Showing papers on "Red blood cell published in 1989"

Erythrocyte membrane lipid peroxidation and glycosylated hemoglobin in diabetes.

Abstract: Erythrocytes of diabetic patients have abnormal membrane properties. We examined in vivo membrane lipid peroxidation in erythrocytes of diabetic subjects and its possible relationship with hyperglycemia. Lipid peroxidation was assessed in fresh, untreated erythrocytes by quantitating thiobarbituric acid reactivity and an adduct of phospholipids and malonyldialdehyde (MDA), an end product of lipid peroxidation, with thin-layer chromatography of lipid extract of diabetic erythrocytes. There was a significantly increased membrane lipid peroxidation in diabetic erythrocytes compared with nondiabetic erythrocytes. The degree of membrane lipid peroxidative damage in erythrocytes was significantly correlated with the level of glycosylated hemoglobin, an index of mean glucose level for the preceding 3-4 mo. This suggests that peroxidation of membrane lipids and accumulation of MDA occurs in erythrocytes of diabetic patients.

The behavior of sonicated albumin microbubbles within the microcirculation: a basis for their use during myocardial contrast echocardiography.

Abstract: The purpose of this study was to determine whether the behavior of sonicated albumin microbubbles accurately mimics red blood cell flow in the microcirculation and is thus consistent with their use as in vivo tracers of red blood cell flow during myocardial contrast echocardiography. Accordingly, microbubbles prepared from fluorescein-conjugated albumin and fluorescently labeled red blood cells were injected intravascularly in eight golden hamsters. Their intravascular distribution, velocities, arteriolar-to-venular transit and flux ratios at branch points were determined in the microcirculation of the cheek pouch. Albumin microbubbles (mean diameter, 4.9 +/- 3.6 microns) and red blood cells displayed a similar frequency of distribution across the arteriolar lumen (33% in the central 20% of the arterioles), and their arteriolar velocities were also similar (2.5 +/- 0.7 mm/sec and 2.3 +/- 0.7 mm/sec,p = NS). The mean velocities of microbubbles correlated well with those of red blood cells at baseline and after adenosine application (r = 0.97 and r = 0.89, respectively), as did the calculated maximum velocity (r = 0.98 and r = 0.80, baseline and adenosine, respectively). The velocity profiles across the lumen of the vessels for albumin microbubbles and red blood cells were similar at baseline and after adenosine-induced velocity changes. The flux ratios at branch points also correlated well (r = 0.92, p less than 0.001). Arteriolar-to-venular transit times of albumin microbubbles were similar to those of red blood cells in vessels ranging in size from 22 microns to 45 microns. We conclude that the behavior of albumin microbubbles in the microcirculation mimics that of red blood cells and supports their use as intravascular tracers of red blood cell flow during myocardial contrast echocardiography.

Structure and Function of the Red Blood Cell Anion Transport Protein

Abstract: PHYSIOLOGICAL FUNCTION . Role of Chloride-Bicarbonate Exchange in Carbon Dioxide Transport .. Other Substrates of Red Cell Band 3 . . Transport Functions Other Than Anion Exchange . . Band 3 Homologs in Nonerythroid Cells ..

Control and coordination of gas transfer in fishes

Abstract: Recent developments pertaining to the control and coordination of gas transfer in fishes have been reviewed. Gill ventilatory water flow can markedly affect blood respiratory and blood acid–base status. Although arterial oxygen content traditionally has been considered the predominant factor controlling ventilation, we present evidence for additional involvement of both blood acid–base status and circulating catecholamines. An analysis of the independent effects of blood oxygen content, acid–base status, and catecholamines in controlling ventilation is confounded by the interrelationships among these variables. It is likely, however, that each factor is involved to some extent in ventilatory control in fishes. Blood oxygen transport is affected by the carrying capacity of the blood and red blood cell chemical status. Blood oxygen-carrying capacity is increased during periods of stress by adrenergic release of red blood cells from the spleen. Concurrently, adrenergic stimulation of red blood cell Na+–H+ ex...

In vivo regeneration of red cell 2,3-diphosphoglycerate following transfusion of DPG-depleted AS-1, AS-3 and CPDA-1 red cells.

Abstract: Regeneration of 2,3-diphosphoglycerate (DPG) was determined following transfusion of DPG-depleted group O red cells into group A recipients. Blood from five donors was stored in the adenine-containing solutions CPDA-1, AS-1 or AS-3 for 35 d at 4 degrees C. Post-transfusion red cell DPG and ATP were measured in separated group O red cells over a 7 d period. The studies confirmed rapid in vivo DPG regeneration with greater than or equal to 50% of the maximum level being achieved within 7 h. An average of 95% of the recipients' pre-transfusion DPG level was achieved by 72 h and by 7 d mean (+/- SEM) DPG levels relative to recipient's pre-transfusion DPG averaged 84% (+/- 13%), 92% (+/- 17%) and 84% (+/- 21%) for CPDA-1, AS-1 and AS-3 red cells, respectively. Results were comparable to those previously reported for blood stored in ACD for 15-20 d (Valeri & Hirsch, 1969; Beutler & Wood, 1969). The immediate regeneration rate, V, closely approximated first order regeneration kinetics with AS-3 red cells exhibiting double the rate of CPDA-1 red cells (P less than 0.001). AS-1 red cells exhibited an intermediate rate of regeneration which was not significantly different compared to either CPDA-1 or AS-3 (P greater than 0.05). V exhibited a significant (P less than 0.05) positive correlation with ATP levels 5-7 h post-infusion. ATP regeneration of the infused cells was rapid with a mean increase of 1.2 mumol/g Hb above post-storage levels being achieved 1 h following transfusion.

Effect of hydroxyurea on the rheological properties of sickle erythrocytes in vivo.

Abstract: We have monitored the rheological effects of hydroxyurea (HU) on erythrocytes obtained from two patients with severe sickle cell anemia who were enrolled in a therapeutic trial of this drug. Erythrocyte membrane stability and whole cell and membrane deformability of red cells from treated and untreated patients and normal controls were determined in room air using an ektacytometer--a laser viscodiffractometer. The percentage of dense cells was quantitated by centrifugation on a discontinuous Stractan density gradient. F reticulocytes (FR), absolute F reticulocytes (AFR), and F cells (FC) were determined by single-cell radial immunolgic assays. After 1 year of treatment with HU, there was a significant increase in the level of hemoglobin (Hb) F, FR, AFR, and FC. The degree of anemia remained the same, but there was significant increase in the mean cell volume (MCV) and a significant decrease in the mean corpuscular Hb concentration (MCHC). Whole cell deformability improved by twofold, but membrane stability remained within normal limits. The hydration status of sickle erythrocytes improved as was indicated by a change toward normal in gradient osmotic ektacytometry, an increase in RBC K+ content, a decrease in percent of dense cells, and a decrease in the MCHC. The data indicate that, in addition to its effect on the production of Hb, F, HU has a salutary effect on whole cell deformability and on the hydration status of sickle erythrocytes. Determination of the rheological properties of erythrocytes may be of value in monitoring the response to HU.

Relationship between the membrane inhibitor of reactive lysis and the erythrocyte phenotypes of paroxysmal nocturnal hemoglobinuria.

Abstract: Susceptibility to hemolysis initiated by activated cobra venom factor (CoF) complexes is a characteristic that distinguishes the most complement-sensitive type III erythrocytes of paroxysmal nocturnal hemoglobinuria (PNH) from the intermediately sensitive type II and the normally sensitive type I cells. Recently we isolated a membrane constituent from normal erythrocytes that inhibits CoFBb-initiated hemolysis, and this protein was designated membrane inhibitor of reactive lysis (MIRL). To investigate the molecular basis of the variability in complement sensitivity among PNH erythrocytes, the surface expression of MIRL and decay accelerating factor (DAF) on the three phenotypes of PNH was quantified immunochemically. Both complement regulatory proteins were markedly deficient on the erythrocytes from a patient with predominately type III cells. The erythrocytes from patients with a majority of either type II or I cells were also significantly deficient in both MIRL and DAF. While cytofluorometric analysis confirmed the quantitative deficiencies, segregation of erythrocytes into discrete subpopulations that expressed either no MIRL or normal amounts of MIRL was not observed. The results of immunoprecipitation studies were consistent with quantitative, but not qualitative abnormalities of MIRL and DAF. Selective removal of the sensitive erythrocytes indicated that approximately 20% of the normal amount of MIRL is sufficient to protect cells from CoF-initiated lysis. These studies suggest that relatively subtle quantitative differences in membrane complement regulatory proteins underlie the variability in complement sensitivity of PNH erythrocytes.

Heterogeneity of the Ro/SSA antigen. Different molecular forms in lymphocytes and red blood cells.

Abstract: Ro(SSA) is an intracellular ribonucleoprotein against which autoantibodies are found in a portion of patients with Sjogren's syndrome and systemic lupus erythematosus. A form of Ro(SSA) is described in red blood cells that shares a line of identity with purified Ro(SSA) from bovine spleen and human lymphocytes in counterimmunoelectrophoresis, but has different molecular properties. Ro(SSA) from red blood cells exists in association with only two small RNAs as opposed to four in other cell types, as determined by RNA extraction of protein A-assisted immunoprecipitates. In addition to the common 60-kD Ro(SSA) protein, Western blot analysis revealed an additional 52-kD protein in lymphocytes and a 54-kD protein in red blood cells. The 60-kD form of Ro(SSA) in red cells was found to be antigenically distinct from that in the lymphocyte, because sera were identified that bound each exclusively. Finally, a rabbit antibovine Ro(SSA) serum distinguished red cell from lymphocyte Ro(SSA). These results suggest two distinctive populations of Ro(SSA) proteins and distributions of Ro(SSA) RNAs in the lymphocyte and red blood cell.

Initial stages of influenza hemagglutinin-induced cell fusion monitored simultaneously by two fluorescent events: cytoplasmic continuity and lipid mixing.

Abstract: We have monitored the mixing of both aqueous intracellular and membrane-bound fluorescent dyes during the fusion of human red blood cells to influenza hemagglutinin-expressing fibroblasts using fluorescence spectroscopy and low light, image-enhanced video microscopy. The water-soluble fluorescent dye, N-(7-nitrobenzofurazan-4-yl)taurine, was incorporated into intact human red blood cells. The fluorescence of the dye in the intact red blood cell was partially quenched by hemoglobin. The lipid fluorophore, octadecylrhodamine, was incorporated into the membrane of the same red blood cell at self-quenching concentrations (Morris, S. J., D. P. Sarkar, J. M. White, and R. Blumenthal. 1989. J. Biol. Chem. 264: 3972-3978). Fusion, which allowed movement of the water-soluble dye from the cytoplasm of the red blood cell into the hemagglutinin-expressing fibroblasts, and movement of octadecylrhodamine from membranes of red blood cell to the plasma membrane of the fibroblasts, was observed by fluorescence microscopy as a spatial relocation of dyes, and monitored by spectrofluorometry as an increase in fluorescence. Upon lowering the pH below 5.4, fluorescence increased after a delay of about 30 s at 37 degrees C, reaching a maximum within 3 min. The kinetics, pH profile, and temperature dependence were similar for both fluorescent events measured simultaneously, indicating that influenza hemagglutinin-induced fusion rapidly establishes bilayer continuity and exchange of cytoplasmic contents.

Methods and devices for the separation of plasma or serum from whole blood

Abstract: A device and method for permitting the separation of plasma or serum from whole blood. The device comprises a matrix of hydrophilic sintered porous material to which at least one red blood cell agglutinating agent has been applied. According to a first method of using the device, a sample of whole blood is applied to a first end of the matrix and the red blood cells within the sample come in contact with the agglutinating agents present in the matrix. The blood cells agglutinate, and are entrapped in the interstices of the matrix, while substantially blood-cell-free serum or plasma accumulates near the outlet of the device. A filter means in liquid receiving relationship with the matrix functions to wick the serum or plasma from the matrix. According to an alternative aspect of the invention, a filter means in liquid receiving relationship with the outlet of the matrix functions to retain any blood cells which pass through the matrix is the filter means wicks the plasma or serum from the matrix. Additional agglutinating agents may be incorporated within the filter means to facilitate retention of blood cells which pass through the matrix.

Anticancer activities of adenine nucleotides in mice are mediated through expansion of erythrocyte ATP pools.

Abstract: ATP and AMP exhibit significant anticancer activities against established footpad CT26 colon adenocarcinoma in CB6F1 mice. Adenosine, inorganic phosphate, and inorganic pyrophosphate were without such effects under identical conditions. Daily intraperitoneal injections of adenine nucleotides in large volumes of saline, starting after the tumors became palpable, resulted in inhibition of tumor growth and a few "cures." The treatment was not toxic to the host as determined by changes in body weights. Weight loss observed in animals upon progression of the fast-growing CT26 tumors was slowed markedly in adenine nucleotide-treated mice. The inhibition of weight loss in tumor-bearing mice was shown to be neither the cause nor the effect of the inhibition of tumor growth. Intraperitoneal injections of AMP or ATP but not of adenosine yielded expansions of erythrocyte ATP pools in host animals. The expanded erythrocyte ATP pools are stable over a period of hours, while slowly releasing micromolar amounts of ATP into the blood plasma compartment, leading to several-fold increases in plasma (extracellular) ATP levels. Based on previous studies in which 1-5 microM extracellular ATP effectively inhibited the growth of a variety of tumor cells in several in vitro systems, it is suggested that similar levels of ATP in blood plasma account for the anticancer activities observed in a murine host.

The micronucleus test and erythropoiesis. Effects of erythropoietin and a mutagen on the ratio of polychromatic to normochromatic erythrocytes (P/N ratio)

Abstract: It is considered that a decrease of the ratio of polychromatic erythrocytes (PCE) to normochromatic erythrocytes (NCE) (P/N) in the micronucleus test is an indicator of bone marrow toxicity induced by mutagens. However, the exact meaning of fluctuation in the P/N ratio is not yet known. We have studied this point by counting the total number of erythrocytes and nucleated cells in the bone marrow following various treatments. The P/N ratio decreased gradually with time after administration of mitomycin C. Our data suggest that the decrease of P/N ratio was attributable to an increase in the numbers of the denominator, i.e. NCE, caused by rapid differentiation and multiplication or denucleation of erythroblasts which remained in the bone marrow instead of entering the peripheral blood stream. A decrease of P/N ratio was also observed in the early phase after administration of erythropoietin, an agent which induces differentiation and multiplication of erythroblasts. This phenomenon might result from an increase of PCE delivery into the blood circulation. However, following the initial decrease, the P/N ratio increased gradually 48 h after administration of erythropoietin. It is supposed that this increase probably resulted from an increase in PCE in the bone marrow due to the direct effects of erythropoietin on erythropoiesis. The drastic change in erythropoiesis in the bone marrow induced by either mutagen or erythropoietin treatment will affect the fluctuations of the P/N ratio or the number of micronucleated erythrocytes per non-micronucleated erythocytes in the micronucleus test. This contrasts with the original explanation for such fluctuations which attributed them to replenishment of the marrow by peripheral blood.

Characterization of a modified red cell membrane protein expressed on erythrocytes infected with the human malaria parasite Plasmodium falciparum: possible role as a cytoadherent mediating protein.

Abstract: Infections with the human malaria Plasmodium falciparum are characterized by the retention of parasitized erythrocytes in tissue capillaries and venules. Erythrocytes containing trophozoites and schizonts attach to the endothelial cells that line these vessels by means of structurally identifiable excrescences present on the surface of the infected cell. Such excrescences, commonly called knobs, are visible by means of scanning or transmission electron microscopy. The biochemical mechanisms responsible for erythrocyte adherence to the endothelial cell are still undefined. In an attempt to identify the cytoadhesive molecule on the surface of the infected cell, we have prepared monoclonal antibodies to knob-bearing erythrocytes infected with the FCR-3 strain of P. falciparum. One of these monoclonal antibodies, designed 4A3, is an IgM that reacts (by means of immunofluorescence) with the surface of unfixed erythrocytes bearing mature parasites of the knobby line; it does not react with knobless lines or uninfected erythrocytes. By immunoelectron microscopy the monoclonal antibody 4A3 was localized to the knob region. In an in vitro cytoadherence assay, the monoclonal antibody partially blocked the binding of knob-bearing cells (FCR-3 strain) to formalin-fixed amelanotic melanoma cells. The monoclonal antibody was used to immunoprecipitate a protein from extracts of knobby erythrocytes that had been previously surface iodinated. By a two-dimensional peptide mapping technique, the antigen recognized by the monoclonal antibody was found to be structurally related to band 3 protein, the human erythrocyte anion transporter.

Monoclonal antibodies to the membrane domain of the human erythrocyte anion transport protein: localization of the C-terminus of the protein to the cytoplasmic side of the red cell membrane and distribution of the protein in some human tissues

Abstract: (1) We have prepared murine monoclonal antibodies to the membrane domain of the human erythrocyte anion transport protein (band 3). (2) All of these antibodies react with regions of the protein located at the cytoplasmic surface of the red cell. (3) One of the antibodies reacts with an epitope present on a cytoplasmic loop of the protein located between the C-terminus and a point 168 amino acids from the C-terminus. The other antibodies recognize different epitopes on the C-terminal tail of the protein and the sequences likely to be involved in these epitopes are defined. (4) Our results show that the C-terminus of the red-cell anion transport protein is located on the cytoplasmic side of the red-cell membrane. (5) None of the antibodies inhibited sulphate exchange transport when introduced into resealed red-cell membranes; however, the bivalent form of one of the antibodies reduced the inhibitory potency of 4-acetamido-49-isothiocyanatostilbene disulphonate on sulphate exchange transport in resealed erythrocyte membranes. (6) Immunostaining of human kidney sections with the antibodies showed strong staining of the basolateral membrane of some but not all of the epithelial cells of distal tubules and the initial connecting segment of collecting tubules. With human liver, only the haematopoeitic cells of fetal liver reacted with all the antibodies.

Role of chloride in potassium transport through a K-Cl cotransport system in human red blood cells

Abstract: In this paper, we report experiments demonstrating the coupling of Cl and K movements in a volume-dependent K-Cl cotransport system in human red blood cells. We show that an outwardly directed Cl gradient can promote net K efflux against an inwardly directed K gradient at constant membrane potential. Red blood cell membrane potential was kept constant by using anions that are not transported through the K-Cl cotransport system but that are more permeable than Cl (NO3 and SCN). Under these conditions, when the activities of band 3 (capnophorin)-mediated anion exchange and of the carbonic anhydrase have been inhibited, it is possible to maintain a Cl gradient at constant membrane potential. Similar data were obtained in human red blood cells (least dense fraction from normal subjects and whole blood from patients with homozygous hemoglobin S disease), in rabbit red blood cells, and in low-K sheep red blood cells. These data confirm that the volume-dependent Cl-dependent K movement in these cells operates through coupled K-Cl cotransport.