Molecular cloning and characterization of a sodium-pump ATPase of the moss Physcomitrella patens

Physcomitrella patens grew slowly at 600 mm Na+, pH 6.0, affected by the low water potential but without signs of suffering Na+ toxicity. At pH 8.0, tolerance seemed to be lower but it grew at 200 mm Na+, again without signs of Na+ toxicity. The resistance of Physcomitrella cells to the toxic effects of Na+ can be accounted for by their capacity to keep high K+:Na+ ratios and to extrude Na+ by a system that is not dependent on DeltapH. Physcomitrella expresses two P-type ATPases similar in sequence to fungal ENA-type Na+-ATPases. A functional study in yeast demonstrated that one of these ATPases, PpENA1, is an Na+-pump. We also found that P. patens has a plant-type SOS1 Na+/H+ antiporter. We discuss that Na+-ATPases existed in early land plants but that they were lost during the evolution of bryophytes to flowering plants.     

Voltage gated sodium channels as drug discovery targets

Voltage-gated sodium (Na_V) channels are a family of transmembrane ion channel proteins. They function by forming a gated, water-filled pore to help establish and control cell membrane potential via control of the flow of ions between the intracellular and the extracellular environments. Blockade of Na_Vs has been successfully accomplished in the clinic to enable control of pathological firing patterns that occur in a diverse range of conditions such as chronic pain, epilepsy, and cardiac arrhythmias. First generation sodium channel modulator drugs, despite low inherent subtype selectivity, preferentially act on over-excited cells which reduces undesirable side effects in the clinic. However, the limited therapeutic indices observed with the first generation demanded a new generation of sodium channel inhibitors. The structure, function and the state of the art in sodium channel modulator drug discovery are discussed in this chapter.     

Micellization of conjugated chenodeoxy- and ursodeoxycholates and solubilization of cholesterol into their micelles: comparison with other four conjugated bile salts species

Micelle formations of sodium glyco- and taurochenodeoxycholate (NaGCDC and NaTCDC) and sodium glyco- and tauroursodeoxycholates (NaGUDC and NaTUDC) was studied at 308.2 K for their critical micelle concentrations at various NaCl concentrations by pyrene fluorescence probe, and the degree of counterion binding to micelle was determined using the Corrin-Harkins plots. The degree of counterion binding was found to be 0.37-0.38 for chenodeoxycholate conjugates, while the determination of the degree was quite difficult for ursodeoxycholate conjugates. The change of I1/I3 values on the fluorescence spectrum with the conjugate bile salt concentration suggested two steps for their bile salt aggregation. The first step is a commencement of smaller aggregates, the first cmc, and the second one is a starting of stable aggregates, the second cmc. The aggregation number was determined at 308.2 K and 0.15 M NaCl concentration by static light scattering: 16.3 and 11.9 for sodium NaGCDC and NaTCDC, and 7.9 and 7.1 for NaGUDC and NaTUDC, respectively. The solubilization of cholesterol into the bile salt micelles in the presence of coexisting cholesterol phase and the maximum additive concentration (MAC) of cholesterol was determined against the bile salt concentration. The standard Gibbs energy change for the solubilization was evaluated, where the micelles were regarded as a chemical species. The solubilization was stabilized in the order of NaGUDC approximately = NaTUDC < NaTC < NaGC < NaTCDC < NaGCDC < NaTDC < NaGDC, where the preceding results were taken into the order.     

Dietary salt intake and its relevance to ionic regulation in freshwater salmonids

Dietary sodium intake for freshwater salmonids feeding in the wild (invertebrate diet) or in captivity (pellet diet) was calculated and compared with published branchial sodium influx values. Dietary sodium intake (mmol kg⁻¹ per month) increases from winter minimum values of 5 and 30-40 to reach maximum values in summer of 175 and 240 for invertebrate and pellet diet, respectively. In summer, dietary sodium intake for fish feeding in the wild was of the same magnitude as branchial sodium influx. The implications of dietary sodium intake for sodium balance in freshwater fish are discussed.     

Effects of inclusion level and source of dietary sodium on performance and meat characteristics of broiler chickens

The effect of different concentrations of dietary Na from three Na salts (NaCl, NaHCO₃ and Na₂SO₄) was assessed in two experiments carried out on broiler chickens aged from 1 to 35 days. In Exp. 1, diets were supplemented with 0.15, 0.20 and 0.25% Na, which increased the average Na content of the diets to 0.19, 0.24 and 0.30% respectively. In Exp. 2, the amounts of selected Na salts (NaCl and Na₂SO₄) were reduced and the estimated Na contents of experimental diets amounted to 0.10, 0.13, 0.15 and 0.19%. In view of the risk factors for the development of foot pad dermatitis, our aim was to find an optimum source of Na and to keep dietary Na intake at the minimum level sufficient to support normal growth and acceptable slaughter quality. The present results suggest that the amount of Na required for the undisturbed growth of broilers and adequate feed conversion is not less than 0.15% of additional Na in the starter period (1–14 d), and not less than 0.11% in the grower period (until day 35). Higher dietary Na levels did not lead to further production advantages, and were found to increase the moisture content of droppings. Dry matter concentration of excreta was also affected by Na source. In comparison with NaHCO₃, Na₂SO₄ seemed to be a better alternative for NaCl. Na₂SO₄ also tended to surpass NaHCO₃ as a dietary alternative for NaCl in terms of feed utilisation during the starter period. The applied additional Na levels (0.25 and 0.15%) and Na sources had no effect on the sensory profile and composition of breast meat.     

Role of the kdr and super-kdr sodium channel mutations in pyrethroid resistance: correlation of allelic frequency to resistance level in wild and laboratory populations of horn flies (Haematobia irritans)

The kdr and super-kdr point mutations found in the insect sodium channel gene are postulated to confer knockdown resistance (kdr) to pyrethroids. Using an allele-specific PCR assay to detect these mutations in individual horn flies, Haematobia irritans (L.), we determined the allelic frequency of the kdr and super-kdr mutations in several wild and laboratory populations. Wild populations with very similar allelic frequencies had resistance levels that ranged widely from 3- to 18-fold relative to a susceptible population. Conversely, the kdr allele frequency in a lab population with 17-fold resistance was nearly double that found in a heavily pressured wild population with 18-fold resistance. We conclude that, although the kdr mutation confers significant levels of pyrethroid resistance, a substantial component of resistance in insecticidally pressured populations is conferred by mechanisms that are PBO-suppressible. High super-kdr allele frequencies were detected in two resistant lab populations, but in wild populations with equivalent resistance the super-kdr allele frequency was very low. Interestingly, in over 1200 individuals assayed, the super-kdr mutation was never detected in the absence of the kdr mutation.     

Determination of montelukast sodium in human plasma by column-switching high-performance liquid chromatography with fluorescence detection

MK-0476 (montelukast sodium) is a potent and selective cysteinyl leukotriene receptor antagonist that is being investigated in the treatment of asthma. A simple and sensitive method for the determination of MK-0476 in human plasma was developed using column-switching high-performance liquid chromatography (HPLC) with fluorescence detection. A plasma sample was injected directly onto the HPLC system consisting of a pre-column (Capcell pak MF) and an analytical column (Capcell pak C18) which were connected with a six-port switching valve. The column eluate was monitored with a fluorescence detector (excitation at 350 nm; emission at 400 nm). The calibration curve was linear in a concentration range of 1–500 ng ml⁻¹ for MK-0476 in human plasma. The intra-day coefficients of variation of all concentrations within the range was less than 9.2%, and the intra-day accuracy values were between 97.2 and 114.6%. This method was used to measure the plasma concentration of MK-0476 following oral administration of the drug in humans.     

Development and validation of a sensitive liquid chromatographic-tandem mass spectrometric method for the determination of cromolyn sodium in human plasma

Cromolyn sodium is a safe compound with potent anti-allergic properties when used locally or topically. Clinical data from systemic exposure is not available because of the poor GI absorption when given orally. In order to evaluate a new approach to enhance the absorption and bioavailability of cromolyn sodium, a sensitive assay was needed to support an oral-dose study in humans. This paper describes a liquid chromatographic-tandem mass spectrometric (LC-MS-MS) method for the analysis of cromolyn sodium in human plasma. The method consists of a two-step extraction with subsequent analysis using a high-performance liquid chromatography electrospray tandem mass spectrometer system. The compounds were eluted isocratically on a C(18) column followed by a backflush. The total run time is 6 min. The standard curve of cromolyn sodium was over the range of 0.313 to 750 ng/mL with a lower limit of quantitation (LLOQ) of 0.313 ng/mL when 0.5 mL of plasma was used for analysis. The percent coefficient of variation (C.V.) for accuracy and precision (inter-assay and intra-assay) was less than 15% over the validated concentration range and the coefficients of determination, r(2), were >0.991577. The method is simple, sensitive, and selective, and has been successfully utilized for oral cromolyn sodium clinical studies.     

Validation of a high-performance liquid chromatography assay for urinary nedocromil sodium following oral and inhaled administration

The validation of a solid-phase extraction and an ion pair high-performance liquid chromatographic assay for the determination of nedocromil sodium (NCS) in urine samples following oral and inhaled administration to healthy volunteers is described. NCS and its internal standard sodium cromoglycate (SCG) were extracted from urine samples using solid-phase extraction and then quantified using high-performance liquid chromatography (HPLC). A 25-cm C₈ Spherisorb 5 μm stationary phase with a mobile phase containing a long alkyl chain ion-pair reagent (methanol–0.045 M phosphate buffer–0.05 M dodecyl triethyl ammonium phosphate; 550:447.6:2.4, v/v) was used. The mean (S.D.) intra-day accuracy and precision of the HPLC assay was 99.9 (1.6) and 7.05 (4.9)%, respectively. These values for the inter-day data were 102.4 (4.07) and 10.5 (2.7)%, respectively, over the concentration range investigated. The method described permits the detection of NCS in human urine at concentrations as low as 0.04 μg ml⁻¹ where the signal-to-noise ratio is greater than 3:1. In 10 healthy volunteers a significantly greater amount of NCS was excreted in the urine following inhalation than after oral dosing (p<0.001). The mean (S.D.) amount of NCS renally excreted at 0.5, 1.0 and 24 h following inhalation of four 2-mg doses of NCS from a metered dose inhaler (MDI) was 0.513 (0.24), 1.163 (0.49) and 4.00 (1.73)% of the nominal dose. Similar values after oral administration of 8 mg of NCS were 0.026 (0.03), 0.079 (0.06) and 0.930 (0.74)%, respectively.     

Expedient liquid chromatographic method with fluorescence detection for montelukast sodium in micro-samples of plasma

This study describes an expedient assay for the analysis of the asthma medication, montelukast sodium (Singulair, MK-0476), in human plasma samples. After a simple extraction of the plasma, the drug and internal standard, quinine bisulfate, were measured by HPLC. The chromatographic system consisted of a single pump, a refrigerated autosampler, a C₈ 4-μm particle size radial compression cartridge at 40°C and a fluorescence detector with the excitation and emission wavelengths set at 350 and 400 nm, respectively. The mobile phase which was delivered at 1.0 ml/min, was prepared by adding 200 ml of 0.025 M sodium acetate, pH adjusted to 4.0 with acetic acid, to 800 ml of acetonitrile, with 50 μl triethylamine. With a run time of only 10 min per sample, this assay had an overall recovery of >97% with a detection limit of 1 ng/ml. The inter- and intra-run relative standard deviations at 0.05, 0.2 and 1.0 μg/ml were all <9.2%, while the analytical recovery at the same concentrations were within 7.7% of the amount added.     

钠离子通道研究进展

细胞电活动是生命现象的基本特征之一,而离子通道是其结构和功能的基础。因此,研究离子通道作用机制具有重大的理论和现实意义。许多神经系统和心血管疾病,如多发性硬化症、癫痫、脑溢血、神经痛、Brugada综合征(BrS)、进行性心脏传导缺陷(PCCD)和原发性心室纤颤(IVF)等,都与钠离子通道氨基酸序列和结构发生的改变有关。该文对钠离子通道的研究进展进行综述。     

Implication of ENaC in salt-sensitive hypertension

Arterial blood pressure is critically dependent on sodium balance. The kidney is the key player in maintaining sodium homeostasis. Aldosterone-dependent epithelial sodium transport in the distal nephron is mediated by the highly selective, amiloride-sensitive epithelial sodium channel (ENaC). Direct evidence that dysfunction of ENaC participates in blood pressure regulation has come from the molecular analysis of two human genetic diseases, Liddle’s syndrome and pseudohypoaldosteronism type 1 (PHA-1). Both, increased sodium reabsorption despite low aldosterone levels in Liddle’s patients and decreased sodium reabsorption despite high aldosterone levels in PHA-1 patients, demonstrated that ENaC is an effector for aldosterone action. Gene-targeting and classical transgenic technology enable the generation of mouse models for these diseases and the analysis of the involvement of the epithelial sodium channel (ENaC) in the progress of these diseases. A first mouse model using ENaC transgenic knockout mice [ENaC(−/−)Tg] mimicked several clinical features of PHA-1, like salt-wasting, metabolic acidosis, high aldosterone levels, growth retardation and increased early mortality. Such mouse models will be necessary in testing the involvement of genetic and/or environmental factors like salt-intake in hypertension.     

3D Flexible Carbon Felt Host for Highly Stable Sodium Metal Anodes

Sodium (Na) metal, which possesses a high theoretical capacity and the lowest electrochemical potential, is regarded as a promising anode material for Na–metal batteries. However, both Na dendrite growth and large volume change in cycling have severely impeded its practical applications. This study demonstrates that a 3D flexible carbon (C) felt which is already commercialized in large‐scale can be employed as a host for prestoring Na via a melt infusion strategy, through which a Na/C composite anode is obtained. The resulting anode exhibits a stable voltage profile and a small hysteresis over 120 cycles in carbonate‐based electrolytes in symmetrical cells owing to the fact that the metallic Na is confined in a conductive carbon felt host, which increases the Na⁺ deposition sites to lower the effective current density and render a uniform Na nucleation, restricting the dimension change in electrochemical cycling. More importantly, effective inhibition of Na dendrite growth and large volume change is achieved. When coupled with a Na_0.67Ni_0.33Mn_0.67O₂ cathode, the Na/C composite demonstrates a good suitability in full cells. This work provides an alternative option for the fabrication of stable Na metal anodes, which is of great significance for the practical applications of Na metal anodes in high‐energy‐density batteries.     

Domesticated cultivation of alkali-tolerant sulfate-reducing bacteria using hydrogen and carbon dioxide as energy and carbon sources

Abstract

The ability of sulfate-reducing bacteria (SRB) to transform sodium sulfate into sodium carbonate has been investigated. The goal was to adapt and cultivate SRB to survive in a high-alkaline and high-salt environment. First, several strains of Gram-positive SRB were isolated and purified from sludge samples and used as initial strains; they were straight to curved rods belonging to the genus Clostridium. The isolates were acclimatized by increasing the concentration of sodium bicarbonate in the medium in a stepwise manner. Hydrogen and carbon dioxide were used as the electron donor and carbon source, respectively. Repeated semi-continuous cultivations created isolates that were able to reduce sulfate in media containing 1% w/v Na₂SO₄ and saturated with NaHCO₃.     

Sodium currents in toad cardiac pacemaker cells

Cells in the pacemaker region of toad (Bufo marinus) sinus venosus had spontaneous rhythmic action potentials. The rate of firing of action potentials, the rate of diastolic depolarization and the maximum rate of rise of action potentials were reduced by TTX (10 nm to 1 m). Currents were recorded with the whole cell, tight seal technique from cells enzymatically dissociated from this region. Cells studied were identified as pacemaker cells by their characteristic morphology, spontaneous rhythmic action potential activity that could be blocked by cobalt but not by TTX and lack of inward rectification. When calcium, potassium and nonselective cation currents (I_f) activated by hyperpolarization were blocked, depolarization was seen to generate transient and persistent inward currents. Both were sodium currents: they were abolished by tetrodotoxin (10 to 100 nm), their reversal potential was close to the sodium equilibrium potential and their amplitude and reversal potential were influenced as expected for sodium currents when extracellular sodium ions were replaced with choline ions. The transient sodium current was activated at potentials more positive than –40 mV while the persistent sodium current was obvious at more negative potentials. It was concluded that, in toad pacemaker cells, TTX-sensitive sodium currents contributing both to the upstroke of action potentials and to diastolic depolarization may play an important role in setting heart rate.We thank the Australian National Heart Foundation for their support. D.A.S. is an NHMRC Senior Research Officer.     

The study of sodium channels involved in pain responses using specific modulators

Voltage-gated sodium channels(VGSCs) are transmembrane proteins responsible for generation and conduction of action potentials in excitable cells.Physiological and pharmacological studies have demonstrated that VGSCs play a critical role in chronic pain associated with tissue or nerve injury.Many long-chain peptide toxins(60-76 amino acid residues) purified from the venom of Asian scorpion Buthus martensii Karsch(BmK) are investigated to be sodium channel-specific modulators.The α-like neurotoxins that can ...     

On the Thermodynamics,the Role of the Carbon Cathode,and the Cycle Life of the Sodium Superoxide (NaO2) Battery

Batteries based on the cell reaction between alkali metals and oxygen are highly attractive for energy storage due to their superior theoretical energy density. However, despite continuous progress, fundamental challenges in the further development of these cell systems remain. Understanding the oxygen electrode reaction and improving cycle life, while at the same time maximizing the practical energy density, are some of the most important issues that need to be addressed. Here, the product formation in aprotic sodium‐oxygen cells is studied and it is shown how cycle life and practical capacities can be improved. Different cell reactions (leading to either NaO₂ or Na₂O₂ as discharge products) have recently been reported. To understand whether the carbon structure or the local current density has any influence on the product stoichiometry or the cell performance, several carbon materials with a broad range in properties are tested. Phase‐pure NaO₂ is always found as a discharge product, but capacities range from 300 to values as high as 4000 mAh g(C)^?1 depending on the type of carbon. More importantly, the cycle life of Na/O₂ cells can be largely improved by shallow cycling, steadily yielding capacities of 1666 mAh g(C)^?1 for at least 60 cycles using a Ketjen black carbon electrode.     

Production of both interstitial and basement membrane procollagens by fibroblastic WI-38 cells from human embryonic lung

The synthesis and secretion of type IV procollagen, in addition to that of procollagen types I and III, was detected in cells derived from human embryonic lung (WI-38) by immunofluorescence, metabolic labeling, immunoprecipitation, collagenase digestion and the characteristic polypeptide sizes of both intact procollagen type IV chains and their initial pepsin-resistant fragments as determined by polyacrylamide gel electrophoresis. Locally obtained human embryonic lung cells secreted the same procollagens, but neither embryonic nor adult human skin fibroblasts were found to secrete type IV procollagen in amounts detectable by the same methods.     

Salt sensitivity and low discrimination between potassium and sodium in bean plants

Bean plants (Phaseolus vulgaris) were very sensitive to moderate concentrations of NaCl, showing a dramatic decrease in their K⁺ content in the presence of this salt. Increasing the KCl content of the nutrient medium released the inhibitory effect of NaCl by increasing the K⁺ content of the plants. Likewise moderate concentrations of KCl were toxic for bean plants because they produced a large K⁺ loading. NaCl partially released this toxicity by inhibiting the K⁺ loading. When compared to the moderately salt tolerant sunflower plants (Helianthus annuus), bean plants showed a lower capacity to discriminate between K⁺ and Na⁺, at high Na⁺ levels, and an uncontrolled K⁺ uptake at moderate concentrations of K⁺. It is concluded that this low capacity of discrimination of the K⁺ uptake system of bean plants in presence of Na⁺ can account for by the NaCl sensitivity of bean plants.