scholarly journals Hemoglobin Charge Dependence on Hemoglobin Concentration in Vitro

1971 ◽  
Vol 57 (3) ◽  
pp. 283-289 ◽  
Author(s):  
C. M. Gary-Bobo ◽  
A. K. Solomon
Keyword(s):  
External Solution ◽  
Hemoglobin Concentration ◽  
Bulk Water ◽  
Red Cells ◽  
A Value ◽  
Charge Dependence ◽  
Hemoglobin Molecule ◽  
Sigmoid Curve ◽  
Human Red Cells

Studies have been made of the dependence of the charge of the hemoglobin molecule on hemoglobin concentration in the concentration range between 3 and 11 mmolal. The charge has been determined by measuring the distribution of 42K between a hemoglobin solution in a cellophane bag and an external solution. The pH was 6.6, the K concentration was 10 mM, and the temperature was 4°C. The charge decreased along a sigmoid curve from a value of +3 in the most dilute solutions to a value of +0.5 in the most concentrated solutions. The results were in excellent agreement with earlier studies of Gary-Bobo and Solomon in which Cl distribution was measured between human red cells and external solutions and thus give added support to the conclusion that the apparent anomalous osmotic behavior of human red cells may be attributed to concentration-dependent changes in the hemoglobin net charge. The present findings also support the view that the water in the red cell is solvent water for K and Cl and differs in no quantitatively important respect from bulk water in free solution.

In Vitro Assay of Platelet Adhesiveness with Washed and Tanned Human Red Cells

1968 ◽  
Vol 20 (03/04) ◽  
pp. 384-396 ◽  
Author(s):  
G Zbinden ◽  
S Tomlin
Keyword(s):  
Platelet Adhesion ◽  
Sodium Citrate ◽  
Blood Platelets ◽  
In Vitro Assay ◽  
Red Cells ◽  
Platelet Adhesiveness ◽  
Vitro Assay ◽  
In Vitro System ◽  
Human Red Cells

SummaryAn in vitro system is described in which adhesion of blood platelets to washed and tannic acid-treated red cells was assayed quantitatively by microscopic observation. ADP, epinephrine and TAME produced a reversible increase in platelet adhesiveness which was antagonized by AMP. With Evans blue, polyanetholsulfonate, phthalanilide NSC 38280, thrombin and heparin at concentrations above 1-4 u/ml the increase was irreversible. The ADP-induced increase in adhesiveness was inhibited by sodium citrate, EDTA, AMP, ATP and N-ethylmaleimide. EDTA, AMP and the SH-blocker N-ethylmaleimide also reduced spontaneous platelet adhesion to red cells. No significant effects were observed with adenosine, phenprocoumon, 5-HT, phthalanilide NSC 57155, various estrogens, progestogens and fatty acids, acetylsalicylic acid and similarly acting agents, hydroxylamine, glucose and KCN. The method may be useful for the screening of thrombogenic and antithrombotic properties of drugs.

scholarly journals Reticulocyte Survival in Sickle Cell Anemia: Effect of Cyanate

Blood ◽  
1972 ◽  
Vol 40 (5) ◽  
pp. 733-739 ◽  
Author(s):  
Blanche P. Alter ◽  
Yuet Wai Kan ◽  
David G. Nathan
Keyword(s):  
Cell Survival ◽  
Sickle Cell ◽  
Fetal Hemoglobin ◽  
Red Cells ◽  
Red Cell ◽  
Hemoglobin Molecule ◽  
Red Cell Survival ◽  
And Control

Abstract Cyanate prevents sickling in vitro and apparently prolongs the survival of 51Cr-tagged sickle erythrocytes in vivo. Cautious interpretation is required because the effects of cyanate on 51Cr binding to sickle and fetal hemoglobin-containing red cells are unknown, and comparison of the effect of cyanate on sickle red cell survival to control red cell survival must be performed sequentially. We have studied the survival of sickle reticulocytes utilizing radioactive amino acids that are incorporated into hemoglobin. Two informed adult patients with sickle cell disease were studied. In each study, two 50-ml samples of blood were incubated separately with 14C- and 3H-leucine for 2 hr, after which 50 mM cyanate was added to one aliquot for 1 hr. The cells were then washed and reinfused. Frequent venous samples were obtained, and the specific activities of 14C and 3H in the hemoglobin were followed. The t ½ of the carbamylated cells was tripled, but remained below normal. This method provides a generally useful measurement of the influence of drugs bound to red cells on reticulocyte lifespan. The labels are incorporated into the hemoglobin molecule of the reticulocyte, and simultaneous comparison of the survivals of the same cohort of drug-treated and control cells is achieved.

scholarly journals Determination of Equivalent Pore Radius for Human Red Cells by Osmotic Pressure Measurement

1960 ◽  
Vol 44 (1) ◽  
pp. 1-18 ◽  
Author(s):  
David A. Goldstein ◽  
A. K. Solomon
Keyword(s):  
Experimental Data ◽  
Pore Radius ◽  
Initial Rate ◽  
Equilibrium Concentration ◽  
External Solution ◽  
Red Cells ◽  
Cell Swelling ◽  
Zero Time ◽  
Human Red Cells

A new method has been developed to measure the equivalent pore radius in cellular membranes, and has been applied to human red cells. When red cells are suddenly introduced into a non-isosmolar concentration of non-lipid-soluble non-electrolyte molecules, water will enter or leave the cell. The rate of cell swelling or shrinking is determined and extrapolated to zero time to give the initial rate of volume change. By suitable adjustment of the concentration of the external solution the initial rate may be brought to zero. The transient equilibrium concentration, determined by interpolation from experimental data, gives a measure of Staverman's reflection coefficient, σ. The zero time method has enabled us to determine σ for nine permeant molecules. σ is directly related to the equivalent pore radius; the experimental data lead to a value of 4.2 Å for the equivalent pore radius in man, in good agreement with the previous figure of 3.5 Å given by Paganelli and Solomon. The zero time method offers a number of advantages over previous methods for determination of this parameter. It requires no measurement of the rate of water entrance into the cell, and is essentially independent of the kinetics of cell swelling. It may be applied to a variety of living cells so that many additional membranes may now be characterized in terms of their equivalent pore radius.

scholarly journals Antisickling effect of tellurite: a potent membrane-acting agent in vitro

Blood ◽  
1984 ◽  
Vol 64 (1) ◽  
pp. 305-307 ◽  
Author(s):  
T Asakura ◽  
Y Shibutani ◽  
MP Reilly ◽  
RH DeMeio
Keyword(s):  
Hemoglobin Concentration ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Cell Swelling ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Potassium Tellurite ◽  
New Type ◽  
Acting Agent

Abstract Potassium tellurite (K2TeO3) was found to be a potent antisickling agent that inhibited red cell sickling at concentrations less than 10 mumol/L. The inhibitory effect depended on the incubation time, with the effect increasing with longer incubation periods. Because tellurite causes swelling of red cells, and because the antisickling effect of tellurite correlated with the degree of red cell swelling, the antisickling effect of tellurite is assumed to be due to the decreased mean cell hemoglobin concentration. Swelling of red cells by tellurite was accelerated by the addition of reduced glutathione. Tellurite appears to be a new type of antisickling agent that interacts with the red cell membrane.

scholarly journals Some Interrelationships of Blood and the Fava Bean Principle in Vitro

Blood ◽  
1952 ◽  
Vol 7 (7) ◽  
pp. 721-728 ◽  
Author(s):  
WILLIAM P. CREGER ◽  
HOUGHTON GIFFORD
Keyword(s):  
Animal Species ◽  
Gamma Globulin ◽  
Test Tube ◽  
Red Cells ◽  
Red Cell ◽  
Fava Bean ◽  
Increased Susceptibility ◽  
Human Red Cells

Abstract 1. Saline suspensions of human red cells, as well as those of several animal species, were agglutinated by normal saline extracts of the Fava bean. 2. This agglutination was potentiated in titer 100-fold in a medium of 10 per cent acacia, as a diluent. 3. The inhibition of the hemagglutination action of the Fava bean extract by human serum was apparently attributable to the gamma globulin fraction. 4. The Fava bean principle could be transferred from cell to cell, as shown by heat-elution and acacia technics. 5. Fava-sensitized red cells did not exhibit increased susceptibility in the test tube to complement, hemolysin, or osmotic or mechanical fragility. 6. The mechanism of in vivo red cell destruction in Favism is as yet unknown, but a special immunologic susceptibility to the action of the bean’s principle is suspected in certain persons. 7. It is suggested that the relation of acacia to Fava-sensitized red cells may form the basis of a diagnostic test for Favism in the early, acute stages of the disease.

Influence of Cell Age and Ketoaminic Linkage on Rapid Glycosylation of Hemoglobin in Human Red Cells In Vitro

1985 ◽  
Vol 17 (04) ◽  
pp. 201-204 ◽  
Author(s):  
R. Cordera ◽  
G. Andraghetti ◽  
R. Bonadonna ◽  
R. Gherzi ◽  
M. Maiello ◽  
...  
Keyword(s):  
Red Cells ◽  
Cell Age ◽  
Human Red Cells

Calmodulin carboxylmethyl ester formation in intact human red cells and modulation of this reaction by divalent cations in vitro

FEBS Letters ◽  
1982 ◽  
Vol 147 (1) ◽  
pp. 125-130 ◽  
Author(s):  
L. Runte ◽  
H.-L. Jürgensmeier ◽  
C. Unger ◽  
H.D. Söling
Keyword(s):  
Divalent Cations ◽  
Red Cells ◽  
Ester Formation ◽  
Human Red Cells
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