Peripheral Blood Erythrocytes Research Articles

Clinical and laboratory findings characterizing the need for systemic corticosteroids and nonsteroidal systemic therapies, and the predicted outcomes in cutaneous polyarteritis nodosa: A single-centre retrospective analysis

Abstract Background Cutaneous polyarteritis nodosa (cPN) is a necrotizing arteritis of medium-sized vessels limited to the skin. Because of its rarity and the diversity of its clinical manifestations, there is no consensus treatment. Moreover, there are no established indicators that predict disease severity or its outcome. Objectives To investigate clinico-laboratory features that predict patients requiring systemic therapy, including corticosteroids, to control the disease activity. Methods Thirty-six cPN patients who had not received systemic corticosteroids at the initial visit were retrospectively analysed by correlating the treatment and its response with clinico-laboratory findings. Results The major medications administered were antiplatelet agents (63.9%), vasodilators (38.9%), and prednisolone (PSL) (36.1%). In all, 23 cases achieved remission without PSL; 5 were managed with compression therapy alone or even observation; 18 received antiplatelet monotherapy or combined with vasodilator/dapsone; 13 required PSL; 10 achieved remission with PSL monotherapy or PSL and single/multiple medications and 3 with PSL and multiple drugs failed to achieve remission and underwent limb amputation. There were more skin ulcers and an elevated peripheral white blood cell (WBC) count and erythrocyte sedimentation rate (ESR) before corticosteroid induction in patients requiring PSL. Three cases with treatment failure had a markedly elevated ESR (>50). Conclusions More than half of cPN can achieve remission without corticosteroids; an elevated WBC and the presence of skin ulcers predict the need for PSL; a high ESR before corticosteroid induction predicts treatment resistance, even with PSL.

The relationship between disease activity with pan-immune-inflammatory value and systemic immune-inflammation index in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disease. Immune system cells have an important role in RA. Our aim was to investigate the relationship between disease activity, systemic immune-inflammation index (SII), and pan-immune-inflammation value (PIV) levels in RA patients. We planned to investigate whether these 2 measurements have an advantage over each other. About 67 patients diagnosed with RA and 49 healthy controls included in this study. RA was diagnosed based on 2010 ACR classification criteria. In this cross-sectional study, peripheral blood tests, C-reactive protein (CRP), hemogram, and erythrocyte sedimentation rate levels were noted after the physical examination of all participants. PIV was calculated with the formula: (neutrophil count × platelet count × monocyte count) / lymphocyte count. SII was calculated as follows: (neutrophil count × monocytes count) / lymphocyte count. The disease activity score 28 (DAS28) were noted in patients with RA. CRP values of active RA group were significantly higher than remission RA and control groups (P < .001), control and remission RA groups were similar (P = .86). PIV and SII are significantly higher in active RA than remission RA and control (P < .001, P < .001) higher in remission RA than control (P < .001, P < .001). Receiver operating characteristic curve analysis in predicting remission compared to the control group, CRP was not significant, PIV and SII was significant and PIV has higher sensitivity and sensitivity, a PIV value of > 217.31 have sensitivity 75.0% and specificity 85.7%. CRP, PIV, and SII are statistically significant in predicting active RA compared to the remission RA and control group. Our findings show that PIV, and SII are easy, inexpensive and reliable markers predicting remission in RA patients. CRP was not significant compared to remission RA and control group, PIV and SII was significant and PIV has higher sensitivity and specificity than SII in the remission group in RA. Patients with high disease activity, PIV, SII, and CRP levels were effective in showing disease activity compared to RA remission group and healthy controls.

Description of an intramonocytic haemoparasite, Hepatozoon lainsoni sp. nov. (Apicomplexa: Adeleorina: Hepatozoidae), infecting Ameiva ameiva lizard (Reptilia: Squamata: Teiidae) in northern Brazil.

Haemogregarine (Apicomplexa: Adeleorina) parasites are considered to be the most common and widespread haemoparasites in reptiles. The genus Hepatozoon (Apicomplexa: Adeleorina: Hepatozoidae) can be found parasitizing a broad range of species and, in reptiles, they infect mainly peripheral blood erythrocytes. The present study detected and characterized a haemogregarine isolated from the lizard species, Ameiva ameiva, collected from the municipality of Capanema, Pará state, north Brazil. Blood smears and imprints from lungs, brain, heart, kidney, liver, bone marrow and spleen were observed using light microscopy and the parasite was genetically identified by molecular analysis. Morphological, morphometric and molecular data were obtained. Parasite gamonts were found in 49.5% (55/111) of the blood smears from A. ameiva, and were characterized as oval, averaging 12.0 ± 0.8 × 5.9 ± 0.6 μm2 in size, which displaced the nuclei of parasitized monocytes laterally. Parasite forms resembling immature gamonts were observed in the spleen and bone marrow of the lizards. Furthermore, phylogenetic analyses of 18S rRNA sequences did not reveal gene similarity with other Hepatozoon spp. sequences from reptiles. Thus, morphological and molecular analyses have identified a new species of Hepatozoon parasite, Hepatozoon lainsoni sp. nov., which infects monocytes of the A. ameiva lizard.

Alteration in the number, morphology, function, and metabolism of erythrocytes in high-altitude polycythemia.

Introduction: High-altitude polycythemia (HAPC) is a common chronic high-altitude disease characterized by significantly increased erythrocyte, hemoglobin (Hb), and hematocrit values and decreased arterial oxygen saturation. The mechanisms underlying HAPC development are unclear; we aimed to investigate this in an HAPC rat model. Methods: Twelve Sprague-Dawley rats were divided into control and HAPC groups. The HAPC group was exposed to hypobaric hypoxia. This HAPC model was assessed using routine blood tests and blood gas analyses. Bone marrow, peripheral blood reticulocytes (RETs), and peripheral blood erythrocyte apoptosis were measured using flow cytometry. Erythrocyte osmotic fragility (EOF) tests were conducted. Abnormal erythrocytes were counted using electron microscopy. Plasma-free hemoglobin, 5'-nucleotidase (CD73), adenosine, erythrocyte cytosolic adenosine, sphingosine-1-phosphate (S1P), and 2,3-bisphosphoglycerate (BPG) levels were measured using enzyme-linked immunosorbent assays. Erythrocyte metabolic pathway-related protein [adenosine A2B receptor (ADORA2B), erythrocyte equilibrative nucleoside transporter 1 (eENT1), sphingosine kinase 1 (SPHK1), phospho-SPHK1, bisphosphoglycerate mutase (BPGM), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)] levels were assessed by Western blotting. Results: The HAPC rat model was successfully established (Hb > 210g/L). Indices of bone marrow and peripheral blood RET proportions were significantly higher in the HAPC than the control group (p = 0.04 and p < 0.001, respectively). The proportion of peripheral blood erythrocytes in early apoptosis was significantly lower in the HAPC than the control group (p < 0.001). Vesicular erythrocyte and acanthocyte proportions were significantly higher in the HAPC than the control group (p < 0.001 and p = 0.019, respectively). The EOF tests revealed that 50% erythrocyte hemolysis occurred at 4.0-4.5 and 4.5-5.0g/L NaCl in the control and HAPC groups, respectively. Plasma-free hemoglobin, CD73, adenosine, erythrocyte cytosolic adenosine, S1P, and 2,3-BPG levels and ADORA2B, eENT1, phospho-SPHK1, S1P, BPGM, and GAPDH erythrocyte expression levels (all p ≤ 0.02) were significantly higher in the HAPC than the control group. Conclusion: In model rats, an HAPC-related erythrocyte increase was associated with enhanced bone marrow hematopoietic function and reduced erythrocyte apoptosis, whereas numerous abnormal erythrocytes, increased EOF, and reduced hemolysis resistance were associated with erythrocyte metabolism. CD73/adenosine/S1P/2,3-BPG and eENT1/adenosine/BPGM/2,3-BPG metabolic pathways in erythrocytes were activated in HAPC rats, facilitating oxygen release. These findings further reveal the intrinsic HAPC mechanism and forms a basis for future development of preventive and therapeutic strategies for HAPC.

Detection of early-stage gastrointestinal cancers by micronuclei DNA from erythrocytes.

382 Background: Gastrointestinal (GI) cancers account for over a third of cancer deaths. However, their early detection remains a challenge. Micronuclei (MN) are extranuclear bodies containing damaged chromosome segments indicative of genomic instability. Increased MN frequency in hematopoietic cells is observed in patients with solid tumors. We have developed a method (WO2021/228246 A1) for purifying and characterizing micronuclei DNA (mnDNA) in erythrocytes from peripheral blood. Here, we evaluated the potential of mnDNA for early-stage GI cancer detection, including colorectal (CC) and gastric (GC) cancers. Methods: Peripheral blood (1 ml) was collected from healthy donors (HD) and cancer patients mnDNA isolation and purification from erythrocytes. Participants were randomly divided into training, validation and independent test cohorts in a 7:2:1 ratio, maintaining similar cancer types, CC/GC/HD ratio, gender and age distribution. Distinct mnDNA features between cancer patients and HDs were identified using sequencing data. Logistic regression algorithms were applied using these features for cancer detection. Results: The study enrolled 1046 participants, including 419 HDs, 366 CC and 261 GC cases. Of these, over half were diagnosed with early-stage diseases; TNM stage I accounted for 38.7%, stage II for 22.8%, stage III for 30.5% and stage IV for 8.0%. The cancer detection model with mnDNA features yielded an overall accuracy of 84.8%, with a sensitivity of 84.1% and specificity of 85.7%. For individual caner types, sensitivity was 89.2% for CC and 76.9% for GC. For early-stage (stage I-II) CC and GC, the model demonstrated sensitivities of 88.9% and 76.5%, respectively, at 85.7% specificity. Conclusions: Unlike cell-free DNA, mnDNA are chromosomal fragments in the cytoplasm. The abundance of erythrocytes in peripheral blood provides an easily accessible source for mnDNA enrichment. This pilot study illustrates the potential of mnDNA as a tool for early GI cancer detection, contributing to large-scale efforts towards developing an effective GI cancer screening test.

Novel betulin derivatives as multidrug reversal agents targeting P-glycoprotein

Chemotherapy is a powerful means of cancer treatment but its efficacy is compromised by the emergence of multidrug resistance (MDR), mainly linked to the efflux transporter ABCB1/P-glycoprotein (P-gp). Based on the chemical structure of betulin, identified in our previous work as an effective modulator of the P-gp function, a series of analogs were designed, synthesized and evaluated as a source of novel inhibitors. Compounds 6g and 6i inhibited rhodamine 123 efflux in the P-gp overexpressed leukemia cells, K562/Dox, at concentrations of 0.19 µM and 0.39 µM, respectively, and increased the intracellular accumulation of doxorubicin at the submicromolar concentration of 0.098 µM. Compounds 6g and 6i were able to restore the sensitivity of K562/Dox to Dox at 0.024 µM and 0.19 µM, respectively. Structure–activity relationship analysis and molecular modeling revealed important information about the structural features conferring activity. All the active compounds fitted in a specific region involving mainly transmembrane helices (TMH) 4–6 from one homologous half and TMH 7 and 12 from the other, also showing close contacts with TMH 6 and 12. Compounds that bound preferentially to another region were inactive, regardless of their free energy of binding. It should be noted that compounds 6g and 6i were devoid of toxic effects against peripheral blood mononuclear normal cells and erythrocytes. The data obtained indicates that both compounds might be proposed as scaffolds for obtaining promising P-gp inhibitors for overcoming MDR.

Quantitative and Qualitative Composition of the Peripheral Blood of Fish in the Gradient of Long-Term Radiation Exposure

The paper deals with analysis of hematological parameters of the common rudd (&lt;i&gt;Scardinius erythrophthalmus &lt;/i&gt;L.) and Prussian carp (&lt;i&gt;Carassius gibelio&lt;/i&gt; Bloch) from the most radionuclide-contaminated water bodies of the Chernobyl Exclusion Zone. Data on absolute and relative leukocyte composition and morphological disorders of erythrocytes in peripheral blood were obtained. The leukocyte fraction of both fish species similarly responded to the absorbed dose rate. In the doses' range of 5.1-54.1 &amp;mu;Gy/h, reactive changes of the compensatory nature were recorded with increase of the leukocytes content at the expense of lymphocyte and granulocyte fractions. Under the higher doses (up to 84.5 &amp;mu;Gy/h), hematopoiesis was damaged with sharp decrease of the leukocytes' content and significant changes in leukograms, which was also confirmed by the hematological indices' values. The morphological disorders in erythrocytes increased along with absorbed dose rate.

Investigations on the genotoxic potential of styrene in Fischer 344 rats using multiple endpoints.

Genotoxicity of styrene monomer was evaluated in male Fischer 344 rats using the alkaline comet assay for DNA damage, micronucleus assay for cytogenetic damage and the Pig-a assay for gene mutations. In a dose range finding (DRF) study, styrene was administered by oral gavage in corn oil for 28 consecutive days at 0, 100, 500, and 1000 mg/kg/day. The bioavailability of styrene was confirmed in the DRF by measuring its plasma levels at approximately 7- or 15-min following dosing. The 1000 mg/kg/day group exceeded the maximum tolerated dose based on body weight and organ weight changes and signs of central nervous system depression. Based on these findings, doses of 0, 100, 250, and 500 mg/kg/day (for 28 or 29 days) were selected for the genotoxicity assays. Animals were sacrificed 3-4 h after treatment on Day 28 or 29 for assessing various genotoxicity endpoints. Pig-a mutant frequencies and micronucleus frequencies were determined in peripheral blood erythrocytes. The comet assay was conducted in the glandular stomach, duodenum, liver, lung, and kidney. These studies were conducted in accordance with the relevant OECD test guidelines. Oral administration of styrene did not lead to genotoxicity in any of the investigated endpoints. The adequacy of the experimental conditions was assured by including animals treated by oral gavage with the positive control chemicals ethyl nitrosourea and ethyl methane sulfonate. Results from these studies supplement to the growing body of evidence suggesting the lack of in vivo genotoxic potential for styrene.

Changes in peripheral blood parameters of lambs with spontaneous eimeriosis

In order to study the morphological and biochemical parameters of the blood of lambs affected by the causative agent of eimeria, an experimental group of animals of the Romanov breed, two months old, was formed. The lambs were spontaneously infected with the causative agent of eimeria, which was confirmed by laboratory and clinical studies. The second group of animals was free from eimeria and was kept separately. According to the results of blood tests of sick lambs, a decrease in the number of peripheral blood erythrocytes by 29.66 %, an increase in the total number of leukocytes by 47.12 % compared to the blood parameters of the control group of animals was revealed. A change in leukogram parameters was also noted: an increase in the number of eosinophils by 55.43 %, young neutrophils by 75 %. At the same time, there was a decrease in the number of rod-shaped neutrophils and an increase in the number of segmented neutrophils by 8.7 and 29.51 %, respectively. A decrease in the number of lymphocytes and monocytes by 31.68 and 12 %, respectively, was registered. Biochemical changes in the blood serum of sick lambs also underwent changes. A decrease in the content of total protein by 36.06 %, albumin by 13.09 % was established. An increase in the content of globulins and dysproteinemia was noted. Hemoglobin content was also reduced compared to blood parameters of healthy lambs by 32.25 %, as well as glucose content by 27.83 %. The activity level of ALT and AST increased by 43.43 and 33.15 %, respectively, as well as a slight decrease in GGT activity. The difference was statistically significant (Р &lt; 0.01 and Р &lt; 0.05). Differences in the blood parameters of sick lambs have been established and indicate a syndrome of the body's general inflammatory reaction, anemia, damage to the alimentary canal by the causative agent of eimeria and intoxication.

Diabetes-correcting and antioxidant effects of grape pomace extract rich in natural complex of polyphenols

Background. The positive health effects of polyphenols have led to an increased scientific interest in these natural compounds over the past decade. Many studies confirm the effectiveness of polyphenols as additional therapy in diabetes, especially due to the sugar-lowering effect of polyphenols. The aim of the research was to investigate the morphological and functional state of peripheral blood erythrocytes and the indices of oxidative stress in the liver of rats with experimental diabetes and after the administration of grape pomace extract rich in natural complex of polyphenols. Materials and Methods. We obtained grape pomace extract, which contains a variety of polyphenolic compounds. Rats of the following groups were used in the experiments: control animals, animals treated with grape pomace extract rich in natural complex of polyphenols for 14 days, animals with streptozotocin-induced diabetes mellitus, and animals with streptozotocin-induced diabetes mellitus treated with grape pomace extract rich in natural complex of polyphenols for 14 days. The number of erythrocytes and reticulocytes, the concentration of hemoglobin and glycated hemoglobin were determined in the peripheral blood of rats. The activities of catalase, superoxide dismutase and the content of thiobarbituric acid-reactive substance and carbonyl groups of proteins were determined in the liver tissues of rats. Results. The study has shown an increase in the number of erythrocytes and the level of hemoglobin, a decrease in the level of glycated hemoglobin and the number of reticulocytes in the peripheral blood of rats after administration of grape pomace extract rich in natural complex of polyphenols to rats with experimental diabetes. A decrease in the content of thiobarbituric acid-reactive substance and the content of carbonyl groups of proteins of neutral and basic character and an increase in the activity of catalase and superoxide dismutase in liver tissues were found under the same conditions. Conclusions. The results indicate that the extract of the natural complex of polyphenols is capable of correcting the morphological and functional state of erythrocytes, as well as improving the activity of antioxidant enzymes and the content of marker molecules of oxidative stress in hepatocytes of rats under experimental diabetes mellitus.

Adhesion of Immunoglobulins to Band3 Promotes Increased Erythrocyte Sedimentation Rate in Multiple Myeloma

An increase in ESR can be seen in approximately 85% of patients with MM [1]. Therefore, ESR is often used as one of the sensitive indicators of active MM [1]. It is now widely accepted that the increase in ESR is related to the adhesion of immunoglobulins secreted by myeloma cells to erythrocytes [2, 3]. However, this explanation is descriptive and speculative and has not been confirmed by experimental data [3]. We applied peripheral blood erythrocyte samples from MM patients and used immunofluorescence/immunoprecipitation tandem protein analysis/molecular docking to determine the molecular mechanism by which all types of MM immunoglobulins can target and adhere to erythrocyte membrane Band3. This result, not only helps to explain the molecular mechanism of ESR acceleration in MM, but will also advance our understanding of the pathophysiologic changes that occur in ESR. At the same time, it may provide a new way to target protection of normal erythrocytes and targeted removal of erythrocyte-bound immunoglobulins.

A multiscale study of the effects of a diet containing CdSe/ZnS-COOH quantum dots on Salmo trutta fario L.: Potential feed-related nanotoxicity

Quantum dots (QDs) receive widespread attention in industrial and biomedical fields, but the risks posed by the use of nanoparticles to aquatic organisms and the associated toxicological effects are still not well understood. In this study, effects of the 7-day dietary exposure of Salmo trutta fario L. juveniles to CdSe/ZnS-COOH QDs were evaluated at molecular, cellular, physiological and whole-organism levels. Fish feeding with QDs-contaminated feed resulted in an increased somatic index of the liver, a genotoxic effect on peripheral blood erythrocytes, altered enzyme activity and decreased MDA level. Furthermore, Cd levels in the gills and liver tissues of the exposed fish were found to be significantly higher than in those of the control fish. Alpha diversity indexes of the gut microbiota of the QDs-exposed S. trutta fario L. individuals exhibited a decreasing trend. The principal coordinate analysis (PCoA) showed that the gut microbiota of the control fish was significantly different from that of the fish exposed to QDs (p < 0.05). Additionally, the linear discriminant analysis (LDA) performed using an effect size (LEfSe) algorithm unveiled 19 significant taxonomic differences at different taxonomic levels between the control group and the QDs-exposed group. In the QDs-exposed group, the relative abundance of the genus Citrobacter (Proteobacteria phylum) in the gut microbiota was found to be significantly increased whereas that of the genus Mycoplasma (Tenericutes phylum) significantly decreased compared to the control group. In summary, QDs-contaminated diet affects the gut microbiota of fish by significantly changing the relative abundance of some taxa, potentially leading to dysbiosis. This, together with morphophysiological, cytogenetic and biochemical changes, poses a risk to fish health.

Assessment of the water quality and toxicity effects on zebrafish (Danio rerio) of a stream near a phosphorus chemical plant in Guizhou Province, southwestern China.

To reveal the influence of the phosphorus chemical industry (PCI) on regional water environmental quality and safety, the water quality and ecotoxicological effects of a stream near a phosphorus chemical plant (PCP) in Guizhou Province, southwestern China, were investigated based on water samples collected from the stream. The results showed that the average concentrations of NH3-N, TN, P, F-, Hg, Mn, and Ni were 3.14mg/L, 30.09mg/L, 3.34mg/L, 1.18mg/L, 1.06μg/L, 45.82μg/L, and 11.30μg/L, respectively. The overall water quality of the stream was in the heavily polluted category, and NH3-N, TN, P, F-, and Hg were the main pollution factors. The degree of pollution was in the order of rainy period > transitional period > dry period, and the most polluted sample site was 1100m from the PCP. After 28 days of exposure to stream water, there was no significant change in the growth parameters of zebrafish. The gills of zebrafish showed a small amount of epithelial cell detachment and a small amount of inflammatory cell infiltration, and the liver tissue displayed a large amount of hepatocyte degeneration with loose and lightly stained cytoplasm. Compared with the control group, the%DNA intail, tail length, tail moment, and olive tail moment were significantly increased (p < 0.05), indicating that the water sample caused DNA damage in the peripheral blood erythrocytes of zebrafish. The stream water in the PCI area was found to be polluted and exhibited significant toxicity to zebrafish, which could pose a threat to regional ecological security.

PIG-A gene mutation as a mutagenicity biomarker among coke oven workers.

PIG-A gene mutations can be detected in humans, and PIG-A assays can potentially predict the risk of exposure to carcinogens. However, extensive, population-based studies to validate this are lacking. We studied a cohort of occupational coke oven workers with chronic high exposure to carcinogenic polycyclic aromatic hydrocarbons, which are well-studied genotoxins classified by the IARC as carcinogenic to humans. Peripheral blood erythrocytes of workers were assessed for gene mutations using a PIG-A assay, and chromosome damage using the cytokinesis-block micronucleus test with lymphocytes. Two sample populations from a non-industrialized city and new employees in industrial plants were selected as controls. We observed a significantly elevated PIG-A mutation frequency (MF) and increased frequencies of micronuclei (MN) and nuclear buds (NBUDs) in coke oven workers, compared with levels in the control groups. We found that the coke oven workers with different lengths of service had a relatively high mutation frequency. Overall, the study findings showed that occupational exposure of coke oven workers increases the genetic damage and the PIG-A MF could be a potential biomarker for risk assessment of carcinogen exposure.

The Diagnostic Potential of the Human Blood Microbiome: Are We Dreaming or Awake?

Human blood has historically been considered a sterile environment. Recently, a thriving microbiome dominated by Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes phyla was detected in healthy blood. The localization of these microbes is restricted to some blood cell populations, particularly the peripheral blood mononuclear cells and erythrocytes. It was hypothesized that the blood microbiome originates from the skin-oral-gut axis. In addition, many studies have evaluated the potential of blood microbiome dysbiosis as a prognostic marker in cardiovascular diseases, cirrhosis, severe liver fibrosis, severe acute pancreatitis, type 2 diabetes, and chronic kidney diseases. The present review aims to summarize current findings and most recent evidence in the field.

Green Synthesis of Selenium Nanoparticles Using Luffa cylindrica and its Biocompatibility Studies for Potential Biomedical Applications

Selenium nanoparticles (SeNPs), due to their unique properties, have attracted researchers’ attention. Though SeNPs have been used for wide applications, the chemically synthesized one lacks stability due to aggregation, and it releases toxic byproducts. These drawbacks can be overcome by producing SeNPs using natural sources as reducing and capping agents. Luffa cylindrica is an immense source of phytochemical compounds reported for its potential therapeutical value towards cancer, asthma, and sinusitis. In the current study, we have synthesized SeNPs using leaf extract of L. cylindrica and evaluated its biocompatibility and haemocompatibility using peripheral blood mononuclear cells and erythrocytes respectively. The formation of SeNPs was confirmed by a color change from greenish yellow to ruby red during 6 h incubation at 40[Formula: see text]C and further confirmed by the maximum absorbance at 266[Formula: see text]nm and 380[Formula: see text]nm in the UV–Vis spectrum. The fingerprint regions of the Fourier-transform infrared spectroscopy spectrum between 1500[Formula: see text]cm[Formula: see text] and 500[Formula: see text]cm[Formula: see text] revealed the presence of phytoconstituents of L. cylindrica. The particle size analysis showed a size range of 100[Formula: see text]nm and zeta potential of −13.6 mV. Scanning electron micrograph showed flower-shaped surface morphology with a size range of 100[Formula: see text]nm. The erythrocytes treated with higher concentrations of LC-SeNPs showed less than 5% lysis compared to the positive control. Similarly, in the apoptosis assay, 80.45% of cells remained viable after being treated with LC-SeNPs, which is comparable with that of untreated control. Since the synthesized SeNPs possess biocompatibility and are less cytotoxic, they could be used in cardiac tissue engineering applications. However, further in vitro and in vivo studies are required to confirm its role in cardiac tissue engineering.

A novel enzyme-linked immunostaining technique based on silk membrane for the prenatal detection of fetomaternal haemorrhage.

Objective: Developing a simple, rapid, reliable, sensitive, and cost-effective method for prenatal detection of fetomaternal haemorrhage by combining multi-aperture silk membrane with enzyme-linked immunosorbent assay (ELISA), which does not require any complicated instruments and can be visually colored, so as to provide a new method for clinical detection of fetomaternal haemorrhage. Methods: As a carrier, a chemically treated silk membrane was used to immobilize anti-A/anti-B antibody reagent. PBS washed slowly after vertically dropping red blood cells. After adding biotin-labeled anti-A/anti-B antibody reagent, PBS is slowly washed, enzyme-labeled avidin is added, and TMB is used for color development after washing. Results: When there were both anti-A and anti-B fetal erythrocytes in pregnant women's peripheral blood, the final color was dark brown. When there are no anti-A and anti-B fetal red blood cells in pregnant women's peripheral blood, the final color development results do not change, which corresponds to the color of chemically treated silk membrane. Conclusion: The new enzyme-linked immunosorbent assay (ELISA) based on a silk membrane can distinguish fetal red blood cells from maternal red blood cells prenatally and can be used for prenatal detection of fetomaternal haemorrhage.

THE MORFOFUNCTIONAL STATE OF PERIPHERAL ERYTHROCYTES AT CHRONIC HEART FAILURE AND VITAMIN D DEFICIENCY IN

The aim of the work To study the effect of vitamin D deciency in blood plasma on the morphofunctional state of peripheral blood erythrocytes after exercise in patients with chronic heart failure. Methods: scanning electron microscopy of peripheral blood erythrocytes, enzyme immunoassay to determine the level of vitamin D. Results. A morphological study revealed heterogeneity of erythrocyte subpopulations in 90% of patients with CHF and vitamin D deciency. In peripheral blood, in addition to normocytes with sufcient hemoglobinization, subpopulations of macro- and microcytes were recorded. In most patients with vitamin D deciency, several types of morphological changes in the shape of erythrocytes were revealed, manifested in the form of echino-, acantho- and stomatocytosis, which indicates destabilization of the erythrocyte's membrane. After a 6-minute walk test in CHF patients with vitamin D deciency, the number of discocytes decreases, the ability to deform decreases, lytic cells were recorded with a reduced number of normocytes. The Conclusion: identied morphobiochemical changes indicate the destructurization of the erythrocyte membrane components and have negative consequences, taking into account the likely impact on the somatic health of patients who, in addition to changes characteristic of CHF, receive signicant stress during physical exertion against the background of hypovitaminosis. D.

Влияние КВЧ-излучения на частоту микроядер и ядерных аномалий в эритроцитах личинок амфибий

The level of technogenic non-ionizing electromagnetic radiations of various frequencies constantly grows in the biosphere. Therefore, studies of these radiations influence on biological organisms are very actual. The purpose of the work was to study the consequences of prolonged exposure to EHF radiation on the stability of the genome of anurans. Larvae of the green toad (Bufo viridis) were subjected to EMR at four different EHF frequencies (34.52, 34.67, 34.79, and 35.04 GHz with an energy flux density of 4.88, 5.33, 4.94, and 3.76 mW/cm2, respectively) for 120 hours. After irradiation the frequencies of micronuclei (MN) and nuclear anomalies (NA) in peripheral blood erythrocytes of tadpoles were studied. EHF radiation with a frequency of 34.52 GHz induced a small and statistically insignificant increase in the frequency of MN and NA. Exposure to EHF radiation at frequencies of 34.67, 34.79, and 35.04 GHz caused a statistically significant increase in the frequency of MN and NA in tadpole erythrocytes. A statistically significant increase of the frequency of MN and NA in anuran larvae erythrocytes after exposure to EHF radiation proves a certain danger of an increasing level of EMR in the biosphere, as well as the need for further studies of the effect of non-ionizing radiation on the genomes of lower vertebrates.

Peculiarities of phospholipiid changes in erythrocyte membranes in parturient women with COVID-19-associated community-acquired pneumonia

Aim. To evaluate the phospholipid composition of erythrocyte membranes in parturient women who had COVID-19-associated community-acquired pneumonia (CAP) in the third trimester.Materials and methods. The material for the study was erythrocytes of peripheral blood of 65 parturient women diagnosed with COVID-19, moderate/severe course, CAP of viral etiology (main group). Patients of the main group, depending on the severity of CAP, were divided into two subgroups: subgroup 1 – moderate course of pneumonia (n=33), subgroup 2 – severe course of pneumonia (n=32). The control group consisted of 35 healthy parturient women. The quantitative composition of phospholipids was studied by two-dimensional thin-layer chromatography according to Kirchner.Results. In subgroup 1, the concentration of phosphatidylethanolamine and phosphatidylcholine in erythrocyte membranes was below the standard values by 38% and 29%, respectively (p&lt;0.001), in subgroup 2, these indicators decreased by 32% and 48%, respectively (p&lt;0.001). At the same time, a significant increase in the concentration of lysophosphatidylcholine was found in patients of subgroup 1 by 92% (p&lt;0.001) and in patients of subgroup 2 by 110% (p&lt;0.001), compared with the group of healthy individuals. In addition, structural changes in the lipid bilayer of erythrocyte membranes under conditions of COVID-19associated CAP were characterized by a pronounced increase in the concentration of minor fractions of phospholipids: phosphatidylserine and phosphatidylinositol in subgroup 1 by 63% and 53%, respectively (p&lt;0.001), in subgroup 2 by 79% and 68%, respectively (p&lt;0.001), compared with similar indicators in the control group.Conclusion. With COVID19-associated CAP in maternity women, structural disorganization of the phospholipid components of erythrocyte membranes is determined, manifested by a decrease in the concentration of phosphatidylethanolamine and phosphatidylcholine with a simultaneous increase in the level of lysophosphatidylcholine, phosphatidylserine and phosphatidylinositol. These disorders increase with increasing severity of pulmonary inflammation. The revealed changes in the lipid spectrum of peripheral blood and the composition of erythrocyte membrane phospholipids in COVID-19-associated CAP indicate the need to develop methods for their correction.