequate model to study the role and pharmacological properties of this receptor during neuronal differentiation.
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The Blastocladiella emersonii life cycle presents a number of drastic biochemical and morphological changes, mainly during two cell differentiation stages: germination and sporulation. To investigate the transcriptional changes taking place during the sporulation phase, which culminates with the production of the zoospores, motile cells responsible for the dispersal of the fungus, microarray experiments were performed. Among the 3,773 distinct genes investigated, a total of 1,207 were classified as differentially expressed, relative to time zero of sporulation, at at least one of the time points analyzed. These results indicate that accurate transcriptional control takes place during sporulation, as well as indicating the necessity for distinct molecular functions throughout this differentiation process. The main functional categories overrepresented among upregulated genes were those involving the microtubule, the cytoskeleton, signal transduction involving Ca(2+), and chromosome organization. On the other hand, protein biosynthesis, central carbon metabolism, and protein degradation were the most represented functional categories among downregulated genes. Gene expression changes were also analyzed in cells sporulating in the presence of subinhibitory concentrations of glucose or tryptophan. Data obtained revealed overexpression of microtubule and cytoskeleton transcripts in the presence of glucose, probably causing the shape and motility problems observed in the zoospores produced under this condition. In contrast, the presence of tryptophan during sporulation led to upregulation of genes involved in oxidative stress, proteolysis, and protein folding. These results indicate that distinct physiological pathways are involved in the inhibition of sporulation due to these two classes of nutrient sources.
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Incubation of T. cruzi epimastigotes with the lectin Cramoll 1,4 in Ca(2+) containing medium led to agglutination and inhibition of cell proliferation. The lectin (50 A mu g/ml) induced plasma membrane permeabilization followed by Ca(2+) influx and mitochondrial Ca(2+) accumulation, a result that resembles the classical effect of digitonin. Cramoll 1,4 stimulated (five-fold) mitochondrial reactive oxygen species (ROS) production, significantly decreased the electrical mitochondrial membrane potential (Delta I(m)) and impaired ADP phosphorylation. The rate of uncoupled respiration in epimastigotes was not affected by Cramoll 1,4 plus Ca(2+) treatment, but oligomycin-induced resting respiration was 65% higher in treated cells than in controls. Experiments using T. cruzi mitochondrial fractions showed that, in contrast to digitonin, the lectin significantly decreased Delta I(m) by a mechanism sensitive to EGTA. In agreement with the results showing plasma membrane permeabilization and impairment of oxidative phosphorylation by the lectin, fluorescence microscopy experiments using propidium iodide revealed that Cramoll 1,4 induced epimastigotes death by necrosis.
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Metal cation toxicity to basidiomycete fungi is poorly understood, despite its well-known importance in terrestrial ecosystems. Moreover, there is no reported methodology for the routine evaluation of metal toxicity to basidiomycetes. In the present study, we describe the development of a procedure to assess the acute toxicity of metal cations (Na(+), K(+), Li(+), Ca(2+), Mg(2+), Co(2+), Zn(2+), Ni(2+), Mn(2+), Cd(2+), and Cu(2+)) to the bioluminescent basidiomycete fungus Gerronema viridilucens. The method is based on the decrease in the intensity of bioluminescence resulting from injuries sustained by the fungus mycelium exposed to either essential or nonessential metal toxicants. The assay described herein enables LIS to propose a metal toxicity series to Gerronenia viridilucens based on data obtained from the bioluminescence intensity (median effective concentration [EC50] values) versus metal concentration: Cd(2+) > Cu(2+) > Mn(2+) approximate to Ni(2+) approximate to Co(2+) > Zn(2+) > Mg(2+) > Li(+) > K(+) approximate to Na(+) > Ca(2+), and to shed some li-ht on the mechanism of toxic action of metal cations to basidiomycete fungi. Environ. Toxicol. Chem. 2010;29:320-326. (C) 2009 SETAC
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An all-in-one version of a capacitively coupled contactless conductivity detector is introduced. The absence of moving parts (potentiometers and connectors) makes it compact (6.5 cm(3)) and robust. A local oscillator, working at 1.1 MHz, was optimized to use capillaries of id from 20 to 100 lam. Low noise circuitry and a high-resolution analog-to-digital converter (ADC) (21 bits effective) grant good sensitivities for capillaries and background electrolytes currently used in capillary electrophoresis. The fixed frequency and amplitude of the signal generator is a drawback that is compensated by the steady calibration curves for conductivity. Another advantage is the possibility of determining the inner diameter of a capillary by reading the ADC when air and subsequently water flow through the capillary. The difference of ADC reading may be converted into the inner diameter by a calibration curve. This feature is granted by the 21-bit ADC, which eliminates the necessity of baseline compensation by hardware. In a typical application, the limits of detection based on the 3 sigma criterion (without baseline filtering) were 0.6, 0.4, 0.3, 0.5, 0.6, and 0.8 mu mol/L for K(+), Ba(2+), Ca(2+), Na(+), Mg(2+), and Li(+), respectively, which is comparable to other high-quality implementations of a capacitively coupled contactless conductivity detector.
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The prion protein (PrP(C)) is highly expressed in the nervous system, and its abnormal conformer is associated with prion diseases. PrP(C) is anchored to cell membranes by glycosylphosphatidylinositol, and transmembrane proteins are likely required for PrP(C)-mediated intracellular signaling. Binding of laminin (Ln) to PrP(C) modulates neuronal plasticity and memory. We addressed signaling pathways triggered by PrP(C)-Ln interaction in order to identify transmembrane proteins involved in the transduction of PrP(C)-Ln signals. The Ln gamma 1-chain peptide, which contains the Ln binding site for PrP(C), induced neuritogenesis through activation of phospholipase C (PLC), Ca(2+) mobilization from intracellular stores, and protein kinase C and extracellular signal-regulated kinase (ERK1/2) activation in primary cultures of neurons from wild-type, but not PrP(C)-null mice. Phage display, coimmunoprecipitation, and colocalization experiments showed that group I metabotropic glutamate receptors (mGluR1/5) associate with PrP(C). Expression of either mGluR1 or mGluR5 in HEK293 cells reconstituted the signaling pathways mediated by PrP(C)-Ln gamma 1 peptide interaction. Specific inhibitors of these receptors impaired PrP(C)-Ln gamma 1 peptide-induced signaling and neuritogenesis. These data show that group I mGluRs are involved in the transduction of cellular signals triggered by PrP(C)-Ln, and they support the notion that PrP(C) participates in the assembly of multiprotein complexes with physiological functions on neurons.-Beraldo, F. H., Arantes, C. P., Santos, T. G., Machado, C. F., Roffe, M., Hajj, G. N., Lee, K. S., Magalhaes, A. C., Caetano, F. A., Mancini, G. L., Lopes, M. H., Americo, T. A., Magdesian, M. H., Ferguson, S. S. G., Linden, R., Prado, M. A. M., Martins, V. R. Metabotropic glutamate receptors trans-duce signals for neurite outgrowth after binding of the prion protein to laminin gamma 1 chain. FASEB J. 25, 265-279 (2011). www.fasebj.org
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Polysaccharide natural seed coat from the tree Magonia pubescens, in the form of hydrogel was used to remove metals in aqueous solution. Swelling tests indicate that seed coat presents hydrogel behavior, with maximum water absorption of 292 g water/g. Adsorption experiments performed using Na(+), Mg(2+), K(+), Ca(2+), Cr(3+), Fe(3+) and Zn(2+) demonstrated that the polysaccharide structure has a high capacity to extract these ions from the aqueous solution. Scanning electron microscopy revealed significant morphological changes of the material before and after water contact. Differential scanning calorimetry measurements indicate a signal shift of the water evaporation temperature in the material with adsorbed zinc. X-ray photoelectron spectroscopy analysis combined with theoretical studies by the density functional theory and on Hartree-Fock (HF) level evidence that the metallic ions were adsorbed through coordination with hydroxyl groups of polysaccharide. In the case of Zn(2+) the lowest HF energy was observed for the tetracoordination mode, where Zn(2+) is coordinated by two hydroxyl groups and two water molecules.
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The aim of this study was to develop a fast capillary electrophoresis method for the determination of inorganic cations (Na(+), K(+), Ca(2+), Mg(2+)) in biodiesel samples, using barium (Ba(2+)) as the internal standard. The running electrolyte was optimized through effective mobility curves in order to select the co-ion and Peakmaster software was used to determine electromigration dispersion and buffer capacity. The optimum background electrolyte was composed of 10 mmol L(-1) imidazole and 40 mmol L(-1) of acetic acid. Separation was conducted in a fused-silica capillary (32 cm total length and 23.5 cm effective length, 50 mu m I.D.), with indirect UV detection at 214 nm. The migration time was only 36 s. In order to obtain the optimized conditions for extraction, a fractional factorial experimental design was used. The variables investigated were biodiesel mass, pH, extractant volume, agitation and sonication time. The optimum conditions were: biodiesel mass of 200 mg, extractant volume of 200 mu L. and agitation of 20 min. The method is characterized by good linearity in the concentration range of 0.5-20 mg kg(-1) (r > 0.999), limit of detection was equal to 0.3 mg kg(-1), inter-day precision was equal to 1.88% and recovery in the range of 88.0-120%. The developed method was successfully applied to the determination of cations in biodiesel samples. (c) 2010 Elsevier B.V. All rights reserved.
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The interaction of emeraldine base (PANI-EB) with silver and gold colloids was probed by using Surface-Enhanced Resonance Raman Scattering (SERRS) at 3 different exciting radiations. Due to the great sensitivity of SERRS technique the detection limit of PANI-EB concentration was ca. 2 x 10(-7) mol L(-1) in Ag and Au colloidal suspensions. The UV-vis-NIR spectra of metal colloids in function of PANI-EB concentrations showed that gold colloids present a higher degree of aggregation than silver colloids. SERRS of PANI-EB on metal colloids allowed the study of the polymeric species formed primarily on the metallic surface. The polymer formed after the adsorption of PANI-EB on metallic nanoparticles is strongly dependent on the nature of the metal colloids. The oxidation of PANI-EB to pernigraniline occurred for silver colloids, while a doping process of PANI-EB on Au nanoparticles was evidenced through the observation of the characteristic SERRS spectrum of emeraldine salt at 1064nm.
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Anneberg är ett område i Danderyds kommun där det skall beredas plats för ett nytt bostadsområde. Området skall bebyggas med flerbostadshus, gruppbostäder och ett sjukhem. Denna förstudie beskriver översiktligt 3 systemförslag som kan användas för uppvärmning av husen i bostadsområdet Anneberg. Målsättningen är att presentera uppvärmningssystem som visar hur solenergi kan användas för att öka värmepumpsystemens värmefaktor.Systemen modellerades i TRNSYS och systemfunktionen samt energiflöden simulerades. Simulerade prestanda för tre olika typer av uppvärmningssystem redovisas. System A är ett vanligt värmepumpsystem med borrhål och värmepump placerad i ett flerfamiljshus av typ 3. System B liknar system A, men har kompletterats med en glasad solfångare för varmvattenberedning. System C är en lösning som kan tillämpas för större byggnader eller för ett område med flera byggnader. Systemet har ett gemensamt värmelager och ett kulvertsystem som förbinder byggnaderna med värmelagret. I varje ansluten byggnad installeras sedan en värmepump och en oglasad solfångare.Simuleringsresultatet redovisas som en värmefaktor för systemets fem första driftår. System A får en värmefaktor på mellan 2,3 och 2,7 för de första 5 driftåren. System B får en värmefaktor på mellan 3,4 och 3,7 och system C får en värmefaktor på mellan 4,0 och 4,5. Studien visar att det går att öka värmefaktorn på en värmepumpanläggning från ca 2,5 upp till 4 eller 4,5 genom att komplettera anläggningen med solfångare och värmelager. Detta innebär att elförbrukningen minskar från att vara ca 40 % av värmebehovet ned till under 25 % av värmebehovet. Det bör således finnas en potential för att komplettera värmepumpanläggningar med solvärme. Vilket utförande som kan bli ekonomiskt intressant kan inte bedömas i denna förstudie. I förstudien visas enbart resultatet för tre enstaka systemutföranden. Inga parametervariationer (tex solfångaryta, antal borrhål och avstånd mellan borrhålen) är utförda. En sådan systemoptimering bör göras med förstudien som utgångsläge.
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A proteína S100B pertence à família S100 de proteínas ligantes de Ca+2. Sua expressão dá-se primariamente em astrócitos, os quais também secretam esta proteína, exercendo um papel trófico sobre as células vizinhas. A adição de S100B tem promovido a sobrevivência de neurônios em cultura e, recentemente, tem sido proposto um papel protetor da S100B contra a excitoxicidade. Neste trabalho investigamos a liberação de S100B na presença de alta concentração de glutamato. A secreção de S100B em astrócitos de ratos em cultura foi quantificada, pelo método de ELISA, durante 24 horas após uma privação de soro de 30 minutos (condição estimulada) ou não (condição basal). A integridade dos astrócitos foi analisada por ensaios de exclusão de azul de tripan e medida da LDH. Glutamato (1 mM) não teve efeito sobre a secreção basal de S100B, mas diminuiu a liberação 1h depois da privação de soro. A privação de soro que estimulou a liberação de S100B foi dependente de síntese protéica e reduzida por Rp-AMPc e H-89, sugerindo o envolvimento da via AMPc/PKA, possivelmente sobre o elemento sensível ao AMPc no gene de S100B. Além disso, a privação de soro foi acompanhada por um aumento transitório do conteúdo intracelular de AMPc. Nossos resultados sugerem que o proposto papel neurotrófico da S100B, pelo menos em cultura de astrócitos hipocampais, poderia estar prejudicado por altos níveis de glutamato. Não está claro ainda se o efeito do glutamato é medeado por receptores. Na tentativa de investigar o mecanismo envolvido usamos inibidores do transporte de glutamato e agonistas de glutamato. Os resultados indicam que ambos receptores metabotrópicos do Grupo I/II e transportadores de glutamato estão envolvidos no decréscimo da secreção de S100B induzida pelo glutamato.
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Nesta tese da especialidade de Hidrogeologia, é apresentado um primeiro levantamento vulcano-estratigráfico da ilha da Madeira. Os trabalhos de campo permitiram definir, até agora, sete unidades geológicas principais, descritas da mais antiga para a mais recente: 1- Complexo Vulcânico Antigo (CA) 2- Calcários Marinhos dos Lameiros - S. Vicente (CM) 3- Depósito Conglomerático-Brechóide (CB) 4- Complexo Vulcânico Principal (CP) 5- Complexo Vulcânico S. Roque/Paul (SRP) 6- Episódios Vulcânicos Recentes (VR) 7- Depósitos de Vertente (dv), Fajãs (fj), Quebradas (q), Depósitos de Enxurrada Recentes (dr), Areias de Praia (ap), Dunas Fósseis (df), Terraços (t), e Aluviões (a) A caracterização climática da ilha da Madeira foi feita com base nos dados recolhidos em 27 pontos de observação climatológicos, dos quais 14 são estações meteorológicas e 13 são postos udométricos, o que representa uma densidade de 1 ponto de observação/27 km2. Constatou-se que a variação da precipitação com a altitude não era linear e dependia da orientação das vertentes. Efectuou-se o balanço hídrico sequencial diário, com base nos valores de precipitação média diária, registados no Paul da Serra, durante os últimos 15 anos hidrológicos. Pretendeu-se quantificar a precipitação oculta na vegetação típica da ilha da Madeira, de modo a poder avaliar a contribuição daquele tipo de precipitação para os recursos hídricos subterrâneos, e, ainda, determinar qual o potencial daquele recurso natural, como contribuição importante às fontes tradicionais de abastecimento de água à ilha. Para o efeito, procedemos a dois tipos principais de medição: medição directamente sob a vegetação de altitude da Madeira, o urzal, que se desenvolve entre os 1200 e os 1600 m de altitude, e construção de aparelhos constituídos por obstáculos artificiais, de modo a interceptarem as gotículas de água contidas no nevoeiro. Pela sua importância, os resultados obtidos justificam o desenvolvimento futuro deste estudo. A caracterização hidrodinâmica das formações da ilha da Madeira baseou-se nas observações feitas no interior das galerias e túneis em escavação, na análise dos registos de caudais de galerias e nascentes e na interpretação de ensaios de bombeamento dos furos de captação. Os dados evidenciam a grande heterogeneidade e anisotropia características do meio vulcânico. As transmissividades vão desde 11 m2/d até 25 766 m2/d. A caracterização hidrogeoquímica permitiu identificar um grupo de 5 águas termais, emergentes em falhas, no Complexo Antigo, com características muito próprias, bastante distintas das restantes, que representam, quer em quantidade, quer em volume de caudais captados, a grande maioria das águas da ilha da Madeira. O principal fenómeno mineralizador das águas é a hidrólise de minerais silicatados, verificando-se, nas águas dos furos, nas das nascentes de altitude e nas situadas próximo do litoral, o efeito da contaminação de sais de origem marinha. A partir dos dados hidrogeológicos obtidos, foi possível elaborar um modelo conceptual de funcionamento hidrogeológico para a ilha da Madeira, que, apesar de possuir alguns elementos comuns, é, no seu conjunto, diferente dos modelos conhecidos para outras ilhas vulcânicas, nomeadamente, Canárias, Reunião, Havai e Polinésia Francesa.
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A 140,0 kDa lectin was purified and characterized from the mushroom Clavaria cristata. The purification procedures from the crude extract of the mushroom comprised gel filtration chromatography on Sephacryl s200 and ion exchange on Resource Q column. The purified lectin agglutinated all types of human erythrocytes with preference for trypsinized type O erythrocytes. The haemagglutinating activity is dependent of Ca 2+ ions and was strongly inhibited by the glycoprotein bovine submaxillary mucin (BSM) up to the concentration of 0, 125 mg/mL. The C. cristata lectin (CcL) was stable in the pH range of 2,5-11,5 and termostable up to 80 °C. CcL molecular mass determined by gel filtration on a Superose 6 10 300 column was approximately 140,3 kDa. SDS polyacrilamide gel electrophoresis revealed a single band with a molecular mass of approximately 14,5 kDa, when the lectin was heated at 100 ⁰C in the presence or absence of β-mercaptoethanol. CcL induced activation of murine peritoneal macrophages in vitro resulting in the release of nitric oxide (NO), reaching the maximum production at 24 h. In experimental paw oedema model in mice, CcL showed proinflammatory activity being able to induce oedema formation. Cell viability of HepG2, MDA 435 e 3T3 cell lines was examined after 72 h of incubation with CcL in different concentrations (0,5-50 μg/mL). CcL inhibited HepG2 cells growth with an IC50 value of 50 μg/mL. In the present work, the observed immunomodulatory and antiproliferative effects indicate CcL as a possible immunomodulator compound, interfering in the macrophages immune response, taking possible anti-parasitic, anti-tumoral effects or diagnostic and/or therapeutic