Lactate and pyruvate gradients between red blood cells and plasma during acute asphyxia

1964 ◽  
Vol 19 (6) ◽  
pp. 1100-1104 ◽  
Author(s):  
Salha S. Daniel ◽  
Hisayo O. Morishima ◽  
L. Stanley James ◽  
Karlis Adamsons
Keyword(s):  
Sodium Lactate ◽  
Red Cells ◽  
Red Cell ◽  
Cell Ratio ◽  
Lactate And Pyruvate ◽  
The Mean ◽  
Endogenous Release ◽  
Pyruvate Ratio

The rate of equilibration of lactate and pyruvate between plasma and red cells has been studied during asphyxia and following addition of sodium lactate in vivo and in vitro. In the resting, well-oxygenated guinea pig, the mean plasma/red cell ratio of lactate was 1.55 and that of pyruvate 2.47. During asphyxia, the plasma/red cell ratio of lactate rose and that of pyruvate fell, indicating a delay in equilibration. Incomplete equilibration affected particularly the lactate/pyruvate ratio in the two compartments. Infused neutral sodium lactate penetrated the red cells at a rate comparable to that observed following endogenous release of lactic acid during acute asphyxia. In vitro at pH 6.8@#X2013;7.4 at 38 C, the time to 50% equilibration of lactate between plasma and cells of human blood was less than 2 min. It is concluded that during acute asphyxia and resuscitation whole blood values of lactate and pyruvate do not bear a constant relationship to those of plasma. lactate/pyruvate ratio Submitted on March 16, 1964

Nitrite and Red Cell Function in Carp: Control Factors for Nitrite Entry, Membrane Potassium Ion Permeation, Oxygen Affinity and Methaemoglobin Formation

1990 ◽  
Vol 152 (1) ◽  
pp. 149-166 ◽  
Author(s):  
FRANK B. JENSEN
Keyword(s):  
Cell Function ◽  
Oxygen Affinity ◽  
Red Cells ◽  
Red Cell ◽  
In Vitro Experiments ◽  
K Uptake ◽  
The Mean ◽  
K Release

Red cell function was studied in carp by a combination of in vivo and in vitro experiments with nitrite as the perturbing agent. In vivo accumulation of nitrite caused a marked increase in the red cell methaemoglobin content, and reduced the mean cellular volume. The oxygen affinity of unoxidized haemoglobin was strongly decreased, partly as result of the elevated concentration of cellular nucleoside triphosphates and haemoglobin associated with red cell shrinkage. Red cell pH was unchanged compared to controls, but reduced when referred to constant extracellular pH and O2 saturation. The mean cellular K+ content decreased, reflecting a K+ loss from the red cells during their shrinkage. This K+ loss contributed significantly to the large plasma hyperkalaemia during nitrite exposure. In vitro experiments revealed that nitrite influx into deoxygenated red cells was much larger than into oxygenated red cells. Nitrite permeation of the red cell membrane was not inhibited by DIDS and did not change extracellular pH. Methaemoglobin (MetHb) formation was more pronounced in deoxygenated blood than in oxygenated blood, but quasi-steady states were reached, reflecting a balance between nitrite-induced MetHb formation and the action of MetHb reductase. Red cells incubated in the oxygenated state released K+, whereas a net K+ uptake occurred in deoxygenated cells. Nitrite did not change the K+ loss from oxygenated cells, but shifted the K+ uptake in deoxygenated cells to a pronounced K+ release by the time high MetHb levels were reached. Both types of red cell K+ release were inhibited by DIDS and appeared to occur via a route involving Band 3. The data are consistent with the hypothesis that a significant DIDS-sensitive K+ efflux from the red cells occurs whenever a large fraction of the haemoglobin molecules assumes an R-like quaternary structure.

LACK OF EFFECT OF CORTICOSTEROIDS ON THE IN VIVO DISAPPEARANCE OF SULFHYDRYL-INHIBITED AND ANTIBODY-COATED ERYTHROCYTES

1964 ◽  
Vol 47 (3_Suppl) ◽  
pp. S28-S36
Author(s):  
Kailash N. Agarwal
Keyword(s):  
Red Cells ◽  
List Type ◽  
Red Cell

ABSTRACT Red cells were incubated in vitro with sulfhydryl inhibitors and Rhantibody with and without prior incubation with prednisolone-hemisuccinate. These erythrocytes were labelled with Cr51 and P32 and their disappearance in vivo after autotransfusion was measured. Prior incubation with prednisolone-hemisuccinate had no effect on the rate of red cell disappearance. The disappearance of the cells was shown to take place without appreciable intravascular destruction.

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals The relationship between in vivo generated hemoglobin skeletal protein complex and increased red cell membrane rigidity

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1427-1431 ◽  
Author(s):  
N Fortier ◽  
LM Snyder ◽  
F Garver ◽  
C Kiefer ◽  
J McKenney ◽  
...  
Keyword(s):  
Protein Complex ◽  
Sodium Dodecyl ◽  
Red Cells ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Cellular Dehydration ◽  
Thalassemia Minor ◽  
Membrane Rigidity

Abstract In vitro induced oxidative damage to normal human RBCs has previously been shown to result in increased membrane rigidity as a consequence of the generation of a protein complex between hemoglobin and spectrin. In order to determine if in vivo generated hemoglobin-spectrin complexes may play a role in increased membrane rigidity of certain pathologic red cells, we measured both these parameters in membranes prepared from hereditary xerocytosis (Hx), sickle cell disease (Sc), and red cells from thalassemia minor (beta thal). Membranes were prepared from density-fractionated red cells, and membrane deformability was measured using an ektacytometer. Hemoglobin-spectrin complex was determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel analysis, as well as by Western blot analysis using a monoclonal antibody against the beta- subunit of hemoglobin. For these three types of pathologic red cells, progressive cellular dehydration was associated with increased membrane rigidity and increased content of hemoglobin-spectrin complex. Moreover, the increase in membrane rigidity appeared to be directly related to the quantity of hemoglobin-spectrin complex associated with the membrane. Our findings imply that hemoglobin-spectrin complex is generated in vivo, and this in turn results in increased membrane rigidity of certain pathologic red cells. The data further suggest that oxidative crosslinking may play an important role in the pathophysiology of certain red cell disorders.

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.

SECRETION OF PROGESTERONE BY HUMAN OVARIES PERFUSED IN VITRO

1975 ◽  
Vol 79 (1) ◽  
pp. 111-121 ◽  
Author(s):  
H. Fukunishi ◽  
H. Mickan ◽  
J. Zander
Keyword(s):  
Ovarian Function ◽  
Sodium Lactate ◽  
Bicarbonate Buffer ◽  
Ldh Activity ◽  
Progesterone Secretion ◽  
Experimental Approaches ◽  
Lactate And Pyruvate ◽  
Human Ovaries

ABSTRACT A system for the perfusion of isolated human ovaries is described. Krebs-Ringer bicarbonate buffer (pH 7.4) containing glucose, sodium lactate and pyruvate was perfused at a flow rate of 5–8 ml/min at 37°C through the ovarian artery. Oxygen uptake, LDH-activity, pH and progesterone secretion were determined as parameters of viability of the perfused organ. Our results demonstrate that a relatively simple system used for perfusion was able to preserve ovarian function for at least two hours. This experimental design should not be regarded as physiological but in addition to other experimental approaches it might provide information on the physiology of the human ovary. Though lower values of progesterone secretion than in vivo were found, a correlation of secretion and functional stage of the ovaries could be demonstrated.

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393 ◽  
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

Abstract The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals Near-normal circulatory survival of rabbit red cells exposed to high levels of Ca and ionophore in vitro

Blood ◽  
1985 ◽  
Vol 66 (1) ◽  
pp. 220-223 ◽  
Author(s):  
RM Bookchin ◽  
EF Jr Roth ◽  
VL Lew
Keyword(s):  
Adenosine Triphosphate ◽  
Indirect Evidence ◽  
Red Cells ◽  
Ionophore A23187 ◽  
Red Cell ◽  
L Cells

Abstract The belief is widely held, on the basis of indirect evidence, that a substantial, even brief elevation of red cell Ca content must result in a marked shortening of circulatory survival. To test this notion directly, we exposed rabbit red cells in vitro to the ionophore A23187 and Ca so as to produce sustained uniform cell Ca levels of 40 to 360 mumol/L cells for one to 60 minutes, and compared the survival of the Ca-loaded cells in vivo with that of ionophore-treated controls, simultaneously, in the same rabbits. Despite marked reductions in cell adenosine triphosphate and dehydration of the Ca-exposed cells prior to reinfusion, the majority of cells, all of which had experienced these high cytoplasmic Ca levels, showed normal or near-normal survival in the circulation.

scholarly journals Enhanced survival of sickle erythrocytes upon treatment with glyceraldehyde

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 544-546 ◽  
Author(s):  
LJ Benjamin ◽  
JM Manning
Keyword(s):  
Life Span ◽  
Lysine Residue ◽  
Red Cell ◽  
Hemoglobin Tetramer ◽  
Sickle Erythrocytes ◽  
The Mean

Abstract Glyceraldehyde has been demonstrated to be an antisickling agent in vitro. In the present investigation, chromium-51 red cell studies were used to investigate the life span in vivo of sickle erythrocytes after treatment with glyceraldehyde in vitro. The mean survival (T1/2) of control cells was 5.8 +/- 1.6 days, whereas cells treated with 10 mmol/L or 20 mmol/L glyceraldehyde survived 9.0 +/- 1.4 (P less than .05) and 11.3 +/- 0.8 (P less than .002) days, respectively. The extent of modification by glyceraldehyde was 0.4 to 1.0 lysine residue per hemoglobin tetramer. These studies demonstrate not only a prolongation of the life span of sickle erythrocytes by treatment with glyceraldehyde but also the absence of any deleterious effects that would be revealed by this study.

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.

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