Osmotic Resistance Of Erythrocytes Research Articles

BIOPHYSICAL MARKERS OF ISCHEMIC STROKE

An important and influential factor that directly affects the severity of ischemic stroke (IS) and determines its outcome is the functional state of the membrane-receptor complex (MRC) of cells. One of the most important criteria for assessing this parameter is the β‑adrenergic activity of cytoplasmic membranes (β‑ARM), which plays a leading role in the pathogenesis of IS. The article presents the results of a comprehensive study of the peculiarities of changes in the structural and functional characteristics of brain tissue and β‑adrenoceptors in the acute period of IS to optimize diagnosis. Measurement of changes in the osmotic resistance of erythrocytes (ORE) under the action of β‑blockers was determined by photoelectron colorimetry. The work was based on the materials of a comprehensive examination of 350 patients with the new-onset IS on the 1st, 10th and 21st day of the disease. The severity of the condition and the degree of neurological deficit were objectified using the stroke scale of the National Institutes of Health (NIHSS) with a score in the first hours of the disease, in the dynamics of treatment on the 10th and 21st day. All patients were divided into 2 clinical groups: 1st (n = 183) – patients in moderate severity condition (mean score on the NIHSS scale 11.74 ± 0.33); 2nd (n = 167) – patients in severe condition (mean score on the NIHSS scale 24.06 ± 0.29). As a result of the study, the indicators of β‑ARM of the control group were within normal limits (15.3 ± 4.4 SU), which corresponds to normal β‑ARM. In patients with moderate IS, the indicator exceeded the control values by 1.97 times, which is typical for the average degree of β‑ARM (21–40 SU), and in severe – 2.8 times and was characterized by a low degree of β‑ARM (> 41 SU). An unfavorable sign for the prognosis of the acute period of IS was represented by a further increase in the levels of β‑ARM in the dynamics, which was observed in clinically severe patients. Keywords adrenergic activity, ischemia, erythrocyte, receptor.

Possibilities of target neurotrophic therapy of ischemic stroke

The study aimed to comprehensively investigation the features of changes in the structural and functional characteristics of the brain tissue, cytokine profile, and β-adrenergic reception in the acute period of ischemic stroke (IS) to optimize treatment. Materials and methods. EHF dielectrometry was used to measure the complex dielectric conductivity (CDC) of peripheral blood erythrocytes in patients with IS. Changes in the osmotic resistance of erythrocytes (ORE) under the action of β-adrenergic blockers (β-AB) were determined by photoelectron colourimetry. Plasma levels of interleukin (IL)-6 and tumour necrosis factor (TNF)-α were assessed using an enzyme-linked immunosorbent assay. The basis of the work was the materials of a comprehensive examination of 350 patients with the first in life IS in the dynamics of treatment with human cryopreserved cord blood serum (CCBS). Results. In patients with IS, from the first hours of the development of the disease, there is a sharp increase in the levels of proinflammatory cytokines IL-6 and TNF-α in the blood serum (by 9.3 and 3.9 times, respectively). At the onset of IS, there is a significant increase in the level of β-ARM by 2.4 times as compared with the control and a decrease in CDC by 10.0 % after exposure to an adrenaline solution. The maximum levels of β-ARM (42.43 ± 3.64 CU) are observed in patients with initially severe disease. The established direct correlations between plasma levels of IL-6, TNF-α and β-ARM (r 0.73; p < 0.05 and r = +0,86; p < 0.05, respectively); IL-6, TNF-α and total clinical score on the NIHSS scale (r = +0.895; p < 0.05 and r = +0.9; p < 0.05, respectively). Conclusions. The study has demonstrated the positive immunomodulatory and membrane-protective effects of human CCBS in the acute period of IS. Stabilization of the absolute values of CDC indicated changes in the levels of cell hydration, causing the activation of not only the membrane receptor complex (MRC) of erythrocytes but also an increase in the functional characteristics of the sympathoadrenal system (SAS). The use of CCBS caused a more significant and rapid decrease in the concentrations of the central proinflammatory cytokines IL-6 and TNF-α, which indicated the regulatory effect of the drug in suppressing the local inflammatory response initiated by hypoxia.

Determination of osmotic resistance of erythrocytes using flow cytometry in patients with hereditary spherocytosis

To diagnose hereditary spherocytosis (HS), an osmotic resistance test is used to assess the severity of hemolysis in a series of hypotonic sodium chloride solutions, which demands intensive and time consuming workload and has low sensitivity and specificity. New methods for evaluating osmotic resistance based on flow cytometry were proposed, but they are not sufficiently standardized. Objective: development and testing of the method for determining the osmotic resistance of red blood cells (RBCs) using flow cytometry in patients with HS. Material and methods. In 2017-2019, 33 patients with HS and 30 children in the control group were examined. The method included the study of blood preparations in phosphate-buffered saline (PBS) and their serial dilutions in distilled water using flow cytometry, both in fresh blood and after 24-hour incubation. The proportion of preserved (not hemolized) RBCs was evaluated. As reference intervals, 5 and 95 percentiles of the control group indicators were used. Results and discussion. Among people with HS, before incubation, pathological changes in at least two dilutions were detected in 81.8 % of patients; after incubation, pathological changes were detected additionally in 4 out of 6 patients. Diagnostic sensitivity of the test is 93.9 %, specificity 83.3 %, positive and negative predictive values 86.1 and 92.6 %, respectively. It was found that the method with proposed reference intervals allows evaluating the osmotic resistance of erythrocytes fast and objectively, and can be used for differential diagnosis in hereditary spherocytosis.

Study of general toxic effect of bacteriosens in rodents and large laboratory animals

Purpose of the study . The objectives of the research are to evaluate the potential toxicity of Bacteriosens when administered to animals once only («acute» toxicity) or repeatedly («chronic» toxicity) and to study its biocompatibility with blood components. Materials and methods . Bacteriosens is the medicine on the basis of photoactive compound of mezo -tetra (3-pyridyl) bacteriochlorin with λ max of 747 nm. Ex vivo study of its biocompatibility with blood components included the evaluation of the impact of the solution on the coagulability of non-stabilized venous blood. The in vitro method of detecting erythrocyte osmotic resistance to hemolytic ability of Bacteriosens was used. The «acute» toxicity was studied in F 1 hybrid mice (CBA x C57Bl/ 6j ) and non-bred rats, male and female, after single intravenous and intraperitoneal injections of Bacteriosens. The «chronic» toxicity was studied in non-bred rats, male and female, and Soviet Chinchilla rabbits after multiple intravenous injection of the drug (over the period of 14 days). Time period before toxicity-induced death of animals, clinical signs of intoxication, and the impact of the medicine on the inner organs and body systems were evaluated using physiologic and pathophysiologic, clinical, and laboratory methods. Results . Incompatibility of non-stabilized rat blood with Bacteriosens after the incubation with the medicine in concentrations varying from 0.5 till 0.005 mg/ml has not been discovered. The aquatic solution of Bacteriosens should be injected into animals and later on into humans slowly or by drip feed in concentrations not exceeding 0.5 mg/ml. A single injection of Bacteriosens in doses from 5.0 to 50.0 mg/kg and from 1.3 to 20.8 mg/kg to mice and rats (male and female), respectively, was tolerated by the animals satisfactorily. No toxicity-induced deaths or skin phototoxicity have been detected. The studied doses of Bacteriosens in mice 25-250 times exceeded the calculated equitherapeutic dose for humans, and in rats, 6.5-104 times. Bacteriosens in total doses from 18.2 to 72.8 mg/kg and from 8.3 to 33 mg/kg administered as multiple intravenous injections for 14 days to rats and rabbits respectively, did not induce the death of the animals and demonstrated no toxic impact on blood, liver, kidneys, heart, hemostasis system, or central nervous system. The drug did not adversely affect carbohydrate and lipid metabolism. Conclusion . Bacteriosens when administered as a single dose or as multiple injections in the range of studied doses did not have adverse toxic effect on mice, rats, and rabbits. The studied doses of Bacteriosens in rats 91-364 times exceeded the calculated equitherapeutic dose for humans, and 41-165 times, in rabbits.

Stability of refrigerated whole blood samples for osmotic fragility test

BackgroundThe osmotic fragility test (OFT), conventionally used for assisting the diagnosis of many erythrocyte disorders, is a manual and time-consuming analysis not daily performed in many medical laboratories. This study was aimed at defining the stability of whole blood samples used for assessing erythrocyte osmotic resistance. MethodsTwenty-one consecutive routine whole blood samples collected into 5.4 mg K2EDTA were tested immediately after collection (day 0) and at different time intervals afterward (day 1, 2, 3, 4, 7, 10 and 14) after storage at 4 °C. The OFT was performed with the Osmored Monotest (1.3% glycerol; Eurospital, Trieste, Italy). Results at the different time points were compared with those obtained at day 0 and with the reference change value (i.e., 33%). ResultsThe median value of both hyperosmolar and hyposmolar resistance increased from baseline, reaching statistical significance at day 7 for hyperosmolar resistance and at day 1 for hyposmolar resistance, respectively. The median relative increase of hemolysis percentage values become greater than the reference change value at day 3 for hyposmolar resistance, while this limit was never overcome for hyperosmolar resistance. A significant inverse association was found between the mean increase in hyperosmolar resistance and the baseline value of hyperosmolar resistance (r = −0.92), mean corpuscular volume (MCV; r = −0.46) or mean corpuscular hemoglobin (MCH; r = −0.44), as well as between the mean increase in hyposmolar resistance and the baseline value of hyposmolar resistance (r = −0.86), or patient age (r = −0.56). ConclusionsThe sample stability seems critical for the OFT. Whole blood specimens should not be stored refrigerated at 4 °C for >2 days before testing.

Влияние локальной холодовой травмы на осмотическую резистентность эритроцитов

Влияние локальной холодовой травмы на осмотическую резистентность эритроцитов

Red blood cells state and calcium content of in the blood plasma during physical activity in marathon runners of various qualifications

The aim of the work is to investigate the morpho-functional state of peripheral blood erythrocytes and the content of Ca2+ in the blood plasma during physical exercise in marathoners of different qualifications.
 Material and methods. The studies were conducted in three groups of marathoners aged 20–25 years old with conditionally high, medium and low performance. Exercise was performed for 90 minutes on a bicycle ergometer with a given power. The study of peripheral blood erythrocytes was performed before and after exercise. The osmotic resistance of erythrocytes, the content of Ca2+ and free fatty acids in the blood were determined. Morphological study of erythrocytes was performed using a scanning electron microscope and stained by standard techniques. The red blood cell form index was determined by a special formula. Changing the content of Ca2+ enhances lipid melting, increases membrane fluidity and increases surface tension, with the result that the discocytes are stretched and thinned, the central pits deepen and the surface roughness increases, the volume and number of conical pores, and peripheral blood erythrocytes density.
 Results. Marathon runners of different groups after physical exercise differed in terms of the frequency of cardiac sweeps, the content of free fatty acids and the osmotic resistance of red blood cells. This is closely correlated with changes in the electrolyte and metric composition of erythrocytes and the presence of their reversible and irreversible forms in the general circulation after exercise.
 Conclutions. Physical activity causes structural and functional reorganization of erythrocytes, which is based on a change in their microelement composition, a decrease in osmotic resistance, mainly in poorly trained marathoners, and the appearance of various forms of erythrocytes against the background of the destruction of degenerative forms and an increase in the active release of young forms, which is a sign of high adaptive capacity in highly skilled athletes.

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.

The erythrocyte osmotic resistance test as screening tool for cholesterol-related lysosomal storage diseases

BackgroundErythrocyte volume regulation and membrane elasticity are essential for adaptation to osmotic and mechanical stress, and life span. Here, we evaluated whether defective cholesterol trafficking caused by the rare lysosomal storages diseases (LSDs), Niemann-Pick type C (NPC) and Lysosomal acid lipase (LAL) deficiency (LALD) impairs these properties. Moreover, we tested whether measurements of cholesterol membrane content and osmotic resistance serve as a screening test for these LSDs. MethodsPatients were genotyped for mutations in NPC1, NPC2, or LIPA genes. We measured LSD plasma biomarkers and LAL activity. Red blood cells (RBC) membrane cholesterol content was evaluated in 73 subjects. Osmotic resistance tests (ORT) were conducted in 121 blood samples from LSD suspected patients and controls. ResultsWe did not find statistically significant differences between RBC cholesterol content between subjects and controls. However, the ORT, particularly at 0.49% (w/v) hypotonic sodium chloride solution, revealed a significant higher osmotic resistance in LSDs patients than in controls. We established a cut-off value of ≤51% of haemolysis with sensibility and specificity values of 80% and 70%, respectively. ConclusionsNPC and LALD do not alter cholesterol content in the RBC membrane but increase osmotic resistance. Therefore, ORT serves as screening test for the studied LSDs.

Erythrocyte osmotic resistance test as a screening tool for lysosomal diseases

Trypanosoma (Herpetosoma) lewisi is a cosmopolitan parasite of rodents strongly linked to the human dispersal of Rattus spp. from Asia to the rest of the world. This species is highly phylogenetically related to trypanosomes from other rodents (T. lewisi-like), and sporadically infects other mammals. T. lewisi may opportunistically infect humans, and has been considered an emergent rat-borne zoonosis associated to poverty. We developed the THeCATL-PCR based on Cathepsin L (CATL) sequences to specifically detect T. (Herpetosoma) spp., and assess their genetic diversity. This method exhibited high sensitivity using blood samples, and is the first molecular method employed to search for T. lewisi in its flea vectors. THeCATL-PCR surveys using simple DNA preparation from blood preserved in ethanol or filter paper detected T. lewisi in Rattus spp. from human dwellings in South America (Brazil and Venezuela), East Africa (Mozambique), and Southeast Asia (Thailand, Cambodia and Lao PDR). In addition, native rodents captured in anthropogenic and nearby human settlements in natural habitats harbored T. (Herpetosoma) spp. PCR-amplified CATL gene fragments (253 bp) distinguish T. lewisi and T. lewisi-like from other trypanosomes, and allow for assessment of genetic diversity and relationships among T. (Herpetosoma) spp. Our molecular surveys corroborated worldwide high prevalence of T. lewisi, incriminating Mastomys natalensis as an important carrier of this species in Africa, and supported its spillover from invader Rattus spp. to native rodents in Brazil and Mozambique. THeCATL-PCR provided new insights on the accurate diagnosis and genetic repertoire of T. (Herpetosoma) spp. in rodent and non-rodent hosts, revealing a novel species of this subgenus in an African gerbil. Phylogenetic analysis based on CATL sequences from T. (Herpetosoma) spp. and other trypanosomes (amplified using pan-trypanosome primers) uncovered rodents harboring, beyond mammal trypanosomes of different subgenera, some species that clustered in the lizard-snake clade of trypanosomes.

Influence of chlorpromazine on the resistance of erythrocytes of rats of different ages to hypertonic conditions

The osmotic stability of native and modified with chlopromazine (CPR) erythrocytes of 1- and 12-month rats to hypertonic conditions in sucrose solutions and hypertonic shock (4.0 M NaCl) has been studied. It has been shown that 2-min incubation of rat erythrocytes of different ages in hypertonic sucrose media does not reveal any differences in the osmotic stability of these cells. In this case, CPR does not affect cell hemolysis. An increase of the incubation time in hypertonic sucrose solutions to 30 minutes allowed detecting a greater osmotic sensitivity of erythrocytes of 1-month animals to the action. In this case, the protective effect of CPR for older age rat erythrocytes (12 months) has been established. It has been found that in 4.0 M NaCl the hemolysis level of animal erythrocytes of both age groups increases with preliminary incubation (2 min) in a sucrose medium with a concentration of 0.7 M and above. With increasing exposure time (30 min) in sucrose hypertonic solutions, the sensibilization of animal erythrocytes of both age groups to the action of hypertonic shock is also intensified. In this study it has been shown that the influence of CPR on the sensitivity of 1-month-old animal erythrocytes to the transfer in 4.0 M NaCl depends on the tonicity and duration of the cell initial incubation in sucrose solutions. Thus, CPR increases the osmotic resistance of erythrocytes, which were preexposed in sucrose solutions at a concentration of 0.6–0.8 M for 2 and 30 min. Some increase of hemolysis level of these animals erythrocytes modified with CPR in 4.0 M NaCl has been observed after 2 min of incubation in sucrose solutions at a concentration of 0.27–0.5 M. The exclusively protective influence of CPR on 12-month-old animal erythrocytes in conditions of hypertonic shock has been revealed. A quantitative estimation of the efficiency of CPR at hypertonic shock (4.0 M NaCl) of different age animal erythrocytes has been carried out by calculation of the antihemolytic activity value (AG). Short-term incubation (2 min) in a sucrose media does not reveal any differences in the values of AG of CPR for erythrocytes of both age groups. For the cells of young rats, increase of AG of CPR is observed at incubation in sucrose medium to 30 min and for erythrocytes of the older group – to 10 min. With an increase in incubation time of up to 60 min the CPR efficiency in hypertonic saline media is reduced for rat cells in both age groups but in varying degrees.

The effect of nanochromium citrate on the osmotic resistance of erythrocyte membranes in rats with the experimental diabetes mellitus

Diabetes mellitus (DM) is a serious medical and social problem because of its wide spread in the world and a steady tendency to increase the number of patients. Aim. To study the effect of nanochromium citrate on osmotic resistance of erythrocyte membranes of animals with diabetes mellitus.Materials and methods. The study was performed on 18-month laboratory male rats. Diabetes mellitus was caused by dexamethasone (0.125 mg/kg, subcutaneously during 14 days). Nanochromiun citrate (NCC) was introduced into the stomach (0.01 mg/kg during 14 days). Permeability of erythrocyte membranes (the erythrocyte osmotic stability) was detected by the erythrocyte resistance in the mixture of isotonic solutions of urea and sodium chloride with their different volumetric composition. The osmotic hemolysis degree (%) was measured by photocolorimetry in relation to the optic density of the standard urea solution. The erythrocyte membrane osmotic resistance (EMOR) – (% of hemolyzed cells) was compared with the similar index in the control intact animals. The glucose level in the blood was measured by a glucometer.Results. In animals with the experimental DM in addition to the elevated glucose level in the blood to 14.2 ± 0.4 mmol/L the percentage of hemolyzed erythrocytes increased by 8.2-2.3 times compared to the control according to the increased concentration of urea in the solution. The increased permeability of erythrocyte membranes resulting in the reduced osmotic resistance of erythrocytes and their hemolysis is indicative of lipid peroxide oxidation (LPO) activation and decrease in activity of the antioxidant defense system in animals with DM. Introduction of NCC in DM reduced the glucose level in the blood to 6.6±0.4 mmol/L and increased EMOR considerably, and it indicates the presence of NCC membrane-protective properties.Conclusions. In case of the experimental (dexamethasone) DM the erythrocyte membrane osmotic resistance decreases, destructive processes are activated, resulting in their accelerated destruction. NCC decreases the glucose level in the blood of animals with DM and increases osmotic resistance of erythrocyte membranes.

The Uremic Toxin Indoxyl Sulfate Accelerates Thrombotic Response after Vascular Injury in Animal Models.

Chronic kidney disease (CKD) patients are at high risk for thrombotic events. Indoxyl sulfate (IS) is one of the most potent uremic toxins that accumulates during CKD. Even though IS is associated with an increased risk for cardiovascular disease, its impact on thrombotic events still remains not fully understood. The purpose of the study was to evaluate the direct effect of IS on thrombotic process. We examined the impact of acute exposure to IS on thrombus development induced by electric current in Wistar rats, intravital thrombus formation after laser-induced injury in the mice endothelium, coagulation profile, clot formation dynamics, platelet aggregations, and erythrocyte osmotic resistance. IS doses: 10, 30 and 100 mg/kg body weight (b.w.) increased weight of thrombus induced by electric current in dose-dependent manner (p < 0.001). Furthermore, two highest IS doses increased laser-induced thrombus formation observed via confocal system (increase in fluorescence intensity and total thrombus area (p < 0.01)). Only the highest IS dose decreased clotting time (p < 0.01) and increased maximum clot firmness (p < 0.05). IS did not affect blood morphology parameters and erythrocyte osmotic resistance, but augmented collagen-induced aggregation. Obtained data indicate that IS creates prothrombotic state and contributes to more stable thrombus formation. Thus, we concluded that IS may be one of crucial uremic factors promoting thrombotic events in CKD patients.

Ultramolecolar diluted homeopathic medicinal products alter human erythrocyte osmotic resistance

Ultramolecolar diluted homeopathic medicinal products alter human erythrocyte osmotic resistance

Hematological changes and resistance of erythrocytes of crimean horses in response to 32 km races

Aim. The aim of the present study was to investigate the alterations of some hematological parameters (hae- matocrit (HCT), haemoglobin concentration (HGB), the count of red blood cells (RBC), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), red cell distribution width (RDW), as well as resistance of erythrocytes to urea and hydrogen peroxide in horses after 32 km endurance race. Methods. Seven horses from Crimea region (Bilohirsk, Crimean region) were involved in this study. Haematological parameters (haematocrit (HCT), haemoglobin concentration (HGB), the count of red blood cells (RBC), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), red cell distribution width (RDW)) were determined with use of hematological and biochemical methods. Blood samples have been investigated by centrifugation at 3,000 g for 15 min. The peroxide and osmotic resistance of erythrocytes were determined spectrophoto- metrically at 540 nm by monitoring the rate of erythrocytes disintegration by hydrogen peroxide. Endurance h orses used in this study are trained and conditioned to perform over long distances at moderate speeds. The prolonged exercises were used in endurance race. The walk about 3 km/h for 20 min, the trot about 7 km/h for 15 min, and the canter about 5 km/h for 15 min) and the walk about 1 km were repeated for 1 h (phase I); rest in an outdoor paddock without access to water for 30 min. And phase II: the walk about 3 km/h for 20 min, the trot about 7 km/h for 15 min, and the canter about 5 km/h for 15 min) and the walk about 1 km was repeated for 1 h. Results. The results of the present study showed that adequate endurance race of low intensity could im- prove oxygen-dependent respiratory function in horses from Crimean region. Furthermore, the non-signifi cant increase of red blood cells indices in endurance horses indicate about good athletic level after 32 km endur- ance ride. Statistically signifi cant differences in the percentage of hemolyzed erythrocytes between pre- and post-ride period were observed and thereby signifying an oxidative stress-dependent impairment of erythrocyte stability. Conclusions. The haematological changes caused by various physical efforts refl ect changes in the functions of different systems and can be used for health control and diagnosis of diseases. It also allow the evaluating the level of sport performance, the accuracy of training, and physiological condition of horses.

Effect of aqueous lyosol of turpentine oil on erythrocyte distribution by osmotic resistance

To study the possibility of using turpentine oil and other turpentines as adjuvants, the effects of aqueous lyosols of turpentine oil on the erythrocyte distribution by osmotic resistance were analyzed. In our study, differential osmoerythrograms obtained by modifying the Idelson’s approach to the erythrocyte osmotic resistance measurement were utilized. It was found that, when turpentine oil content in the lyosol was insufficient for direct hemolysis, four concentration zones with corresponding effects can be distinguished: I— 0.14 g/L. It was assumed that the detected effect will allow controlling the intravascular hemolysis when using terpenoids as adjuvants.

Physical Effects of Buckwheat Extract on Biological Membrane In Vitro and Its Protective Properties.

Buckwheat is a valuable source of many biologically active compounds and nutrients. It has properties that reduce blood cholesterol levels, and so reduces the risk of atherosclerosis, seals the capillaries, and lowers blood pressure. The aim of the study was to determine quantitative and qualitative characteristics of polyphenols contained in extracts from buckwheat husks and stalks, the biological activity of the extracts, and biophysical effects of their interaction with the erythrocyte membrane, treated as a model of the cell. An analysis of the extract’s composition has shown that buckwheat husk and stalk extracts are a rich source of polyphenolic compounds, the stalk extracts showing more compounds than the husk extract. The study allowed to determine the location which incorporated polyphenols occupy in the erythrocyte membrane and changes in the membrane properties caused by them. It was found that the extracts do not induce hemolysis of red blood cells, causing an increase in osmotic resistance of erythrocytes. They affect mainly the hydrophilic region by changing the degree of order of the polar heads of lipids, but do little to change the fluidity of the membrane and its hydration. The results showed also that polyphenolic substances included in the extracts well protect the membranes of red blood cells against oxidation and exhibit anti-inflammatory effect.Electronic supplementary materialThe online version of this article (doi:10.1007/s00232-015-9857-y) contains supplementary material, which is available to authorized users.

Oxidative Stress Biomarkers and Erythrocytes Hemolysis in Well-Trained Equine Athletes Before and After Exercise

The chronic exposure to regular exercise training seems to increase resistance to oxidative stress and improves the antioxidant defense system. The purpose of the present study was to investigate the effect of an exercise test of moderate intensity on oxidative stress biomarkers, antioxidant enzymes activity, and osmotic resistance of erythrocytes in well-trained equine athletes. Eighteen middle-aged horses of Ukrainian warmblood (8.3 ± 1.6 years) and Holsteiner (7.4 ± 1.9 years) breeds were used in this study. All horses have been in regular training for several years. The exercise test induced a significant increase of erythrocyte values, hemoglobin concentration, and hematocrit in horses of both breeds. Regular training induces activation the antioxidant enzymes and thereby can reduce oxidative stress in athletic horses. Our results suggest that the exercise test in horses of both breeds attenuates oxidative stress and accompanied with a significant decrease of lipid peroxidation and oxidatively modified proteins in erythrocytes after exercise. The findings of the present study demonstrated the elevated level of erythrocytes' catalase and glutathione reductase in Ukrainian warmblood horses, as well as decreased level of superoxide dismutase and glutathione reductase in Holsteiner horses reporting changes in levels of exercise-induced oxidative stress biomarkers in horses of both breeds. Statistically significant differences in the percentage of hemolyzed erythrocytes between pre-exercise and postexercise tests were observed and thereby signifying an oxidative stress-dependent impairment of erythrocyte stability. Our data suggest that oxidative stress and enzymatic antioxidant defense biomarkers can be used for the monitoring of fitness level, health benefits, and performance of equine athletes.

Increase in erythrocyte osmotic resistance following polyunsaturated fatty acids (PUFA) supplementation in show jumper horses

Osmotic fragility test has been used to study the influence of different factors on the osmotic properties of cell membranes. Considering the importance of polyunsaturated fatty acids (PUFA) in biological functions we aimed to investigate the effects of dietary PUFA supplementation on erythrocyte osmotic fragility (EOF), blood lactate (BL), hematocrit (Hct), red blood cell (RBC), hemoglobin (Hb), and mean cell volume (MCV) in 10 jumper horses. Two events occurred prior to start supplementing horse's diet and two events occurred after 4 weeks PUFA supplementation. Five horses received the PUFA supplementation (Group A), and five served as controls (Group B). Blood samples were taken before and after each course. The statistical analysis revealed significant increase in BL, Hct, RBC and Hb following exercise (P<0.0001). However, the interaction between exercise and PUFA supplementation (P=0.0083) showed PUFA-supplemented horses having a smaller rise in BL levels (P=0.0107) following exercise. Significant interactions between exercise and PUFA treatment were also found on EOF levels (P<0.05). The hemolysis curves showed PUFA-supplemented horses exhibiting a reduction in EOF compared to controls (P<0.05). Although hemolysis never occurred at 0.9% NaCl concentration, jumping exercise determined an increase in EOF (P=0.0014) at 0.8% NaCl solution. A significant interaction between exercise and PUFA treatment (P=0.0022) was found showing PUFA-supplemented horses having lower EOF (P=0.0015) following exercise. The assessment of EOF is a suitable indicator of athletic performance. The results showed that PUFA supplementation might exert beneficial effects on the horse body system by enhancing the performance in high-level show jumpers.

Thymoquinone Therapy Improves Hyperglycemia, Erythrocyte Indices, Erythropoietin Production and Erythrocyte Osmotic Resistance in Rat Model of Streptozotocin-induced Diabetes

Herein, both antidiabetic and hematoprotective effect of thymoquinone (TQ) therapy was investigated in rat model of streptozotocin (STZ)-induced diabetes. After disease induction, TQ (35 mg/kg/day) was given for 28 days and fasting blood glucose level was weekly measured. At day 29, blood samples, pancreatic and kidney specimens were prepared and screened for: (1) hematological parameters: counts of red blood cells (RBCs), white blood cells (WBCs) and platelets (PLTs), erythrocyte osmotic fragility, normal hemoglobin (Hb) and glycosylated hemoglobin (HbA1c), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC), (2) serum insulin and serum and renal erythropoietin (EPO) levels, (3) serum creatinine and urea levels, (4) concentrations of glutathione (GSH), superoxide dismutase (SOD) and thiobarbituric acid reactive substances (TBARS) in serum, pancreatic and renal tissues, and (5) pancreas histopathology.STZ-injected rats showed injured pancreatic islets accompanied with persistent hyperglycemia, significant decreases in serum insulin, serum and renal EPO, RBCs counts and indices, PLTs counts, and increases in HbA1c, erythrocyte osmotic, WBCs counts, and serum creatinine and urea. However, therapy with