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1.
Sci Rep ; 7(1): 2122, 2017 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-28522840

RESUMO

The physiological effects of caesium (Cs) on living cells are poorly understood. Here, we examined the physiological role of Cs+ on the activity of the potassium transporters in E. coli. In the absence of potassium (K+), Kup-mediated Cs+ uptake partially supported cell growth, however, at a much lower rate than with sufficient K+. In K+-limited medium (0.1 mM), the presence of Cs+ (up to 25 mM) in the medium enhanced growth as much as control medium containing 1 mM K+. This effect depended on the maintenance of basal levels of intracellular K+ by other K+ uptake transporters. Higher amounts of K+ (1 mM) in the medium eliminated the positive effect of Cs+ on growth, and revealed the inhibitory effect of high Cs+ on the growth of wild-type E. coli. Cells lacking Kdp, TrkG and TrkH but expressing Kup grew less well when Cs+ was increased in the medium. A kdp mutant contained an increased ratio of Cs+/K+ in the presence of high Cs+ in the medium and consequently was strongly inhibited in growth. Taken together, under excess Cs+ conditions Kup-mediated Cs+ influx sustains cell growth, which is supported by intracellular K+ supplied by Kdp.


Assuntos
Adenosina Trifosfatases/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Césio/metabolismo , Proteínas de Escherichia coli/metabolismo , Potássio/metabolismo , Transportadores de Cassetes de Ligação de ATP/metabolismo , Proliferação de Células , Escherichia coli/metabolismo , Transporte de Íons , Proteínas de Membrana/metabolismo , Canais de Potássio/metabolismo
2.
J Hazard Mater ; 335: 75-83, 2017 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-28432972

RESUMO

Uranium ore waste has led to soil contamination that may affect both environmental and soil health. To analyze the risk of metal transfer, metal bioavailability must be estimated by measuring biological parameters. Kinetic studies allow taking into account the dynamic mechanisms of bioavailability, as well as the steady state concentration in organisms necessary to take into account for relevant risk assessment. In this way, this work aims to model the snail accumulation and excretion kinetics of uranium (U), cesium (Cs) and thorium (Th). Results indicate an absence of Cs and Th accumulation showing the low bioavailability of these two elements and a strong uranium accumulation in snails related to the levels of soil contamination. During the depuration phase, most of the uranium ingested was excreted by the snails. After removing the source of uranium by soil remediation, continued snails excretion of accumulated uranium would lead to the return of their initial internal concentration, thus the potential trophic transfer of this hazardous element would stop.


Assuntos
Césio/metabolismo , Caracois Helix/metabolismo , Modelos Biológicos , Poluentes Radioativos do Solo/metabolismo , Tório/metabolismo , Urânio/metabolismo , Animais , Biodegradação Ambiental , Disponibilidade Biológica , Césio/isolamento & purificação , França , Poluentes Radioativos do Solo/isolamento & purificação , Tório/isolamento & purificação , Urânio/isolamento & purificação
3.
Sci Rep ; 7: 43170, 2017 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-28230101

RESUMO

Phytoaccumulation is a technique to extract metals from soil utilising ability of plants. Cesium is a valuable metal while radioactive isotopes of cesium can be hazardous. In order to establish a more efficient phytoaccumulation system, small molecules which promote plants to accumulate cesium were investigated. Through chemical library screening, 14 chemicals were isolated as 'cesium accumulators' in Arabidopsis thaliana. Of those, methyl cysteinate, a derivative of cysteine, was found to function within the plant to accumulate externally supplemented cesium. Moreover, metabolite profiling demonstrated that cesium treatment increased cysteine levels in Arabidopsis. The cesium accumulation effect was not observed for other cysteine derivatives or amino acids on the cysteine metabolic pathway tested. Our results suggest that methyl cysteinate, potentially metabolised from cysteine, binds with cesium on the surface of the roots or inside plant cells and improve phytoaccumulation.


Assuntos
Arabidopsis/metabolismo , Césio/metabolismo , Cisteína/análogos & derivados , Cisteína/metabolismo , Raízes de Plantas/metabolismo
4.
FEBS Lett ; 591(5): 718-727, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28134446

RESUMO

The aim of the present study was to investigate the influence of Cs+ on cultured human cells. We find that HeLa cell growth is suppressed by the addition of 10 mm CsCl into the culture media. In the Cs+ -treated cells, the intracellular Cs+ and K+ concentrations are increased and decreased, respectively. This leads to a decrease in activity of the glycolytic enzyme pyruvate kinase, which uses K+ as a cofactor. Cs+ -treated cells show an intracellular pH shift towards alkalization. Based on these results, CsCl presumably suppresses HeLa cell proliferation by inducing an intracellular cation imbalance that affects cell metabolism. Our findings may have implications for the use of Cs+ in cancer therapy.


Assuntos
Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Césio/farmacologia , Cloretos/farmacologia , Glicólise/efeitos dos fármacos , Piruvato Quinase/antagonistas & inibidores , Cátions Monovalentes , Sobrevivência Celular/efeitos dos fármacos , Césio/metabolismo , Meios de Cultura/química , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Potássio/metabolismo , Piruvato Quinase/metabolismo
5.
Elife ; 52016 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-27588351

RESUMO

The dynamics of different ionic currents shape the bursting activity of neurons and networks that control motor output. Despite being ubiquitous in all animal cells, the contribution of the Na(+)/K(+) pump current to such bursting activity has not been well studied. We used monensin, a Na(+)/H(+) antiporter, to examine the role of the pump on the bursting activity of oscillator heart interneurons in leeches. When we stimulated the pump with monensin, the period of these neurons decreased significantly, an effect that was prevented or reversed when the h-current was blocked by Cs(+). The decreased period could also occur if the pump was inhibited with strophanthidin or K(+)-free saline. Our monensin results were reproduced in model, which explains the pump's contributions to bursting activity based on Na(+) dynamics. Our results indicate that a dynamically oscillating pump current that interacts with the h-current can regulate the bursting activity of neurons and networks.


Assuntos
Potenciais de Ação , Geradores de Padrão Central/fisiologia , Trocadores de Sódio-Hidrogênio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Animais , Césio/metabolismo , Coração , Sanguessugas , Monensin/metabolismo , Ionóforos de Sódio/metabolismo
6.
Am J Physiol Endocrinol Metab ; 306(6): E697-706, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24452455

RESUMO

The question whether K⁺ depolarization is an appropriate experimental substitute for the physiological nutrient-induced depolarization of the ß-cell plasma membrane was investigated using primary mouse ß-cells and islets. At basal glucose 40 mM K⁺ induced a massive monophasic response, whereas 15 mM K⁺ had only a minimal insulinotropic effect, even though the increase in the cytosolic Ca²âº concentration ([Ca²âº]i) was not inferior to that by 20 mM glucose. In voltage-clamp experiments, Ca²âº influx appeared as nifedipine-inhibitable inward action currents in the presence of sulfonylurea plus TEA to block compensatory outward K⁺ currents. Under these conditions, 15 mM K⁺ induced prolonged action currents and 40 mM K⁺ transformed the action current pattern into a continuous inward current. Correspondingly, 15 mM K⁺ led to an oscillatory increase and 40 mM K⁺ to a plateau of [Ca²âº]i superimposed on the [Ca²âº]i elevated by sulfonylurea plus TEA. Raising K⁺ to 15 or 40 mM in the presence of sulfonylurea (±TEA) led to a fast further increase of insulin secretion. This was reduced to basal levels by nifedipine or CoCl2. The effects of 15 mM K⁺ on depolarization, action currents, and insulin secretion were mimicked by adding 35 mM Cs⁺ and those of 40 mM K⁺ by adding 35 mM Rb⁺, in parallel with their ability to substitute for K⁺ as permeant cation. In conclusion, the alkali metals K⁺, Rb⁺, or Cs⁺ concentration-dependently transform the pattern of Ca²âº influx into the ß-cell and may thus generate stimuli of supraphysiological strength for insulin secretion.


Assuntos
Sinalização do Cálcio , Membrana Celular/metabolismo , Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Potenciais da Membrana , Potássio/metabolismo , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Células Cultivadas , Césio/metabolismo , Glucose/metabolismo , Hipoglicemiantes/farmacologia , Secreção de Insulina , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/efeitos dos fármacos , Ilhotas Pancreáticas/efeitos dos fármacos , Ilhotas Pancreáticas/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos , Concentração Osmolar , Técnicas de Patch-Clamp , Bloqueadores dos Canais de Potássio/farmacologia , Rubídio/metabolismo , Técnicas de Cultura de Tecidos
7.
J Anim Physiol Anim Nutr (Berl) ; 97(3): 495-501, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22463084

RESUMO

Feed with Ammonium-iron-hexa-cyanoferrate (AFCF; 1250 mg AFCF/kg) was fed between March 2009 and March 2011 to wild boars in a territory of 4.5 km(2) (experimental group, EXP). One hundred and forty similar territories in the same county (500 km(2) , spruce forest, agriculture) served as control (CON). Data for comparison from all territories were available from March 2005 to March 2011. Wild boars could move between, into and from the territories. Lean skeletal muscle meat (500 g) of all wild boars that were killed by humans (hunting and traffic accidents) was investigated for gamma-radiation from (137) Cs with a becquerel monitor with a sodium iodide scintillator crystal (range of detection 20-9999 Bq/kg). The wild boars were weighed, and gender and age were determined. For the analyses of effects, multivariable regression models were fitted with the (137) Cs concentration as response variable. There was a significant difference between the (137) Cs contamination of wild boars from CON (563 ± 932 Bq/kg meat, n = 1253) and EXP (236 ± 276 Bq/kg meat; n = 45). (137) Cs contamination decreased with increasing body weight by -5 Bq/kg meat/kg body weight increase (p < 0.05). Females had higher Bq measurements than males (by +80 Bq/kg meat, p < 0.05). Piglets were lower than adults, but turn-coats higher. From November to May, contamination was higher (by +500 to +600 Bq/kg meat, p < 0.05) than during the rest of the year. In 2010, contamination was higher (by +200 to + 300 Bq/kg meat, p < 0.05) in comparison with the other years under observation. When all covariates were controlled for, the effect of AFCF was highly significant. Interaction analyses showed that the intervention decreased (137) Cs contamination by -500 Bq/kg meat during November to May and by -200 Bq/kg meat during the rest of the year. In summary, AFCF feeding reduces (137) Cs contamination of wild boars living in the wild significantly, particularly during the season from November to May.


Assuntos
Envelhecimento , Peso Corporal , Césio/metabolismo , Ferrocianetos/farmacologia , Estações do Ano , Sus scrofa , Ração Animal , Animais , Quelantes/farmacologia , Acidente Nuclear de Chernobyl , Dieta , Feminino , Alemanha , Masculino , Fatores Sexuais , Fatores de Tempo
8.
Pharm Res ; 29(5): 1404-18, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22146802

RESUMO

PURPOSE: To investigate the potential use of Prussian blue-coated magnetic nanoparticles, termed "Prussian blueberry", to bring about the magnetic elimination of cesium. METHODS: Prussian blueberry were prepared by a layer-by-layer assembly method. The morphology, structure and physical properties of the Prussian blueberry were investigated as was their ability to magnetically eliminate cesium. RESULTS: We confirmed that Prussian blueberry were composed of a magnetite nanoparticle-core and a Prussian blue-shell. Under a magnetic field, Prussian blueberry (5 mg) reduced the cesium concentration of seawater (3 ml) from 150 ppm to about 50 ppm; but regular Prussian blue could not magnetically eliminate cesium. Moreover, Prussian blueberry removed a similar proportion of cesium from a larger volume of seawater, and from fetal bovine serum and cow's milk. CONCLUSIONS: Under a magnetic field, Prussian blueberry was able to rapidly eliminate cesium from seawater and from biological matrices such as serum and milk.


Assuntos
Césio/metabolismo , Ferrocianetos/química , Magnetismo , Nanocompostos , Animais , Carcinógenos Ambientais/química , Bovinos , Césio/análise , Césio/sangue , Césio/química , Radioisótopos de Césio/análise , Radioisótopos de Césio/sangue , Radioisótopos de Césio/química , Radioisótopos de Césio/metabolismo , Estabilidade de Medicamentos , Concentração de Íons de Hidrogênio , Leite/química , Medicina Preventiva , Água do Mar/química , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Fatores de Tempo , Difração de Raios X
9.
J Mol Model ; 17(5): 1091-108, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-20676707

RESUMO

Crown ether architectures were explored for the inclusion of Cs(+) and Sr(2+) ions within nano-cavity of macrocyclic crown ethers using density functional theory (DFT) modeling. The modeling was undertaken to gain insight into the mechanism of the complexation of Cs(+) and Sr(2+) ion with this ligand experimentally. The selectivity of Cs(+) and Sr(2+) ions for a particular size of crown ether has been explained based on the fitting and binding interaction of the guest ions in the narrow cavity of crown ethers. Although, Di-Benzo-18-Crown-6 (DB18C6) and Di-Benzo-21-Crown-7 (DB21C7) provide suitable host architecture for Sr(2+) and Cs(+) ions respectively as the ion size match with the cavity of the host, but consideration of binding interaction along with the cavity matching both DB18C6 and DB21C7 prefers Sr(2+) ion. The calculated values of binding enthalpy of Cs metal ion with the crown ethers were found to be in good agreement with the experimental results. The gas phase binding enthalpy for Sr(2+) ion with crown ether was higher than Cs metal ion. The ion exchange reaction between Sr and Cs always favors the selection of Sr metal ion both in the gas and in micro-solvated systems. The gas phase selectivity remains unchanged in micro-solvated phase. We have demonstrated the effect of micro-solvation on the binding interaction between the metal ions (Cs(+) and Sr(2+)) and the macrocyclic crown ethers by considering micro-solvated metal ions up to eight water molecules directly attached to the metal ion and also by considering two water molecules attached to metal-ion-crown ether complexes. A metal ion exchange reaction involving the replacement of strontium ion in metal ion-crown ether complexes with cesium ion contained within a metal ion-water cluster serves as the basis for modeling binding preferences in solution. The calculated O-H stretching frequency of H(2)O molecule in micro-solvated metal ion-crown complexes is more red-shifted in comparison to hydrated metal ions. The calculated IR spectra can be compared with an experimental spectrum to determine the presence of micro-solvated metal ion-crown ether complexes in extractant phase.


Assuntos
Césio/química , Complexos de Coordenação/química , Éteres de Coroa/química , Estrôncio/química , Biocatálise , Técnicas Biossensoriais/métodos , Césio/metabolismo , Complexos de Coordenação/metabolismo , Éteres de Coroa/metabolismo , Gases , Modelos Moleculares , Conformação Molecular , Teoria Quântica , Soluções , Estrôncio/metabolismo , Termodinâmica , Água/química
10.
J Physiol Sci ; 60(6): 441-6, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20842544

RESUMO

Interaction between the selectivity filter and the adjacent pore helix of voltage-gated K(+) (Kv) channels controls pore stability during K(+) conduction. Kv channels, having their selectivity filter destabilized during depolarization, are said to undergo C-type inactivation. We examined the functionality of a residue at the pore helix of the Kv1.2 channel (V370), which reportedly affects C-type inactivation. A mutation into glycine (V370G) caused a shift in reversal potential from around -72 to -9 mV. The permeability ratios (P(Na)/P(K)) of the wild type and V370G mutant are 0.04 and 0.76, respectively. In the wild-type, P(Rb)/P(K), P(Cs)/P(K) and P(Li)/P(K) are 0.78, 0.10 and 0.05, respectively. Kv1.2 V370G channels had enhanced permeability to Rb(+) and Cs(+) (P(Rb)/P(K) and P(Cs)/P(K) are 1.63 and 1.18, respectively); however, Li(+) permeability was not significantly augmented (P(Li)/P(K) is 0.13). Therefore, in addition to its known effect on pore stability, V370 of Kv1.2 is also crucial in controlling ion selectivity.


Assuntos
Cátions Monovalentes/metabolismo , Ativação do Canal Iônico/genética , Canal de Potássio Kv1.2/genética , Substituição de Aminoácidos , Césio/metabolismo , Humanos , Lítio/metabolismo , Modelos Moleculares , Neoplasias Bucais/metabolismo , Estrutura Secundária de Proteína , Rubídio/metabolismo , Células Tumorais Cultivadas
11.
Biol Trace Elem Res ; 135(1-3): 1-9, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19655100

RESUMO

The knowledge about cesium metabolism and toxicity is sparse. Oral intake of cesium chloride has been widely promoted on the basis of the hypothesis referred to as "high pH cancer therapy", a complimentary alternative medicine method for cancer treatment. However, no properly confirmed tumor regression was reported so far in all probability because of neither theoretical nor experimental grounds for this proposal. The aim of the present review was to resume and discuss the material currently available on cesium salts and their applications in medicine. The presence of cesium in the cell does not guarantee high pH of its content, and there is no clinical evidence to support the claims that cancer cells are vulnerable to cesium. Cesium is relatively safe; signs of its mild toxicity are gastrointestinal distress, hypotension, syncope, numbness, or tingling of the lips. Nevertheless, total cesium intakes of 6 g/day have been found to produce severe hypokalemia, hypomagnesemia, prolonged QTc interval, episodes of polymorphic ventricular tachycardia, with or without torsade de pointes, and even acute heart arrest. However, full information on its acute and chronic toxicity is not sufficiently known. Health care providers should be aware of the cardiac complications, as a result of careless cesium usage as alternative medicine.


Assuntos
Césio/toxicidade , Césio/metabolismo , Césio/uso terapêutico , Cloretos/uso terapêutico , Feminino , Humanos , Concentração de Íons de Hidrogênio , Hipopotassemia/induzido quimicamente , Síndrome do QT Longo/induzido quimicamente , Masculino , Pessoa de Meia-Idade
12.
Nat Struct Mol Biol ; 16(1): 35-41, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19098915

RESUMO

Here we present a detailed characterization of ion binding in the NaK pore using the high-resolution structures of NaK in complex with various cations. These structures reveal four ion binding sites with similar chemical environments but vastly different ion preference. The most nonselective of all is site 3, which is formed exclusively by backbone carbonyl oxygen atoms and resides deep within the selectivity filter. Additionally, four water molecules in combination with four backbone carbonyl oxygen atoms are seen to participate in K(+) and Rb(+) ion chelation, at both the external entrance and the vestibule of the NaK filter, confirming the channel's preference for an octahedral ligand configuration for K(+) and Rb(+) binding. In contrast, Na(+) binding in the NaK filter, particularly at site 4, utilizes a pyramidal ligand configuration that requires the participation of a water molecule in the cavity. Therefore, the ability of the NaK filter to bind both Na(+) and K(+) ions seemingly arises from the ions' ability to use the existing environment in unique ways, rather than from any structural rearrangements of the filter itself.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Canais de Potássio/química , Canais de Potássio/metabolismo , Trifosfato de Adenosina/metabolismo , Proteínas de Bactérias/genética , Sítios de Ligação , Cátions , Césio/metabolismo , Cinética , Modelos Moleculares , Potássio/metabolismo , Canais de Potássio/genética , Ligação Proteica , Conformação Proteica , Rubídio/metabolismo , Deleção de Sequência , Especificidade por Substrato , Água/metabolismo
13.
J Gen Physiol ; 132(5): 537-46, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18955594

RESUMO

Mutations in human bestrophin-1 are linked to various kinds of retinal degeneration. Although it has been proposed that bestrophins are Ca(2+)-activated Cl(-) channels, definitive proof is lacking partly because mice with the bestrophin-1 gene deleted have normal Ca(2+)-activated Cl(-) currents. Here, we provide compelling evidence to support the idea that bestrophin-1 is the pore-forming subunit of a cell volume-regulated anion channel (VRAC) in Drosophila S2 cells. VRAC was abolished by treatment with RNAi to Drosophila bestrophin-1. VRAC was rescued by overexpressing bestrophin-1 mutants with altered biophysical properties and responsiveness to sulfhydryl reagents. In particular, the ionic selectivity of the F81C mutant changed from anionic to cationic when the channel was treated with the sulfhydryl reagent, sodium (2-sulfonatoethyl) methanethiosulfonate (MTSES(-)) (P(Cs)/P(Cl) = 0.25 for native and 2.38 for F81C). The F81E mutant was 1.3 times more permeable to Cs(+) than Cl(-). The finding that VRAC was rescued by F81C and F81E mutants with different biophysical properties shows that bestrophin-1 is a VRAC in S2 cells and not simply a regulator or an auxiliary subunit. F81C overexpressed in HEK293 cells also exhibits a shift of ionic selectivity after MTSES(-) treatment, although the effect is quantitatively smaller than in S2 cells. To test whether bestrophins are VRACs in mammalian cells, we compared VRACs in peritoneal macrophages from wild-type mice and mice with both bestrophin-1 and bestrophin-2 disrupted (best1(-/-)/best2(-/-)). VRACs were identical in wild-type and best1(-/-)/best2(-/-) mice, showing that bestrophins are unlikely to be the classical VRAC in mammalian cells.


Assuntos
Canais de Cloreto , Cloretos/metabolismo , Proteínas de Drosophila , Mesilatos/farmacocinética , Animais , Bestrofinas , Linhagem Celular Transformada , Césio/metabolismo , Canais de Cloreto/efeitos dos fármacos , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Drosophila/genética , Drosophila/metabolismo , Proteínas de Drosophila/efeitos dos fármacos , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Humanos , Canais Iônicos , Transporte de Íons/efeitos dos fármacos , Transporte de Íons/genética , Macrófagos , Camundongos , Camundongos Knockout , Mutagênese Sítio-Dirigida , Interferência de RNA
14.
J Agric Food Chem ; 56(20): 9641-6, 2008 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-18800803

RESUMO

The characteristics of Cs accumulation and localization in edible mushrooms were examined using the mycelia of Pleurotus ostreatus-Y1. Scanning electron microscope images revealed the existence of white spots, and energy dispersive X-ray microanalyzer analysis indicated the presence of larger amounts of Cs and P in these spots in mycelia cultured on medium containing 25 mM CsCl. The (137)Cs activities in the mycelia were approximately 4-6 times higher than those in water used for (137)Cs elution. Higher Cs concentrations in the sediment fraction including vacuolar pellets were obtained compared to the upper fractions. It was observed that yellowish spots caused by the fluorescence of 4',6-diamidino-2-phenylindole (DAPI)-stained polyphosphate were localized in the mycelia. The higher fluorescence intensity of the yellowish-grained spots was measured in comparison with other regions in the mycelium. These results suggested that Cs in the mycelia was trapped by polyphosphate in vacuoles or other organelles.


Assuntos
Césio/metabolismo , Micélio/metabolismo , Pleurotus/metabolismo , Césio/análise , Radioisótopos de Césio/análise , Radioisótopos de Césio/metabolismo , Microanálise por Sonda Eletrônica , Micélio/química , Micélio/ultraestrutura , Pleurotus/química , Pleurotus/ultraestrutura , Polifosfatos/metabolismo
15.
Am J Physiol Heart Circ Physiol ; 294(1): H354-61, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17951372

RESUMO

Inhibition of neuronal nitric oxide synthase (nNOS) in cardiac postganglionic sympathetic neurons leads to enhanced cardiac sympathetic responsiveness in normal animals, as well as in animal models of cardiovascular diseases. We used isolated atria from mice with selective genetic disruption of nNOS (nNOS(-/-)) and their wild-type littermates (WT) to investigate whether sympathetic heart rate (HR) responses were dependent on nNOS. Immunohistochemistry was initially used to determine the presence of nNOS in sympathetic [tyrosine hydroxylase (TH) immunoreactive] nerve terminals in the mouse sinoatrial node (SAN). After this, the effects of postganglionic sympathetic nerve stimulation (1-10 Hz) and bath-applied norepinephrine (NE; 10(-8)-10(-4) mol/l) on HR were examined in atria from nNOS(-/-) and WT mice. In the SAN region of WT mice, TH and nNOS immunoreactivity was virtually never colocalized in nerve fibers. nNOS(-/-) atria showed significantly reduced HR responses to sympathetic nerve activation and NE (P < 0.05). Similarly, the positive chronotropic response to the adenylate cyclase activator forskolin (10(-7)-10(-5) mol/l) was attenuated in nNOS(-/-) atria (P < 0.05). Constitutive NOS inhibition with L-nitroarginine (0.1 mmol/l) did not affect the sympathetic HR responses in nNOS(-/-) and WT atria. The paucity of nNOS in the sympathetic innervation of the mouse SAN, in addition to the attenuated HR responses to neuronal and applied NE, indicates that presynaptic sympathetic neuronal NO does not modulate neuronal NE release and SAN pacemaking in this species. It appears that genetic deletion of nNOS results in the inhibition of adrenergic-adenylate cyclase signaling within SAN myocytes.


Assuntos
Frequência Cardíaca , Óxido Nítrico Sintase Tipo I/metabolismo , Nó Sinoatrial/inervação , Fibras Simpáticas Pós-Ganglionares/enzimologia , Adenilil Ciclases/metabolismo , Animais , Césio/metabolismo , Colforsina/farmacologia , AMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Estimulação Elétrica , Ativadores de Enzimas/farmacologia , Inibidores Enzimáticos/farmacologia , Átrios do Coração/inervação , Frequência Cardíaca/efeitos dos fármacos , Imuno-Histoquímica , Técnicas In Vitro , Masculino , Camundongos , Camundongos Knockout , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I/deficiência , Óxido Nítrico Sintase Tipo I/genética , Nitroarginina/farmacologia , Norepinefrina/farmacologia , Nó Sinoatrial/efeitos dos fármacos , Nó Sinoatrial/metabolismo , Fibras Simpáticas Pós-Ganglionares/efeitos dos fármacos , Simpatomiméticos/farmacologia , Tirosina 3-Mono-Oxigenase/metabolismo
16.
J Pharmacol Exp Ther ; 321(3): 911-20, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17377062

RESUMO

The human ether-a-go-go-related gene (hERG) encodes a channel that conducts the rapidly activating delayed rectifier K(+) current (I(Kr)), which is important for cardiac repolarization. Mutations in hERG reduce I(Kr) and cause congenital long QT syndrome (LQTS). More frequently, common medications can reduce I(Kr) and cause LQTS as a side effect. Protein trafficking abnormalities are responsible for most hERG mutation-related LQTS and are recently recognized as a mechanism for drug-induced LQTS. Whereas hERG trafficking has been studied in recombinant expression systems, there has been no reported study on cardiac I(Kr) trafficking at the protein level. In the present study, we identified that I(Kr) is present in cultured neonatal rat ventricular myocytes and can be robustly recorded using Cs(+) as the charge carrier. We further discovered that 4,4'-(isopropylidenedithio)-bis-(2,6-di-t-butylphenol) (probucol), a cholesterol-lowering drug that induces LQTS, disrupted I(Kr) trafficking and prolonged the cardiac action potential duration. Probucol did not directly block I(Kr). Probucol also disrupted hERG trafficking and did not block hERG channels expressed in human embryonic kidney 293 cells. We conclude that probucol induces LQTS by disrupting ether-a-go-go-related gene trafficking, and that primary culture of neonatal rat cardiomyocytes represents a useful system for studying native I(Kr) trafficking.


Assuntos
Canais de Potássio Éter-A-Go-Go/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Probucol/farmacologia , Potenciais de Ação/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Antiarrítmicos/farmacologia , Linhagem Celular , Membrana Celular/química , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Células Cultivadas , Césio/metabolismo , Canal de Potássio ERG1 , Eletrofisiologia , Canais de Potássio Éter-A-Go-Go/genética , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Miócitos Cardíacos/citologia , Miócitos Cardíacos/fisiologia , Técnicas de Patch-Clamp , Piperidinas/farmacologia , Potássio/metabolismo , Transporte Proteico/efeitos dos fármacos , Piridinas/farmacologia , RNA Interferente Pequeno/genética , Ratos , Ratos Sprague-Dawley , Transfecção
17.
J Biol Chem ; 281(47): 35735-41, 2006 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-16990282

RESUMO

14-3-3 proteins modulate the plant inward rectifier K+ channel KAT1 heterologously expressed in Xenopus oocytes. Injection of recombinant plant 14-3-3 proteins into oocytes shifted the activation curve of KAT1 by +11 mV and increased the tau(on). KAT1 was also modulated by 14-3-3 proteins of Xenopus oocytes. Titration of the endogenous 14-3-3 proteins by injection of the peptide Raf 621p resulted in a strong decrease in KAT1 current (approximately 70% at -150 mV). The mutation K56E performed on plant protein 14-3-3 in a highly conserved recognition site prevented channel activation. Because the maximal conductance of KAT1 was unaffected by 14-3-3, we can exclude that they act by increasing the number of channels, thus ruling out any effect of these proteins on channel trafficking and/or insertion into the oocyte membrane. 14-3-3 proteins also increased KAT1 current in inside-out patches, suggesting a direct interaction with the channel. Direct interaction was confirmed by overlay experiments with radioactive 14-3-3 on oocyte membranes expressing KAT1.


Assuntos
Proteínas 14-3-3/metabolismo , Proteínas de Arabidopsis/fisiologia , Canais de Potássio Corretores do Fluxo de Internalização/fisiologia , Animais , Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismo , Césio/metabolismo , Eletrofisiologia/métodos , Escherichia coli/metabolismo , Ativação do Canal Iônico , Mutação , Oócitos/metabolismo , Técnicas de Patch-Clamp , Canais de Potássio Corretores do Fluxo de Internalização/química , Canais de Potássio Corretores do Fluxo de Internalização/metabolismo , Proteínas Recombinantes/química , Xenopus , Quinases raf/química
18.
Biochem J ; 399(2): 315-23, 2006 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-16803457

RESUMO

The Pgp (P-glycoprotein) multidrug transporter couples ATP hydrolysis at two cytoplasmic NBDs (nucleotide-binding domains) to the transport of hydrophobic compounds. Orthovanadate (V(i)) and fluoroaluminate (AlF(x)) trap nucleotide in one NBD by forming stable catalytically inactive complexes (Pgp-M2+-ADP-X), which are proposed to resemble the catalytic transition state, whereas the complex formed by beryllium fluoride (BeF(x)) is proposed to resemble the ground state. We studied the trapped complexes formed via incubation of Pgp with ATP (catalytically forward) or ADP (reverse) and V(i), BeF(x) or AlF(x) using Mg2+ or Co2+ as the bivalent cation. Quenching of intrinsic Pgp tryptophan fluorescence by acrylamide, iodide and caesium indicated that conformational changes took place upon formation of the trapped complexes. Trapping with V(i) and ATP led to a 6-fold increase in the acrylamide quenching constant, K(SV), suggesting that large conformational changes take place in the Pgp transmembrane regions on trapping in the forward direction. Trapping with V(i) and ADP gave only a small change in quenching, indicating that the forward- and reverse-trapped complexes are different. TNP (trinitrophenyl)-ATP/TNP-ADP interacted with all of the trapped complexes, however, the fluorescence enhancement differed for the trapped states, suggesting a change in polarity in the nucleotide-binding sites. The nucleotide-binding site of the BeF(x)-trapped complex was much more polar than that of the V(i) and AlF(x) complexes. Functionally, all the trapped complexes were able to bind drugs and TNP-nucleotides with unchanged affinity compared with native Pgp.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/química , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Proteínas Associadas à Resistência a Múltiplos Medicamentos/química , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Acrilamida/metabolismo , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Células CHO , Césio/metabolismo , Cricetinae , Cricetulus , Fluorescência , Iodo/metabolismo , Cinética , Conformação Proteica , Trinitrobenzenos/metabolismo , Triptofano/química
19.
J Physiol ; 571(Pt 3): 503-17, 2006 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-16423852

RESUMO

Airway ciliated cells express an ATP-gated P2X receptor channel of unknown subunit composition (P2X(cilia)) which is modulated by Na+ and by long exposures to ATP. P2X(cilia) was investigated by recording currents from freshly dissociated rabbit airway ciliated cells with the patch-clamp technique in the whole-cell configuration. During the initial continuous exposure to extracellular ATP, P2X(cilia) currents gradually increase in magnitude (priming), yet the permeability to N-methyl-D-glucamine (NMDG) does not change, indicating that priming does not arise from a progressive change in pore diameter. Na+, which readily permeates P2X(cilia) receptor channels, was found to inhibit the channel extracellular to the electric field. The rank order of permeability to various monovalent cations is: Li+, Na+, K+, Rb+, Cs+, NMDG+ and TEA+, with a relative permeability of 1.35, 1.0, 0.99, 0.91, 0.79, 0.19 and 0.10, respectively. The rank order for the alkali cations follows an Eisenman series XI for a high-strength field site. Ca2+ has been estimated to be 7-fold more permeant than Na+. The rise in [Ca2+]i in ciliated cells, induced by the activation of P2X(cilia), is largely inhibited by either Brilliant Blue G or KN-62, indicating that P2X7 may be a part of P2X(cilia). P2X(cilia) is augmented by Zn2+ and by ivermectin, and P2X4 receptor protein is detected by immunolabelling at the basal half of the cilia, strongly suggesting that P2X4 is a component of P2X(cilia) receptor channels. Taken together, these results suggest that P2X(cilia) is either assembled from P2X4 and P2X7 subunits, or formed from modified P2X4 subunits.


Assuntos
Cílios/metabolismo , Receptores Purinérgicos P2/metabolismo , Traqueia/metabolismo , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/análogos & derivados , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/farmacologia , Trifosfato de Adenosina/farmacologia , Animais , Cálcio/metabolismo , Césio/metabolismo , Cílios/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Técnicas In Vitro , Canais Iônicos/antagonistas & inibidores , Canais Iônicos/metabolismo , Ivermectina/farmacologia , Masculino , Potenciais da Membrana , Coelhos , Receptores Purinérgicos P2/análise , Receptores Purinérgicos P2/efeitos dos fármacos , Receptores Purinérgicos P2X , Receptores Purinérgicos P2X4 , Receptores Purinérgicos P2X7 , Sódio/metabolismo , Traqueia/efeitos dos fármacos
20.
J Hazard Mater ; 116(1-2): 125-34, 2004 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-15561371

RESUMO

Biosorption batch experiments were conducted to determine the cesium binding ability of native biomass and chemically modified biosorbents derived from marine algae, namely ferrocyanide algal sorbents type 1 and type 2 (FASs1 and FASs2). The applicability of the Langmuir and Freundlich isotherms for representation of the experimental data was investigated. The cesium sorption performances of the various types of sorbents were compared using the maximum capacities (qmax values) obtained from fitting the Langmuir isotherm to the values calculated from the sorption experiments, which FASs type 1 and type 2 showed better sorption performances for cesium. FASs1 and FASs2 derived from formaldehyde and glutaraldehyde crosslinked Padina australis exhibited lower sorption capacities than those prepared from the non-crosslinked one. Most of the cesium ions were bound to FASs1, derived from Sargassum glaucescens and P. australis, in < 2 min and equilibrium reached within the first 30 min of contact. Biosorption of cesium by FASs1 derived from P. australis and Cystoseria indica was constantly occurred at a wide range of pH, between 1 and 10, and the highest removal took place at pH 4. The presence of sodium and potassium at 0.5 and 1mM did not inhibit cesium biosorption by algae biomass. The maximum cesium uptake was acquired using the large particles of FAS2 originated from S. glaucescens (2-4 mm). Desorption of cesium from the metal-laden FASs1 (from P. australis, S. glaucescens and Dictyota indica) was completely achieved applying 0.5 and 1 M NaOH and KOH, although the cesium sorption capacity of the biosorbents (from C. indica and S. glaucescens) decreased by 46-51% after 9 sorption-desorption cycles.


Assuntos
Césio/isolamento & purificação , Césio/metabolismo , Phaeophyceae/química , Rodófitas/química , Purificação da Água/métodos , Adsorção , Biodegradação Ambiental , Biomassa , Biotecnologia/tendências
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