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PubMedDate not supplied

Sequential feline esophageal nutrient blood flow: perfusion measurements in vivo.

The utilization of a catheter semiconductor beta detector (CASRAD) to perform in vivo, sequential, esophageal nutrient blood flow distribution studies in cats is described. A diffusable radioisotope, Rubidium 86 (86Rb), was injected intravenously and the CASRAD placed within the esophageal lumen. The distribution of 86Rb remained stable within the feline esophagus for sufficient time to allow for counting at 1.5 cm levels in the esophagus. The accuracy limitations of the CASRAD system were assessed by comparing the in vivo, recorded distribution of 86Rb with the esophageal distribution of 86Rb as determined by a well-type scintillation counter. The distribution throughout most of the esophagus was similar by both techniques. Sequential esophageal studies performed with the CASRAD showed less variability than existed in the biological differences between cats and the methodology of obtaining and counting tissue by well-type scintillation counter (mean coefficient of variation 11.6% versus coefficient of variation 23.1%).

Animals↗
PubMed1995

Basolateral membrane chloride permeability of A6 cells: implication in cell volume regulation.

The permeability to Cl- of the basolateral membrane (blm) was investigated in renal (A6) epithelial cells, assessing their role in transepithelial ion transport under steady-state conditions (isoosmotic) and following a hypoosmotic shock (i.e. in a regulatory volume decrease, RVD). Three different complementary studies were made by measuring: (1) the Cl- transport rates (delta F/Fo s-1 (x10(-3))), where F is the fluorescence of N-(6-methoxyquinoyl) acetoethyl ester, MQAE, and Fo the maximal fluorescence (x10(-3)) of both membranes by following the intracellular Cl- activities (ai Cl-, measured with MQAE) after extracellular Cl- substitution (2) the blm 86Rb and 36Cl uptakes and (3) the cellular potential and Cl- current using the whole-cell patch-clamp technique to differentiate between the different Cl- transport mechanisms. The permeability of the blm to Cl- was found to be much greater than that of the apical membranes under resting conditions: aiCl- changes were 5.3 +/- 0.7 mM and 25.5 +/- 1.05 mM (n = 79) when Cl- was substituted by NO3(-) in the media bathing apical and basolateral membranes. The Cl- transport rate of the blm was blocked by bumetanide (100 microM) and 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, 50 microM) but not by N-phenylanthranilic acid (DPC, 100 microM). 86Rb and 36Cl uptake experiments confirmed the presence of a bumetanide- and a NPPB-sensitive Cl- pathway, the latter being approximately three times more important than the former (Na/K/2Cl cotransporter). Appli-cation of a hypoosmotic medium to the serosal side of the cell increased delta F/Fo s-1 (x10(-3)) after extracellular Cl- substitution (1.03 +/- 0.10 and 2.45 +/- 0.17 arbitrary fluorescent units s-1 for isoosmotic and hypoosmotic conditions respectively, n = 11); this delta F/Fo s-1 (x10(-3)) increase was totally blocked by serosal NPPB application; on the other hand, cotransporter activity was decreased by the hypoosmotic shock. Cellular Ca2+ depletion had no effect on delta F/Fo s-1 (x10(-3)) under isoosmotic conditions, but blocked the delta F/Fo s-1 (x10(-3)) increase induced by a hypoosmotic stress. Under isotonic conditions the measured cellular potential at rest was -37.2 +/- 4.0 mV but reached a maximal and transient depolarization of -25.1 +/- 3.7 mV (n = 9) under hypoosmotic conditions. The cellular current at a patch-clamping cellular potential of -85 mV (close to the Nernst equilibrium potential for K+) was blocked by NPPB and transiently increased by hypoosmotic shock (≈50% maximum increase). This study demonstrates that the major component of Cl- transport through the blm of the A6 monolayer is a conductive pathway (NPPB-sensitive Cl- channels) and not a Na/K/2Cl cotransporter. These channels could play a role in transepithelial Cl- absorption and cell volume regulation. The increase in the blm Cl- conductance, inducing a depolarization of these membranes, is proposed as one of the early events responsible for the stimulation of the 86Rb efflux involved in cell volume regulation.

Animals↗
PubMed1993

Na+/K+/2Cl- cotransport in medullary thick ascending limb cells: kinetics and bumetanide binding.

We examined the properties of Na+/K+/2Cl- cotransport in cultured mouse mTAL cells with respect to its kinetics, the contribution of K/K exchange to K fluxes mediated by the cotransporter, and [3H]bumetanide binding and turnover numbers in media with varying osmolality. The addition of bumetanide, the replacement of external Na+ or the replacement of external Cl- resulted in an almost identical (approx. 50%) decrease in K+ influx, suggesting that Na(+)-dependent, Cl(-)-dependent, BS K+ influx was a measure of Na+/K+/2Cl- cotransport. The kinetics of the BS K+ influx revealed a high affinity for external Na+ (apparent Km 7 mM) and external K+ (apparent Km 1.3 mM), but a very low affinity for external Cl- (apparent Km 67 mM with a two-site model). Of interest was the finding that none of the K+ (86Rb+) efflux was sensitive to bumetanide, suggesting the absence of cotransport mediated K/K exchange in this cell type. Specific [3H]bumetanide binding was a saturable function of free bumetanide concentration with a Kd of 0.20 microM and maximum binding (Bmax) of 0.63 pmol/mg, or about 53,000 sites per cell. Simultaneous transport and bumetanide binding assays yielded a turnover number of 255 min-1. The omission of external Na+, K+ or Cl- reduced specific [3H]bumetanide binding to values indistinguishable from zero. Changing medium osmolarity resulted in a co-ordinate change in BS K+ influx and bumetanide binding, with a monotonic increase in both transport and bumetanide binding with increase in osmolality from 200 to 400 mosmol/kg. About 85% of the cotransporter sites were located on the apical side, as in the intact mTAL tubule. The simultaneous measurement of BS ion transport and [3H]bumetanide binding in the mTAL cell may provide valuable insights into the regulation of Na+/K+/2Cl- cotransport in this nephron segment.

Animals↗
PubMed1990

Bumetanide reduces insulin release by a direct effect on the pancreatic beta-cells.

The effect of the loop diuretic bumetanide on glucose-induced insulin release, 45Ca2+ uptake, 36Cl- fluxes and 86Rb+ (K+ analogue) efflux was tested in isolated beta-cell-rich mouse pancreatic islets. Low concentrations of bumetanide (0.1-10 microM) reduced glucose-induced insulin release as well as 45Ca2+ uptake. High concentrations (0.5-1 mM) augmented glucose-induced insulin release and an intermediate concentration (100 microM) had no effect. Bumetanide (0.01-1 mM) reduced the islet accumulation of 36Cl-. The net efflux of 36Cl- in the presence of 20 mM D-glucose was reduced by a concentration (10 microM) that lowered glucose-induced insulin release. Bumetanide (10 microM) did not affect the rate coefficient for 36Cl- efflux, which suggests that chloride permeability is not affected. Bumetanide (10 microM) reduced 86Rb+ efflux from preloaded islets. The data show that bumetanide reduces insulin release by a direct effect on pancreatic beta-cells and suggest that this may be due to reduced chloride accumulation by a Na+, K+, Cl- co-transport system. It is suggested that the reduced chloride level is responsible for the decrease in glucose-induced chloride efflux and insulin release.

Animals↗
PubMed1989

Properties of gramicidin A channels in erythrocyte membranes.

Studies for the cation permeability properties of the gramicidin A channel in erythrocyte membranes are presented. It is shown that gramicidin A interacts with the membrane in a cooperative manner, creating aggregates of the antibiotic molecules in the lipid lattice of the membrane. Cationic channels exist in these aggregates with the following order of selectivity: Rb+ greater than Cs+ greater K+ greater than Na+. The cation permeability of the channels depends on the media surrounding the membrane. This finding has been explained on the basis of Hodgkin-Keynes theory for single-file ion diffusion through extra-narrow pores.

Cations↗
PubMed1996

The riminophenazines, clofazimine and B669, inhibit potassium transport in gram-positive bacteria by a lysophospholipid-dependent mechanism.

The effects of the riminophenazine antimicrobial agents clofazimine and B669 as well as those of lysophosphatidylcholine (LPC), on microbial K(+)-transporting systems were investigated in a range of Gram-positive and Gram-negative bacteria using 42K and 86Rubidium (86Rb) as tracers. Exposing the Gram-positive bacteria to 0.1-10 mg/L of the drugs resulted in a dose-related inhibition of uptake of both radiolabelled cations due primarily to the inhibition of their influx which was prevented by pretreating the microorganisms with 25 mg/L alpha-tocopherol (vitamin E) which forms a complex with lysophospholipids. In contrast, Gram-negative bacteria were resistant to riminophenazine-mediated inhibition of K(+)-transport, with only one of four well-characterised K(+)-transport system mutants of Escherichia coli, namely Kup, being affected by the antimicrobial agents. The selective antimicrobial activity of riminophenazines against Gram-positive bacteria is probably achieved by lysophospholipid-mediated inactivation of K(+)-transport, while Gram-negative microorganisms possess several K(+)-transport systems which are either inaccessible and/or insensitive to lysophospholipids. Thus, K(+)-transport systems may represent novel targets for antimicrobial agents.

Adenosine Triphosphate↗
PubMed2001

Perturbation of the pump-leak balance for Na(+) and K(+) in malaria-infected erythrocytes.

In human erythrocytes infected with the mature form of the malaria parasite Plasmodium falciparum, the cytosolic concentration of Na(+) is increased and that of K(+) is decreased. In this study, the membrane transport changes underlying this perturbation were investigated using a combination of (86)Rb(+), (43)K(+), and (22)Na(+) flux measurements and a semiquantitative hemolysis technique. From >15 h postinvasion, there appeared in the infected erythrocyte membrane new permeation pathways (NPP) that caused a significant increase in the basal ion permeability of the erythrocyte membrane and that were inhibited by furosemide (0.1 mM). The NPP showed the selectivity sequence Cs(+) > Rb(+) > K(+) > Na(+), with the K(+)-to-Na(+) permeability ratio estimated as 2.3. From 18 to 36 h postinvasion, the activity of the erythrocyte Na(+)/K(+) pump increased in response to increased cytosolic Na(+) (a consequence of the increased leakage of Na(+) via the NPP) but underwent a progressive decrease in the latter 12 h of the parasite's occupancy of the erythrocyte (36-48 h postinvasion). Incorporation of the measured ion transport rates into a mathematical model of the human erythrocyte indicates that the induction of the NPP, together with the impairment of the Na(+)/K(+) pump, accounts for the altered Na(+) and K(+) levels in the host cell cytosol, as well as predicting an initial decrease, followed by a lytic increase in the volume of the host erythrocyte.

Animals↗
PubMed1989

Cooperative binding of primycin and gramicidin on erythrocyte membranes. A cation transport study.

In this paper the authors present a comparative study of the actions of the antibiotics primycin and gramicidin on the erythrocyte membrane permeability. It has been found that both antibiotics have a nonlinear effect on the membrane permeability. Above a threshold antibiotic concentration, which is characteristic of the type of the antibiotic, the cation permeability of the erythrocyte membranes increases sharply. In the range of nonlinearity the transport-kinetic curves level off before achieving the equilibrium radioactive ion distribution between the extra- and intracellular spaces. A stochastic model of the cooperative and aspecific incorporation of antibiotic molecules into the membrane explains the experimental findings. The authors conclude that membrane permeability increases at the places where two or more antibiotic molecules form aggregates in the membrane.

Anti-Bacterial Agents↗
PubMed1991

Ion transport in epithelial cells from the bovine trachea in culture: effects of dexamethasone and aldosterone.

The ion transport properties of epithelial cells from the bovine trachea grown in culture were investigated. The cellular uptake of 22Na+, 36Cl- and 86Rb+ was measured and the sensitivity of ion uptake to amiloride and bumetanide revealed the presence of the transport pathways that exist in the native tissue. The action of corticosteroids, dexamethasone and aldosterone (0.1 or 1.0 microM), was studied by measuring the short-circuit current and the uptake of Na+ and Cl- following a treatment of 18 h, 2 or 8 days. The results showed that, in each situation, dexamethasone enhanced markedly the amiloride-sensitive current and Na+ uptake. In contrast, aldosterone exerted no detectable effect on the ion transport and electrical properties.

Absorption↗
PubMed1992

A continuous flow technique for analysis of the stoichiometry of the gastric H,K-ATPase.

A continuous flow method was developed for determining the stoichiometry of the gastric proton pump H,K-ATPase in its hydrolysis of ATP, translocation of H+ and the K+ congener 86Rb+. H,K-ATPase-containing vesicles which had been isolated from pig gastric mucosa were incubated at 37 degrees C for 2 h in 150 mM 86RbCl, 0.5 mM EGTA and 3 mM Mes-buffer adjusted to pH 6.1 with Tris, and then applied to a 0.45 micron pore size filter. The immobilized vesicles were superfused with 0.15 mM Mes/Tris buffer, pH 6.1, containing 150 mM choline-Cl and 0.2 mM MgCl2. After the medium was changed to one containing 0.1 mM ATP, the amounts and rates of H+ uptake, 86Rb+ efflux, and ATP hydrolysis were measured in fractions collected after the filter. The initial ratio of transported Rb+ to hydrolysed ATP gave values of 0.96 +/- 0.26 (mean +/- SD, n = 28). The initial ratio of ATP-dependent Rb+ efflux to H+ uptake gave values of 0.92 +/- 0.28 (mean +/- SD, n = 28). The MgATPase activity was measured in vesicles which had been incubated with choline-Cl instead of RbCl. In the initial fractions used for calculation of the stoichiometry, the MgATPase activity was 15.8% +/- 8.7 (mean +/- S.D.) of the maximal ATPase activity obtained with Rb(+)-loaded vesicles. The MgATPase may be an intrinsic activity of the H,K-ATPase. However, whether corrections were made for the MgATPase or not, it had only marginal effects on the calculations of the stoichiometry of the pump.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1992

Spatially dependent deadtime losses in high count rate cardiac PET.

Cardiac PET scans result in nonhomogeneous distributions of activity within the body, which might lead to great variations in singles rates around the detector ring. Conventional deadtime correction algorithms assume that the singles rates are uniform. This paper investigates singles nonuniformities during several typical cardiac scanning protocols (bolus injections of 15O-water and 82Rb, slow infusion of 18F-FDG and static imaging with FDG) and estimates how such nonuniformities might affect quantitative data. Nonuniformity was observed in all studies and was described by an asymmetry index which increased to 58% during bolus water injection, the most inhomogeneous study. These results are valid for any scanner with a ring diameter of approximately 78 cm and are independent of the amount of activity injected. Deadtime losses depend on the amount of activity and on the scanner type. Nonhomogeneities in singles can be shown to produce spatially dependent deadtime correction factors; for our scanner, these were seen to differ by up to 16% from the mean deadtime correction during bolus water injection. To demonstrate the distortions generated by average deadtime correction, the activity distribution during a clinical cardiac study was simulated using a phantom. A simple local deadtime correction and its implementation on our system are described, and the resulting improvements in both absolute and relative quantitation of the phantom study are shown.

Deoxyglucose↗
PubMed1990

Noninvasive assessment of coronary collaterals in man by PET perfusion imaging.

At present, coronary collateralization cannot be identified or assessed noninvasively in patients. In animal studies, coronary collaterals are associated with coronary steal, defined as a regional fall in perfusion during coronary arteriolar vasodilation. To determine the effect of coronary arteriolar vasodilation on collateral bed perfusion in man, myocardial perfusion imaging was performed before and after pharmacologic coronary vasodilation in patients with coronary artery disease (CAD). Regional myocardial activity of 82Rb or 13N ammonia was measured by positron emission tomography (PET) at rest and with intravenous dipyridamole/handgrip stress in 28 patients with angiographic collaterals and in 25 control patients with similar CAD severity by quantitative arteriography. Regional myocardial activity decreased after dipyridamole, indicating coronary steal, in 25 of 28 patients with angiographic collaterals and in only 4 of 25 control patients without angiographic collaterals. These findings suggest that developed collaterals are associated with myocardial steal in patients with CAD, allowing potential use of PET for non-invasive identification of coronary collateralization.

Collateral Circulation↗