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Siberian wildrye (Elymus sibiricus) is a xero-mesophytic forage grass with high nutritional quality and stress tolerance. Among its numerous germplasm resources, some possess superior drought resistance. In this study, we firstly investigated the physiological differences between the leaves of drought-tolerant (DT) and drought-sensitive (DS) genotypes under different field water contents (FWC) in soil culture. The results showed that, under drought stress, DT maintained a lower leaf water potential for water absorption, sustained higher photosynthetic efficiency, and reduced oxidative damage in leaves by efficiently maintaining the ascorbic acid-glutathione (ASA-GSH) cycle to scavenge reactive oxygen species (ROS) compared to DS. Secondly, using RNA sequencing (RNA-seq), we analyzed the gene expression profiles of DT and DS leaves under osmotic stress of hydroponics induced by PEG-6000. Through differential analysis, we identified 1226 candidate unigenes, from which we subsequently screened out 115/212 differentially expressed genes (DEGs) that were more quickly induced/reduced in DT than in DS under osmotic stress. Among them, Unigene0005863 (EsSnRK2), Unigene0053902 (EsLRK10) and Unigene0031985 (EsCIPK5) may be involved in stomatal closure induced by abscisic acid (ABA) signaling pathway. Unigene0047636 (EsCER1) may positively regulates the synthesis of very-long-chain (VLC) alkanes in cuticular wax biosynthesis, influencing plant responses to abiotic stresses. Finally, the contents of wax and cutin were measured by GC–MS under osmotic stress of hydroponics induced by PEG-6000. Corresponding to RNA-seq, contents of wax monomers, especially alkanes and alcohols, showed significant induction by osmotic stress in DT but not in DS. It is suggested that limiting stomatal and cuticle transpiration under drought stress to maintain higher photosynthetic efficiency and water use efficiency (WUE) is one of the critical mechanisms that confer stronger drought resistance to DT. This study provides some insights into the molecular mechanisms underlying drought tolerance in E. sibiricus. The identified genes may provide a foundation for the selection and breeding of drought-tolerant crops. [ABSTRACT FROM AUTHOR]

Abstract Siberian wildrye (Elymus sibiricus) is a xero-mesophytic forage grass with high nutritional quality and stress tolerance. Among its numerous germplasm resources, some possess superior drought resistance. In this study, we firstly investigated the physiological differences between the leaves of drought-tolerant (DT) and drought-sensitive (DS) genotypes under different field water contents (FWC) in soil culture. The results showed that, under drought stress, DT maintained a lower leaf water potential for water absorption, sustained higher photosynthetic efficiency, and reduced oxidative damage in leaves by efficiently maintaining the ascorbic acid-glutathione (ASA-GSH) cycle to scavenge reactive oxygen species (ROS) compared to DS. Secondly, using RNA sequencing (RNA-seq), we analyzed the gene expression profiles of DT and DS leaves under osmotic stress of hydroponics induced by PEG-6000. Through differential analysis, we identified 1226 candidate unigenes, from which we subsequently screened out 115/212 differentially expressed genes (DEGs) that were more quickly induced/reduced in DT than in DS under osmotic stress. Among them, Unigene0005863 (EsSnRK2), Unigene0053902 (EsLRK10) and Unigene0031985 (EsCIPK5) may be involved in stomatal closure induced by abscisic acid (ABA) signaling pathway. Unigene0047636 (EsCER1) may positively regulates the synthesis of very-long-chain (VLC) alkanes in cuticular wax biosynthesis, influencing plant responses to abiotic stresses. Finally, the contents of wax and cutin were measured by GC–MS under osmotic stress of hydroponics induced by PEG-6000. Corresponding to RNA-seq, contents of wax monomers, especially alkanes and alcohols, showed significant induction by osmotic stress in DT but not in DS. It is suggested that limiting stomatal and cuticle transpiration under drought stress to maintain higher photosynthetic efficiency and water use efficiency (WUE) is one of the critical mechanisms that confer stronger drought resistance to DT. This study provides some insights into the molecular mechanisms underlying drought tolerance in E. sibiricus. The identified genes may provide a foundation for the selection and breeding of drought-tolerant crops.

Hereditary spherocytosis in cattle is a disorder resulting from a nonsense mutation in the SLC4A1 gene, which encodes band 3 membrane protein. This membranopathy leads to the production of spherically-shaped RBCs with reduced deformability and a shorter lifespan in circulation. Laboratory diagnosis of hereditary spherocytosis commonly relies on the erythrocyte osmotic fragility test (OFT), which is time- and labour-consuming and can be influenced by preanalytical factors. The aim of the study was to determine the effect of the preanalytical phase on the stability of osmotic fragility and morphological changes in bovine erythrocytes. Blood was collected from the tail vein of Holstein-Friesian cows (n = 20) into tubes with EDTA or lithium heparin. Erythrocyte resistance to haemolysis was measured by the osmotic fragility test immediately after blood collection and 24, 48 and 72 hours after storage at 4C or at room temperature. Thin blood smears were also prepared, stained and examined for cell morphology. The results showed that bovine erythrocyte osmotic fragility remains stable during 24-hour storage at 4C irrespective of the anticoagulant used. This suggests that OFT can be performed the day after blood collection if blood is stored at 4C. Bovine erythrocytes stored at room temperature were also found to have reduced osmotic resistance. However, heparin better preserves the osmotic resistance of bovine erythrocytes at room temperature. Erythrocytes undergo changes during storage. The microscopic examination of cell morphology revealed rapid transformation of bovine discocytes into various stages of echinocytes during the first three days of storage.

BACKGROUND: The triad "insulin resistance, prediabetes, diabetes" is three independent neologies with characteristic features and development. In addition, each are characterized by progression and the possibility of transition from one form to other. Due to the fact that diabetes is one of the common diseases associated with high rates of disability, it is necessary to improve diagnostic methods and educational regimens for successful prevention and treatment of the disease. OBJECTIVE: We investigated Band 3 protein (B3p) level, osmotic resistance of erythrocytes, the total antioxidant activity (TAA) of blood serum, level of HbA1 in group patients with insulin resistance (IR), prediabetes, and Type 2 Diabetes Mellitus (T2DM) and comparative with health control group. METHODS: We used original, accurate research methods that measure the essence of the studied quantities. RESULTS: Disruptions of glucose and insulin homeostasis ay lead to the initiation of oxidative stress (in our study demonstrated by a decrease of TAA of blood serum) increased redox-sensitive PTP activity and aberrant band 3 phosphorylation, potentially leading to reduced erythrocyte deformability. At the same time glycation of Hb during T2DM may affect its cross-link with membrane proteins, in particular with B3p, and although appears to limit its cross-linking and decrease its clusterization ability, induces alterations in the cytoskeletal matrix, and thereby decrease erythrocytes' osmotic resistance making them more susceptible to hemolysis. CONCLUSIONS: The osmotic resistance of the erythrocytes can be used as a sensitive marker for the detection of the early stages of hyperglycemia (prediabetes). This set of clinical trials will make it possible to identify diseases that make up the triad at an early stage. Early detection of disorders and continued research in this direction will help in the development of a diagnostic scheme for the prevention of such patients. Based on our data, research into anti-oxidation drugs is very important. With the help of the array of studies described in the article and antioxidant treatment, the likelihood of successful treatment will increase. [ABSTRACT FROM AUTHOR]

BACKGROUND: The triad "insulin resistance, prediabetes, diabetes" is three independent neologies with characteristic features and development. In addition, each are characterized by progression and the possibility of transition from one form to other. Due to the fact that diabetes is one of the common diseases associated with high rates of disability, it is necessary to improve diagnostic methods and educational regimens for successful prevention and treatment of the disease. OBJECTIVE: We investigated Band 3 protein (B3p) level, osmotic resistance of erythrocytes, the total antioxidant activity (TAA) of blood serum, level of HbA1 in group patients with insulin resistance (IR), prediabetes, and Type 2 Diabetes Mellitus (T2DM) and comparative with health control group. METHODS: We used original, accurate research methods that measure the essence of the studied quantities. RESULTS: Disruptions of glucose and insulin homeostasis ay lead to the initiation of oxidative stress (in our study demonstrated by a decrease of TAA of blood serum) increased redox-sensitive PTP activity and aberrant band 3 phosphorylation, potentially leading to reduced erythrocyte deformability. At the same time glycation of Hb during T2DM may affect its cross-link with membrane proteins, in particular with B3p, and although appears to limit its cross-linking and decrease its clusterization ability, induces alterations in the cytoskeletal matrix, and thereby decrease erythrocytes' osmotic resistance making them more susceptible to hemolysis. CONCLUSIONS: The osmotic resistance of the erythrocytes can be used as a sensitive marker for the detection of the early stages of hyperglycemia (prediabetes). This set of clinical trials will make it possible to identify diseases that make up the triad at an early stage. Early detection of disorders and continued research in this direction will help in the development of a diagnostic scheme for the prevention of such patients. Based on our data, research into anti-oxidation drugs is very important. With the help of the array of studies described in the article and antioxidant treatment, the likelihood of successful treatment will increase. [ABSTRACT FROM AUTHOR]

Hereditary spherocytosis in cattle is a disorder resulting from a nonsense mutation in the SLC4A1 gene, which encodes band 3 membrane protein. This membranopathy leads to the production of spherically-shaped RBCs with reduced deformability and a shorter lifespan in circulation. Laboratory diagnosis of hereditary spherocytosis commonly relies on the erythrocyte osmotic fragility test (OFT), which is time- and labour-consuming and can be influenced by preanalytical factors. The aim of the study was to determine the effect of the preanalytical phase on the stability of osmotic fragility and morphological changes in bovine erythrocytes. Blood was collected from the tail vein of Holstein-Friesian cows (n = 20) into tubes with EDTA or lithium heparin. Erythrocyte resistance to haemolysis was measured by the osmotic fragility test immediately after blood collection and 24, 48 and 72 hours after storage at 4℃ or at room temperature. Thin blood smears were also prepared, stained and examined for cell morphology. The results showed that bovine erythrocyte osmotic fragility remains stable during 24-hour storage at 4℃ irrespective of the anticoagulant used. This suggests that OFT can be performed the day after blood collection if blood is stored at 4℃. Bovine erythrocytes stored at room temperature were also found to have reduced osmotic resistance. However, heparin better preserves the osmotic resistance of bovine erythrocytes at room temperature. Erythrocytes undergo changes during storage. The microscopic examination of cell morphology revealed rapid transformation of bovine discocytes into various stages of echinocytes during the first three days of storage. [ABSTRACT FROM AUTHOR]

Nanoparticles are extremely promising components that are used in diagnostics and medical therapies. Among them, silica nanoparticles are ultrafine materials that, due to their unique physicochemical properties, have already been used in biomedicine, for instance, in cancer therapy. The aim of this study was to investigate the cytotoxicity of three types of nanoparticles ( SiO 2 , SiO 2 - SH , and SiO 2 - COOH ) in relation to red blood cells, as well as the impact of silicon dioxide nanoparticles on biological membranes and liposome models of membranes. The results obtained prove that hemolytic toxicity depends on the concentration of nanoparticles and the incubation period. Silica nanoparticles have a marginal impact on the changes in the osmotic resistance of erythrocytes, except for SiO 2 - COOH , which, similarly to SiO 2 and SiO 2 - SH , changes the shape of erythrocytes from discocytes mainly towards echinocytes. What is more, nanosilica has an impact on the change in fluidity of biological and model membranes. The research gives a new view of the practical possibilities for the use of large-grain nanoparticles in biomedicine. [ABSTRACT FROM AUTHOR]

Oxidative stress and multiple erythrocyte abnormalities have been observed in hypertension. We focused on the effects of angiotensin-converting enzyme 2 (ACE2) inhibition by MLN-4760 inhibitor on angiotensin peptides, oxidative stress parameters, and selected erythrocyte quality markers in spontaneously hypertensive rats (SHR). We also investigated the potential effects of polyphenolic antioxidant taxifolin when applied in vivo and in vitro following its incubation with erythrocytes. SHRs were divided into four groups: control, taxifolin-treated, MLN-4760-treated, and MLN-4760 with taxifolin. MLN-4760 administration increased the blood pressure rise independent of taxifolin treatment, whereas taxifolin decreased it in control SHRs. Body weight gain was also higher in ACE2-inhibited animals and normalized after taxifolin treatment. However, taxifolin did not induce any change in angiotensin peptide concentrations nor a clear antioxidant effect. We documented an increase in Na,K-ATPase enzyme activity in erythrocyte membranes of ACE2-inhibited SHRs after taxifolin treatment. In conclusion, ACE2 inhibition deteriorated some selected RBC properties in SHRs. Although taxifolin treatment did not improve oxidative stress markers, our data confirmed the blood pressure-lowering potential, anti-obesogenic effect, and some "erythroprotective" effects of this compound in both control and ACE2-inhibited SHRs. In vitro investigations documenting different effects of taxifolin on erythrocyte properties from control and ACE2-inhibited SHRs accentuated the irreplaceability of in vivo studies. [ABSTRACT FROM AUTHOR]

Semen cryopreservation is a crucial part of assisted reproductive techniques (ART) in animals, and recently it is gaining more and more attention among cat breeders. Even if fresh semen quality is good, sometimes spermatozoa do not survive freezing. The freezability prediction was widely studied in many species, but not in the domestic cat. The aim of this study was to verify the usefulness of osmotic challenge tests and membrane structure markers (Yo-Pro 1 and Merocyanine 540) for the prediction of the quality of post-thawed feline semen. Semen was collected by urethral catheterization from 22 male cats. After a basic evaluation of semen, 20×106 spermatozoa were cryopreserved; the rest were evaluated by flow cytometry for membrane integrity (SYBR-14/PI), acrosome status (lectin PNA/PI), mitochondrial potential (JC-1) and membrane stability (Yo-Pro 1/M540 staining). Hypo- and hyperosmotic challenge tests were also performed. The thawed samples were evaluated as fresh ones. The Pearson correlation between all parameters in fresh semen and all parameters in cryopreserved spermatozoa was assessed. Although some moderate correlations were found between the results of the osmotic tests and markers of sperm membrane stability (Yo-Pro 1 and Merocyanine 540) and post-thaw semen quality parameters, the predictive value of studied markers was rather weak – no cut-off values could be established and, based on regression models, they explained less than 40 % of variability in post-thaw quality. Our results confirm that cryodamage is a complex matter, in which many different factors play a role, affecting sperm motility and membrane integrity differently. ● Cryopreservation differently affects sperm motility and membrane integrity. ● Post-thaw viability is mostly related to initial sperm membrane characteristics. ● Post-thaw motility is moderately correlated with several different parameters. ● Acrosome damage >10 % can be regarded as an unfavorable prognostic factor. [ABSTRACT FROM AUTHOR]

The aim of the experiment was to highlight the toxicity of the CrVI, administered during three generations, on the haemoglobin and on the osmotic resistance of the erythrocyte membrane in female rats. The determinations have been carried out on rat blood collected from female rats that reached sexual maturity, from F0, F1, F2 generation, exposed to LOAEL, of CrVI, as potassium dichromate, in drinking water (3 months). F1 and F2 were obtained from female rats from generation F0 and respectively F1 exposed for 3 months to chromium dose, mated with male rats exposed to the same dose for three months, before mating. The results of the experiment indicated a high significant decrease of haemoglobin, (p

The present study aimed to evaluate the effects of propolis-iron oxide nanoparticles in eliminating the T. evansi parasite and rehome stasis of deleterious iron status in experimentally infected rabbits. Twenty male rabbits were divided into equal four groups (n=5). the 1st group as Control negative, 2nd control positive, 3rd trypanosomiasis and treated with propolis iron oxide nanoparticles, and 4th trypanosomiasis and treated with diminazene,2nd ,3rd, and 4th groups were inoculated with T. evansi, and were checked for the onset of parasitemia. After 15 of the onset of parasitemia 3rd group was treated with propolis- iron-oxide nanoparticles 30 mg iron /kg BW, and 4th group was treated with diminazene drug with a single dose 3.5 mg/kg BW. The result showed that experimentally infection with T. evansi caused a significant decrease of serum iron and ferritin and a significant increase in total iron-binding capacity and unsaturation iron-binding capacity, as well erythrocytes fragility, bilirubin totally and partially. Treatment with propolis-iron oxide nanoparticles improved iron status parameters to semi-normal values much better than diminazene drug, in addition, reduced the total bilirubin concentration and osmotic fragility of erythrocytes toward a normal state. It can be concluded that the propolis-iron oxide nanoparticles proved successfully rebalancing iron status and eliminating the parasite and making iron available.

Nanoparticles are extremely promising components that are used in diagnostics and medical therapies. Among them, silica nanoparticles are ultrafine materials that, due to their unique physicochemical properties, have already been used in biomedicine, for instance, in cancer therapy. The aim of this study was to investigate the cytotoxicity of three types of nanoparticles (SiO2, SiO2-SH, and SiO2-COOH) in relation to red blood cells, as well as the impact of silicon dioxide nanoparticles on biological membranes and liposome models of membranes. The results obtained prove that hemolytic toxicity depends on the concentration of nanoparticles and the incubation period. Silica nanoparticles have a marginal impact on the changes in the osmotic resistance of erythrocytes, except for SiO2-COOH, which, similarly to SiO2 and SiO2-SH, changes the shape of erythrocytes from discocytes mainly towards echinocytes. What is more, nanosilica has an impact on the change in fluidity of biological and model membranes. The research gives a new view of the practical possibilities for the use of large-grain nanoparticles in biomedicine.

Cultured red blood cells from human induced pluripotent stem cells (cRBC_iPSCs) are a promising source for future concepts in transfusion medicine. Before cRBC_iPSCs will have entrance into clinical or laboratory use, their functional properties and safety have to be carefully validated. Due to the limitations of established culture systems, such studies are still missing. Improved erythropoiesis in a recently established culture system, closer simulating the physiological niche, enabled us to conduct functional characterization of enucleated cRBC_iPSCs with a focus on membrane properties. Morphology and maturation stage of cRBC_iPSCs were closer to native reticulocytes (nRETs) than to native red blood cells (nRBCs). Whereas osmotic resistance of cRBC_iPSCs was similar to nRETs, their deformability was slightly impaired. Since no obvious alterations in membrane morphology, lipid composition, and major membrane associated protein patterns were observed, reduced deformability might be caused by a more primitive nature of cRBC_iPSCs comparable to human embryonic- or fetal liver erythropoiesis. Blood group phenotyping of cRBC_iPSCs further confirmed the potency of cRBC_iPSCs as a prospective device in pre-transfusional routine diagnostics. Therefore, RBC membrane analyses obtained in this study underscore the overall prospects of cRBC_iPSCs for their future application in the field of transfusion medicine. [ABSTRACT FROM AUTHOR]

The damage to erythrocytes during cardiopulmonary bypass (CPB) remains a recent problem. The aim of this research was to study the effect of fructose-1,6-diphosphate on the state of the erythrocyte membrane during CPB and the level of phosphorus in blood as a marker of the energy potential in the cell. Patients were divided into two groups. The control group 1 (Gr 1) consisted of 75 individuals. The group 2 (Gr 2) included patients to whom fructose-1,6-diphosphate (FDP) was administrated according to the developed scheme as follows 10 g of the drug was diluted in 50 ml of a solvent, 5 g of the drug was injected intravenously with the use of perfusor immediately before initiation of CPB at a rate of 10 ml/min and 5 g at the 30th minute of CPB (before the stage of warming) the same way. When comparing two groups the best results in hemolysis (p

Extracellular polymeric substances (EPSs) in excess sludge of wastewater treatment plants are valuable biopolymers that can act as recovery materials. However, effectively concentrating EPSs consumes a significant amount of energy. This study employed novel energy-saving pressure-free dead-end forward osmosis (DEFO) technology to concentrate various biopolymers, including EPSs and model biopolymers [sodium alginate (SA), bovine serum albumin (BSA), and a mixture of both (denoted as BSA-SA)]. The feasibility of the DEFO technology was proven and the largest concentration ratios for these biopolymers were 94.8 % for EPSs, 97.1 % for SA, 97.8 % for BSA, and 98.4 % for BSA-SA solutions. An evaluation model was proposed, incorporating the FO membrane's water permeability coefficient and the concentrated substances' osmotic resistance, to describe biopolymers' concentration properties. Irrespective of biopolymer type, the water permeability coefficient decreased with increasing osmotic pressure, remained constant with increasing feed solution (FS) concentration, increased with increasing crossing velocity in the draw side, and showed little dependence on draw salt type. In the EPS DEFO concentration process, osmotic resistance was minimally impacted by osmotic pressure, FS concentration, and crossing velocity, and monovalent metal salts were proposed as draw solutes. The interaction between reverse diffusion metal cations and EPSs affected the structure of the concentrated substances on the FO membrane, thus changing the osmotic resistance in the DEFO process. These findings offer insights into the efficient concentration of biopolymers using DEFO. • DEFO has high concentration efficiency for biopolymers. • FO membrane permeability and concentrate osmotic resistance model proposed • BSA and SA have coordinated pollution effects increasing osmotic resistance. • Osmotic resistance of EPS is little affected by various factors. • Reverse diffusion metal cation affects concentrated substances' structure. [ABSTRACT FROM AUTHOR]

Trehalose is a non-reducing disaccharide with high stability and strong water absorption properties that can improve the resistance of organisms to various abiotic stresses. Trehalose-6-phosphate synthase (TPS) plays important roles in trehalose metabolism and signaling. In this study, the full-length cDNA of ThTPS was cloned from Tamarix hispida Willd. A phylogenetic tree including ThTPS and 11 AtTPS genes from Arabidopsis indicated that the ThTPS protein had a close evolutionary relationship with AtTPS7. However, the function of AtTPS7 has not been determined. To analyze the abiotic stress tolerance function of ThTPS, the expression of ThTPS in T. hispida under salt and drought stress and JA, ABA and GA3 hormone stimulation was monitored by qRT-PCR. The results show that ThTPS expression was clearly induced by all five of these treatments at one or more times, and salt stress caused particularly strong induction of ThTPS in the roots of T. hispida. The ThTPS gene was transiently overexpressed in T. hispida. Both physiological indexes and staining results showed that ThTPS gene overexpression increased salt and osmotic stress tolerance in T. hispida. Overall, the ThTPS gene can respond to abiotic stresses such as salt and drought, and its overexpression can significantly improve salt and osmotic tolerance. These findings establish a foundation to better understand the responses of TPS genes to abiotic stress in plants. [ABSTRACT FROM AUTHOR]

The effect of local cold injury in rats on the osmotic resistance of erythrocytes in the early and late reactive terms was investigated. Cryoinfluence was performed by pressing the applicator to the skin on the lateral surface of the rat thigh for 30, 60 and 120 s. Afterwards, the blood was collected from the aorta of animals after 1 and 4 hours and after 24 and 48 hours (early and late reactive periods, respectively). The effect of hypotonic solutions on cells was used as a test for osmotic resistance of erythrocytes. It is shown that the osmotic resistance of erythrocytes in the early reactive term increases (compared with control cells). In the late reactive term, the indices of osmotic resistance of erythrocytes after 24 hours approached the control values and after 48 hours they reached the norm. Osmotic resistance of erythrocytes in both reactive terms after local cold injury does not depend on the duration of cryoinfluence (30, 60, 120 s). The findings are considered from the standpoint of adaptive reactions that have an allostatic nature. [ABSTRACT FROM AUTHOR]

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.

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.

The damage to erythrocytes during cardiopulmonary bypass (CPB) remains a recent problem. The aim of this research was to study the effect of fructose-1,6-diphosphate on the state of the erythrocyte membrane during CPB and the level of phosphorus in blood as a marker of the energy potential in the cell. Patients were divided into two groups. The control group 1 (Gr 1) consisted of 75 individuals. The group 2 (Gr 2) included patients to whom fructose-1,6-diphosphate (FDP) was administrated according to the developed scheme as follows 10 g of the drug was diluted in 50 ml of a solvent, 5 g of the drug was injected intravenously with the use of perfusor immediately before initiation of CPB at a rate of 10 ml/min and 5 g at the 30th minute of CPB (before the stage of warming) the same way. When comparing two groups the best results in hemolysis (p<0.01), mechanical (p<0.01). osmotic resistance of erythrocytes (p<0.01), the time of acid hemolysis (p<0.01) and the permeability of the erythrocyte membrane in postperfusion period were in Gr 2. Вefore cardiac surgery hypophosphatemia was detected in 18% out of 150 and in 32% out of 150 patients – a lower limit of normal phosphorus content in the blood. After CPB in Gr 1 phosphorus content in blood was 0.85±0.32 mmol/l and hypophosphatemia was in 53% out of 75 patients. This indicates a pronounced energy deficit in this group. In Gr 2 phosphorus level was 1.7±0.31 mmol/l and there was no hypophosphatemia. As a result, FDP as an endogenous high-energy intermediate metabolite of the glycolytic pathway leads to resistance to hemolysis, protects the erythrocyte membrane from damage and increases the energy potential of the cell during CPB. [ABSTRACT FROM AUTHOR]

The osmotolerant Zygosaccharomyces rouxii is known for its trait to survive in extreme high sugar environments. This ability determines its role in the fermentation process and leads to yeast spoilage in the food industry. However, our knowledge of the gene expression in response to high sugar stress remains limited. Here, we conducted RNA-sequencing (RNA-seq) under different sugar concentrations of the spoilage yeast, Z. rouxii, which exhibit extremely high tolerance to sugar stress. The obtained differentially expressed genes (DEGs) are significantly different to that of the Saccharomyces cerevisiae, which is sensitive to extreme high sugar stress. Most of the DEGs participated in the “glucan synthesis,” “transmembrane transport,” “ribosome,” etc. In this work, we also demonstrated that the gene ZYRO0B03476g (ZrKAR2) encoding Kar2p can significantly affect the growth of Z. rouxii under high sugar stress. In addition, we combined with a previous study on the genome sequence of Z. rouxii, indicating that several gene families contain significantly more gene copies in the Z. rouxii lineage, which involved in tolerance to sugar stress. Our results provide a gene insight for understanding the high sugar tolerance trait, which may impact food and biotechnological industries and improve the osmotolerance in other organisms.

Objective: to carry out a comparative study of structural and functional responses of rats and hamsters RBCs when the animals entered and left an artificial hypobiosis state under conditions of hypothermia-hypoxia-hypercapnia. Methods: RBCs of rats and hamsters: control, in artificial hypobiotic state, 2 and 24 h post-hypobiosis state, and those from winter-hibernating hamsters were investigated. Osmotic resistance was determined by the method of small-angle light scattering. Relative content of main hemoglobin forms (oxy-, deoxy- and metHb) was determined by the method of differential spectrophotometry. Microviscosity of cytosol was evaluated within a range of 37-0°C by EPR spin probe method. Results: the artificial hypobiotic state was characterized by body temperature decreasing (down to 16°C), in the same way for both homoiothermal and heterothermal mammals. This was accompanied by changes of osmotic resistance, redistribution of main hemoglobin forms, cytosol microviscosity in RBCs. Increased osmotic resistance and significantly elevated relative content of oxyHb were revealed in 2 h after hypobiotic state. Modifications of the parameters kept changing up to 24 h post-hypobiotic-state. RBC responses to natural and artificial hypobiosis had common as well as different features. Difference in the reaction of RBC cytosol of homoiotherms and heterotherms, associated with the season, was found. Conclusions: RBCs response in vivo to physiological rearrangements induced by artificial hypobiosis of both heterotherm and homoiotherm mammals. The significant changes of the structural and functional state of erythrocyte kept changing up to 24 h post-hypobiotic-state, while the physiological state of the animals was similar to the control already in 2 hours after hypobiosis. Existence of mammals RBC “aftersensations” in posthypobiotic state allows to consider the model of artificial hypobiosis perspective for clarification of the cellular mechanisms of controlled induction of hypometabolic state.

The osmotolerant Zygosaccharomyces rouxii is known for its trait to survive in extreme high sugar environments. This ability determines its role in the fermentation process and leads to yeast spoilage in the food industry. However, our knowledge of the gene expression in response to high sugar stress remains limited. Here, we conducted RNA-sequencing (RNA-seq) under different sugar concentrations of the spoilage yeast, Z. rouxii , which exhibit extremely high tolerance to sugar stress. The obtained differentially expressed genes (DEGs) are significantly different to that of the Saccharomyces cerevisiae , which is sensitive to extreme high sugar stress. Most of the DEGs participated in the "glucan synthesis," "transmembrane transport," "ribosome," etc. In this work, we also demonstrated that the gene ZYRO0B03476g (ZrKAR2) encoding Kar2p can significantly affect the growth of Z. rouxii under high sugar stress. In addition, we combined with a previous study on the genome sequence of Z. rouxii , indicating that several gene families contain significantly more gene copies in the Z. rouxii lineage, which involved in tolerance to sugar stress. Our results provide a gene insight for understanding the high sugar tolerance trait, which may impact food and biotechnological industries and improve the osmotolerance in other organisms. [ABSTRACT FROM AUTHOR]

We studied the morphological features of the restructuring and osmotic stability of erythrocytes of 30 people with mental disorders and disorders of behavioural reactions caused by 5-7 years consumption of alcohol, the objects of the research being in a state of abstinence during the period of the investigation,. The study of quantitative composition of red blood cells and hemoglobin, as well as their ratio in people with mental and behavioural disorders developed as a result of alcohol consumption showed a decrease in the number of red blood cells and hemoglobin of 42.2% and 25.1%, respectively, indicating the presence in those people of anemic hypoxia. This is closely correlated with a decrease of osmotic resistance of red blood cells to different concentrations of sodium chloride solutions, whereby the state of erythrocyte membranes deteriorates and their resistance to hemolysis decreases. Parallel to this, is a reduction in the size of red blood cells coupled with a variety of changes in their microrelief. Conformational changes of erythrocytes are closely correlated with changes in the activity of enzymes responsible for the biotransformation of xenobiotics, primarily products of lipid peroxidation. It is shown that activity of the erythrocyte antioxidant defense system is sharply reduced as a result of increase in partially oxidized products of metabolism. It was found that the state of red blood cells is a sensitive indicator of the normal course of physiological changes, biochemical and biophysical processes in the bodies of people with mental disorders and behaviour reactions developed due to consumption of alcohol.At the same time, the measurement of the osmotic resistance of red blood cells and changes in the shape of red blood cells connected with this phenomenon is an important research method for studying mechanisms of pathological processes and the action of certain drugs or biologically active substances, expands the possibilities of establishing the depth of destruction of body tissues, and also provides excellent opportunities for researchers to identify the effectiveness of treatment and prevention for people with mental disorders and behaviour reactions caused by consumption of alcohol. This article discusses prospects of studies of the level of glutathione peroxidase activity in the erythrocytes among people with mental and behavioural disorders as a marker of highly specific homeostasis in the human body in response to long-term alcohol consumption, and considers using this index as a criterion for selecting patients for different groups of physical rehabilitation and recreation according to the level of rehabilitation potential.

Background: The triad "insulin resistance, prediabetes, diabetes" is three independent neologies with characteristic features and development. In addition, each are characterized by progression and the possibility of transition from one form to other. Due to the fact that diabetes is one of the common diseases associated with high rates of disability, it is necessary to improve diagnostic methods and educational regimens for successful prevention and treatment of the disease., Objective: We investigated Band 3 protein (B3p) level, osmotic resistance of erythrocytes, the total antioxidant activity (TAA) of blood serum, level of HbA1 in group patients with insulin resistance (IR), prediabetes, and Type 2 Diabetes Mellitus (T2DM) and comparative with health control group., Methods: We used original, accurate research methods that measure the essence of the studied quantities., Results: Disruptions of glucose and insulin homeostasis ay lead to the initiation of oxidative stress (in our study demonstrated by a decrease of TAA of blood serum) increased redox-sensitive PTP activity and aberrant band 3 phosphorylation, potentially leading to reduced erythrocyte deformability. At the same time glycation of Hb during T2DM may affect its cross-link with membrane proteins, in particular with B3p, and although appears to limit its cross-linking and decrease its clusterization ability, induces alterations in the cytoskeletal matrix, and thereby decrease erythrocytes' osmotic resistance making them more susceptible to hemolysis., Conclusions: The osmotic resistance of the erythrocytes can be used as a sensitive marker for the detection of the early stages of hyperglycemia (prediabetes). This set of clinical trials will make it possible to identify diseases that make up the triad at an early stage. Early detection of disorders and continued research in this direction will help in the development of a diagnostic scheme for the prevention of such patients. Based on our data, research into anti-oxidation drugs is very important. With the help of the array of studies described in the article and antioxidant treatment, the likelihood of successful treatment will increase.

Uncaria tomentosa (Willd.) DC is a woody climber species originating from South and Central America that has been used in the therapy of asthma, rheumatism, hypertension, and blood purification. Our previous study showed that U. tomentosa extracts altered human erythrocyte shape, which could be due to incorporation of the compounds contained in extracts into the erythrocyte membrane. The aim of the present study was to determine how the compounds contained in U. tomentosa extracts incorporate into the human erythrocyte membrane. The study has assessed the effect of aqueous and ethanolic extracts from leaves and bark of U. tomentosa on the osmotic resistance of the human erythrocyte, the viscosity of erythrocyte interior, and the fluidity of erythrocyte plasma membrane. Human erythrocytes were incubated with the studied extracts in the concentrations of 100, 250, and 500 µg/mL for 2, 5, and 24 h. All extracts tested caused a decrease in erythrocyte membrane fluidity and increased erythrocyte osmotic sensitivity. The ethanolic extracts from the bark and leaves increased viscosity of the erythrocytes. The largest changes in the studied parameters were observed in the cells incubated with bark ethanolic extract. We consider that the compounds from U. tomentosa extracts mainly build into the outer, hydrophilic monolayer of the erythrocyte membrane, thus protecting the erythrocytes against the adverse effects of oxidative stress.

The problem of iron deficiency is urgent in many countries, regardless of the socio-economic standards of living. Among biomedical problems, the study of biochemical blood parameters is of paramount importance. A great interest in various aspects of this issue is not accidental, since the transport function of the blood largely depends on it and, consequently, the efficient delivery of oxygen, glucose, amino acids, fats, vitamins, mineral salts, hormones, mediators, antibodies and metabolites to the micro-zones in the tissues. The purpose of this article is to study the hematological status and the state of the red blood cell membrane in women of reproductive age suffering from iron deficiency anemia (IDA). Iron deficiency anemia (IDA) is a combination of clinical and hematological symptoms characterized by impaired hemoglobin formation due to iron deficiency in serum and bone marrow, as well as the development of trophic disorders in the organs and tissues. During the study, a complete blood count was performed using an enzyme-linked immunosorbent assay, using the kits produced by JSC Vector-Best, and the Kamyshnikov method. Based on the results of the study of the erythrocyte sorption capacity (ESC) in women of reproductive age with IDA, it was established that this parameter is 1.2 times higher than that in the control group. The development of anemia is associated with significant changes in the osmotic stability of the erythrocytes, which results in an increase in the proportion of both unstable and high-resistance erythrocytes. The scientific novelty of the article is that the authors established the key parameters for iron deficiency anemia detection. [ABSTRACT FROM AUTHOR]

Long-term high fat-carbohydrates diet (HF-CD) contributes to the formation of irreversible changes in the organism that lead to the emergence of civilization diseases. In this study, the impact of three-month high-fat diet on the physical properties of erythrocytes (RBCs) was studied. Furthermore, the biological activity of Cistus incanus L. extracts, plant known with high pro-health potential, in relation to normal and HF-CD RBCs, was determined. Obtained results have shown that, applied HF-CD modified shape, membrane potential and osmotic resistance of erythrocytes causing changes in membrane lipid composition and the distribution of lipids. The impact of HF-CD on physical properties of RBCs along with atherosclerotic lesions of the artery was visible, despite the lack of statistically significant changes in blood morphology and plasma lipid profile. This suggests that erythrocytes may be good markers of obesity-related diseases. The studies of biological activity of Cistus incanus L. extracts have demonstrated that they may ameliorate the effect of HF-CD on erythrocytes through the membrane-modifying and antioxidant activity.

Ethylene-responsive factors (ERFs) comprise a large family of transcription factors in plants and play important roles in developmental processes and stress responses. Here, we characterized a novel AP2/ERF transcription factor, AcERF2, from the halophyte Atriplex canescens (four-wing saltbush, Chenopodiaceae ). AcERF2 was proved to be a transcriptional activator in yeast and localized to the nucleus upon transient expression in Nicotiana benthamiana, indicating its potential role as a transcription factor. Overexpression of AcERF2 driven by a CaMV35S promoter led to decreased accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), and increased antioxidant enzymatic activities, as well as rapid stomatal closure under osmotic treatment in Arabidopsis . Arabidopsis plants overexpressing AcERF2 were hypersensitive to abscisic acid (ABA) during germination, seedling establishment, and primary root elongation, and exhibited significant tolerance to osmotic stress. Furthermore, overexpression of AcERF2 induced transcript accumulation of plant defense-related genes ( PR1 , PR2 , PR5 , ERF1 and ERF3 ) and increased Arabidopsis resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 and the necrotrophic fungal pathogen Botrytis cinerea. These results suggest that AcERF2 may play a positive modulation role in response to osmotic stress and pathogen infection in plants. [ABSTRACT FROM AUTHOR]

During ejaculation and the deposition in the female genital tract, spermatozoa undergo hypo-osmotic stress and need to withstand it for optimal fertility. Resistance to hypo-osmotic stress may be affected by the interaction of the spermatozoa with seminal fluid components. The hypo-osmotic resistance of epididymal and ejaculated spermatozoa from dogs, rams and boars was assessed by flow cytometric measurement of sperm viability after incubation in NaCl solutions with osmolalities ranging from 0 to 300 mmol/kg. The hypotonic resistance of epididymal spermatozoa was greater than those of ejaculated spermatozoa in all three species. Among species comparison revealed that ejaculated spermatozoa from dogs were much more resistant than those from rams and boars as 80.4 ± 5.3%, 56.7 ± 4.7 and 9.6 ± 3.6% of live spermatozoa were observed following exposure to an osmolality of 90 mmol/kg in dogs, rams and boars respectively. This can be explained by the fact that dog, ram and boar differ markedly in composition of the seminal plasma owing to the presence (ram, boar) or absence (dog) of seminal vesicles. Hypotonic resistance of epididymal and ejaculated dog spermatozoa was similar whereas ram and boar spermatozoa showed a marked drop in resistance after ejaculation. The in vitro incubation of boar epididymal spermatozoa with raw seminal plasma or the seminal plasma protein fraction induced a similar loss of resistance, suggesting that seminal proteins are involved in the lack of resistance to hypotonic stress of boar ejaculated spermatozoa. [ABSTRACT FROM AUTHOR]

Corbicula fluminea arrived in the Miño Estuary in 1989 and, from there, colonized more than 150 km upstream. Our aim was to test the capacity of C. fluminea to cope with osmotic stress conditions previously to invade new freshwater habitats through estuaries. Based on previously collected information, the experiment aims to study the response of the species to marine osmotic stress, evaluated by survival and behaviour. Experiments determined the resistance by the species to various levels of osmotic stress, and recovery time after exposure to high salinity levels, representative of the temporal and spatial salinity variation existing in the estuary. Under osmotic stress the semi-maximum response was reached after 19 days exposure. The species tolerance range, measured by individual maintained activity, was at salinity ∼20 when exposed to winter temperatures, while when animals were exposed to summer ones its tolerance was reduced to salinity lower than 15. C. fluminea show a large physiological flexibility to cope with salinity variations in estuaries. In summer, the temperature increases the metabolic rate thus making the species more vulnerable to osmotic stress exposure. These findings are relevant to preventing new invasions through ship ballast waters ensuring complete mortality if individuals are retained for >26 days. [ABSTRACT FROM AUTHOR]

Background Hypophosphatemia in early lactating dairy cows has been implicated as primary cause for postparturient hemoglobinuria in cattle. Decreased availability of phosphorus has been proposed to reduce adenosine triphosphate synthesis of erythrocytes and thereby reduce osmotic resistance of these cells. Hypothesis/Objectives To study the effect of phosphorus depletion on the phosphate concentration ([Pi]) in plasma and erythrocytes and the osmotic resistance of erythrocytes and to determine the association between plasma [Pi] and erythrocyte [Pi]. Animals Ten healthy midlactating dairy cows in their 3rd to 5th lactation. Methods Prospective study. Dietary phosphorus depletion for 5 weeks followed by phosphorus supplementation. Plasma and erythrocyte [Pi] and erythrocyte osmotic resistance were measured. Four cows underwent continuous dextrose infusion at the end of phosphate depletion to exacerbate hypophosphatemia. Results Dietary P depletion resulted in a marked decline of the plasma [Pi] from 4.1 ± 1.0 mg/dL to a nadir of 1.7 ± 0,5 mg/dL, but did not alter erythrocyte [Pi] or osmotic resistance. Similarly, dextrose infusion induced a decline of the plasma [Pi] from 2.4 ± 0.5 mg/dL to 1.5 ± 0.5 mg/dL, but had no effect on erythrocyte [Pi] or osmotic resistance. Conclusions and Clinical Importance In cattle, marked hypophosphatemia induced by dietary P depletion was neither associated with a decline in erythrocyte [Pi] nor with decreased osmotic resistance of erythrocytes. Phosphorus depletion alone is therefore unlikely to cause intravascular hemolysis and the plasma [Pi] is an unreliable index for the intracellular [Pi] of erythrocytes. [ABSTRACT FROM AUTHOR]

The fundamentals of a photometry method for determination of membrane permeability of some fish spermatozoa for water molecules are presented. Osmotic tolerance of carp spermatozoa membranes was studied using EPR-spectroscopy and photometric analysis methods. It was shown that carp spermatozoa look like the ideal osmometers in their reaction on media of different osmolarity. The value of membrane permeability of carp spermatozoa for water molecules was determined. Data obtained can be used in cryobiology for creating cryoprotective media and regimes of fish sperm cryopreservation. [ABSTRACT FROM AUTHOR]

Erythrocyte osmotic fragility (EOF) was carried out in nineteen dogs naturally infected by Leptospira interrogans serovar icterohaemorrhagiae/copenhagi. A decreased EOF was observed, suggesting a modification of erythrocyte components secondary to disturbances that occur during canine leptospirosis, such as renal damage and hepatic disease.A fragilidade osmótica eritrocitária foi estudada em dezenove cães infectados naturalmente pela Leptospira interrogans serovar icterohaemorrhagiae/copenhagi. Observou-se uma redução da fragilidade osmótica eritrocitária, sem a presença de anemia, possivelmente relacionada aos distúrbios hepato-renais que ocorrem nesta patologia.

Neste trabalho descreve-se a técnica do teste de resistência osmótica modificado (SORT modificado) e sua eficiência na avaliação da integridade da membrana do acrossoma do espermatozóide do varrão. Modificaram-se as soluções usadas nos testes de resistência osmótica atuais, eliminando-se também a incubação em solução isosmótica (300 mOsml). Esta foi substituída pela percentagem de NAR das amostras de morfologia espermática.

Objectives Massive transfusion ( MT) can cause severe electrolyte and acid-base disturbances in neonates and infants due to the unphysiological composition of packed red blood cells ( PRBCs). Washing of the PRBCs using Cell Saver systems prior to MT is recommended for this reason. Aim The composition of normal saline (NaCl), the standard wash fluid for Cell Saver systems, is considerably different from that of physiological plasma. The aim of the study presented here was to investigate the effect of washing the PRBCs with a bicarbonate-buffered hemofiltration solution ( BB- HS) in comparison with washing with NaCl and to evaluate the impact on electrolyte concentrations, acid-base balance and the stability of PRBCs. Methods In an experimental in vitro setting, PRBCs were washed with Cell Saver systems prepared with NaCl or BB- HS as washing solutions. Before and after the washing procedure, electrolyte concentrations, acid-base parameters, adenosine triphosphate ( ATP) and free hemoglobin (fHb) concentrations were measured. Results In both groups, the potassium concentrations decreased (baseline: 18.4 ± 5.17 mmol·l−1, end of study: NaCl 2.71 ± 1,81 mmol·l−1, BB- HS 2.50 ± 1.54 mmol·l−1, P < 0.05) while the acid-base balance improved only in the BB- HS-group (baseline: base excess −21.6 ± 3.52 mmol·l−1, end of study: NaCl −30.2 ± 1.42 mmol·l−1, BB- HS −7.51 ± 2.49 mmol·l−1, P < 0.05). Furthermore, markers of erythrocyte stability such as fHb and ATP concentrations were improved in the BB- HS-group. Conclusions Washing of PRBCs with BB- HS rather than NaCl results in a more physiological composition with improvements of electrolyte concentrations, acid-base balance and erythrocyte stability. [ABSTRACT FROM AUTHOR]