Factors Limiting O2 Transfer of Red Blood Cells: Model Analysis Using Results from Stopped-Flow Experiments

2015 ◽  
pp. 11-17 ◽  
Author(s):  
P. Scheid ◽  
C. Hook ◽  
K. Yamaguchi ◽  
J. Piiper
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Model Analysis ◽  
Stopped Flow ◽  
Flow Experiments ◽  
O2 Transfer

Oxygen transfer of red blood cells: experimental data and model analysis

1988 ◽  
Vol 72 (1) ◽  
pp. 65-82 ◽  
Author(s):  
Christian Hook ◽  
Kazuhiro Yamaguchi ◽  
Peter Scheid ◽  
Johannes Piiper
Keyword(s):  
Experimental Data ◽  
Red Blood Cells ◽  
Oxygen Transfer ◽  
Blood Cells ◽  
Model Analysis

Cat carotid body: oxygen consumption and other parameters

1981 ◽  
Vol 50 (1) ◽  
pp. 129-133 ◽  
Author(s):  
W. J. Whalen ◽  
P. Nair ◽  
T. Sidebotham ◽  
J. Spande ◽  
M. Lacerna
Keyword(s):  
Oxygen Consumption ◽  
Red Blood Cells ◽  
Carotid Body ◽  
Blood Cells ◽  
Blood Perfusion ◽  
Stopped Flow ◽  
Solution Flow ◽  
Consumption Rates ◽  
Disappearance Curve ◽  
Nerve Discharge

Cats were anesthetized with pentobarbital, pump ventilated with air, and given heparin, and the carotid body (CB) was vascularly isolated except for the supplying artery. The CB could be normally blood perfused, or alternatively, perfused with Locke's solution; flow of either could be stopped suddenly. Sinus nerve discharge was measured. Tissue oxygen tension (TPO2) in the CB was measured with an O2 microelectrode. Oxygen consumption rates (VO2) calculated from the disappearance curve of O2 during stopped flow were PO2 dependent. When TPO2 was high (100-130 Torr), VO2 (ml.100 g-1.min-1) averaged 1.9 +/- 0.18 (SE) during blood perfusion and either 1.9 +/- 0.1 during perfusion with Locke's solution equilibrated with 25% O2-5% CO2-70% N2 or 1.4 +/- 0.08 when the Locke's solution was equilibrated with air. This significant effect of CO2 could have been due to the delayed onset of sinus nerve discharge when CO2 was not added to the perfusion solution. The number of red blood cells in histological sections from CB frozen during stopped flow of blood was significantly below normal. We concluded that the similarity of the disappearance curves during stopped flow of blood and Locke's solutions was primarily due to the extrusion of red blood cells. In five experiments the broken-off tip of the O2 microelectrode was found in the core of the CB.

scholarly journals Rapid Cl−/HCO3−exchange kinetics of AE1 in HEK293 cells and hereditary stomatocytosis red blood cells

2013 ◽  
Vol 305 (6) ◽  
pp. C654-C662 ◽  
Author(s):  
Etienne Frumence ◽  
Sandrine Genetet ◽  
Pierre Ripoche ◽  
Achille Iolascon ◽  
Immacolata Andolfo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cell ◽  
Red Blood Cells ◽  
Anion Exchange ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Hek293 Cells ◽  
Single Amino Acid ◽  
Stopped Flow ◽  
Wild Type

Anion exchanger 1 (AE1) or band 3 is a membrane protein responsible for the rapid exchange of chloride for bicarbonate across the red blood cell membrane. Nine mutations leading to single amino-acid substitutions in the transmembrane domain of AE1 are associated with dominant hereditary stomatocytosis, monovalent cation leaks, and reduced anion exchange activity. We set up a stopped-flow spectrofluorometry assay coupled with flow cytometry to investigate the anion transport and membrane expression characteristics of wild-type recombinant AE1 in HEK293 cells, using an inducible expression system. Likewise, study of three stomatocytosis-associated mutations (R730C, E758K, and G796R), allowed the validation of our method. Measurement of the rapid and specific chloride/bicarbonate exchange by surface expressed AE1 showed that E758K mutant was fully active compared with wild-type (WT) AE1, whereas R730C and G796R mutants were inactive, reinforcing previously reported data on other experimental models. Stopped-flow analysis of AE1 transport activity in red blood cell ghost preparations revealed a 50% reduction of G796R compared with WT AE1 corresponding to a loss of function of the G796R mutated protein, in accordance with the heterozygous status of the AE1 variant patients. In conclusion, stopped-flow led to measurement of rapid transport kinetics using the natural substrate for AE1 and, conjugated with flow cytometry, allowed a reliable correlation of chloride/bicarbonate exchange to surface expression of AE1, both in recombinant cells and ghosts and therefore a fine comparison of function between different stomatocytosis samples. This technical approach thus provides significant improvements in anion exchange analysis in red blood cells.

scholarly journals Stopped-flow kinetics of oxygen uptake and release by embryonic red blood cells

1989 ◽  
Vol 261 (3) ◽  
pp. 1009-1013 ◽  
Author(s):  
T Brittain ◽  
R Simpson
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Stopped Flow ◽  
Embryonic Cells ◽  
O2 Uptake ◽  
O2 Concentration ◽  
Time Courses ◽  
Early Mouse ◽  
Kinetics Of ◽  
Uptake And Release

The processes of O2 uptake and release by the three embryonic haemoglobins contained within early mouse embryonic red blood cells have been studied using dual-wavelength stopped-flow kinetic spectroscopy. The rate of O2 uptake in the pseudo-spherical, nucleated, embryonic red blood cells exhibits a greater than first-order dependence on O2 concentration. The time courses for the release from the red blood cells into dithionite-containing solutions tends towards a limiting rate at high dithionite concentrations. The rates of both the uptake and release processes observed in the embryonic cells are compared with those previously seen for adult mouse red blood cells. A new mathematical model is described which accurately simulates both uptake and release experimental data for the nucleated embryonic red blood cells.

scholarly journals A Stopped Flow System with Hydrodynamic Injection for Red Blood Cells Osmotic Fragility Test: Possibility for Automatic Screening of Beta-thalassemia Trait

2009 ◽  
Vol 25 (6) ◽  
pp. 819-824 ◽  
Author(s):  
Supada KHONYOUNG ◽  
Supaporn KRADTAP HARTWELL ◽  
Jaroon JAKMUNEE ◽  
Somchai LAPANANTNOPPAKHUN ◽  
Torpong SANGUANSERMSRI ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Flow System ◽  
Osmotic Fragility ◽  
Beta Thalassemia ◽  
Stopped Flow ◽  
Hydrodynamic Injection ◽  
Thalassemia Trait

Hemoglobin crystals of the red blood cells in the human renal cortex: electron microscopic observations of the renal lithiasis

1978 ◽  
Vol 36 (2) ◽  
pp. 480-481
Author(s):  
Kosuke Ueda ◽  
Hiroto Washida ◽  
Nakazo Watari
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Renal Cortex ◽  
Uranyl Acetate ◽  
Osmic Acid ◽  
Renal Lithiasis ◽  
Electron Microscopic ◽  
Hemoglobin C ◽  
First Case ◽  
Ultrathin Sections

IntroductionHemoglobin crystals in the red blood cells were electronmicroscopically reported by Fawcett in the cat myocardium. In the human, Lessin revealed crystal-containing cells in the periphral blood of hemoglobin C disease patients. We found the hemoglobin crystals and its agglutination in the erythrocytes in the renal cortex of the human renal lithiasis, and these patients had no hematological abnormalities or other diseases out of the renal lithiasis. Hemoglobin crystals in the human erythrocytes were confirmed to be the first case in the kidney.Material and MethodsTen cases of the human renal biopsies were performed on the operations of the seven pyelolithotomies and three ureterolithotomies. The each specimens were primarily fixed in cacodylate buffered 3. 0% glutaraldehyde and post fixed in osmic acid, dehydrated in graded concentrations of ethanol, and then embedded in Epon 812. Ultrathin sections, cut on LKB microtome, were doubly stained with uranyl acetate and lead citrate.

Densitometric Identification of Primitive Erythroblasts After Reaction With Diaminobenzidine

1973 ◽  
Vol 31 ◽  
pp. 328-329
Author(s):  
John A. Trotter
Keyword(s):  
Red Blood Cells ◽  
Analytical Method ◽  
Blood Cells ◽  
Guinea Pigs ◽  
Specific Protein ◽  
Visual Examination ◽  
Hemoglobin Synthesis ◽  
Peroxidatic Activity ◽  
Small Density

Hemoglobin is the specific protein of red blood cells. Those cells in which hemoglobin synthesis is initiated are the earliest cells that can presently be considered to be committed to erythropoiesis. In order to identify such early cells electron microscopically, we have made use of the peroxidatic activity of hemoglobin by reacting the marrow of erythropoietically stimulated guinea pigs with diaminobenzidine (DAB). The reaction product appeared as a diffuse and amorphous electron opacity throughout the cytoplasm of reactive cells. The detection of small density increases of such a diffuse nature required an analytical method more sensitive and reliable than the visual examination of micrographs. A procedure was therefore devised for the evaluation of micrographs (negatives) with a densitometer (Weston Photographic Analyzer).

Scanning electron microscopy of erythrophagocytosis by Entamoeba histolytica trophozoites

1992 ◽  
Vol 50 (1) ◽  
pp. 880-881
Author(s):  
Victor Tsutsumi ◽  
Adolfo Martinez-Palomo ◽  
Kyuichi Tanikawa
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Red Blood Cells ◽  
Entamoeba Histolytica ◽  
Causative Agent ◽  
Blood Cells ◽  
Protozoan Parasite ◽  
Complex Phenomenon ◽  
Great Capacity ◽  
Scanning Electron

The protozoan parasite Entamoeba histolytica is the causative agent of amebiasis in man. The trophozoite or motile form is a highly dynamic and pleomorphic cell with a great capacity to destroy tissues. Moreover, the parasite has the singular ability to phagocytize a variety of different live or death cells. Phagocytosis of red blood cells by E. histolytica trophozoites is a complex phenomenon related with amebic pathogenicity and nutrition.

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