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Docosahexaenoic acid (DHA) shows anti-inflammatory/proresolution effects in the brain. Higher red blood cell (RBC) DHA in humans is associated with improved cognitive performance and a lower risk for suicide. Here, we hypothesized that binding to the 18 kDa translocator protein (TSPO), a proxy for microglia levels, will be higher in individuals with low DHA relative to high DHA levels. We also postulated that higher TSPO would predict poor cognitive performance and impaired stress resilience.RBC DHA screening was performed in 320 healthy males. [sup11/supC]PBR28 positron emission tomography was used to measure binding to TSPO in 38 and 32 males in the lowest and highest RBC DHA quartiles. Volumes of distribution expressed relative to total plasma ligand concentration (VsubT/sub) was derived using an arterial input function-based kinetic analysis in 14 brain regions.[sup11/supC]PBR28 VsubT/subwas significantly lower (by 12% and 20% in C/T and C/C rs6971 genotypes) in males with low RBC DHA than in males with high RBC DHA. Regional VsubT/subwas correlated positively and negatively with RBC DHA and serum triglycerides, respectively. No relationships between VsubT/suband cognitive performance or stress resilience measures were present.Contrary to our hypothesis, we found lower TSPO binding in low-DHA than in high-DHA subjects. It is unclear as to whether low TSPO binding reflects differences in microglia levels and/or triglyceride metabolism in this study. Future studies with specific targets are necessary to confirm the effect of DHA on microglia. These results underscore the need to consider lipid parameters as a factor when interpreting TSPO positron emission tomography clinical findings.

Circulating tumor cells (CTCs), as important liquid biopsy target, can provide valuable information for cancer progress monitoring and individualized treatment. However, current isolation platforms incapable of balancing capture efficiency, specificity, cell viability, and gentle release have restricted the clinical applications of CTCs. Herein, inspired by the structure and functional merits of natural membrane interfaces, we established an antibody-engineered red blood cell (RBC-Ab) affinity interface on microfluidic chip for high-performance isolation and release of CTCs. The lateral fluidity, pliability, and anti-adhesion property of the RBC microfluidic interface enabled efficient CTCs capture (96.5%), high CTCs viability (96.1%), and high CTCs purity (average 4.2-log depletion of leukocytes). More importantly, selective lysis of RBCs by simply changing the salt concentration was utilized to destroy the affinity interface for efficient and gentle release of CTCs without nucleic acid contamination. Using this chip, CTCs were successfully detected in colon cancer samples with 90% sensitivity and 100% specificity (20 patients and 10 healthy individuals). After the release process, KRAS gene mutations of CTCs were identified from all the 5 cancer samples, which was consistent with the results of tissue biopsy. We expect this RBC interface strategy will inspire further biomimetic interface construction for rare cell analysis.

The aim of this study was to identify the effect of inclusion of defatted black soldier fly larvae (Def-BSFL) meal as a protein source on the performance and blood plasma constituents of broiler chickens. A total of 360-day-old chicks were assigned into four dietary groups, which included four different levels of Def-BSFL meal namely control (0% BSFL), T1(4% BSFL), T2 (8% BSFL) and T3 (12% BSFL) for six weeks experimental feeding period. At the end of the experiment, the blood samples of three birds from each treatment were collected to measure plasma constituents. Birds fed control and T1 diets demonstrated higher feed intake during the finisher stage compared with T2 and T3 diets. The heaviest weight for the 6-week feeding trial was recorded at T1 (1043.8 ± 65.9 g). Birds fed T1 (1.1 ± 0.0) and T3 (0.9 ± 0.1) diets displayed lower feed conversion ratio during the finisher stage than those fed control (1.7 ± 0.1) and T2 (1.8 ± 0.3) diets. Birds fed the control diet demonstrated the highest red blood cell with mean and standard deviation of 7.5 ± 0.34, whereas those fed the T2 diet showed the highest haemoglobin levels with mean and standard deviation of 15.8 ± 0.24. Birds fed T1, T2, and T3 diets exhibited a higher number (P

Background Tibetan pigs have exhibited unique characteristics from low‐altitudes pigs and adapted well to the Qinghai‐Tibet Plateau. Objectives The current study was undertaken to investigate the hypoxic adaptation of heart in Tibetan pigs. Methods The hearts of Tibetan pigs and Landrace pigs raised at high or low altitudes were compared using 3D casting technology, scanning electron microscopy and real‐time quantitative PCR (qRT‐PCR). Results We found that the ratio of the major axis to the minor axis and the density of the heart were significantly higher in Tibetan pigs than in Landrace pigs (p, Tibetan pigs have exhibited unique characteristics from low‐altitudes pigs and have adapted well to the Qinghai‐Tibet Plateau, but how this occurs is incompletely understood. We found that Tibetan pigs had larger diameters and higher densities of arterioles than Landrace pigs (p

The major transmembrane protein of the red blood cell, known as band 3, AE1, and SLC4A1, has two main functions: 1) catalysis of Cl−/[Formula: see text] exchange, one of the steps in CO2 excretion, and 2) anchoring the membrane skeleton. This review summarizes the 150-year history of research on red cell anion transport and band 3 as an experimental system for studying membrane protein structure and ion transport mechanisms. Important early findings were that red cell Cl− transport is a tightly coupled 1:1 exchange and band 3 is labeled by stilbenesulfonate derivatives that inhibit anion transport. Biochemical studies showed that the protein is dimeric or tetrameric (paired dimers) and that there is one stilbenedisulfonate binding site per subunit of the dimer. Transport kinetics and inhibitor characteristics supported the idea that the transporter acts by an alternating access mechanism with intrinsic asymmetry. The sequence of band 3 cDNA provided a framework for detailed study of protein topology and amino acid residues important for transport. The identification of genetic variants produced insights into the roles of band 3 in red cell abnormalities and distal renal tubular acidosis. The publication of the membrane domain crystal structure made it possible to propose concrete molecular models of transport. Future research directions include improving our understanding of the transport mechanism at the molecular level and of the integrative relationships among band 3, hemoglobin, carbonic anhydrase, and gradients (both transmembrane and subcellular) of [Formula: see text], Cl−, O2, CO2, pH, and nitric oxide (NO) metabolites during pulmonary and systemic capillary gas exchange.

Anemia is a common complication of chronic kidney disease, affecting the quality of life of patients. Among various factors, such as iron and erythropoietin deficiency, reduced red blood cell (RBC) lifespan has been implicated in the pathogenesis of anemia. However, mechanistic data on in vivo RBC dysfunction in kidney disease are lacking. Herein, we describe the development of chronic kidney disease-associated anemia in mice with proteinuric kidney disease resulting from either administration of doxorubicin or an inducible podocin deficiency. In both experimental models, anemia manifested at day 10 and progressed at day 30 despite increased circulating erythropoietin levels and erythropoiesis in the bone marrow and spleen. Circulating RBCs in both mouse models displayed altered morphology and diminished osmotic-sensitive deformability together with increased phosphatidylserine externalization on the outer plasma membrane, a hallmark of RBC death. Fluorescence-labelling of RBCs at day 20 of mice with doxorubicin-induced kidney disease revealed premature clearance from the circulation. Metabolomic analyses of RBCs from both mouse models demonstrated temporal changes in redox recycling pathways and Lands’ cycle, a membrane lipid remodeling process. Anemic patients with proteinuric kidney disease had an increased proportion of circulating phosphatidylserine-positive RBCs. Thus, our observations suggest that reduced RBC lifespan, mediated by altered RBC metabolism, reduced RBC deformability, and enhanced cell death contribute to the development of anemia in proteinuric kidney disease.

Inhibition of tight junction formation between two malaria parasite proteins, apical membrane antigen 1 and rhoptry neck protein 2, crucial for red blood cell invasion, prevents the disease progression. In this work, we have utilized a unique approach to design a chimeric peptide, prepared by fusion of the best features of two peptide inhibitors, that has displayed parasite growth inhibition, in-vitro, with nanomolar IC50, which is hundredfold better than any of its parent peptides. Further, to gain structural insights, we computationally modeled the hybrid peptide on its receptor.

The impact of exposure to different sizes of particulate matter (PM1, PM2.5, PM7, and PM10) was evaluated in Babylon concrete plant workers who had been exposed to concrete dust for at least 10 years. The effects of these particles on the hematological parameters, malondialdehyde (MDA) levels, and antioxidant enzymes (catalase and glutathione peroxidase ) were examined. The results exhibited that the levels of PM2.5 and PM10 were higher than the acceptable limits approved by the National Ambient Air Quality Standards (NAAQS). The blood parameters, namely white blood cells (WBC), red blood cell (RBC) and platelets counts, demonstrated non-significant differences between workers exposed to the PM as compared to the control group. However, differentiated white blood cells count revealed a significant increase of polymorphonuclear leukocytes (PMN) in exposed workers in comparison with the control group. However, both MDA and glutathione peroxidase showed a highly significant increase in the workers ass compared to the control group. Thus, we may conclude that the concrete plant workers are exposed to a higher risk of oxidative stress that could lead to alterations in hematological parameters, enzymatic activities, and MDA level.

Background. In view of the worsening forecast for global temperature rise worldwide, it seems relevant to study the effects of abnormal heat waves on systemic regulatory processes in people with chronic diseases, in particular coronary artery disease (CAD).Aims. This study aimed to investigate the effect of hyperthermia on oxidative stress parameters in patients with various severity of CAD and in healthy subjects.Materials and methods. We studied the level of malonic dialdehyde (MDA) and the activity of Cu,Zn-containing superoxide dismutase (Cu,Zn-SOD) in healthy subjects under conditions of 30-day long simulated hyperthermia and in patients with different severity of CAD after the summer heat wavesResults. We revealed signs of oxidative stress in healthy volunteers during model hyperthermia that manifested as an increase in content of MDA in blood plasma. At the same time we observed increasing activity of Cu,Zn-SOD in erythrocytes that utilizes reactive oxygen species. The increase of Cu,Zn-SOD activity started with a certain latency what also can be explained by de novo enzyme biosynthesis induction. We also studied oxidative stress parameters in patients at high and moderate cardiovascular risk according to the SCORE risk chart with uncomplicated CAD course and in patients with complicated CAD with severe coronary damage according to angiography during the summer heat waves. We observed accumulation of MDA in blood plasma and increasing activity of erythrocyte Cu,Zn-SOD in patients with uncomplicated CAD. At the same time we noted that accumulation of MDA in blood plasma was not followed by any increase in activity of red blood cell Cu,Zn-SOD in patients with severe complicated CAD. This fact indicates dysregulation of free radical processes in patients with severe course of CAD during the heat waves.Conclusions. The dysregulation of free-radical processes in patients with a severe clinical course of CAD has been revealed.

BACKGROUND Both M and N alleles encode antigens on Glycophorin A (GPA), a red blood cell (RBC) surface sialoglycoprotein. Interaction between RBC GPA and leukocyte surface lectins may downregulate their activation. The current study investigates if RBC autoantibodies against GPA, such as auto-anti-M/N, prime an activated phenotype in peripheral blood leukocytes. METHODS Leukocyte activation was assessed in whole blood from patients with auto-anti-GPA (anti-M/N) and compared to those with allo-anti-M/N and healthy subjects. Control samples from healthy subjects with no antibodies incubated in vitro with either anti-GPA or anti-Rh were analyzed for neutrophil and monocyte surface activation marker expression, reactive oxygen species (ROS) content, and formation of aggregates with RBCs. Samples incubated with an IgG1 isotype antibody served as controls. RESULTS Ex vivo, neutrophil CD66b and monocyte CD63 surface expression was increased in patients with auto-anti-M/N compared to those with allo anti-M/N (p = .1757; p = .0698) and to healthy subjects (p = .0186; p = .013). In vitro, neutrophil CD66b and monocyte CD63 surface expression was increased following incubation with anti-GPA compared to anti-Rh (p = .0003; p = .0328) and isotype control (p = .000; p = .0062). Intracellular ROS content increased in both neutrophils and monocytes incubated with anti-GPA compared to anti-Rh (p = .0012; p = .0693) and isotype control (p = .001; p = .0021). Percentage of neutrophil-RBC aggregates was decreased when incubated with anti-GPA compared to isotype control (p

The aim of this research was to study the effect of red blood cell system disorders on the further development and productivity of calves that had had bronchopneumonia. The study included 170 Holstein heifers at the age of 180-195 days: healthy heifers (n = 92) and heifers with moderate bronchopneumonia (n = 78). All animals had been under constant clinical supervision for 27 months. Blood sampling was performed in 10 animals from each group at the beginning of the experiment and on day 22, and also when they reached a body weight of 380 kg (the first artificial insemination), 30 days before the planned calving date, and on days 7, 90, 180 of lactation. The blood samples obtained were examined by a hematological analyzer, and the content of fetal hemoglobin was determined by Singer’s method. It was shown that after completion of the course of treatment the clinical symptoms of bronchopneumonia disappeared in calves, but microcytic hypochromic anemia appeared. Further, in these animals an increase was registered in the age of fruitful fertilization by 10.8% and of the insemination index by - 36.7%. After calving and during lactation, the severity of hypochromia decreased, but there was a tendency for macrocytosis, which, combined with increased anisocytosis, indicated hypoplastic anemia. This form of anemia occurred against the background of pathological residual changes after pneumonia due to the depletion of the compensatory potential of the bone marrow and its functional overload during lactation. The operational anemia revealed in lactating cows caused a decrease in milk productivity by 23.2-26.7% (P

Hemolytic disease of the fetus and newborn (HDFN) carries significant fetal mortality risks. Although anti-D as a source of HDFN has been prevented for decades using D-specific immunoglobulin to prevent alloimmunization between fetus and mother, minor blood groups may still result in disease, with potentially disastrous consequences if left untreated. Strategies such as intrauterine transfusion, early delivery, and vigilant serologic monitoring of fetal anemia have been the standards of care for alloimmunized patients, but beyond this not much more is possible. Mothers with rare phenotypes who are alloimmunized against extremely common red blood cell antigens may find access to rare antigen-negative blood units limited. This case study presents a healthy G10P6 woman with known anti-U presenting for treatment via intrauterine transfusion in the second trimester and follows the patient through successful delivery. Difficulties in obtaining rare blood for the patient because of concomitant delivery events involving 2 patients with anti-U at the facility opened discussions about the difficulties of and alternatives to intrauterine transfusion where rare blood phenotypes are involved.

A method for the ABO and Rhesus (Rh) blood group typing from individual erythrocytes is proposed in this study. Blood-group-specific antibodies immobilized to gold nanoparticles (BG-AuNP) were utilized for the identification of blood groups from individual erythrocytes by objective-type dark-field microscopy (OTDFM). The scattering of free BG-AuNP and their Brownian motion as well as BG-AuNP attached on erythrocytes is easily observed by OTDFM. The strong scattering intensity caused by BG-AuNP packing-enhanced nanoscattering (PENS) on erythrocytes is first demonstrated. PENS combined with OTDFM allows us to identify blood groups within 5 s for all blood group antigens including A, B, D, C, c, E, and e. This was immediately identified by mixing with BG-AuNP without any washing step or waiting for hemoagglutination. Therefore, PENS combined with OTDFM demonstrates feasibility and advantages for use in emergency transfusions where the blood group of patients is unknown. Moreover, matching RhD+ in the case of emergency transfusions may also be beneficial in reducing the shortage of RhD- red blood cell concentrate in the case of a population with a high frequency in RhD-.

Background Although precursor-targeted immune-mediated anemia (PIMA) is thought to be caused by immune targeting of erythroid precursors (nucleated RBCs, nRBCs), its pathogenesis is unknown. Immunoglobulin G (IgG) or phosphatidylserine (PS) may promote nRBC destruction in PIMA. Hypothesis Dogs with PIMA have increased nRBC IgG and PS, and dogs with immune-mediated hemolytic anemia (IMHA) have increased RBC PS compared to healthy dogs. Animals Blood from 20 healthy dogs and from dogs with IMHA (11) or other (non-IMHA) conditions (9), and marrow aspirates with or without blood from 10 healthy dogs and from dogs with PIMA (17) or other (non-IMHA, non-PIMA) conditions (7). Methods Marrow nRBC stages were separated by density gradient. Flow cytometry was used to assess the percentage of RBCs or nRBCs with increased IgG or PS. Results Red blood cell (RBC) IgG positivity was increased in 9/11 IMHA dogs and 0/9 non-IMHA dogs. Red blood cell PS positivity was increased in 10/11 IMHA dogs and 2/9 non-IMHA dogs. Five of 17 PIMA dogs had increased nRBC IgG positivity in mid- or late-stage fractions, whereas all 7 non-PIMA dogs were negative. Mid- and late-stage erythroid precursor PS was significantly higher in PIMA dogs compared to healthy dogs. Five of 14 PIMA dogs had increased RBC IgG positivity. Conclusions Immunoglobulin G and PS may promote destruction of nRBCs in PIMA dogs; PS may promote destruction of RBCs in IMHA dogs.

Background The current assessment of nutritional status and diagnosis of essential fatty acids deficiency (EFAD) utilizes the analysis of long-chain fatty acids (LCFAs) in serum or plasma; however, these concentrations do not represent habitual LCFA intake. LCFAs in red blood cells (RBCs) are less prone to intra-individual variability and exclude the need for fasting, which is unrealistic in pediatric populations. Our study objective was to characterize the RBC LCFA profiles in pediatric and adult reference populations and establish age-specific reference intervals (RIs). Methods Twenty-one LCFAs in RBCs were measured in 523 pediatric and adult controls by gas chromatography-mass spectrometry. Model-based clustering was used to identify possible age subgroups. After removing outliers by the Tukey method, initial age subgroups were then compared using the Harris-Boyd method in an iterative manner. RIs (95%), with confidence intervals (90%), in the final age groups were established using parametric or non-parametric statistics. Results Our data showed heterogeneous changes in the concentrations of most LCFAs and the EFAD biomarkers (mead acid, Triene/Tetraene ratio) during infancy. Model-based clustering identified six initial age subgroups per fatty acid, on average. Our application of the iterative Harris-Boyd method decreased the average number of age groups to three per fatty acid, with 13 total unique age cut-offs. Finally, using these age groups, we established age-specific RIs for 21 fatty acids, six group totals, and the Triene/Tetraene ratio. Conclusion Our study revealed significant age-dependent changes in RBC fatty acid profiles warranting separate pediatric and adults RIs. Model-based clustering and the iterative application of the Harris-Boyd method were successfully used to establish RBC fatty acid RIs for an objective assessment of long-term nutritional status in pediatric and adult populations.

Membrane proteins tend to interact with each other in the cell membranes to form protein clusters and perform the corresponding physiological functions. However, because channel proteins are involved in many biological functions, their distribution and nano-organization in these protein clusters are unclear. To study the distribution patterns and relationships between the different channel proteins, we identified the locations of glucose transporter 1 (Glut1) and Band3 (anion transporter 1) precisely in the topography of the cytoplasmic side of the human red blood cell (hRBC) membranes using combined atomic force microscopy (AFM) and single-molecule localization microscopy (SMLM). The AFM results revealed that membrane proteins interacted with each other and aggregated into protein islands. The SMLM results showed that Glut1 and Band3 tended to form protein clusters in the hRBC membranes, and there was a strong colocalization between the two proteins. The results of the combined AFM and SMLM method indicated that the protein clusters of Glut1 and Band3 were mainly located in the protein islands of topography, and the protein islands in topography also interacted with each other to assemble into larger protein clusters or functional microdomains.

Objective The development of antibodies against red blood cell (RBC) antigens is one of the most significant adverse effects of chronic blood transfusions. This study aimed to estimate the frequency and causes of RBC immunization in patients with major thalassemia. Methods The findings could help determine the limitation of current practices and help future developments in the selection of suitable blood units for the transfusion support of patients with thalassemia. Results The alloimmunization rate was detected in 52 of 650 patients with major thalassemia (8%). The most frequent antibodies were against K (50%), D (26%), and E (15.4%). Sex, age, and splenectomy did not have any impact on the immunization rate. Immunization was lower in patients who had received only leuko-reduced blood units. Conclusion It is recommended that antibody screening before the first transfusion and extended RBC matching particularly for the most frequent antigens (ABO/Rh/Kell) should be performed for patients with thalassemia.

During the vertebrate stage of the Plasmodium life cycle, obligate intracellular malaria parasites establish a vacuolar niche for replication, first within host hepatocytes at the pre-patent liver-stage and subsequently in erythrocytes during the pathogenic blood-stage. Survival in this protective microenvironment requires diverse transport mechanisms that enable the parasite to transcend the vacuolar barrier. Effector proteins exported out of the vacuole modify the erythrocyte membrane, increasing access to serum nutrients which then cross the vacuole membrane through a nutrient-permeable channel, supporting rapid parasite growth. This review highlights the most recent insights into the organization of the parasite vacuole to facilitate the solute, lipid and effector protein trafficking that establishes a nutrition pipeline in the terminally differentiated, organelle-free red blood cell.

Erythrocyte enucleation is thought to have evolved in mammals to support their energetic cost of high metabolic activities. However, birds face similar selection pressure yet possess nucleated erythrocytes. Current hypotheses on the mammalian erythrocyte enucleation claim that the absence of cell organelles allows erythrocytes to 1) pack more hemoglobin into the cells to increase oxygen carrying capacity and 2) decrease erythrocyte size for increased surface area-to-volume ratio, and improved ability to traverse small capillaries. In this article, we first empirically tested current hypotheses using both conventional and phylogenetically informed analysis comparing literature values of mean cell hemoglobin concentration (MCHC) and mean cell volume (MCV) between 181 avian and 194 mammalian species. We found no difference in MCHC levels between birds and mammals using both conventional and phylogenetically corrected analysis. MCV was higher in birds than mammals according to conventional analysis, but the difference was lost when we controlled for phylogeny. These results suggested that avian and mammalian erythrocytes may employ different strategies to solve a common problem. To further investigate existing hypotheses or develop new hypothesis, we need to understand the functions of various organelles in avian erythrocytes. Consequently, we covered potential physiological functions of various cell organelles in avian erythrocytes based on current knowledge, while making explicit comparisons with their mammalian counterparts. Finally, we proposed by taking an integrative and comparative approach, using tools from molecular biology to evolutionary biology, would allow us to better understand the fundamental physiological functions of various components of avian and mammalian erythrocytes.

Ancylostoma ceylanicum is a common zoonotic nematode that inhabits the small intestine of humans, dogs, and cats. Saposin-like proteins (SLPs) have hemolytic and antibacterial activities and could be used as diagnostic or vaccine candidates. To explore the biological functions of Ancylostoma ceylanicum SLP (Ace-SLP-1), cDNA-encoding Ace-SLP-1 mature peptide was cloned into prokaryotic expression vector pET-28a and transformed into Escherichia coli BL21 (DE3) to induce expression. After incubation of canine red blood cell suspension with different concentrations of recombinant Ace-SLP-1, the supernatant was separated to measure OD value and calculate the hemolysis rate. The different concentrations of recombinant protein were co-cultured with E. coli and Enterococcus faecalis, and colony-forming units (CFU) were determined by the plate counting method. Peripheral blood mononuclear cells (PBMCs) from healthy dogs were incubated with different concentrations of recombinant Ace-SLP-1, and the cytokine expression was evaluated by relative quantitative PCR. Our results showed that the hemolytic activity of Ace-SLP-1 increased with the increase in protein concentration from 25 to 100 μg/mL. The recombinant protein had no antibacterial activity against the two kinds of bacteria but could stimulate the secretion of cytokines (IL-4, IL-10, IL-12, and IL-13) in canine PBMCs. These data suggest that Ace-SLP-1 is involved in hookworm blood-feeding and survival and has good immunogenicity, supporting its potential as a diagnostic and vaccine target molecule.

Despite the recent advancements in transfusion medicine, red blood cell (RBC) alloimmunization remains a challenge for multiparous women and chronically transfused patients. At times, diagnostic laboratories depend on difficult‐to‐procure rare reagent RBCs for the identification of different alloantibodies in such subjects. We have addressed this issue by developing erythroblasts with custom phenotypes (Rh null, GPB null and Kx null/Kell low) using CRISPR/Cas9 gene‐editing of a human induced pluripotent stem cell (hiPSC) parent line (OT1‐1) for the blood group system genes: RHAG, GYPB and XK. Guide RNAs were cloned into Cas9‐puromycin expression vector and transfected into OT1‐1. Genotyping was performed to select puromycin‐resistant hiPSC KOs. CRISPR/Cas9 gene‐editing resulted in the successful generation of three KO lines, RHAG KO, GYPB KO and XK KO. The OT1‐1 cell line, as well as the three KO hiPSC lines, were differentiated into CD34+CD41+CD235ab+ hematopoietic progenitor cells (HPCs) and subsequently to erythroblasts. Native OT1‐1 erythroblasts were positive for the expression of Rh, MNS, Kell and H blood group systems. Differentiation of RHAG KO, GYPB KO and XK KO resulted in the formation of Rh null, GPB null and Kx null/Kell low erythroblasts, respectively. OT1‐1 as well as the three KO erythroblasts remained positive for RBC markers—CD71 and BAND3. Erythroblasts were mostly at the polychromatic/ orthochromatic stage of differentiation. Up to ~400‐fold increase in erythroblasts derived from HPCs was observed. The availability of custom erythroblasts generated from CRISPR/Cas9 gene‐edited hiPSC should be a useful addition to the tools currently used for the detection of clinically important red cell alloantibodies.

The objective of this randomized, placebo-controlled, double-blinded field trial was to investigate the effects of oral administration of purple coneflower (Echinacea purpurea L. (EP)) on performance, health and immune parameters in calves. Calves (n = 27) were enrolled to three groups (9 calves per group): 0.5 g EP/calf per day (ECL), 5 g EP/calf per day (ECH) or placebo. Calves were vaccinated with Bluetongue-Virus (BTV) serotype 4 vaccine to investigate EPs effects on seroconversion. Clinical and performance parameters, inter alia body weight, health and milk intake were recorded for 57 days. Blood samples were analyzed for BTV antibodies and IgG by ELISA, white and red blood cell counts by flow cytometry and mRNA abundance of various inflammatory markers in leukocytes (IL-1β, IL-8, tumor necrosis factor α (TNFα), cyclooxygenase 2 (Cox-2) and prostaglandin E synthase) was studied. The findings demonstrated no differences between groups regarding performance parameters. In all groups, calves suffered from diarrhea for a minimum of 2 days, but EP reduced the number of diarrhea days by 44% in ECL and increased the body temperature. Interestingly, ECL resulted in an increased number of respiratory disease days during the follow-up period. EP did not change blood cell and IgG counts, whereas eosinophil granulocytes were reduced in ECL. Decreased levels of hemoglobin and hematocrit were found in ECH. Prostaglandin E synthase levels in leukocytes were higher in ECL and ECH, whereas no differences were obtained for IL-1β, IL-8, TNFα and Cox-2. Due to the unexpected occurrence of BTV seropositive calves before the first vaccination, 13 calves were excluded from the evaluation on seroconversion and no statistical analyses could be performed regarding antibody production. BTV-4 antibodies were not produced in 4 placebo-calves, whereas 4 of 5 and 1 of 6 ECL- and ECH-calves produced antibodies. Further investigations are needed to draw final conclusions on mode of action and efficacy of EP in calves.

Sickle cell disease (SCD) poses a major public health challenge in sub-Saharan Africa, including Nigeria. Blood transfusion is a mainstay in SCD treatment. Erythrocyte alloimmunization is known to complicate the transfusional care of patients with SCD. Immune alloantibodies are associated with hemolytic transfusion reactions and transfusion refractoriness. We aimed to determine the prevalence, specificities, and clinical associations/risk factors of immune erythrocyte alloantibodies among adult patients with SCD compared with healthy blood donors in Lagos, Nigeria, through a cross-sectional study. All participants were interviewed using a structured questionnaire to obtain details on bio-data, hemoglobin phenotype, blood transfusion history, and SCD history where relevant. Blood specimens obtained from each participant were subjected to antibody screening/identification using tube agglutination method. The mean age of the SCD participants and healthy blood donors was 27.92 and 29.04 years, respectively. The majority (72.5%) of the SCD participants had received at least 1 unit of red blood cell (RBC) transfusion in their lifetime, compared with only 7.5 percent of blood donors. Six SCD participants (7.5%) tested positive for atypical erythrocyte alloantibodies, with none among blood donors. Most of the antibodies (75%) belonged to the Rh blood group system. The most frequent antibody was anti-E, followed by anti-C and anti-D. Advancing age (30 years or more), recent transfusions (last 4 weeks), higher transfusion rates, and established renal disease were significantly associated with alloimmunization (p values of 0.026, 0.043, 0.002, and 0.043, respectively). This study suggests blood transfusion as a strong risk factor for RBC alloimmunization in SCD patients. Extended RBC phenotyping is recommended for all patients with SCD, especially those receiving regular transfusions. Sickle cell disease (SCD) poses a major public health challenge in sub-Saharan Africa, including Nigeria. Blood transfusion is a mainstay in SCD treatment. Erythrocyte alloimmunization is known to complicate the transfusional care of patients with SCD. Immune alloantibodies are associated with hemolytic transfusion reactions and transfusion refractoriness. We aimed to determine the prevalence, specificities, and clinical associations/risk factors of immune erythrocyte alloantibodies among adult patients with SCD compared with healthy blood donors in Lagos, Nigeria, through a cross-sectional study. All participants were interviewed using a structured questionnaire to obtain details on bio-data, hemoglobin phenotype, blood transfusion history, and SCD history where relevant. Blood specimens obtained from each participant were subjected to antibody screening/identification using tube agglutination method. The mean age of the SCD participants and healthy blood donors was 27.92 and 29.04 years, respectively. The majority (72.5%) of the SCD participants had received at least 1 unit of red blood cell (RBC) transfusion in their lifetime, compared with only 7.5 percent of blood donors. Six SCD participants (7.5%) tested positive for atypical erythrocyte alloantibodies, with none among blood donors. Most of the antibodies (75%) belonged to the Rh blood group system. The most frequent antibody was anti-E, followed by anti-C and anti-D. Advancing age (30 years or more), recent transfusions (last 4 weeks), higher transfusion rates, and established renal disease were significantly associated with alloimmunization (p values of 0.026, 0.043, 0.002, and 0.043, respectively). This study suggests blood transfusion as a strong risk factor for RBC alloimmunization in SCD patients. Extended RBC phenotyping is recommended for all patients with SCD, especially those receiving regular transfusions.

The Inb antigen of the Indian blood group system is a high-prevalence antigen. The presence of alloanti-Inb in a recipient may pose a problem in finding compatible blood for transfusion. The aim of this study was to screen blood donors for Inb and to include individuals found to be In(b-) in our rare donor registry. To save resources, a unique study design was constructed. Blood group O donors were tested for Inb because their red blood cell (RBC) units could serve recipients across all ABO groups. EDTA blood samples were used for serologic and genomic testing. These samples were first tested serologically for Ina, and samples typed as In(a+) were then tested both serologically and molecularly for Ina and Inb to find homozygous IN*01/01 [i.e., the predicted In(b-) phenotype]. A cost-conservative approach in using recycling of antibody was adopted to economize available resources. Of 6300 donors, 196 donor samples typed as In(a+) and were also found to be In(b+) when tested by serologic and genomic methods. Although none of the donors typed as In(b-), the statistical analysis suggests the expected prevalence for this rare phenotype to be 0.02 percent among the total number of donors tested. In conclusion, this report presents a unique cost-conservative approach using limited reagents to screen a large number of donors for the rare In(b-) phenotype. The Inb antigen of the Indian blood group system is a high-prevalence antigen. The presence of alloanti-Inb in a recipient may pose a problem in finding compatible blood for transfusion. The aim of this study was to screen blood donors for Inb and to include individuals found to be In(b–) in our rare donor registry. To save resources, a unique study design was constructed. Blood group O donors were tested for Inb because their red blood cell (RBC) units could serve recipients across all ABO groups. EDTA blood samples were used for serologic and genomic testing. These samples were first tested serologically for Ina, and samples typed as In(a+) were then tested both serologically and molecularly for Ina and Inb to find homozygous IN*01/01 [i.e., the predicted In(b–) phenotype]. A cost-conservative approach in using recycling of antibody was adopted to economize available resources. Of 6300 donors, 196 donor samples typed as In(a+) and were also found to be In(b+) when tested by serologic and genomic methods. Although none of the donors typed as In(b–), the statistical analysis suggests the expected prevalence for this rare phenotype to be 0.02 percent among the total number of donors tested. In conclusion, this report presents a unique cost-conservative approach using limited reagents to screen a large number of donors for the rare In(b–) phenotype.

Cyanide induces acute lethal poisoning resulting from inhibition of cytochrome c oxidase located in the complex IV (Complex IV) of mitochondria. However, current therapies for cyanide poisoning using hydroxocobalamin and nitrous acid compounds remain a clinical issue. Here, we show that liposome-encapsulated methemoglobin (metHb@Lipo), nanosized biomimetic red blood cells, replicate the antidotal mechanism of nitrous acid compounds against cyanide poisoning, achieving superior efficacy and fast action with no adverse effects. The structure of metHb@Lipo, which consists of concentrated methemoglobin in its aqueous core and a lipid membrane resembling the red blood cell membrane, provides favorable characteristics as a cyanide antidote, such as binding properties and membrane permeability. Upon cyanide exposure, metHb@Lipo maintained the mitochondrial function in PC12 cells, resulting in a cell viability comparable to treatment with nitrous acid compounds. In a mouse model of cyanide poisoning, metHb@Lipo treatment dramatically improved mortality with a rapid recovery from the symptoms of cyanide poisoning compared to treatment with nitrous acid compounds. Furthermore, metHb@Lipo also possesses satisfactory pharmacokinetic properties without long-term bioaccumulation and toxicity. Our findings showed a novel concept to develop drugs for cyanide poisoning and provide a promising possibility for biomimetic red blood cell preparations for pharmaceutical applications.

The study was carried out to evaluate the impact of seasonal variations on the hematological responses of goats in Malaysia. Fifty-two goats (24 bucks and 28 does), aged between 1 to 2 years old were used in the study. The goats were fed on commercial concentrate and forage, access to mineral salt and water was ad libitum. Blood samples were obtained from each goat in January, May and September to analyze hematology blood constitute during the study. The results showed that red blood cell (RBC) levels in Anglo-Nubian females rose dramatically in January and May while the Saanen breed remains without diversity. In all seasons, high hemoglobin (Hb) concentration of Anglo-Nubian males was unlike Saanen breed. For Saanen females, mean corpuscular volume (MCV) increased significant-ly, whereas in Saanen males, mean corpuscular hemoglobin (MCH) and MCH concentration (MCHC) were increased. High packed cell volume (PCV) concentration of Anglo females compared to others during September. In conclusion, variations in breed and sex of goats reflect the distinct ability of trop-ical changes that have been formed during seasonal effects on hematological responses between the two breeds of goats.

Genotyping can be used to identify rare blood group antigens and to solve suspected blood group discrepancies, particularly when serologic methods are limited. Unfortunately, only a few such studies have been performed in Pakistan. The present study was conducted to determine the frequency of Dombrock blood group alleles by genotyping samples from blood donors from the north of Pakistan. Blood samples were taken with consent from 300 blood donors; DNA was extracted and tested for DO*01 and DO*02 alleles by sequence-specific primer polymerase chain reaction (PCR-SSP), followed by gel electrophoresis. Allele frequencies were calculated. The observed and expected genotype frequencies were compared using the χ2 test. The allele frequencies for DO*01 and DO*02 were 0.40 and 0.60, respectively. Genotype frequencies were in Hardy-Weinberg equilibrium. This study in Pakistani blood donors provides Dombrock blood group allele frequencies by PCR-SSP. This approach is efficient and economical and can be applied in developing countries. The findings can contribute to the development of in-house red blood cell panels, identification of rare blood types, and establishment of a national rare blood donor program. Genotyping can be used to identify rare blood group antigens and to solve suspected blood group discrepancies, particularly when serologic methods are limited. Unfortunately, only a few such studies have been performed in Pakistan. The present study was conducted to determine the frequency of Dombrock blood group alleles by genotyping samples from blood donors from the north of Pakistan. Blood samples were taken with consent from 300 blood donors; DNA was extracted and tested for DO*01 and DO*02 alleles by sequence-specific primer polymerase chain reaction (PCR-SSP), followed by gel electrophoresis. Allele frequencies were calculated. The observed and expected genotype frequencies were compared using the χ2 test. The allele frequencies for DO*01 and DO*02 were 0.40 and 0.60, respectively. Genotype frequencies were in Hardy-Weinberg equilibrium. This study in Pakistani blood donors provides Dombrock blood group allele frequencies by PCR-SSP. This approach is efficient and economical and can be applied in developing countries. The findings can contribute to the development of in-house red blood cell panels, identification of rare blood types, and establishment of a national rare blood donor program.

This study investigated the hemato- and genotoxic effects of formaldehyde (FA) and the possible mitigating role of hesperidin (HP) and N-acetylcysteine (NAC), each alone and in combination. Sixty-four adult male albino rats were divided into eight equal groups; the study was conducted for 8 weeks; Group I (negative control: received no medication), Group II (positive control: received distilled water), Group III (received HP 50 mg/kg/day), Group IV (received NAC 50 mg/kg/day), Group V (received FA 10 mg/kg/day), Group VI (FA + HP), Group VII (FA + NAC), and Group VIII (FA + HP + NAC). Groups VI, VII, VIII received the same previously mentioned doses and for the same duration. All treatments were given by intraperitoneal administration. At the end of the study, complete blood count, oxidative stress, histopathological changes, immunohistochemical staining of inducible nitric oxide synthase, and proliferating cell nuclear antigen and genotoxicity by comet assay in the bone marrow of treated rats were assessed. FA administration caused significant hematotoxicity represented by elevated white blood cell numbers and serum malondialdehyde levels and reduced red blood cell numbers, platelets, and serum superoxide dismutase values. Histologically, it induced an increase in fat cell numbers in bone marrow tissue with a widening of marrow spaces and decreased cellularity of hematopoietic cells, megakaryocytes, and granulocytes. FA exposure significantly decreased immunoreactivity for proliferating cell nuclear antigen, whereas the immunoreactivity for inducible nitric oxide synthase was increased. Genotoxicity, as measured by comet assay, revealed a significant increase in comet% and tail length in FA-treated group when compared with other groups. The cotreatment with HP and NAC revealed their ability to protect against hematological changes, oxidative damage, histopathological, and immunohistochemical changes, and genotoxicity induced by FA.

Outcomes following administration of very-low-dose recombinant activated factor VIIa (vld-rFVIIa) for cardiac surgical bleeding remain debatable. We sought to determine the association of vld-rFVIIa and adverse surgical outcomes. Retrospective, cohort matching of patients undergoing cardiac surgery who received vld-rFVIIa (median 13.02 μg/kg) for perioperative bleeding were matched to cardiac surgical patients who had bleeding and received standard of care for bleeding without Factor VIIa administration. Of the 362 matched patients (182 in each group), patients who received rFVIIa required significantly less red blood cell transfusions [median 3 units (range 0--60, IQR = 4 units) versus 4 units (range 2-34, IQR = 4 units); P = 0.0004], decreased length of hospital stay (median 8 versus 9 days; P = 0.0158) and decreased renal risk (P

The expression of GGCT (γ-glutamyl cyclotransferase) is upregulated in various human cancers. γ-glutamyl cyclotransferase enzyme activity was originally purified from human red blood cells (RBCs), but the physiological function of GGCT in RBCs is still not clear. Here we reported that Ggct deletion in mice leads to splenomegaly and progressive anaemia phenotypes, due to elevated oxidative damage and the shortened life span of Ggct-/- RBCs. Ggct-/- RBCs have increased reactive oxygen species (ROS), and are more sensitive to H2 O2 -induced damage compared to control RBCs. Glutathione (GSH) and GSH synthesis precursor l-cysteine are decreased in Ggct-/- RBCs. Our study suggests a critical function of Ggct in RBC redox balance and life span maintenance through regulating GSH metabolism.

OBJECTIVES To assess the associations between B-cell activating factor (BAFF) and alloimmunisation in multi-transfused thalassemia. BACKGROUND Red blood cell (RBC) alloimmunisation is a complication of multi-transfused thalassemia. BAFF is promoting B cells that produce alloantibodies. METHODS/MATERIALS Multi-transfused thalassemia, 15 years or older, were recruited in the cohort study. Alloantibodies and BAFF levels were analysed. RESULTS Of 114 patients, the overall prevalence of RBC alloimmunisation was 29.8%. The most common alloantibodies were anti-E, anti-Mia and anti-c. BAFF levels were different among the three groups; the patients with baseline alloantibodies (median ± interquartile range 1251 ± 474 pg/ml), without alloantibodies (1098 ± 453) and healthy controls (719 ± 306), p 25 years old patients (vs. the

Cytidine monophosphate-Nacetylneuraminic acid (Neu5Ac) hydroxylase (CMAH) and glycoprotein, alpha1, 3-galactosyltransferase (GGTA1) double knockout (DKO) pig models were produced to reduce immune reaction for xenotransplantation. However, the role of Neu5Gc and α-Gal in pigs has not been fully elucidated and it is necessary to consider the after-effect of inactivation of GGTA1 and CMAH in pigs. Hematological profiles of DKO pigs were analyzed through complete blood count (CBC). Histology of liver and spleen of DKO were investigated, and lectin blotting and mass spectrometry (MS) were performed to explore glycosylation changes in red blood cell (RBC) membranes of DKO pigs. DKO pigs showed common clinical signs such as weakness (100%), dyspnea (90%) and constipation (65%). DKO pigs revealed a significant decrease in RBC, hemoglobin (HGB) and hematocrit (HGB), and an increase in white blood cell (WBC), lymphocyte (LYM), monocyte (MON), and erythrocyte mean corpuscular volume (MCV). DKO piglets showed swollen liver and spleen, and exhibited raised deposition of hemosiderin and severe bleeding. Lectin assay and MS proved variations in glycosylation on RBC membranes. GGTA1/CMAH DKO pigs developed pathological features which are similar to anemic symptoms, and the variations in glycosylation on RBC membranes of DKO pigs may be attributed to the pathologies observed.

INTRODUCTION A Sysmex XN-series hematology analyzer (Sysmex), the next generation up from the Sysmex XE-series, can provide information regarding malaria infection in the form of a parasitic red blood cell (pRBC) flag. This study aimed to determine the usefulness of the pRBC flag for early detection and follow-up in patients infected with Plasmodium vivax. METHODS A total of 221 patients with fever for whom CBC and malaria microscopy had been requested were analyzed. Sixty-seven individuals were diagnosed with P vivax infection, and 154 were diagnosed with other febrile diseases. The sensitivity and specificity of the pRBC flag for malaria parasite detection and the relationship between parasite density and presence of the pRBC flag were determined. The concordance rate between malaria microscopy and pRBC flag in 147 follow-up cases was calculated. RESULTS The pRBC flag was detected in 56 of 67 malaria patients (sensitivity, 83.6%; specificity, 100%). The patients with the pRBC flag at initial diagnosis revealed significantly higher parasite density than the patients without the pRBC flag (P

Ouled Djellal sheep is the most dominant breed in Algeria due to its adapta-tion to harsh steppe environment. Hematological parameters are good indi-cators of animal health, but their values are breed-specific, and they are affected by age and gender. Therefore, this study aimed to determine the hematological profile of healthy Ouled Djellal lambs and ewe lambs reared traditionally under semi-arid zones of Algeria during the growing peri-od. Blood samples were taken from twenty-four Ouled Djellal lambs (12 males, 12 females) at 42, 63, 84 and 105 days of age to evaluate the count of White Blood Cells, Lymphocyte, Monocyte, Granulocyte, Red Blood Cell and total Platelet, the percentage of Lymphocyte, Monocyte and Granulocyte, Hemoglobin concentration, Hematocrit, Mean Corpuscular Volume, Mean Corpuscular Hemoglobin and Mean Corpuscular Hemoglobin Concentration by usual known methods. Both male and female lambs did not show signifi-cant variations in mean values of White Blood Cells, Lymphocyte count, Red Blood Cells, Hemoglobin, Mean Corpuscular Hemoglobin concentration, per-centage of Monocyte, Monocyte count, Gran percentage and Granulocyte count from 42ͭ ͪ day to 105ͭ ͪ days. Mean values of percentage of Lymphocyte, Hematocrite in lambs, Mean Corpuscular Hemoglobin in ewe lambs, and Mean Corpuscular Volume, Platelets levels in both lambs and ewe lambs indicated also variations during this period of growth. Significant Pearson's correlations were found between some hematological parameters in Ouled Djellal lambs and/or ewe lambs. Therefore, these findings provide usual val-ues, which can be considered for assessing the physiological, nutritional and health status of these animals.

Sickle cell anemia (SCA) is characterized by decreased red blood cell (RBC) deformability due to polymerization of deoxygenated hemoglobin, leading to abnormal mechanical properties of RBC, increased cellular adhesion, and microcirculatory obstruction. Prior work has demonstrated that NO• influences RBC hydration and deformability and is produced at a basal rate that increases under shear stress in normal RBC. Nevertheless, the origin and physiological relevance of nitric oxide (NO•) production and scavenging in RBC remains unclear. We aimed to assess the basal and shear-mediated production of NO• in RBC from SCA patients and control (CTRL) subjects. RBCs loaded with a fluorescent NO• detector, DAF-FM (4-Amino-5-methylamino- 2',7'-difluorofluorescein diacetate), were imaged in microflow channels over 30-min without shear stress, followed by a 30-min period under 0.5Pa shear stress. We utilized non-specific nitric oxide synthase (NOS) blockade and carbon monoxide (CO) saturation of hemoglobin to assess the contribution of NOS and hemoglobin, respectively, to NO• production. Quantification of DAF-FM fluorescence intensity in individual RBC showed an increase in NO• in SCA RBC at the start of the basal period; however, both SCA and CTRL RBC increased NO• by a similar quantity under shear. A subpopulation of sickle-shaped RBC exhibited lower basal NO• production compared to discoid RBC from SCA group, and under shear became more circular in the direction of shear when compared to discoid RBC from SCA and CTRL, which elongated. Both CO and NOS inhibition caused a decrease in basal NO• production. Shear-mediated NO• production was decreased by CO in all RBC, but was decreased by NOS blockade only in SCA. In conclusion, total NO• production is increased and shear-mediated NO• production is preserved in SCA RBC in a NOS-dependent manner. Sickle shaped RBC with inclusions have higher NO• production and they become more circular rather than elongated with shear.

This report is part of a series reporting the GRADE review performed by the 2018-2020 French Working Party on maternal red blood cell alloimmunisation. This report focusses on the clinical significance in obstetrics, as published in the scientific literature, of the rare RH antibodies, variants and antigens (i.e. excluding conventional RH1 trough RH8 antigens, RH12, RH22 and RH27, which are discussed in other reports of this series). Extremely severe or severe haemolytic disease of the fetus and the newborn (HDFN), leading to death or requiring transfusions, have been reported for: anti-RH1 (-D) associated with DVI, DBT and DIVb phenotypes, RHD*12.04 (DOL4), RHD*03.03 (DIIIc), RHD*D-CE(2-5)-D, RHD*01EL.31 (RHD*148+1T), anti-RH9 (-CX), anti-RH11 (-EW), anti-RH17 (-Hr0), anti-RH18 (-Hr), anti-RH19 (-hrS), anti-RH23 (-DW), anti-RH29 ("total" Rh), anti-RH30 (-Goa), anti-RH32, anti-RH34 (-HrB), anti-RH36 (-Bea), anti-RH40 (-Tar), anti-RH46 (-Sec), anti-RH48 (-JAL), anti-RH54 (DAK), and antibodies to high prevalence antigens such as those associated with RHCE*02.08.02 (RHCE*CW-RHD(6-10)), RHCE*03N.01 (RHCE*cEMI). HDFN of moderate, mild or undetailed severity have been reported for: anti-RH1 associated with DHar, DIIIa and DIVa phenotypes, RHD*01EL.08 (RHD*486+1A),RHD*01EL.44 (RHD*D-CE(4-9)-D),RHD*25 (DNB), anti-RH20 (-VS), anti-RH31 (-hrB), anti-RH37 (-Evans), ani-RH42, anti-RH49 (-STEM), anti-RH51 (-MAR), anti-RH55 (-LOCR), anti-RH58 (-CELO). Positive direct antiglobulin test in the newborn but no clinically significant HDFN has been reported for anti-RH1 (-D) associated with RHD*10.05 (DAU5), RHD*12.02 (DOL2). Because so many specificities are associated with severe HDFN in the RH system, all RH antibodies should be considered as potentially able to cause HDFN, even if none has been reported yet.

Hematopoiesis is a mechanism of multifunctional adaptation in vertebrates, including fish. Adaptive capabilities of populations are studied for a species of genus Salvelinus with high ecological plasticity. Char Salvelinus malma is represented in Kamchatka by the anadromous, river, brook, and several lake forms. Peripheral blood cells are investigated for this species juveniles and adults from the Raduga River in the lower Kamchatka River basin. The fish sampled in August 2013 were divided into two groups: sexually mature individuals (10 specimens) and juveniles (10 specimens). The body length (AC) of mature chars (mean ± standard deviation, range in brackets) was 338.90 ± 12.64 (304.0–380.0) mm, the body weight — 437.40 ± 57.74 (235.0–735.0) g, their sex ratio was close to 1 : 1. The length of juvenile chars was 150.50 ± 9.14 (104.0–195.0) mm, the weight — 33.90 ± 6.19 (10.1–68.8) g. Blood was sampled from the caudal vessel of alive fish immediately after catching, the blood smears were dried and fixed with May-Grunwald dye and then stained with azure-eosin. Composition of leukocytes and erythrocytes was determined on the stained blood smears. The blood cells were classified according to Ivanova (1983) and Zhiteneva et al. (1989). Statistical significance of the blood cells composition differences between fish groups was assessed using the Mann-Whitney U-test, after checking for normal distribution of characteristics (Kolmogorov-Smirnov test). The erythroid row of S. malma was represented by proliferating (erythroblasts, basophilic and polychromatophilic normoblasts), maturing (oxyphilic normoblasts) and mature erythrocytes, the leukocyte row — by blast cells, agranulocytes and granulocytes. The juveniles were characterized by intense proliferative processes (great number of immature erythrocytes, large and medium lymphocytes, presence of blast cells) and appearance of poikilocytes; the sexually mature chars were distinguished by greater number of granulocytes, mainly due to stab and segmented neutrophils (p < 0.01). Parasitological analysis showed different infestation of the juvenile and adult chars by pathogenic helminths (Anisakis simplex, Eubothrium salvelini, Neoechinorhynchus salmonis) that influenced on their blood. Broad adaptive capabilities of S. malma hematopoiesis provided by wide morphological and functional diversity of blood cells are concluded that maintains homeostasis of the species. Blood indices for char from the Raduga River can be used to form reference intervals for monitoring physiological state of this species.

Thrombosis is a major complication of cardiovascular disease, leading to myocardial infarction, acute ischemic stroke, or venous thromboembolism. Thrombosis occurs when a thrombus forms inside blood vessels disrupting blood flow. Developments in thrombectomy to remove thrombi from vessels have provided new opportunities to study thrombus composition which may help to understand mechanisms of disease and underpin improvements in treatments. We aimed to review thrombus compositions, roles of components in thrombus formation and stability, and methods to investigate thrombi. Also, we summarize studies on thrombus structure obtained from cardiovascular patients and animal models. Thrombi are composed of fibrin, red blood cells, platelets, leukocytes, and neutrophil extracellular traps. These components have been analyzed by several techniques, including scanning electron microscopy, laser scanning confocal microscopy, histochemistry, and immunohistochemistry; however, each technique has advantages and limitations. Thrombi are heterogenous in composition, but overall, thrombi obtained from myocardial infarction are composed of mainly fibrin and other components, including platelets, red blood cells, leukocytes, and cholesterol crystals. Thrombi from patients with acute ischemic stroke are characterized by red blood cell- and platelet-rich regions. Thrombi from patients with venous thromboembolism contain mainly red blood cells and fibrin with some platelets and leukocytes. Thrombus composition from patients with myocardial infarction is influenced by ischemic time. Animal thrombosis models are crucial to gain further mechanistic information about thrombosis and thrombus structure, with thrombi being similar in composition compared with those from patients. Further studies on thrombus composition and function are key to improve treatment and clinical outcome of thrombosis.

Units of red blood cell (RBC) concentrates with rare phenotypes are typically not included in method validation studies for cryopreservation processes; rather, they are reserved for patients with rare blood needs. Some rare RBC phenotypes may demonstrate membrane abnormalities, like acanthocytosis as observed for RBCs with the McLeod phenotype, and are specifically banked for these rare attributes; however, the impact that rare RBC phenotypes have on post-thaw quality has not been well studied. To evaluate how a rare RBC phenotype is affected by the cryopreservation process, 4 RBC units, cryopreserved in 1993 using manual methods, were selected for evaluation. These RBCs included one with the McLeod phenotype and three with phenotypes not known to cause significant membrane changes. Post-thaw, an altered deglycerolization protocol, implemented to reduce supernatant glycerol after cryopreservation, was used before processing RBCs on an automated closed system (ACP 215; Haemonetics, Boston, MA) to accommodate the use of a closed system cell processor not available when the RBC units were previously cryopreserved. RBC quality was tested at 24 hours, 7 days, and 14 days post-deglycerolization. Before deglycerolization, an extracted sample from the thawed glycerolized RBC unit was used to obtain genetic material for phenotype confirmation. Genotyping confirmed the McLeod phenotype. When comparing McLeod with non-McLeod units, RBCs from the McLeod donor exhibited acanthocytosis, higher rigidity, and lower morphology scores than RBCs from the non-McLeod units post-deglycerolization. Hemolysis, however, was comparable across all 4 units, meeting regulatory standards. Therefore, McLeod RBCs can withstand cryopreservation, suggesting that units from these donors, glycerolized using older methods, can be deglycerolized using the ACP 215 and stored hypothermically for 14 days. It was also determined that genotyping can be performed on non-leukocyte-reduced cryopreserved RBCs, allowing for confirmation of genetic profiles of donor units banked before the implementation of molecular methods. Units of red blood cell (RBC) concentrates with rare phenotypes are typically not included in method validation studies for cryopreservation processes; rather, they are reserved for patients with rare blood needs. Some rare RBC phenotypes may demonstrate membrane abnormalities, like acanthocytosis as observed for RBCs with the McLeod phenotype, and are specifically banked for these rare attributes; however, the impact that rare RBC phenotypes have on post-thaw quality has not been well studied. To evaluate how a rare RBC phenotype is affected by the cryopreservation process, 4 RBC units, cryopreserved in 1993 using manual methods, were selected for evaluation. These RBCs included one with the McLeod phenotype and three with phenotypes not known to cause significant membrane changes. Post-thaw, an altered deglycerolization protocol, implemented to reduce supernatant glycerol after cryopreservation, was used before processing RBCs on an automated closed system (ACP 215; Haemonetics, Boston, MA) to accommodate the use of a closed system cell processor not available when the RBC units were previously cryopreserved. RBC quality was tested at 24 hours, 7 days, and 14 days post-deglycerolization. Before deglycerolization, an extracted sample from the thawed glycerolized RBC unit was used to obtain genetic material for phenotype confirmation. Genotyping confirmed the McLeod phenotype. When comparing McLeod with non-McLeod units, RBCs from the McLeod donor exhibited acanthocytosis, higher rigidity, and lower morphology scores than RBCs from the non-McLeod units post-deglycerolization. Hemolysis, however, was comparable across all 4 units, meeting regulatory standards. Therefore, McLeod RBCs can withstand cryopreservation, suggesting that units from these donors, glycerolized using older methods, can be deglycerolized using the ACP 215 and stored hypothermically for 14 days. It was also determined that genotyping can be performed on non-leukocyte–reduced cryopreserved RBCs, allowing for confirmation of genetic profiles of donor units banked before the implementation of molecular methods.

The Kidd-null phenotype, Jk(a-b-), is rare, and a patient with this phenotype may develop anti-Jk3, a red blood cell (RBC) antibody reactive with a domain common to both Jka and Jkb. Like other antibodies to high-prevalence antigens, the presence of this antibody poses challenges in the immunohematologic evaluation of these patients. Thoughtful laboratory testing is necessary to resolve the antibody specificity and to reveal other underlying antibodies. Moreover, the rarity of the Kidd-null phenotype makes finding blood donors difficult for those who need transfusion and have developed anti-Jk3. This review describes methods used in identifying anti-Jk3 in four pregnant patients. Blood bank records were retrospectively reviewed to illustrate the common approach in anti-Jk3 identification. In all cases, pertinent blood bank history was gathered, and extended RBC phenotyping was performed, followed by adsorption studies and testing of selected RBCs. Underlying antibodies were found in two of the cases. This review also reiterates some common challenges encountered with Kidd antibody analysis and highlights the importance of patient ethnic ancestry and obtaining accurate patient transfusion history.The Kidd-null phenotype, Jk(a–b–), is rare, and a patient with this phenotype may develop anti-Jk3, a red blood cell (RBC) antibody reactive with a domain common to both Jka and Jkb. Like other antibodies to high-prevalence antigens, the presence of this antibody poses challenges in the immunohematologic evaluation of these patients. Thoughtful laboratory testing is necessary to resolve the antibody specificity and to reveal other underlying antibodies. Moreover, the rarity of the Kidd-null phenotype makes finding blood donors difficult for those who need transfusion and have developed anti-Jk3. This review describes methods used in identifying anti-Jk3 in four pregnant patients. Blood bank records were retrospectively reviewed to illustrate the common approach in anti-Jk3 identification. In all cases, pertinent blood bank history was gathered, and extended RBC phenotyping was performed, followed by adsorption studies and testing of selected RBCs. Underlying antibodies were found in two of the cases. This review also reiterates some common challenges encountered with Kidd antibody analysis and highlights the importance of patient ethnic ancestry and obtaining accurate patient transfusion history.

Maternal antibody-mediated fetal red blood cell destruction secondary to non-D Rh system antibodies is a significant cause of hemolytic disease of the fetus and newborn. Here, we report a rare case of severe perinatal hemolytic disease associated with maternal antibody to the e antigen. In addition to severe anemia, the infant developed hyperbilirubinemia. Resolution of the infant's anemia and hyperbilirubinemia occurred after treatment with phototherapy, intravenous immunoglobulin, and transfusion. Maternal antibody-mediated fetal red blood cell destruction secondary to non-D Rh system antibodies is a significant cause of hemolytic disease of the fetus and newborn. Here, we report a rare case of severe perinatal hemolytic disease associated with maternal antibody to the e antigen. In addition to severe anemia, the infant developed hyperbilirubinemia. Resolution of the infant’s anemia and hyperbilirubinemia occurred after treatment with phototherapy, intravenous immunoglobulin, and transfusion.

Background Patients with sickle cell disease (SCD) are frequent recipients of red blood cell (RBC) transfusions and are at risk for RBC alloimmunization. RBC alloimmunization is diagnosed by identifying RBC alloantibodies as part of pre-transfusion testing, but this testing fails to detect alloantibodies that have evanesced. It may be beneficial to screen for new RBC alloantibody development after transfusion before possible antibody evanescence. Study design and methods Our institution started a new initiative for episodically transfused patients with SCD to obtain at least one antibody screen 2-6 months after transfusion as part of their clinical care. A database was created to prospectively track all transfused patients for 1 year and their post-transfusion antibody screen results. Patients received prophylactically CEK-matched RBC units. Results During the study year, 138 patients with SCD received a total of 242 RBC transfusions. Patients with a history of an RBC alloantibody (n = 13, 9.4%) had previously received more RBC units than non alloimmunized patients (median 11 vs. 2 RBC units, p = .0002). A total of 337 post-transfusion antibody screens were obtained in 127 patients (92.0%) with 110 patients (79.7%) having at least one antibody screen 2-6 months post-transfusion. With this prospective testing, two new RBC alloantibodies (anti-C and -M) were identified in two patients. Conclusion It is feasible to test for new RBC alloantibody development in most episodically transfused patients with SCD as part of their routine care. The yield of this screening appears low with CEK matching, but it could still provide important information for individual patients.

Band 3 (anion exchanger 1; AE1) is the most abundant membrane protein in red blood cells, which in turn are the most abundant cells in the human body. A compelling model posits that, at high oxygen saturation, the N-terminal cytosolic domain of AE1 binds to and inhibits glycolytic enzymes, thus diverting metabolic fluxes to the pentose phosphate pathway to generate reducing equivalents. Dysfunction of this mechanism occurs during red blood cell aging or storage under blood bank conditions, suggesting a role for AE1 in the regulation of the quality of stored blood and efficacy of transfusion, a life-saving intervention for millions of recipients worldwide. Here we leveraged two murine models carrying genetic ablations of AE1 to provide mechanistic evidence of the role of this protein in the regulation of erythrocyte metabolism and storage quality. Metabolic observations in mice recapitulated those in a human subject lacking expression of AE11-11 (band 3 Neapolis), while common polymorphisms in the region coding for AE11-56 correlate with increased susceptibility to osmotic hemolysis in healthy blood donors. Through thermal proteome profiling and crosslinking proteomics, we provide a map of the red blood cell interactome, with a focus on AE11-56 and validate recombinant AE1 interactions with glyceraldehyde 3-phosphate dehydrogenase. As a proof-of-principle and to provide further mechanistic evidence of the role of AE1 in the regulation of redox homeo stasis of stored red blood cells, we show that incubation with a cell-penetrating AE11-56 peptide can rescue the metabolic defect in glutathione recycling and boost post-transfusion recovery of stored red blood cells from healthy human donors and genetically ablated mice.

Since its discovery in 1994, leptin continues to have new potential physiological roles uncovered, including a role in the regulation of blood flow. Leptin's role in regulating blood flow is not completely understood. Red blood cell (RBC)-derived ATP is a recognized stimulus of blood flow, and multiple studies suggest that C-peptide, a hormone secreted in equimolar amounts with insulin from the pancreatic β-cells, can stimulate that release when delivered by albumin and in combination with Zn2+. Here, we report leptin delivers C-peptide and Zn2+ to RBCs in a saturable and specific manner. We labeled leptin with technetium-99 m (99mTc) to perform binding studies while using albumin to block the specific binding of 99mTc-leptin in the presence or absence of C-peptide. Our results suggest that leptin has a saturable and specific binding site on the RBC ((Kd = 1.79 ± 0.46) × 10-7 M) that is statistically equal to the binding affinity in the presence of 20 nM C-peptide ((Kd = 2.05 ± 0.20) × 10-7 M). While the binding affinity between leptin and the RBC did not change with C-peptide, the moles of bound leptin did increase with C-peptide, suggesting a separate binding site on the cell for a leptin/C-peptide complex. The RBC-derived ATP increased in the presence of a leptin/C-peptide/Zn2+ addition, in a concentration-dependent manner. Control RBCs ATP release increased (71 ± 5.6%) in the presence of C-peptide and Zn2+, which increased further to (94 ± 5.6%) in the presence of Zn2+, C-peptide, and leptin.

Herein, Vernonia amygdalina stem ethanolic extract, VASEE, was elucidated for its antibacterial activity in vitro by disk-diffusion technique and, in vivo effect on some hematological parameters of normal and monosodium glutamate, MSG-intoxicated rats. For in vivo study, 20 male albino rats assigned into five groups (A, B, C, D, and E) were, for 14 days, respectively administered MSG (8000 mg/kg body weight, bw), VASEE (200 mg/kg bw), Control (Vital feed and tap water), MSG (8000 mg/kg bw + VASEE, 200 mg/kg bw), and MSG (8000 mg/kg bw + VASEE, 400 mg/kg bw). In vitro, the VASEE-related activity against the tested bacterial pathogens was significant (P < 0.05), dose dependent and comparable to that by the standard antibacterial drug, Ciprofloxacin. In vivo, VASEE compared to control and MSG groups improved (P < 0.05) and, notably at the highest tested dose modulated the monosodium glutamate intoxication-related effect on, the studied hematology of rats. Thus, VASEE exhibited potent activity against the tested bacterial pathogens, improved, and potentially modulated MSG-intoxication-related effect on, the rats’ hematological functions. The study underscored a promising antibacterial application of the extract of hitherto wasted bitter leaf stem that could offer novel therapeutic benefits on the hematology of especially MSG-intoxicated rats, warranting further studies. Keywords: Hematology, intoxication, bitter leaf stem, packed cell volume, Red blood cell

An experiment was carried out to determine the growth performance and haematological indices of Red Sokoto goats placed on oil palm leaf meal (OPLM)-wheat offal based diet. Six goats aged 12-18 months, weighing 15 – 20kg were randomly assigned to three treatments,comprising two animals each in a Completely Randomized Design. Each animal received 1kg of the diet daily for 56 days and weighed weekly. Results showed that final body weight was significant (P

The present investigation was conducted to estimate the acute toxicity of Thymus linearis plant extract, its effect on hemato-biochemical parameters and behavioural response in the golden mahseer (Tor putitora). The phytochemical composition present in T. linearis plant extrat were Alkaloids, Flavonoids, Phenols and Tannin. The fishes were subjected to eight different concentrations of T. linearis leaves extract (8.25, 8.50, 8.75, 9.00, 9.25, 9.50 and 9.75 mg/kg) and the control group without plant extract for 96-h LD50 study. The mortality was recorded every 24 h post-treatment. Minimum mortality was recorded in the 8.25 mg/kg, whereas 100% mortality was recorded in the 9.75 mg/kg T. linearis extract after 96-h periods. The LD50 was estimated by probit analysis, and the value of T. linearis at 96 h was found to be 8.71 mg/kg for golden mahseer. A non-lethal dose of 1/10th of 96-h LD50 value (0.87 mg/kg) was taken for the sublethal study. After 96 h, the red blood cell (RBC), white blood cell (WBC), haemoglobin (Hb), packed cell volume (PCV) and blood glucose were measured. RBC (×106/mm3), Hb (%) and PCV (%) significantly decreased at 8.25, 8.50, 8.75, 9.00 mg/kg, but WBC and blood glucose significantly increase at 8.25, 8.50, 8.75, 9.00 mg/kg of T. linearis plant extract. The observations on behaviour response of golden mahseer were also recorded. In the present study, the acute toxicity of wild ajwain was more significant than short-term toxicity. The mortality rate was very high during the study period of T. linearis exposure.

BACKGROUND AND PURPOSE: Previous studies have successfully created blood clot analogs for in vitro endovascular device testing using animal blood of various species. Blood components vary greatly among species; therefore, creating clot analogs from human blood is likely a more accurate representation of thrombi formed in the human vasculature. MATERIALS AND METHODS: Following approval from the Mayo Clinic institutional review board, human whole-blood and platelet donations were obtained from the blood transfusion service. Twelve clot analogs were created by combining different ratios of red blood cells + buffy coat, plasma, and platelets. Thrombin and calcium chloride were added to stimulate coagulation. Clot composition was assessed using histologic and immunohistochemical staining. To assess the similarities of mechanical properties to patient clots, 3 types of clot analogs (soft, elastic, and stiff) were selected for in vitro thrombectomy testing. RESULTS: The range of histopathologic compositions produced is representative of clots removed during thrombectomy procedures. The red blood cell composition ranged from 8.9% to 91.4%, and fibrin composition ranged from 3.1% to 53.4%. Platelets (CD42b) and von Willebrand Factor ranged from 0.5% to 47.1% and 1.0% to 63.4%, respectively. The soft clots had the highest first-pass effect and successful revascularization rates followed by the elastic and stiff clots. Distal embolization events were observed when clot ingestion could not be achieved, requiring device pullback. The incidence rate of distal embolization was the highest for the stiff clots due to the weak clot/device integration. CONCLUSIONS: Red blood cell–rich, fibrin-rich, and platelet-rich clot analogs that mimic clots retrieved from patients with acute ischemic stroke were created in vitro. Differing retrieval outcomes were confirmed using in vitro thrombectomy testing in a subset of clots. This work was supported by the National Institutes of Health grant No. R01 NS105853, the European Regional Development Fund and Science Foundation Ireland grant No. 13/RC/2073, and Enterprise Ireland (IP20190865). peer-reviewed

The pluripotent stem cells which are present in the bone marrow renew by its own and differentiate into mature cells. These stem cells undergo division by the presence of erythropoietin, where the nucleus is extruded out from the cell during the end of differentiation, thereby retaining cytoplasmic RNA to form a reticulocyte. The reticulocyte i s a precursor to red blood cell and on losing the RNA it matures into a Red Blood Cell. The present study aimed to analyze the correlation between the automated histogram patterns along with morphological features of RBC‘s prepared from peripheral smear examination in different types of anemia. viz., MCV, MCH, MCHC & RWD- CV.