ABSTRACT
BACKGROUND AND AIMS: Appliance of electric pulses induces red blood cells (RBCs) membrane poration, membrane aminophospholipid perturbation and alteration of the normal flip-flop process, resulting in various shape changes of the RBCs. We studied morphological and water permeability changes of RBCs bombarded with electrons in an alternating current circuit. METHODS: We used three venous blood samples of 100 mL and an alternating current device. The harvested blood was divided into four experimental sets to be used for various exposure times: 0 hours (control RBCs), 0.5h, 3h and 6h (electric-stimulated RBCs). Following the electric current each of the four sets were further divided into three samples: one for the assessment of the echinocytes/RBCs ratio, another for the electron microscopy study of ultrastructural changes induced by the alternating electrical current and a larger third one for determining water permeability of RCBs by 1H-NMR spectroscopy and morphological measurements. RESULTS: There is a small but statistically significant effect of the RBC exposure to alternating electric current on cell diameters. Exposure to electric current is positively and strongly correlated with the percentage of echinocytes in a duration-dependent manner. There is a strong and statistically significant correlation between electric current exposure and permeability to water as measured by 1H-NMR spectroscopy. CONCLUSION: Following interactions between electric current and RBC membrane, certain modifications were observed in the erythrocyte structure. We attribute the increased cell size to a higher permeability to water and a decreased tonicity. This leads to the transformation of the RBCs into echinocytes.
ABSTRACT
As part of a programme of comparative measurements of Pd (diffusional water permeability) the RBCs (red blood cells) from dingo (Canis familiaris dingo) and greyhound dog (Canis familiaris) were studied. The morphologies of the dingo and greyhound RBCs [examined by light and SEM (scanning electron microscopy)] were found to be very similar, with regard to aspect ratio and size; the mean diameters were estimated to be the same (approximately 7.2 microm) for both dingo and greyhound RBCs. The water diffusional permeability was monitored by using an Mn2+-doping 1H NMR technique at 400 MHz. The Pd (cm/s) values of dingo and greyhound RBCs were similar: 6.5 x 10(-3) at 25 degrees C, 7.5 x 10(-3) at 30 degrees C, 10 x 10(-3) at 37 degrees C and 11.5 x 10(-3) at 42 degrees C. The inhibitory effect of a mercury-containing SH (sulfhydryl)-modifying reagent PCMBS (p-chloromercuribenzene sulfonate) was investigated. The maximal inhibition of dingo and greyhound RBCs was reached in 15-30 min at 37 degrees C with 2 mmol/l PCMBS. The values of maximal inhibition were in the range 72-74% when measured at 25 degrees C and 30 degrees C, and approximately 66% at 37 degrees C. The lowest value of Pd (corresponding to the basal permeability to water) was approximately 2-3 x 10(-3) cm/s in the temperature range 25-37 degrees C. The Ea,d (activation energy of water diffusion) was 25 kJ/mol for dingo RBC and 23 kJ/mol for greyhound RBCs. After incubation with PCMBS, the values of Ea,d increased, reaching 46-48 kJ/mol in the condition of maximal inhibition of water exchange. The electrophoretograms of membrane polypeptides of the dingo and greyhound RBCs were compared and seen to be very similar. We postulate that the RBC parameters reported in the present study are characteristic of all canine species and, in particular in the two cases presented here, these parameters have not been changed by the peculiar Australian habitat over the millennia (as in the case of the dingo) or over shorter time periods, decades or centuries (as in the case of the domestic greyhound).
