scholarly journals Kinetics and stoichiometry of Na-dependent Li transport in human red blood cells.

1978 ◽  
Vol 72 (2) ◽  
pp. 249-265 ◽  
Author(s):  
B Sarkadi ◽  
J K Alifimoff ◽  
R B Gunn ◽  
D C Tosteson
Keyword(s):  
Transport System ◽  
Red Cells ◽  
Normal Male ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Human Red Blood Cells ◽  
Na Efflux ◽  
Tightly Coupled ◽  
Male Subjects ◽  
Human Red Cells

This paper describes the kinetics and stoichiometry of a tightly coupled Na-Li exchange transport system in human red cells. The system is inhibited by phloretin and furosemide but not by ouabain. Li influx by this system increases and saturates with increasing concentrations of external Li and internal Na and is inhibited competitively by external Na. Comparable functions relate Li efflux and Na efflux to internal and external Li and Na concentrations. Analysis of these relations yields the following values for the ion concentrations required to half-maximally activate the transport system: internal Na and Li 9.0 and 0.5 mM, respectively, external Na and Li 25 and 1.5 mM, respectively. The system performs a 1:1 exchange of Na and Li moving in opposite directions across the red cell membrane. We found no evidence for a simultaneous transport of more than one Na and Li by the system. The maximum transport rate of Na-dependent Li transport varied between 0.1 and 0.37 mmol/(liter of cells X h) in the red cells of the five normal male subjects studied. No significant variations between individual subjects were observed for bicarbonate-stimulated Li transport and for the residual Li fluxes which occur in the absence of bicarbonate and in the presence of ouabain plus phloretin.

scholarly journals Lithium transport pathways in human red blood cells.

1978 ◽  
Vol 72 (2) ◽  
pp. 233-247 ◽  
Author(s):  
G N Pandey ◽  
B Sarkadi ◽  
M Haas ◽  
R B Gunn ◽  
J M Davis ◽  
...  
Keyword(s):  
External Medium ◽  
Red Cells ◽  
Red Cell Membrane ◽  
Transport Pathways ◽  
Human Red Blood Cells ◽  
Lithium Transport ◽  
Ca Ions ◽  
Dominant Cation ◽  
Human Red Cells ◽  
In Cells

In human red cells, Li is extruded against its own concentration gradient if the external medium contains Na as a dominant cation. This uphill net Li extrusion occurs in the presence of external Na but not K, Rb, Cs, choline, Mg, or Ca, is ouabain-insensitive, inhibited by phloretin, and does not require the presence of cellular ATP. Li influx into human red cells has a ouabain-sensitive and a ouabain-insensitive but phloretin-sensitive component. Ouabain-sensitive Li influx is competitively inhibited by external K and Na and probably involves the site on which the Na-K pump normally transports K into red cells. Ouabain does not inhibit Li efflux from red cells containing Li concentrations below 10 mM in the presence of high internal Na or K, whereas a ouabain-sensitive Li efflux can be measured in cells loaded to contain 140 mM Li in the presence of little or no internal Na or K. Ouabain-insensitive Li efflux is stimulated by external Na and not by K, Rb, Cs, choline, Mg, or Ca ions. Na-dependent Li efflux does not require the presence of cellular ATP and is inhibited by phloretin, furosemide, quinine, and quinidine. Experiments carried out in cells loaded in the presence of nystatin to contain either only K or only Na show that the ouabain-insensitive, phloretin-inhibited Li movements into or out of human red cells are stimulated by Na on the trans side and inhibited by Na on the cis side of the red cell membrane. The characteristics of the Na-dependent unidirectional Li fluxes and uphill Li extrusion are similar, suggesting that they are mediated by the same Na-Li countertransport system.

scholarly journals Quantitative immunoferritin microscopy of Fya, Fyb, Jka, U, and Dib antigen site numbers on human red cells

Blood ◽  
1980 ◽  
Vol 56 (6) ◽  
pp. 969-977 ◽  
Author(s):  
SP Masouredis ◽  
E Sudora ◽  
L Mahan ◽  
EJ Victoria
Keyword(s):  
Distribution Patterns ◽  
Red Cells ◽  
Diverse Group ◽  
Red Cell ◽  
Human Erythrocyte Membrane ◽  
Red Cell Membrane ◽  
Quantitative Estimates ◽  
Equilibrium Binding ◽  
Antigen Site ◽  
Human Red Cells

Abstract The Fya, Fyb, Jka, U, and Dib antigen site numbers and ultrastructural distribution patterns on the human erythrocyte membrane were determined using quantitative immunoferritin microscopy. For homozygous antigen- positive red cells, the average number of determinants per red cell was about 14,000 for Jka, 17,000 for Fya and Fyb, 19,000 for Dib, and 23,000 for the U antigen, assuming that the equilibrium binding observed represented 80% saturation of the accessible antigen sites. The site numbers for this group of antigens were less than that for the Rh antigens, but considerably more than the Kell and Cellano antigens. The technique used was capable of demonstrating a twofold difference in antigen density between heterozygous and homozygous Fy (a+) red cells. More than 85% of the Fya and Fyb antigen sites were lost following pretreatment of the red cells with papain, consistent with the serologic lability of the Fy antigens following proteolysis. The ferritin distribution observed following conjugate staining of antibody- sensitized ghost membranes was similar for all five antigens studied and showed a random, clustered ferritin pattern. Although the quantitative estimates are valid, the remarkable similarity in antigen distribution pattern for this diverse group of antigens, as well as other considerations, suggest that the findings with ghose membranes probably do not reflect faithfully the antigen arrangement on the intact red cell membrane.

scholarly journals Quantitative immunoferritin microscopy of Fya, Fyb, Jka, U, and Dib antigen site numbers on human red cells

Blood ◽  
1980 ◽  
Vol 56 (6) ◽  
pp. 969-977
Author(s):  
SP Masouredis ◽  
E Sudora ◽  
L Mahan ◽  
EJ Victoria
Keyword(s):  
Distribution Patterns ◽  
Red Cells ◽  
Diverse Group ◽  
Red Cell ◽  
Human Erythrocyte Membrane ◽  
Red Cell Membrane ◽  
Quantitative Estimates ◽  
Equilibrium Binding ◽  
Antigen Site ◽  
Human Red Cells

The Fya, Fyb, Jka, U, and Dib antigen site numbers and ultrastructural distribution patterns on the human erythrocyte membrane were determined using quantitative immunoferritin microscopy. For homozygous antigen- positive red cells, the average number of determinants per red cell was about 14,000 for Jka, 17,000 for Fya and Fyb, 19,000 for Dib, and 23,000 for the U antigen, assuming that the equilibrium binding observed represented 80% saturation of the accessible antigen sites. The site numbers for this group of antigens were less than that for the Rh antigens, but considerably more than the Kell and Cellano antigens. The technique used was capable of demonstrating a twofold difference in antigen density between heterozygous and homozygous Fy (a+) red cells. More than 85% of the Fya and Fyb antigen sites were lost following pretreatment of the red cells with papain, consistent with the serologic lability of the Fy antigens following proteolysis. The ferritin distribution observed following conjugate staining of antibody- sensitized ghost membranes was similar for all five antigens studied and showed a random, clustered ferritin pattern. Although the quantitative estimates are valid, the remarkable similarity in antigen distribution pattern for this diverse group of antigens, as well as other considerations, suggest that the findings with ghose membranes probably do not reflect faithfully the antigen arrangement on the intact red cell membrane.

scholarly journals The State of Water in Human and Dog Red Cell Membranes

1970 ◽  
Vol 55 (4) ◽  
pp. 451-466 ◽  
Author(s):  
F. L. Vieira ◽  
R. I. Sha'afi ◽  
A. K. Solomon
Keyword(s):  
Activation Energy ◽  
Cell Membrane ◽  
Apparent Activation Energy ◽  
Water Diffusion ◽  
Red Cells ◽  
Red Cell ◽  
Kcal Mole ◽  
Red Cell Membrane ◽  
Temperature Range ◽  
Human Red Cells

The apparent activation energy for the water diffusion permeability coefficient, Pd, across the red cell membrane has been found to be 4.9 ± 0.3 kcal/mole in the dog and 6.0 ± 0.2 kcal/mole in the human being over the temperature range, 7° to 37°C. The apparent activation energy for the hydraulic conductivity, Lp, in dog red cells has been found to be 3.7 ± 0.4 kcal/mole and in human red cells, 3.3 ± 0.4 kcal/mole over the same temperature range. The product of Lp and the bulk viscosity of water, η, was independent of temperature for both dog and man which indicates that the geometry of the red cell membrane is not temperature-sensitive over our experimental temperature range in either species. In the case of the dog, the apparent activation energy for diffusion is the same as that for self-diffusion of water, 4.6–4.8 kcal/mole, which indicates that the process of water diffusion across the dog red cell membrane is the same as that in free solution. The slightly, but significantly, higher activation energy for water diffusion in human red cells is consonant with water-membrane interaction in the narrower equivalent pores characteristic of these cells. The observation that the apparent activation energy for hydraulic conductivity is less than that for water diffusion across the red cell membrane is characteristic of viscous flow and suggests that the flow of water across the membranes of these red cells under an osmotic pressure gradient is a viscous process.

Cyclic Loading on the Red Cell Membrane in a Shear Flow: A Possible Cause of Hemolysis

1985 ◽  
Vol 107 (2) ◽  
pp. 91-95 ◽  
Author(s):  
H. Niimi ◽  
M. Sugihara
Keyword(s):  
Cyclic Loading ◽  
Cell Membrane ◽  
Shear Flow ◽  
Red Cells ◽  
Red Cell ◽  
Two Dimensional ◽  
Shear Forces ◽  
Red Cell Membrane ◽  
Possible Cause ◽  
Human Red Cells

The fluid force acting on single human red cells in a high shear flow was analyzed. A two-dimensional elliptical microcapsule as a model of the deformed red cells was adopted to numerically calculate the distributions of the shear forces on both sides of the cell membrane. It is theoretically shown that the cell membrane undergoes an unsteady cyclic loading under the rotational motion around the interior. The mechanism leading to blood cell trauma is examined by repeatedly loading the continuously moving cell membrane.

scholarly journals Reflection coefficients of permeant molecules in human red cell suspensions.

1975 ◽  
Vol 66 (2) ◽  
pp. 251-265 ◽  
Author(s):  
J D Owen ◽  
E M Eyring
Keyword(s):  
Molar Volume ◽  
Cell Suspensions ◽  
Red Cells ◽  
Stopped Flow ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Experimental Procedure ◽  
Rapid Flow ◽  
Coefficient Omega ◽  
Human Red Cells

The Staverman reflection coefficient, sigma for several permeant molecules was determined in human red cell suspensions with a Durrum stopped-flow spectrophotometer. This procedure was first used with dog, cat, and beef red cells and with human red cells. The stopped-flow technique used was similar to the rapid-flow method used by those who originally reported sigma measurements in human red cells for molecules which rapidly penetrate the red cell membrane. The sigma values we obtained agreed with those previously reported for most of the slow penetrants, except malonamide, but disagreed with all the sigma values previously reported for the rapid penetrants. We were unable to calculate an "equivalent pore radius" with our sigma data. The advantages of our equipment and our experimental procedure are discussed. Our sigma data suggest that sigma is indirectly proportional to the log of the nonelectrolyte permeability coefficient, omega. Since a similar trend has been previously shown for log omega and molar volume of the permeant molecules, a correlatioo was shown between sigma and molar volume suggesting the membrane acts as a sieve.

scholarly journals Red Cell Membrane Permeability Deduced from Bulk Diffusion Coefficients

1974 ◽  
Vol 64 (6) ◽  
pp. 706-729 ◽  
Author(s):  
W. R. Redwood ◽  
E. Rall ◽  
W. Perl
Keyword(s):  
Cell Membrane ◽  
Membrane Permeability ◽  
Diffusion Coefficients ◽  
Bulk Diffusion ◽  
Red Cells ◽  
Cell Membrane Permeability ◽  
Red Cell ◽  
Cell Column ◽  
Red Cell Membrane ◽  
One Dimensional

The permeability coefficients of dog red cell membrane to tritiated water and to a series of[14C]amides have been deduced from bulk diffusion measurements through a "tissue" composed of packed red cells. Red cells were packed by centrifugation inside polyethylene tubing. The red cell column was pulsed at one end with radiolabeled solute and diffusion was allowed to proceed for several hours. The distribution of radioactivity along the red cell column was measured by sequential slicing and counting, and the diffusion coefficient was determined by a simple plotting technique, assuming a one-dimensional diffusional model. In order to derive the red cell membrane permeability coefficient from the bulk diffusion coefficient, the red cells were assumed to be packed in a regular manner approximating closely spaced parallelopipeds. The local steady-state diffusional flux was idealized as a one-dimensional intracellular pathway in parallel with a one-dimensional extracellular pathway with solute exchange occurring within the series pathway and between the pathways. The diffusion coefficients in the intracellular and extracellular pathways were estimated from bulk diffusion measurements through concentrated hemoglobin solutions and plasma, respectively; while the volume of the extracellular pathway was determined using radiolabeled sucrose. The membrane permeability coefficients were in satisfactory agreement with the data of Sha'afi, R. I., C. M. Gary-Bobo, and A. K. Solomon (1971. J. Gen. Physiol. 58:238) obtained by a rapid-reaction technique. The method is simple and particularly well suited for rapidly permeating solutes.

Rate of bicarbonate-chloride exchange in human red cells at 37 degrees C

1976 ◽  
Vol 40 (5) ◽  
pp. 707-714 ◽  
Author(s):  
R. A. Klocke
Keyword(s):  
Continuous Flow ◽  
Co2 Concentration ◽  
Carbonic Anhydrase Activity ◽  
Chloride Ions ◽  
Red Cell ◽  
Chloride Permeability ◽  
Red Cell Membrane ◽  
Chloride Exchange ◽  
Rapid Reaction ◽  
Human Red Cells

The rate of exchange of bicarbonate and chloride ions across the red cell membrane was studied in a continuous flow rapid reaction apparatus at 37 degrees C. A transmembrane gradient both ions was produced by mixture of cells suspended in a solution of one ion with an isosmotic solution of the other ion. Carbonic anhydrase activity was inhibited by acetazolamide to prevent changes in CO2 concentration during the experiments. Chloride and bicarbonate efflux from cells were studied in separate experiments at each experimental pH. Using a least squares technique, values of chloride and bicarbonate permeabilities were fitted to each pair of independent experiments. Chloride permeability averaged 1.1 (+/- 0.2 SD) X 10–4 cm/s and was not affected by change in pH. Recovered bicarbonate permeabilities varied widely, always remaining at least fivefold greater than chloride permeability. While bicarbonate permeability could not be accurately characterized, it appears to be greater than chloride permeability. Analysis of CO2 transfer with the estimated permeabilities indicates that the bicarbonate-chloride exchange by itself probably does not limit CO2 transfer.

scholarly journals Genetic variants of human red-cell membrane sialoglycoprotein β. Study of the alterations occurring in the sialoglycoprotein-β gene

1988 ◽  
Vol 250 (2) ◽  
pp. 407-414 ◽  
Author(s):  
M J Tanner ◽  
S High ◽  
P G Martin ◽  
D J Anstee ◽  
P A Judson ◽  
...  
Keyword(s):  
Genetic Variants ◽  
Southern Blotting ◽  
Protein Product ◽  
Red Cells ◽  
The Other ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Initiation Of Translation ◽  
Beta Gene ◽  
Human Red Cell Membrane

We have studied the DNA of individuals who express an altered sialoglycoprotein beta on their red cells by using Southern blotting with sialoglycoprotein-beta cDNA probes. Individuals of the Leach phenotype do not express any beta (sialoglycoprotein beta) or gamma (sialoglycoprotein gamma) on their red cells, and we show that about 7 kb of DNA, including the 3′ end of the beta gene, is deleted in this DNA. Any protein product of this gene is likely to lack the membrane-associating domain of beta. We have also examined the DNA of two types of other individuals (Yus-type and Gerbich-type) who have red cells that lack beta and gamma, but contain abnormal sialoglycoproteins related to beta. These two types of DNA contain different internal deletions of about 6 kb in the beta gene. We suggest that these deletions result from the presence of two different sets of internal homology in the beta gene, and on this basis we propose structures for the abnormal Yus-type and Gerbich-type sialoglycoproteins which are consistent with the other evidence that is available. We provide evidence that beta and gamma are products of the same gene and suggest a possible mechanism for the origin of gamma based on leaky initiation of translation of beta mRNA.

Oxygen Therapeutics: Pursuit of an Alternative to the Donor Red Blood Cell

2007 ◽  
Vol 131 (5) ◽  
pp. 734-741
Author(s):  
Paul M. Ness ◽  
Melissa M. Cushing
Keyword(s):  
Blood Donation ◽  
Blood Substitute ◽  
Blood Substitutes ◽  
Red Cells ◽  
Phase Iii ◽  
Red Cell ◽  
Oxygen Carriers ◽  
Term Survival ◽  
Phase Iii Clinical Trials ◽  
Human Red Blood Cells

Abstract Context.—There is no true substitute for the many functions of human red blood cells, and synthetic products will not replace the need for blood donation in the foreseeable future. Hemoglobin-based oxygen carriers have many characteristics that would serve as a useful adjunct to red cells in clinical settings. Over time, these technologies have the potential to dramatically reshape the practice of transfusion medicine. Objective.—To review the characteristics and potential utility of hemoglobin-based oxygen carriers and perfluorocarbon-based oxygen carriers. Several hemoglobin-based oxygen carriers are under study in phase III clinical trials. Novel uses for synthetic oxygen therapeutics are emphasized. Data Sources.—All published reports with the key words oxygen therapeutics, blood substitutes, and red cell substitutes from 1933 until March 2006 were searched through Medline. Significant findings were synthesized. Conclusions.—Recognition of the true impact of red cell substitutes is still several years away. The most compelling products, hemoglobin-based oxygen carriers, have potential use in trauma, providing immediate oxygen-carrying support in the face of alloantibodies or autoantibodies, and in other clinical situations in which long-term survival of red cells is not essential. In the interim, efforts should be focused on enhancing the current blood supply system while supporting ongoing and planned blood substitute research efforts, including trials assessing novel clinical indications for these products.

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