Kinetics of O2 uptake and release by human erythrocytes studied by a stopped-flow technique

1985 ◽  
Vol 58 (4) ◽  
pp. 1215-1224 ◽  
Author(s):  
K. Yamaguchi ◽  
D. Nguyen-Phu ◽  
P. Scheid ◽  
J. Piiper

The kinetics of O2 uptake into and release from human erythrocytes was investigated at 37 degrees C by a stopped-flow technique. From the time course of O2 saturation (SO2) change a specific transfer conductance of erythrocytes for O2 (GO2) was calculated. The following results were obtained: 1) GO2 decreased in the course of O2 uptake, but initial GO2 was nearly independent of SO2 at which uptake started; 2) addition of albumin to the medium reduced GO2; 3) increasing dithionite concentration in the medium in O2-release experiments progressively enhanced GO2, which became virtually constant for nearly the entire course of release; and 4) O2 uptake and O2 release (without dithoite) in the same SO2 range yielded very similar GO2. These results suggested that O2 uptake and release were importantly limited by diffusion through the external medium and that in the SO2 range between 0.3 and 0.8, chemical reaction exerted little limiting effect. Since O2 release at the highest dithionite concentration (40 mmol/l) appeared to be virtually unlimited by external diffusion, GO2 measured under these conditions, averaging 8.7 ml X min-1 X Torr-1 X ml erythrocytes-1, was considered to mainly reflect intracellular diffusion limitation. The corresponding specific transfer conductance for O2 transfer in whole blood (hematocrit, 0.45) is 3.9 ml X min-1 X Torr-1 X ml blood-1.

1989 ◽  
Vol 261 (3) ◽  
pp. 1009-1013 ◽  
Author(s):  
T Brittain ◽  
R Simpson

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.


1989 ◽  
Vol 260 (1) ◽  
pp. 171-176 ◽  
Author(s):  
T Brittain ◽  
R Simpson

Ligand uptake and release by the haemoglobin contained within adult mouse erythrocytes was studied by using dual-wavelength stopped-flow techniques. The rate of O2 uptake is very much lower than that expected for an equivalent concentration of haemoglobin in free solution. The O2-concentration-dependence found in uptake experiments is greater than first-order. CO uptake shows the same pattern of reactivity as does O2, but the associated rates of uptake are lower and the concentration-dependence of the CO rates is first-order. O2 release from the adult erythrocytes was measured by stopped-flow mixing with Na2S2O4. Under these circumstances the deoxygenation of intracellular haemoglobin shows accelerating time courses. The apparent rate-constant-dependence on dithionite concentration shows a rate limit at high reductant concentrations. Computer simulations of both ligand uptake and release processes were carried out by using a three-dimensional model. The simulations clearly indicate that in rapid-mixing experiments the rather slow experimentally observed O2 uptake rate is due to rate-limiting diffusion through an extracellular stagnant solvent layer. In the case of O2 release, however, the major rate-controlling process is the rate of O2 dissociation from the haemoglobin molecules, which accelerates during the deoxygenation process.


1976 ◽  
Vol 3 (2) ◽  
pp. 141 ◽  
Author(s):  
WS Chow ◽  
AB Hope

The fluorescence quenching of certain fluorescent, weakly ionizing amines has been used to estimate the pH difference (ΔpH) between the intrathylakoid spaces of chloroplasts and the external medium under various conditions. The amines N-(1-naphthyl)ethylenediamine and 9-aminoacridine gave consistent and similar estimates of ΔpH provided the concentration ranges 0.25 - 2 μM and 0.05 - 0.5 μM, respectively, were not exceeded. Values for ΔpH of up to 4 units were estimated for control chloroplasts. Only a small decrease in ΔpH was caused by FCCP and DCMU concentrations that decreased photophosphorylation rates to 10-20% of control. The kinetics of the onset and decay of ΔpH, and of the uptake and release of protons by the chloroplasts, were followed after the sudden application or withdrawal of actinic light. These kinetics suggest a model for the ionic relations of stripped chloroplasts with internal buffering capacity, [α], and dissociation constant, Ka, as parameters. At pH8, [α] is about 80 mol m-3 and Ka is 0.07 mol m-3 (pKa = 4.15). The model is successful in predicting the relation between [H+]I (the internal proton concentration) and ΔH+o (the light-driven uptake of protons from the external medium), as well as the size of ΔH+o , but needs extending to deal with published experiments on the light-driven translocation of chloride ions.


1988 ◽  
Vol 64 (1) ◽  
pp. 451-459 ◽  
Author(s):  
R. L. Hughson ◽  
D. L. Sherrill ◽  
G. D. Swanson

The constancy of the time course (i.e., dynamic linearity) of the O2 uptake (VO2) response to exercise was examined by testing the law of superposition on data from impulse and step work rate forcings. Two impulses (10 s at a 235-W increase above a 25-W base line, I-235; and 5 s at a 475-W increase above a 25-W base line, I-475), four steps (ST) (25-65 W, ST1; 65-105 W, ST2; 25-105 W, ST3; and 25-145 W, ST4), and the corresponding off-transient responses were performed six to eight times by each of five subjects. The integrated area (G) of the VO2 response for I-235 was similar to that of ST1 and ST2 (P greater than 0.05); the I-475 G was significantly greater (P less than 0.05). The time constant of VO2 during the step function on-transient response for the second exponential component was significantly faster for ST1 and significantly slower for I-235 and I-475 than for ST2, ST3, and ST4 (P less than 0.05). However, I-235 and I-475 time constants for VO2 were not different from the ST off-transient values. Attempts to superimpose the integral of the impulse on the ST data showed that the early rapid increase in VO2 in the ST was underpredicted by the impulse and that the impulse response lagged behind the ST at all points before steady state. It can be concluded that VO2 kinetics failed the test of superposition and are therefore described by a nonlinear dynamic system.


1998 ◽  
Vol 37 (03) ◽  
pp. 107-112 ◽  
Author(s):  
I. Lauer ◽  
M. Bähre ◽  
E. Richter ◽  
B. Melier

Summary Aim: In 214 patients with benign thyroid diseases the time-course of urinary iodine excretion (UIE) was investigated in order to identify changes after radioiodine therapy (RITh). Method: UIE was measured photometrically (cerium-arsenite method) and related to urinary creatinine on the first and last day of the radioiodine test and then three days, seven days, four weeks, and six months after 1311 administration. Results: As compared with the level found immediately before radioiodine therapy, median UIE had almost doubled four weeks after therapy and was still significantly elevated six months after therapy. This increase correlated significantly with the target volume as measured by scintigraphy and sonography. Conclusions: The persistent elevation of UIE for months after RITh is a measure of treatment-induced damage to thyrocytes. Therefore, in view of the unfavourable kinetics of iodine that follow it, RITh should if possible be given via a single-dose regime.


1984 ◽  
Vol 160 (3) ◽  
pp. 877-892 ◽  
Author(s):  
G Lamson ◽  
M E Koshland

The time course of differentiative events in the pentamer IgM response was examined by following the expression of J chain and mu chain RNA and their protein products in mitogen-stimulated lymphocytes. The analyses showed that the shift to mus RNA synthesis begins shortly after stimulation and precedes proliferation of the cells and any increase in mu RNA levels. In contrast, expression of J chain RNA and the amplification of J chain and mus message are late events that coincide with a phase of rapid proliferation and with the secretion of pentamer IgM antibody. The kinetics of J and mu chain RNA expression observed in normal lymphocytes were supported by analyses of lymphoid cell lines. B lymphomas were found to display the RNA pattern characteristic of early-activated lymphocytes, i.e., a partial shift to mus RNA production and no J chain RNA, whereas IgM-secreting lines resembled late-activated lymphocytes in their expression of high levels of both mus and J chain mRNA. Moreover, the kinetics of J and mus chain RNA expression correlates with the sequential action of B cell lymphokines in the induction of the pentamer IgM response. This correlation suggests that the successive differentiative changes are triggered by successive membrane stimuli.


2000 ◽  
Vol 29 (7) ◽  
pp. 487-498 ◽  
Author(s):  
E. Błachut-Okrasińska ◽  
E. Bojarska ◽  
A. Niedźwiecka ◽  
L. Chlebicka ◽  
E. Darżynkiewicz ◽  
...  

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