Figure 1.
Elongation index (EI)-shear stress (SS) curves for RBC in suspending medium containing the four alcohols at different concentrations.
(A) Methanol. (B) Ethanol. (C) Propanol. (D) Butanol. The data are means ± SE from 10 experiments on blood samples from different donors. Note that error bars often lie within the symbols and therefore are not visible. The curves representing the EI-SS relations for RBC suspensions containing different concentrations of alcohols were significantly different from each other for all for alcohols as determined by two-way ANOVA (P<0.001).
Figure 2.
Half-maximal shear stress (SS1/2) and maximum elongation index (EImax) calculated by Lineweaver-Burk approach.
The curves presented in Figure 1 were used in calculus. (A) SS1/2 for RBC exposed to ethanol; (B) EImax for RBC exposed to ethanol; (C) SS1/2 for RBC exposed to propanol; (D) EImax for RBC exposed to propanol. The data are means ± SE from 10 experiments on blood samples from different donors. Difference from control without alcohol were tested by one-way ANOVA: * P<0.05; ** P<0.01.
Figure 3.
Effect of alcohol concentration on SS1/2/EImax parameters.
The effects were determined using the EI-SS curves presented in Figure 1. SS1/2/EImax is inversely proportional to RBC deformability. The data are means ± SE from 10 experiments on blood samples from different donors. Statistical significance of the differences in comparison to control (no alcohol) are: * P<0.05 and ** P<0.01, as tested by one-way ANOVA.
Figure 4.
Alcohol concentrations at which the SS1/2/EImax parameter reached a minimum (i.e., maximal RBC deformability).
The minimal concentrations were determined using the fitted curves presented in Figure 3.
Figure 5.
Effect of alcohol concentration on elongation indexes (EI) at 100 Pa SS during a 300 s period of shearing.
(A) Methanol. (B) Ethanol. (C) Propanol. (D) Butanol. EI values are presented as % of the initial value. Each curve represents the mean from 10 experiments. Standard Error (SE) varied between 0.05 and 2.12, and showed increase with the duration of SS (error bars not shown for clarity). The curves were compared to control using two-way ANOVA, with significant differences (P<0.0001) observed for all four concentrations of methanol, ethanol at 0.687 and 1.202 M, both concentrations of propanol, and the single concentration of butanol.
Figure 6.
Changes of SS1/2/EImax following the application of 100 Pa SS for 300 s.
(A) Methanol. (B) Ethanol. (C) Propanol. (D) Butanol. Data are means + SE from 10 experiments on blood samples from different donors. Differences between values that were obtained before (white bars) and after (black bars) the exposure to 100 Pa SS, as determined by the means of two-way ANOVA: * P<0.05; ** P<0.01; *** P<0.001.
Figure 7.
The effects of alcohols on RBC membrane fluidity at various concentrations as determined by EPR spectroscopy.
(A) The order parameter (S) of 5-DS-labeled RBC. S for 2 M methanol was 0.729 ± 0.007 (not shown). (B) Rotational correlation time (τ) of 16-DS-labeled RBC. τ for 2 M methanol was 18.20 ± 0.88** (10-10 s) (not shown). Data are mean ± SD from 4 experiments on blood samples from different donors. Statistically significant differences compared to control (no alcohol) using one-way ANOVA: * P<0.05, ** P<0.01.
Figure 8.
Hemolysis (%) plotted against alcohol concentration for four alcohols with different molecular sizes.
Data are mean ± SE from 7 experiments on blood samples from different donors. Statistical significance from control (no alcohol) tested by one-way ANOVA: * P<0.05; ** P<0.01.
Figure 9.
Light microscopy images of RBC in the presence of alcohols.
The cells were observed in dilute (1:200), fresh, wet-mount, unstained preparations in which cell shape was unaffected by the "glass slide" effect (i.e., crenation due to alkaline conditions at glass surface).