THE IN VITRO METABOLISM OF ERYTHROCYTES FROM NEWBORN INFANTS

1960 ◽  
Vol 38 (1) ◽  
pp. 727-738 ◽  
Author(s):  
A. Zipursky ◽  
T. LaRue ◽  
L. G. Israels
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Membrane Permeability ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Newborn Infants ◽  
Phosphate Esters ◽  
C Storage ◽  
Rapid Fall

In the red blood cells of the newborn there is a rapid fall in non-hydrolyzable phosphate during in vitro incubation. This difference appears to be due to a decreased rate of synthesis of 2,3-diphosphoglyceric acid in the erythrocyte of the newborn. In addition the incorporation of P32 orthophosphate into the red blood cell is slower in the newborn than in the adult. During 4 °C storage of blood of adults and newborns there is a progressive fall in phosphate esters which is similar in both groups.The erythrocytes of the newborn contain more potassium and water than those of adults. During storage at 4 °C the cells of the newborn lose potassium more rapidly than those of the adult. This may be related to differences in membrane permeability.

THE IN VITRO METABOLISM OF ERYTHROCYTES FROM NEWBORN INFANTS

1960 ◽  
Vol 38 (7) ◽  
pp. 727-738 ◽  
Author(s):  
A. Zipursky ◽  
T. LaRue ◽  
L. G. Israels
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Membrane Permeability ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Newborn Infants ◽  
Phosphate Esters ◽  
C Storage ◽  
Rapid Fall

In the red blood cells of the newborn there is a rapid fall in non-hydrolyzable phosphate during in vitro incubation. This difference appears to be due to a decreased rate of synthesis of 2,3-diphosphoglyceric acid in the erythrocyte of the newborn. In addition the incorporation of P32 orthophosphate into the red blood cell is slower in the newborn than in the adult. During 4 °C storage of blood of adults and newborns there is a progressive fall in phosphate esters which is similar in both groups.The erythrocytes of the newborn contain more potassium and water than those of adults. During storage at 4 °C the cells of the newborn lose potassium more rapidly than those of the adult. This may be related to differences in membrane permeability.

scholarly journals Vesiculation of Red Blood Cells in the Blood Bank: A Multi-Omics Approach towards Identification of Causes and Consequences

Proteomes ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 6
Author(s):  
Joames K. Freitas Leal ◽  
Edwin Lasonder ◽  
Vikram Sharma ◽  
Jürgen Schiller ◽  
Giuseppina Fanelli ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Blood Bank ◽  
Cell Aging ◽  
Lipid Organization ◽  
Membrane Components

Microvesicle generation is an integral part of the aging process of red blood cells in vivo and in vitro. Extensive vesiculation impairs function and survival of red blood cells after transfusion, and microvesicles contribute to transfusion reactions. The triggers and mechanisms of microvesicle generation are largely unknown. In this study, we combined morphological, immunochemical, proteomic, lipidomic, and metabolomic analyses to obtain an integrated understanding of the mechanisms underlying microvesicle generation during the storage of red blood cell concentrates. Our data indicate that changes in membrane organization, triggered by altered protein conformation, constitute the main mechanism of vesiculation, and precede changes in lipid organization. The resulting selective accumulation of membrane components in microvesicles is accompanied by the recruitment of plasma proteins involved in inflammation and coagulation. Our data may serve as a basis for further dissection of the fundamental mechanisms of red blood cell aging and vesiculation, for identifying the cause-effect relationship between blood bank storage and transfusion complications, and for assessing the role of microvesicles in pathologies affecting red blood cells.

scholarly journals IN VITRO INTERACTIONS BETWEEN OXYGEN AND CARBON DIOXIDE TRANSPORT IN THE BLOOD OF THE SEA LAMPREY (PETROMYZON MARINUS)

1992 ◽  
Vol 173 (1) ◽  
pp. 25-41 ◽  
Author(s):  
R. A. Ferguson ◽  
N. Sehdev ◽  
B. Bagatto ◽  
B. L. Tufts
Keyword(s):  
Carbon Dioxide ◽  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Blood Cells ◽  
Sea Lamprey ◽  
Carbon Dioxide Transport ◽  
Dissociation Curves

In vitro experiments were carried out to examine the interactions between oxygen and carbon dioxide transport in the blood of the sea lamprey. Oxygen dissociation curves for whole blood obtained from quiescent lampreys had Hill numbers (nH) ranging from 1.52 to 1.89. The Bohr coefficient for whole blood was -0.17 when extracellular pH (pHe) was considered, but was much greater (-0.63) when red blood cell pH (pHi) was considered. The pHi was largely dependent on haemoglobin oxygen- saturation (SO2) and the pH gradient across the red blood cell membrane was often reversed when PCO2 was increased and/or SO2 was lowered. The magnitude of the increase in pHi associated with the Haldane effect ranged from 0.169 pH units at 2.9 kPa PCO2 to 0.453 pH units at a PCO2 of 0.2 kPa. Deoxygenated red blood cells had a much greater total CO2 concentration (CCO2) than oxygenated red blood cells, but the nonbicarbonate buffer value for the red blood cells was unaffected by oxygenation. Plasma CCO2 was not significantly different under oxygenated or deoxygenated conditions. Partitioning of CO2 carriage in oxygenated and deoxygenated blood supports recent in vivo observations that red blood cell CO2 carriage can account for much of the CCO2 difference between arterial and venous blood. Together, the results also suggest that oxygen and carbon dioxide transport may not be tightly coupled in the blood of these primitive vertebrates. Finally, red cell sodium concentrations were dependent on oxygen and carbon dioxide tensions in the blood, suggesting that sodium-dependent ion transport processes may contribute to the unique strategy for gas transport in sea lamprey blood.

In vitro analysis of volume and pH regulation in the red blood cells of a primitive air-breathing fish, the bowfin, Amia calva

1994 ◽  
Vol 72 (2) ◽  
pp. 280-286 ◽  
Author(s):  
B. L. Tufts ◽  
R. C. Drever ◽  
B. Bagatto ◽  
B. A. Cameron
Keyword(s):  
Rainbow Trout ◽  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Disulfonic Acid ◽  
Osmotic Swelling ◽  
Ph Range ◽  
Amia Calva

In the bowfin (Amia calva), a decrease in extracellular pH in vitro was associated with an increase in the water content and chloride concentration in the red blood cells that could be inhibited by the anion-exchange blocker, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS). After a step increase in CO2 tension, the extracellular total CO2 concentration was also significantly reduced by DIDS. Finally, over most of the experimental pH range, the red blood cell pH observed in the presence of DIDS was significantly elevated compared with that of controls. Taken together, these results indicate that as in most other fishes, chloride–bicarbonate exchange is clearly present and functional in bowfin red blood cells. Moreover, within the physiological pH range, ion movements across the anion exchanger have a marked influence on both the volume and the pH of bowfin red blood cells. In sharp contrast to the rainbow trout (Oncorhynchus mykiss), catecholamines had no effect on the volume, pH, or intracellular sodium concentration of red blood cells in the bowfin. Following osmotic swelling, rainbow trout red blood cells were able to regulate their volume back to control levels within 2 h. In the bowfin, however, there was no regulation of red blood cell volume after osmotic swelling. Thus, in contrast to many other fishes examined to date, it would appear that in the bowfin, the physiological mechanisms involved in the adrenergic response and in the regulatory volume decrease after osmotic swelling may be less active or possibly even absent in the red blood cells.

Lactate interferes with ATP release from red blood cells

2007 ◽  
Vol 292 (6) ◽  
pp. H3038-H3042 ◽  
Author(s):  
Michael D. Rozier ◽  
Vincent J. Zata ◽  
Mary L. Ellsworth
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Oxygen Supply ◽  
Atp Release ◽  
Serum Lactate ◽  
Atp Production ◽  
Lactate Levels

Upon exposure to low Po2, the red blood cells of most species, including humans, release increased amounts of ATP that ultimately serves as a regulator of vascular tone matching oxygen supply with demand. In pathological conditions such as malaria and sepsis, a maldistribution of perfusion exists with its severity often correlated with the extent of elevation of serum lactate frequently in the absence of an alteration in pH. We hypothesized that the increased levels of lactate might impair the ability of red blood cells to appropriately respond to conditions of low Po2, thus preventing its important blood flow regulatory role. Using an in vitro system and rabbit red blood cells, we evaluated the capacity of cells incubated with lactate to release increased amounts of ATP in response to acute exposure to low Po2. We found that in the presence of lactate, the red blood cells did not release ATP. However, when sodium dichloroacetate, a drug used clinically to lower blood lactate levels, was added, ATP release was restored to levels that were not different from that of control cells (no lactate), even though intracellular levels of ATP were not. These results support the presence of a distinct flow regulatory pool of ATP within the red blood cell that can be independently regulated, and that lactate interferes with the ATP production within this pool, thereby diminishing the amount of ATP available for release on exposure to low Po2. Therefore, if lactate levels can be reduced, the vascular regulatory capacity of the red blood cell should be restored, thus enabling the appropriate matching of oxygen supply with oxygen demand.

scholarly journals Quantitation of immunoglobulin associated with senescent erythrocytes from the rabbit

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1096-1099 ◽  
Author(s):  
GL Dale ◽  
RB Daniels
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Antibody Binding ◽  
Biochemical Processes ◽  
Senescent Erythrocytes

The biochemical processes that determine the lifespan of mammalian erythrocytes are unknown; one prominent theory suggests that antibody binding to the senescent red blood cell identifies it for removal from the circulation. To address this question, we have used a newly developed procedure for the isolation of aged erythrocytes that involves the biotinylation of rabbit red blood cells, in vivo aging of these cells, and the eventual in vitro recovery of the aged red blood cells by their affinity for an avidin support. Erythrocytes isolated with this method were found to have near-normal levels of cell- associated Ig throughout their 60-day lifespan. These data suggest that IgG accumulation is not part of the normal senescence process for erythrocytes in rabbits.

scholarly journals Validation of Daraex to Resolve Daratumumab-Induced Interferences in Pre-Transfusion Screen Tests

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 4983-4983
Author(s):  
Maria Tenorio ◽  
Gemma Moreno Jiménez ◽  
Valentín García Gutiérrez ◽  
Ana Jiménez ◽  
Maria Jesús Blanchard ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Research Funding ◽  
Screen Test ◽  
New Method ◽  
Negative Controls

Daratumumab is a CD38-directed antibody increasingly used for the treatment of adult patients with multiple mieloma. The membrane of red blood cells express CD38 and thus samples from patients treated with daratumumab show agglutination in red blood cell antibody screen tests performed prior to transfusion. This interference hinders the detection of red blood cell alloantibodies. Published literature has described a method to eliminate CD38 in red blood cells with DTT (Chapuy, 2016). This technique is cumbersome, requires positive and negative controls as DTT destroys Kell antigens and can produce in vitro hemolysis. The increasing number of multiple myeloma patients treated with daratumumab poses the need for a simple and straightforward technique with applicability in standard transfusion centers. DaraEx (Inno-Train) is a new anti-CD38 neutralizing agent that overcomes daratumumab-induced interferences detected in pre-transfusion tests without the major drawbacks associated with the DTT technique. Our aim was to validate and implement DaraEx as the method of choice to solve daratumumab interferences detected in pre-transfusion screen tests in a tertiary care center. A two-step approach using in vitro and in vivo samples was designed to validate the new method. First, we compared DaraEx efficacy in vitro to the reference DTT method in two samples spiked with daratumumab to achieve a concentration of 10mg/mL (Sample A: serum from a patient without known red blood cell alloantibodies; Sample B: serum from a patient with alloantibody anti-c). Red blood cells in the screen test (3 red blood cell screen; ID-DiaCell I-II-III) as well as positive (E+ red blood cells) and negative controls (K+ red blood cells) were treated with DTT 0.2M solution for 30 minutes at 37ºC and then washed four times with saline. In parallel, red blood cells in the screen test were incubated during 30 minutes at room temperature in a shaker (600rpm) with DaraEx. Red blood cells treated with each of these methods were used for indirect antiglobulin test with our gel card system (BioRad; IH-1000). Preference of method in terms of time needed and result interpretation was evaluated by three hematologists specialized in blood banking and four different technicians. Secondly, we tested pre-transfusion samples from patients treated with daratumumab with the DaraEx technique to check in vivo efficacy. There was a 100% concordance between both techniques (DDT reference method and DaraEx new method) in both in vitro samples. All hematologists and technicians found the DaraEx technique less cumbersome in terms of processing and time to result (2 hours with DTT versus 1 hour with DaraEx) and the interpretation straightforward. Twelve samples with daratumumab-induced interference in pre-transfusion screen tests belonging to 5 patients were tested between January and July 2019. All the interferences detected resolved with DaraEx regardless of time from last daratumumab administration (range: 7-145 days; mean: 57 days). Figure 1 shows screen test with and without treatment with DaraEx in a patient sample. In our experience, DaraEx technique is a simple, fast and efficacious method, regardless of time from last daratumumab administration, to resolve interferences secondary to daratumumab administration without the major disadvantages associated with DTT. Figure 1 Disclosures García Gutiérrez: Pfizer: Honoraria, Research Funding; Incyte: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; BMS: Honoraria, Research Funding.

scholarly journals Quantitation of immunoglobulin associated with senescent erythrocytes from the rabbit

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1096-1099 ◽  
Author(s):  
GL Dale ◽  
RB Daniels
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Antibody Binding ◽  
Biochemical Processes ◽  
Senescent Erythrocytes

Abstract The biochemical processes that determine the lifespan of mammalian erythrocytes are unknown; one prominent theory suggests that antibody binding to the senescent red blood cell identifies it for removal from the circulation. To address this question, we have used a newly developed procedure for the isolation of aged erythrocytes that involves the biotinylation of rabbit red blood cells, in vivo aging of these cells, and the eventual in vitro recovery of the aged red blood cells by their affinity for an avidin support. Erythrocytes isolated with this method were found to have near-normal levels of cell- associated Ig throughout their 60-day lifespan. These data suggest that IgG accumulation is not part of the normal senescence process for erythrocytes in rabbits.

scholarly journals Molecular Characterization and Immunological Evaluation of Truncated Babesia microti Rhoptry Neck Protein 2 as a Vaccine Candidate

2021 ◽  
Vol 12 ◽  
Author(s):  
Yu chun Cai ◽  
Chun li Yang ◽  
Wei Hu ◽  
Peng Song ◽  
Bin Xu ◽  
...  
Keyword(s):  
Immune Response ◽  
Blood Cell ◽  
Western Blot ◽  
Red Blood Cells ◽  
Cell Invasion ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Babesia Microti ◽  
Rhoptry Neck Protein

Babesia microtiis a protozoan that infects red blood cells. Babesiosis is becoming a new global threat impacting human health. Rhoptry neck proteins (RONs) are proteins located at the neck of the rhoptry and studies indicate that these proteins play an important role in the process of red blood cell invasion. In the present study, we report on the bioinformatic analysis, cloning, and recombinant gene expression of two truncated rhoptry neck proteins 2 (BmRON2), as well as their potential for incorporation in a candidate vaccine for babesiosis. Western blot and immunofluorescence antibody (IFA) assays were performed to detect the presence of specific antibodies against BmRON2 in infected mice and the localization of N-BmRON2 inB. microtiparasites.In vitroexperiments were carried out to investigate the role of BmRON2 proteins during theB. microtiinvasion process andin vivoexperiments to investigate immunoprotection. Homologous sequence alignment and molecular phylogenetic analysis indicated that BmRON2 showed similarities with RON2 proteins of otherBabesiaspecies. We expressed the truncated N-terminal (33–336 aa, designated rN-BmRON2) and C-terminal (915–1171 aa, designated rC-BmRON2) fragments of the BmRON2 protein, with molecular weights of 70 and 29 kDa, respectively. Western blot assays showed that the native BmRON2 protein is approximately 170 kDa, and that rN-BmRON2 was recognized by serum of mice experimentally infected withB. microti.Immunofluorescence analysis indicated that the BmRON2 protein was located at the apical end of merozoites, at the opposite end of the nucleus.In vitrored blood cell invasion inhibition studies withB. microtirBmRON2 proteins showed that relative invasion rate of rN-BmRON2 and rC-BmRON2 group is 45 and 56%, respectively. Analysis of the host immune response after immunization andB. microtiinfection showed that both rN-BmRON2 and rC-BmRON2 enhanced the immune response, but that rN-BmRON2 conferred better protection than rC–BmRON2. In conclusion, our results indicate that truncated rhoptry neck protein 2, especially its N-terminal fragment (rN-BmRON2), plays an important role in the invasion of host red blood cells, confers immune protection, and shows good potential as a candidate vaccine against babesiosis.

Effect of aggregation and shear rate on the dispersion of red blood cells flowing in venules

2002 ◽  
Vol 283 (5) ◽  
pp. H1985-H1996 ◽  
Author(s):  
Jeffrey J. Bishop ◽  
Aleksander S. Popel ◽  
Marcos Intaglietta ◽  
Paul C. Johnson
Keyword(s):  
Shear Rate ◽  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Cell Aggregation ◽  
Radial Position ◽  
Red Blood Cell Aggregation ◽  
Blood Cell Aggregation

Previous in vitro studies of blood flow in small glass tubes have shown that red blood cells exhibit significant erratic deviations in the radial position in the laminar flow regime. The purpose of the present study was to assess the magnitude of this variability and that of velocity in vivo and the effect of red blood cell aggregation and shear rate upon them. With the use of a gated image intensifier and fluorescently labeled red blood cells in tracer quantities, we obtained multiple measurements of red blood cell radial and longitudinal positions at time intervals as short as 5 ms within single venous microvessels (diameter range 45–75 μm) of the rat spinotrapezius muscle. For nonaggregating red blood cells in the velocity range of 0.3–14 mm/s, the mean coefficient of variation of velocity was 16.9 ± 10.5% and the SD of the radial position was 1.98 ± 0.98 μm. Both quantities were inversely related to shear rate, and the former was significantly lowered on induction of red blood cell aggregation by the addition of Dextran 500 to the blood. The shear-induced random movements observed in this study may increase the radial transport of particles and solutes within the bloodstream by orders of magnitude.

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