scholarly journals Furosemide inhibition of chloride transport in human red blood cells.

1976 ◽  
Vol 68 (6) ◽  
pp. 583-599 ◽  
Author(s):  
P C Brazy ◽  
R B Gunn

The chloride self-exchange flux across the human red cell membrane is rapidly and reversibly inhibited by 10(-4) M furosemide, a potent chloruretic agent. Furosemide reduces the chloride flux at all chloride concentrations and increases the cellular chloride concentration at which the flux is half-maximum. Kinetic analysis of the flux measurements made at several furosemide and chloride concentrations yields a pattern of mixed inhibition with a dissociation constant for the inhibitor-transport mechanism complex of 5 X 10(-5) M. From this pattern of inhibition and other observations, including that the percent inhibition is independent of pH (range 5.6-8.9), we conclude that the anionic form of furosemide interacts primarily with the chloride transport mechanism at a site separate from both the transport site and the halide-reactive modifier site.

1984 ◽  
Vol 83 (5) ◽  
pp. 683-701 ◽  
Author(s):  
P A Knauf ◽  
F Y Law ◽  
T Tarshis ◽  
W Furuya

External N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate (NAP-taurine) inhibits human red cell chloride exchange by binding to a site that is distinct from the chloride transport site. Increases in the intracellular chloride concentration (at constant external chloride) cause an increase in the inhibitory potency of external NAP-taurine. This effect is not due to the changes in pH or membrane potential that usually accompany a chloride gradient, since even when these changes are reversed or eliminated the inhibitory potency remains high. According to the ping-pong model for anion exchange, such transmembrane effects of intracellular chloride on external NAP-taurine can be explained if NAP-taurine only binds to its site when the transport site is in the outward-facing (Eo or EClo ) form. Since NAP-taurine prevents the conformational change from EClo to ECli , it must lock the system in the outward-facing form. NAP-taurine can therefore be used just like the competitive inhibitor H2DIDS (4,4'-diisothiocyano-1,2- diphenylethane -2,2'-disulfonic acid) to monitor the fraction of transport sites that face outward. A quantitative analysis of the effects of chloride gradients on the inhibitory potency of NAP-taurine and H2DIDS reveals that the transport system is intrinsically asymmetric, such that when Cli = Clo, most of the unloaded transport sites face the cytoplasmic side of the membrane.


1984 ◽  
Vol 247 (3) ◽  
pp. C247-C259 ◽  
Author(s):  
M. A. Milanick ◽  
R. B. Gunn

Sulfate influx into human red blood cells was measured at 0 and 22 degrees C at several fixed external pH values between 3 and 10. These cells had normal internal pH and chloride concentrations so that sulfate influx was not limited by the efflux half-cycle reactions. The flux was a Michaelis-Menten function of sulfate concentration at each pH with K1/2SO4 = 4-10 mM. External protons activated influx 100-fold at a single site with a pK = 5.9 at 22 degrees C and 5.5 at 0 degrees C. This pK is similar to the value 5.99 +/- 0.3 for external proton binding to the sulfate-loaded transporter at 0 degrees C (J. Gen. Physiol. 79: 87-114, 1982). The flux was stilbene sensitive even in valinomycin-treated cells and was independent of membrane potential. This proton-activated influx appears to be proton-sulfate cotransport. At high pH there was a proton-independent flux that was membrane potential and stilbene sensitive. This proton-insensitive flux appears to be SO4(2-)/Cl- exchange or net sulfate influx. The sulfate influx over the entire pH range may be described in terms of an equation for the sum of the influxes through these two pathways on band 3.


1976 ◽  
Vol 67 (2) ◽  
pp. 223-234 ◽  
Author(s):  
M Dalmark

Chloride self-exchange was determined by measuring the rate of 36Cl efflux from human red blood cells at pH 7.2 (0 degrees C) in the presence of fluoride, bromide, iodide, and bicarbonate. The chloride concentration was varied between 10--400 mM and the concentration of other halides and bicarbonate between 10--300 mM. Chloride equilibrium flux showed saturation kinetics. The half-saturation constant increased and the maximum flux decreased in the presence of halides and bicarbonate: the inhibition kinetics were both competitive and noncompetitive. The competitive and the noncompetitive effects increased proportionately in the sequence: fluoride less than bromide less than iodide. The inhibitory action of bicarbonate was predominantly competitive. The noncompetitive effect of chloride (chloride self-inhibition) on chloride transport was less dominant at high inhibitor concentrations. Similarly, the noncompetitive action of the inhibitors was less dominant at high chloride concentrations. The results can be described by a carrier model with two anion binding sites: a transport site, and a second site which modifies the maximum transport rate. Binding to both types of sites increases proportionately in the sequence: fluoride less than chloride less than bromide less than iodide.


1993 ◽  
Vol 101 (5) ◽  
pp. 715-732 ◽  
Author(s):  
P K Gasbjerg ◽  
J Funder ◽  
J Brahm

Irreversible inhibition, 99.8% of control values for chloride transport in human red blood cells, was obtained by well-established methods of maximum covalent binding of 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). The kinetics of the residual chloride transport (0.2%, 106 pmol.cm-2 x s-1) at 38 degrees C, pH 7.2) was studied by means of 36Cl- efflux. The outside apparent affinity, expressed by Ko1/2,c, was 34 mM, as determined by substituting external KCl by sucrose. The residual flux was reversibly inhibited by a reexposure to DIDS, and by 4,4'-dinitrostilbene-2,2'-disulfonate (DNDS), phloretin, salicylate, and alpha-bromo-4-hydroxy-3,5-dinitroacetophenone (Killer III) (Borders, C. L., Jr., D. M. Perez, M. W. Lafferty, A. J. Kondow, J. Brahm, M. B. Fenderson, G. L. Breisford, and V. B. Pett. 1989. Bioorganic Chemistry. 17:96-107), to approximately 0.001% of control cells, which is a flux as low as in lipid bilayers. The reversible DIDS inhibition of the residual chloride flux depended on the extracellular chloride concentration, but was not purely competitive. The half-inhibition concentrations at [Cl(o)] = 150 mM in control cells (Ki,o) and covalently DIDS-treated cells (Ki,c) were: DIDS, Ki,c = 73 nM; DNDS, Ki,o = 6.3 microM, Ki,c = 22 microM; phloretin, Ki,o = 19 microM, Ki,c = 17 microM; salicylate, Ki,o = 4 mM, Ki,c = 8 mM; Killer III, Ki,o = 10 microM, Ki,c = 10 microM.


1973 ◽  
Vol 61 (2) ◽  
pp. 185-206 ◽  
Author(s):  
Robert B. Gunn ◽  
Mads Dalmark ◽  
D. C. Tosteson ◽  
J. O. Wieth

The efflux of chloride-36 from human erythrocytes under steady-state conditions is a saturable process that is competitively inhibited by bicarbonate and noncompetitively inhibited by acetate. This chloride self-exchange flux is reversibly dependent on the pH of the medium between 5.7 and 9.6 with a maximum flux at pH 7.8. The increase in chloride flux between pH 5.7 and 7.8 is inexplicable by the fixed charge hypothesis. The interpretations are made that chloride transport in human erythrocytes is carrier mediated, that bicarbonate utilizes the same transport mechanism, and that the mechanism can be titrated with hydrogen ions into less functional forms for chloride transport.


2010 ◽  
Vol 45 (4) ◽  
pp. 451-461 ◽  
Author(s):  
Nandana Perera ◽  
Bahram Gharabaghi ◽  
Peter Noehammer ◽  
Bruce Kilgour

Abstract Occurrence of increasing chloride concentrations in urban streams of cold climates, mainly due to road salt application, has raised concerns on its adverse effects on aquatic and terrestrial ecosystems. Therefore, there is a need for a better understanding of processes associated with road salt application and subsequent discharge into the environment in order to develop management practices to minimize detrimental effects of chlorides. The chloride mass analysis for the Highland Creek watershed based on four years of hourly monitoring data indicates that approximately 60% of the chlorides applied on the watershed enter streams prior to subsequent salting period, 85% of which occurs during the period between November and March. Contribution of private de-icing operations on chloride mass input within Highland Creek watershed was estimated to be approximately 38%, indicating its significance in overall chloride mass balance. Salt application rates, as well as chloride output in the streams, vary spatially based on land use, influencing chloride concentrations in surface waters. The estimated groundwater chloride concentration of 275 mg/L indicates that some aquatic organisms in Highland Creek would potentially be at risk even outside the winter period under dry weather flow conditions.


Materials ◽  
2021 ◽  
Vol 14 (14) ◽  
pp. 3975
Author(s):  
Magdalena German ◽  
Jerzy Pamin

Reinforced concrete structures can be strongly damaged by chloride corrosion of reinforcement. Rust accumulated around rebars involves a volumetric expansion, causing cracking of the surrounding concrete. To simulate the corrosion progress, the initiation phase of the corrosion process is first examined, taking into account the phenomena of oxygen and chloride transport as well as the corrosion current flow. This makes it possible to estimate the mass of produced rust, whereby a corrosion level is defined. A combination of three numerical methods is used to solve the coupled problem. The example object of the research is a beam cross-section with four reinforcement bars. The proposed methodology allows one to predict evolving chloride concentration and time to reinforcement depassivation, depending on the reinforcement position and on the location of a point on the bar surface. Moreover, the dependence of the corrosion initiation time on the chloride diffusion coefficient, chloride threshold, and reinforcement cover thickness is examined.


1989 ◽  
Vol 26 (11) ◽  
pp. 2186-2193 ◽  
Author(s):  
Jacques Locat ◽  
Pierre Gélinas

The results of an extensive hydrogeological investigation of the effects of de-icing road salts on Highway 55 near Trois-Rivières-Ouest indicate that a salt lens with chloride concentrations exceeding 800 mg/L exists below the highway. Maximum chloride concentration at the nearby pumping wells, not exceeding 140 mg/L, is reached only in late summer, whereas the maximum chloride infiltration follows the spring snowmelt. About 1 year's worth of road salts is retained in the unsaturated zone. The salt lens, in the upper part of the aquifer beneath the highway, has developed to a thickness of 8 m and a width of 400 m and constitutes a linear source of salts for the aquifer. The shape of this lens is distorted by the action of the pumping wells, and the lens is partly depleted by the end of the summer. Because of the particular characteristics of the aquifer at the site studied and the exploitation methods, no long-term threat to the water quality is foreseen.


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