Hemolysis In Hypotonic Solutions Research Articles

PB2419 PEPTIDE CORRECTION OF OSMOTIC RESISTANCE DISORDERS IN INFLAMMATION

Background:The development of inflammation is accompanied by a change in the rheology of the blood ‐ an increase in viscosity, aggregation, a decrease in the osmotic resistance of red blood cells, which indicates a change in the structure of their membrane. This leads to a change in the functional properties of red blood cells and impaired microcirculation. The search for new anti‐inflammatory drugs, devoid of side effects is relevant. Promising may be endogenous regulatory peptides of the glyproline family, which have a prophylactic and therapeutic effect in various pathological conditions.Aims:The aim of this study was to investigate changes in the osmotic resistance of erythrocytes in the dynamics of experimental peritonitis with the introduction of the peptide Pro‐Gly‐Pro (PGP).Methods:A pro‐inflammatory agent was used to simulate experimental peritonitis ‐ 40% sodium thioglyaolate solution 4 g/kg, administered intraperitoneally. The erythrocyte osmotic resistance (ORE) was determined 30 minutes, 2 and 18 hours after the induction of inflammation. The evaluation of the wholesale electricity market was carried out to determine the degree of hemolysis in hypotonic solutions of sodium chloride. The light transmittance was recorded on a photoelectrocalorimeter at a wavelength of 580 nm. All experiments were performed in accordance with the ethical principles and regulatory documents recommended by the ESF and the declaration of a humane attitude towards animals.Results:The development of inflammation is accompanied by a decrease in the osmotic resistance of erythrocytes. Statistically significant differences in the wholesale electricity market from control were observed 2 hours after the administration of the thioglycolate. The introduction of PGP peptide at a dose of 3.7 μm/kg before the induction of inflammation reduced pathological changes in the ORE. The percentage of hemolyzed cells decreased, the differences were significant in solutions with salt content of 0.4% and 0.3%. The peptide, introduced against the background of developed inflammation (1 hour and 45 minutes after the introduction of thioglycolate) did not have a corrective effect on the ORE.Summary/Conclusion:The results indicate the ability of PGP to exert a preventive protective effect on the erythrocyte membrane, preventing a decrease in the ORE.

Effects of dietary levels of vitamin A on growth, hematology, immune response and resistance of Nile tilapia (Oreochromis niloticus) to Streptococcus iniae

This study was conducted to evaluate the effect of supplemental levels of vitamin A (0, 2500, 5000, 10,000, and 20,000IU/kg diet) on the growth performance, hematology, immune response and resistance of Nile tilapia, Oreochromis niloticus to Streptococcus iniae challenge. Each diet was fed to Nile tilapia (initial weight, 5.26±0.10g) in quadruplicate aquaria to apparent satiation twice daily for 10 weeks. The results indicate that vitamin A is a dietary essential for Nile tilapia. After 2 weeks of feeding, fish fed the vitamin A unsupplemented diet exhibited significantly lowest weight gain, feed intake and feed efficiency ratio than those of other treatments. Dietary vitamin A levels had no effect on survival. Other gross deficiency signs observed during the 10-week trial were hemorrhages, operculum deformity, fin erosion, darker body coloration and lethargy. Significantly lower hepatosomatic index and hematological parameters (except white blood cell count) were also recorded in fish fed the vitamin A unsupplemented diet. The resistance of erythrocytes to hemolysis in hypotonic solutions was lowest in fish fed vitamin A unsupplemented diet and increased with increasing vitamin A supplementation. A supplemental level of vitamin A of 3910IU/kg diet was sufficient to prevent these deficiency signs in juvenile Nile tilapia. Serum protein, lysozyme activity and superoxide anion production were enhanced by supplementation of vitamin A, whilst serum total immunoglobulin and spontaneous complement activity were not affected by dietary treatments. Dietary supplementation of vitamin A had no effect on the resistance of Nile tilapia to S. iniae infection challenge and antibody production against the same bacterium.

Interaction between plant polyphenols and the erythrocyte membrane.

The purpose of these studies was to determine the effect of polyphenols contained in extracts from apple, strawberry and blackcurrant on the properties of the erythrocyte membrane, treated as a model of the biological membrane. To this end, the effect of the substances used on hemolysis, osmotic resistance and shape of erythrocytes, and on packing order in the hydrophilic region of the erythrocyte membrane was studied. The investigation was performed with spectrophotometric and fluorimetric methods, and using the optical microscope. The hemolytic studies have shown that the extracts do not induce hemolysis at the concentrations used. The results obtained from the spectrophotometric measurements of osmotic resistance of erythrocytes showed that the polyphenols contained in the extracts cause an increase in the resistance, rendering them less prone to hemolysis in hypotonic solutions of sodium chloride. The fluorimetric studies indicate that the used substances cause a decrease of packing order in the hydrophilic area of membrane lipids. The observations of erythrocyte shapes in a biological optical microscope have shown that, as a result of the substances’ action, the erythrocytes become mostly echinocytes, which means that the polyphenols of the extracts localize in the outer lipid monolayer of the erythrocyte membrane. The results obtained indicate that, in the concentration range used, the plant extracts are incorporated into the hydrophilic area of the membrane, modifying its properties.

Increasing Fish Oil Levels in Commercial Diets Influences Hematological and Immunological Responses of Channel Catfish, Ictalurus punctatus

Abstract Growth performance, immune responses and disease resistance were studied in juvenile channel catfish, Ictalurus punctatus, fed a commercial diet (35.3% crude protein and 5.6% lipid) supplemented with menhaden fish oil at levels of 0, 3, 6, and 9% for 15 wk. Dietary fish oil levels did not significantly influence growth performance of catfish. Fatty acid compositions of whole‐body and liver reflected dietary fatty acid composition. No differences were found in hematological values, except that fish fed the 9% fish oil diet had significantly lower hematocrit. The resistance of erythrocytes to hemolysis in hypotonic solutions increased with increasing fish oil levels and the highest resistance was seen with the 9% fish oil diet. Fish fed 6 and 9% added fish oil diets had significantly higher serum protein levels than that of control fish. Serum lysozyme activity of fish fed 3 and 6% added fish oil diet was significantly higher than that of the control. Complement activity and chemotaxis ratio significantly decreased in fish fed diets with 6 or 9% added fish oil. The 3% added fish oil diet, however, had significantly highest natural hemolytic complement activity. Mortality from Edwardsiella ictaluri 14 d postchallenge and antibody titers to E. ictaluridid not differ among treatments.

Hematological Effects of Chlorine Dioxide on In Vitro Exposure in Mouse, Rat and Human Blood and on Subchronic Exposure in Mice

Hematological effects of chlorine dioxide (ClO2) and its metabolites were investigated. In vitro exposure of mouse, rat and human blood cells to ClO2 and the reduction by-product, chlorite (ClO2-) resulted in the formation of methemoglobin, a decrease in the activities of glucose-6-phosphate dehydrogenase (G-6-PD) and glutathione peroxidase (GPX) and in the content of reduced glutathione (GSH), and an increase in hydrogen peroxide (H2O2) formation and hemolysis. The H2O2 formation and hemolysis induced by ClO2 and ClO2 - in mouse blood cells were the highest among cells tested, and human blood cells were more resistant to the oxidative stress than rat and mouse blood cells. Both compounds also showed more toxic responses to E. coli mutants lacking production of catalase DSH19 (katEG), superoxide dismutase DSH56 (sodAB) and both of them DSH67 (katEG sodAB) than the wild strain DSH7 by Kat-sod assay, as a biological detection method for reactive oxygen species, suggesting the production of H2O2 and superioxide anion. For subchronic study of ClO2, mice received drinking water containing 100, 1000, 1500 or 2000mg/l ClO2 in the presence of the stabilizer, 1200mg/l of sodium bicarbonate ad libitum for 30, 60 or 90days. Statistically significant hematological changes were observed in animals exposed to more than 1000mg/l ClO2, which showed augmented G-6-PD activity in erythrocytes and increased resistance to hemolysis in hypotonic solution. The results of this study therefore indicate that ClO2 acutely causes hematotoxicity toward mice by producing reactive oxygen species and by weakening the protection systems to oxidative stress in erythrocytes, although the latter may be induced by long term exposure, while humans appear to be more resistant to this hematotoxicity.

Photometric determination of phenomenological correlation between osmotic behavior and hemolysis of red blood cells.

The osmotic behavior of red blood cells from a human and from several other mammalian species was studied by photometric measurements. When red blood cells were suspended in sodium chloride solutions with various osmotic concentrations, the optical density at 620 nm was reciprocally related to the relative volume of the red cells. Thus, we evaluated the osmotic volume changes in the red cells from optical density measurements. The Boyle-van't Hoff relation was applicable to the osmotic behavior of red cells which responded as a complete osmometer in hypertonic and slightly hypotonic (lower than about 240 mOsm) solutions. Also, we examined the rheological correlation between osmotic volume changes and hemolysis. Osmotic hemolysis occurred corresponding to breakdown of the Boyle-van't Hoff relation in hypotonic solutions. The critical osmotic concentration for the breakdown of the Boyle-van't Hoff relation was that for osmotic hemolysis. In Na2SO4 solutions, although the critical osmotic concentration shifted towards a smaller value, the critical volume for the breakdown of the Boyle-van't Hoff relation and for osmotic hemolysis was maintained at a constant value, indicating that the onset of osmotic hemolysis depends exclusively upon the critical volume. In the samples from a human, the critical volume for the onset of hemolysis was estimated to be 1.25 +/- 0.05 in the ratio to the normal volume in iso-osmotic solution. From these obtained results, it is suggested that the red cell behaves in hypotonic solutions as a viscoelastic body of the type represented by the Voigt model, and the viscoelastic breakdown of the membrane results in osmotic hemolysis in hypotonic solution.

Participation of band 3 protein in hypotonic hemolysis of human erythrocytes.

The study was designed to show whether band 3, anion exchange protein in human erythrocytes, is responsible for hemolysis in hypotonic solution. From ESR spectra of the spin labeled erythrocytes, it was found that the change of the osmotic condition had no effect on membrane fluidity at pH 7.4. At pH 6.0, however, the fluidity at the central part of the lipid bilayer was dependent on osmotic stress. In this case, the fluidity change as a function of KCl concentration was similar to the hemolysis curve. From circular dichroism (CD) spectra of erythrocyte ghosts, it was found that the conformational and/or associative changes of the membrane proteins correlated with the hypotonic hemolysis. In contrast to the osmotic stress, sodium dodecyl sulfate (SDS) changed the fluidity of the membranes but not the conformation of the membrane proteins regardless of the medium pH. In this case, the changes in the fluidity corresponded to the hemolysis curves in SDS solutions. Further, by using anion transport inhibitors, 4,4'-diisothiocyanostilbene-2,2'-disulfonate, eosin 5-isothiocyanate and eosin-5-maleimide, a correlation between the effect on the hypotonic hemolysis and the conformational and/or associative change of band 3 was suggested. These results demonstrate that band 3 is responsible for the hypotonic hemolysis.

Age Dependent Changes in Osmotic Hemolysis of Human Erythrocytes

The osmotic fragility curve and the rate of hemolysis in hypotonic solution were investigated for individuals of different age. It was found that in older individuals the erythrocyte is more fragile with a broader and more asymmetric distribution of fragilities, while the rate of hemolysis is significantly slower for older individuals. The increase in fragility with age can be attributed to the increase in cellular volume, while the decrease in rate of hemolysis with age can be attributed to change in the properties of the erythrocyte membrane. All the changes with respect to individual age cannot be explained by a change in the distribution of cellular ages, and some changes in the formation of erythrocytes and/or the plasma environment of the erythrocyte must take place.

OBSERVATIONS ON THE EFFECT OF IRRADIATION IN CHRONIC ACQUIRED HEMOLYTIC ANEMIA EXHIBITING HEMOLYTIC ACTIVITY FOR TRANSFUSED ERYTHROCYTES

Abstract The history and course of a middle-aged woman with a chronic acquired hemolytic anemia have been presented. Splenectomy resulted in a complete remission of the disease for 18 months before a relapse occurred which led eventually to a fatal termination. The original cause of the abnormal hemolytic process is not known, but the onset of the relapse was closely associated with the resumption of gold therapy. The acceleration of the hemolytic process was associated with an increasing tendency toward spherocytosis and susceptibility of the erythrocytes to hemolysis in hypotonic solution. It is demonstrated that transfused normal erythrocytes shared in the hemolytic process and were eliminated from the circulation at an abnormal rate although no atypical isohemolysin was demonstrated by in vitro tests. An attempt was made to alter the rate of hemolysis in several ways. The injection of thorium dioxide was followed by signs of transitory slowing of red cell destruction. Injections of Congo red were apparently without effect. Irradiation of the mediastinal and periaortic nodes was begun to ascertain if the production of a hemolytic antibody in lymphatic tissue could be altered. Irradiation was followed by a fall in leukocyte and lymphocyte counts in the peripheral blood. Transfusions were then followed by partial remissions in the hemolytic process for varying periods of time in two instances. Irradiation of a second patient with a chronic hemolytic anemia of similar character produced a fall in leukocyte count and lymphocyte count, but in the absence of transfusions no slowing of the hemolytic process occurred. No definite conclusions can be drawn from these fragmentary observations on the effect of irradiation, but further investigation as to the character of the hemolytic substance and possible methods of modifying its production are indicated when the opportunity is afforded.

The resistance of red blood cells to hemolysis in hypotonic solutions of sodium chloride

The resistance of red blood cells to hemolysis in hypotonic solutions of sodium chloride

Osmotic properties of the erythrocyte V. The rate of hemolysis in hypotonic solutions of electrolytes

Osmotic properties of the erythrocyte V. The rate of hemolysis in hypotonic solutions of electrolytes

Osmotic properties of the erythrocyte III. The applicability of osmotic laws to the rate of hemolysis in hypotonic solutions of non-electrolytes

Osmotic properties of the erythrocyte III. The applicability of osmotic laws to the rate of hemolysis in hypotonic solutions of non-electrolytes

The influence of temperature on hemolysis in hypotonic solutions

Hemolysis in hypotonic solutions is progressively increased as the temperature is decreased from thirty seven degrees centigrade to five degrees centigrade. In order to bring out this fact, that modification of Hamburger's method for testing the resistance of erythrocytes, which was introduced by Theobald Smith, was used. The solutions were brought to the required temperature and then the corpuscles were added. The differences are present both at the points of beginning and complete hemolysis, but are only well marked at the intermediate points. This accounts for results obtained by Hamburger (1887 and 1903) who held that temperature within these limits was without influence.The effect of temperature is the same whether sodium chloride or cane-sugar is used to give tonicity to the fluid. The corpuscles of the horse, rabbit, guinea-pig, calf, and sheep are equally affected. The differences become well marked after a few minutes—exposure to the different temperatures, and thereafter bringing them to one tem...