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. 2022 Feb 23;13:837926. doi: 10.3389/fphys.2022.837926

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Copyright © 2022 Maruyama, Hieda, Mawatari and Fujino.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Time-dependent derangement of erythrocytes exposed to 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH). Symbols and bars indicate mean ± SD (n = 6). (A) The filterability of erythrocytes exposed to 50 mM AAPH at 36°C showed time-dependent reduction (), which was marked at incubation time segment of 40–60 min after starting incubation, whereas the filterability of erythrocyte suspension without exposure to AAPH remained at the preincubation levels around 80% (◯). (B) Time-dependent changes of the mean corpuscular volume (MCV; fl) of erythrocytes with and without exposure to 50 mM AAPH. MCV is calculated automatically by hemocytometer as a surrogate of cell size. Erythrocytes exposed to 50 mM AAPH showed a sigmoidal increase in MCV (), which was evident 60 min after starting incubation at 36°C. Erythrocytes without exposure to AAPH (◯) showed no discernible changes in MCV. (C) Time-dependent increases in methemoglobin formation of erythrocytes (%) with and without exposure to 50 mM AAPH. Methemoglobin was assayed by standard spectrophotometric methods using absorbance differences of hemolysate in the presence and the absence of potassium cyanide and/or potassium ferricyanide at the wavelength of 630 nm. Erythrocytes exposed to AAPH showed a time-dependent sigmoidal increase in methemoglobin formation (), which was evident 60 min after starting incubation at 36°C. Erythrocytes without exposure to AAPH (◯) showed slight methemoglobin formation due to natural oxidation. (D) Hemolytic time course of erythrocyte suspension (%) exposed to 50 mM AAPH. Hemolysis was quantified by the absorbance of hemoglobin at 540 nm in the supernatant, and percent hemolysis (%) was calculated with comparison to complete hemolysis using distilled water. Erythrocytes exposed to AAPH () showed time-dependent hemolysis, which was evident 60 min after starting incubation at 36°C, whereas erythrocyte suspension without exposure to AAPH (◯) showed no evident hemolysis [cited from Odashiro et al. (2014) with permission].

Inline graphic — Time-dependent derangement of erythrocytes exposed to 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH). Symbols and bars indicate mean ± SD (n = 6). (A) The filterability of erythrocytes exposed to 50 mM AAPH at 36°C showed time-dependent reduction (), which was marked at incubation time segment of 40–60 min after starting incubation, whereas the filterability of erythrocyte suspension without exposure to AAPH remained at the preincubation levels around 80% (◯). (B) Time-dependent changes of the mean corpuscular volume (MCV; fl) of erythrocytes with and without exposure to 50 mM AAPH. MCV is calculated automatically by hemocytometer as a surrogate of cell size. Erythrocytes exposed to 50 mM AAPH showed a sigmoidal increase in MCV (), which was evident 60 min after starting incubation at 36°C. Erythrocytes without exposure to AAPH (◯) showed no discernible changes in MCV. (C) Time-dependent increases in methemoglobin formation of erythrocytes (%) with and without exposure to 50 mM AAPH. Methemoglobin was assayed by standard spectrophotometric methods using absorbance differences of hemolysate in the presence and the absence of potassium cyanide and/or potassium ferricyanide at the wavelength of 630 nm. Erythrocytes exposed to AAPH showed a time-dependent sigmoidal increase in methemoglobin formation (), which was evident 60 min after starting incubation at 36°C. Erythrocytes without exposure to AAPH (◯) showed slight methemoglobin formation due to natural oxidation. (D) Hemolytic time course of erythrocyte suspension (%) exposed to 50 mM AAPH. Hemolysis was quantified by the absorbance of hemoglobin at 540 nm in the supernatant, and percent hemolysis (%) was calculated with comparison to complete hemolysis using distilled water. Erythrocytes exposed to AAPH () showed time-dependent hemolysis, which was evident 60 min after starting incubation at 36°C, whereas erythrocyte suspension without exposure to AAPH (◯) showed no evident hemolysis [cited from Odashiro et al. (2014) with permission].