Morphological Characteristics of the Blood Cells and Spermatozoa in Chinese Monals

The Effects of Splenectomy and Autologous Spleen Transplantation on Complete Blood Count and Cell Morphology in a Porcine Model

Most hematology analyzers today are capable of performing white blood cell differential analysis but only limited red blood cell (RBC) differential parameters are available. Because of incomplete morphological information on RBC abnormalities, 5-10% of samples in hematology laboratories routinely undergo smear review. A more complete automated RBC differential capability is desired. Abbott CELL DYN 4000 analyzer was modified to perform three-dimensional (3D) RBC differential analysis (RBC/diff) on cell-by-cell basis at 488 nm as well as at 633 nm. Immature RBCs and all nucleated cells are labeled with a fluorescent nuclear stain, prior to RBC/diff using light loss and forward scatter (FSC) signal or two FSC signals and a third side scatter signal, projecting the cytogram onto a precalibrated 3D surface containing grid lines of volume (V) and hemoglobin concentration (HC), to determine the V and HC of a cell. Abnormally shaped RBCs (AbnRBC) are quantitated by the distance of each event to the 3D surface. A total of 154 normal and 484 clinical samples with various hemoglobinopathies have been analyzed. The 3D cytogram and the bivariate distribution of V and HC of individual event provided information to extract quantitative data on V, HC, schistocytes, and AbnRBC of mature and immature RBCs. The accuracy level of the 3D data is difficult to assess because of the semiquantitative nature of smear review, the only available reference method. The 3D RBC cytograms provide a wealth of morphologic information useful for diagnosis and treatment of patients. With powerful computer programs available today, it is evident that the rate of smear review can be significantly reduced as a result.

Introduction: In amphibians, blood may act as a hematopoietic tissue. However, the knowledge concerning hematological features is scarce, there is not much information that allows an analysis about the possible explanations of this physiological feature. Objective: This study aimed to evaluate the relationship between immature red blood cells (RBCs) mitosis and the presence of blood parasites in amphibians. Methods: We sampled 116 amphibians (31 species) in six Colombian localities. Blood was taken by cardiac puncture or maxillary vein puncture. Smears were prepared, fixed, and Giemsa stained for microscopical analysis. The variables analyzed were the percentage of immature RBCs, mitotic cells in peripheral blood, and blood parasite infection. Data were analyzed using Wilcoxon's rank test and exact Fisher statistical tests. Results: Sixty-two individuals showed mitosis in peripheral blood, and these mitotic RBCs shared morphological features with immature RBCs. Overall, parasite prevalence was 30.1 %, distributed as follows: Trypanosoma (24.1 %), Hepatozoon-like (6 %), Dactylosoma (4.3 %), Karyolysus-like (0.9 %), and Filarioidea (2.6 %). A positive association between the percentage of immature RBCs and the presence of mitotic RBCs was found, and also between the blood parasite infection and the percentage of immature RBCs. Conclusions: In this study, we found that the presence of blood parasites, immature RBCs, and RBCs mitosis are frequent events in amphibians' peripheral blood, and our analysis suggests an association between those features. Thus, the release of immature RBCs and the mitosis of those cells in peripheral blood may be a physiological response to blood parasite infection. Further studies characterizing hematology in amphibians and wildlife, in general, are desirable.

We analysed the rates of histone deacetylation in chicken mature and immature red blood cells. A multiplicity of deacetylation rates was observed for the histones and these rates may be subdivided into two major categories based on the extent of histone acetylation. In one set of experiments, cells were labelled with [3H]acetate in the presence of the deacetylase inhibitor n-butyrate, thereby accumulating radiolabel in the hyperacetylated forms of the histone. These hyperacetylated forms are deacetylated rapidly. [3H]Acetate-labelled tetra-acetylated H4 (H4Ac4) in mature cells was deacetylated with an initial half-life (t1/2) of approximately 5 min (time required for the removal of one-half of the labelled acetyl groups). In immature cells, all [3H]acetate-labelled H4Ac4 was deacetylated with a t1/2 of approximately 5 min. Erythrocytes were also labelled with [3H]acetate for extended periods in the absence of the deacetylase inhibitor. During this period, radiolabel accumulated predominantly in the mono- and di-acetylated forms of the histone. Using this protocol, the rate of deacetylation of H4Ac1 was observed to be approximately 145 min for mature cells, and approximately 90 min for immature cells, demonstrating that the less extensively acetylated histone is deacetylated slowly. These results are discussed in the context of the rates of histone acetylation in chicken red blood cells described in the companion paper [Zhang & Nelson (1988) Biochem. J. 250, 233-240].

Assessment of Novel Erythroid Signaling Pathways Modulating Hydroxyurea-Mediated Reduction of Vaso-Occlusive Adhesive Interactions in Early Precursors of Sickle Red Blood Cells (RBCs)

Red blood cells are flexible and oval biconcave disks, they lack a cell nucleus and their disk diameter is 7–10 μm. Approximately 2.4 million new erythrocytes are produced per second and circulate for about 100–120 days in the body. Red blood cells are cells present in blood in order to transport oxygen, the aging red blood cell undergoes changes making it susceptible to selective recognition by macrophages and subsequent phagocytosis. The cytoplasm of erythrocytes is rich in hemoglobin, an iron-containing biomolecule that can bind oxygen and is responsible for the red colour of the cells. Immature red blood cells are lacking the red hemoglobin pigment so these cells are often shades of grayish blue, only mature cells are red.A glycophorin is a sialoglycoprotein of the membrane of a red blood cell, that contains N-acetylneuraminic acid. Ions such as Na+ and Ca2+ can diffuse rapidly through it and can be for 60% it`s negative charge of the plasma membrane. Typical human red blood cell has a diameter of approximately 6.2–8.2 μm, a thickness - 2 μm, circumference – 76-110 μm, speed no more than 2 cm/ sec that is enough to transport oxygen from hemoglobin toward myoglobin. Listed features are changed depending on the lifespan of red blood cells: 1. Decreases the percentage of hemoglobin content, within the part of it is broken down. 2. Changes occurs in the activities electronic change in oxidation and restoration of Fe. 3. As erythrocyte ages, it undergoes changes in its plasma membrane, in particular sialic acid activity. 4. Erythrocyte membrane becomes inflexible, less elastic and rough. 5. Worn-out red blood cells (100-120 day) have a limited functional significance. As a result of this research, both functional and physical indicators are strictly differentiated regarding to human age: 1. The length of erythrocyte life in the elderly (70-75) is twice longer than in younger people (25-30). 2. Red blood cells are remarkably deformable in younger than in elderly people. 3. Because of decreased deformability of the red blood cell, they have trouble to squeeze through capillaries which is the reason of hemodynamics local violations.

More stomatocytes, spherocytes, and erythrocytes with spicules and protrusions are found in blood from newborn infants than in blood from adults. The objective of this study was to measure volume, surface area, membrane rigidity, and viscosity of red blood cells (RBC) with irregular shapes in preterm and full-term neonates. RBC from 13 full-term and 12 preterm (25-36 wk of gestation) infants were studied by means of a micropipette system. In full-term neonates and in preterm infants, 19 and 26% of RBC, respectively, showed shapes different from those of discocytes. Keratocytes, spherocytes, akanthocytes (in full-term infants), and elliptocytes (in preterm infants) had smaller volumes than did discocytes (p < 0.05). Spherocytes showed decreased and elliptocytes had increased excess surface area and swelling capacity compared with discocytes, indicating both volume and membrane loss for spherocytes and only volume loss for elliptocytes. Immature RBC and knizocytes had similar large volumes and surface areas, suggesting that knizocytes in neonates are relatively young RBC. Increased membrane rigidity was found only in stomatocytes. Membrane viscosity was decreased in immature RBC and knizocytes and increased in stomatocytes, keratocytes, akanthocytes, and elliptocytes when compared with discocytes. There were inverse relationships between RBC volume and membrane rigidity (r=-0.42) and between RBC volume and membrane viscosity (r=-0.72). This suggests that volume loss of neonatal RBC is associated with impaired membrane deformability. Moreover, we conclude that the geometry and membrane deformability of some of the irregularly shaped neonatal RBC resemble properties of RBC in hemolytic anemias (e.g. spherocytosis).

After completing this article, readers should be able to: 1. Describe alterations of red blood cells due to bacterial toxins. 2. Describe physical alterations of neutrophils during activation. 3. Delineate the role of red and white blood cells in the impairment of microcirculation and organ damage in sepsis. 4. List drugs that may inhibit neutrophil activation and improve their deformability and their actions. Sepsis remains a major cause of morbidity and mortality in neonates, particularly among preterm infants. (1) Impaired microcirculatory blood flow plays a pivotal role in the development of clinical manifestations and organ dysfunctions in severe sepsis and septic shock. (2)(3)(4) If not corrected, microcirculatory dysfunction can progress to organ dysfunction and subsequently to organ failure and death. Restoration of microcirculatory dysfunction is, therefore, an important step in preventing long-term sequelae (including brain damage) and death of the patient. Both red and white blood cells must deform to pass through narrow channels whose diameters are less than those of the cells. (5) Impaired deformability of red and white blood cells may contribute to impaired microcirculatory blood flow in septicemia. The membranes of neonatal red blood cells (RBCs) deform more in response to a given force than adult cells and are, therefore, more flexible. (6)(7) On the other hand, neonatal RBCs are larger and require higher pressures to enter filter pores and micropipettes that have diameters below the resting cellular diameters. (8)(9) The larger volume and increased deformability of neonatal RBCs are responsible for another favorable property, the increased Fahraeus and Fahraeus-Lindqvist effect (ie, hematocrit and viscosity reduction when going from 500- to 50-mcm tubes). (10) These favorable flow properties of neonatal RBCs suggest that their increased cellular deformability is a prerequisite for adequate flow in arterioles and capillaries. We have shown that loss of …

IntroductionHemodynamically significant patent ductus arteriosus (hsPDA) in preterm infants is associated with increased mortality and morbidity. Early and accurate identification of infants at risk for PDA is critical for improving outcome. Perinatal inflammation is a known risk factor of PDA. CD71+ erythroid cells (CECs) are immature red blood cells with immunomodulatory roles and show variable abundance in preterm infants. We aimed to examine the proportion of CECs in preterm infants and investigate association with PDA.MethodsIn this prospective study, we evaluated the association between CEC levels and the development of hsPDA in a cohort of 108 preterm infants born before 37 weeks of gestation. CEC levels were quantified by flow cytometry within 12 hours of birth. The presence of hsPDA was assessed by echocardiography between days 3 and 7 after birth.ResultsCEC levels showed a significant inverse relationship with gestational age (CEC(%) = -0.4419 × Gestational Age + 18.7029, p<0.001). CEC levels were also inversely correlated with hemoglobin, red blood cells, white blood cells, and platelets (all p < 0.05). Infants who developed hsPDA had significantly higher CEC levels (7.5 ± 2.8% vs. 4.4 ± 1.7%, p < 0.0001), together with lower values of these hematologic parameters than infants without hsPDA. Multivariable logistic regression identified elevated CECs levels (odds ratio: 2.048, 95% CI: 1.326–3.162, p = 0.0012) and lower white blood cell count (odds ratio: 0.782, 95% CI: 0.620–0.986, p = 0.038) as independent predictors of hsPDA. Additionally, CECs levels were inversely correlated with hemoglobin, red blood cells, white blood cells, and platelets (p<0.05).DiscussionThese results suggest that early measurement of CEC levels can identify preterm infants at high risk for hsPDA prior to echocardiographic diagnosis. As a potential predictive biomarker, CECs may improve risk stratification and guide timely intervention, ultimately enhancing clinical outcomes. Further research is needed to elucidate the mechanisms linking CECs to hsPDA.

Scanning Electron Microscopy (SEM) is utilized to elucidate the morphological variation in red blood cells. Hemoglobin concentration in human blood is in the range of 13.0 g/dl -11.5 g/dl in healthy adults. Hemoglobin concentration in anaemic red blood is below the lower limit of the normal range. Sometime the nature of the abnormal shape of the blood cell determines the cause of disease. Normally there occurs a variation in the diameter of the red blood cell for different types of disease. Increased variation of size in blood cell is termed anisocytosis [1]. The classification of anaemia by the size of blood cell is logical i.e. common morphological abnormality of human blood cell [2]. Sometime Cancer cells of different types are analogous with anaemic cells. So for the present study cells are observed under ZEISS SEM with different magnification and applied potential of kV range. Thus the diameters of red blood cells in SEM have been compared with red blood cells photographed with light microscope. Diseased blood cells are observed overlapped with each other with increasing diameter. Keywords : SEM, Anaemia, Anisocytosis, Hemoglobin, Cell-morphology, RBC,Cancer)

Blood cell morphology and count are not uniform across species. Recently, between-species comparisons revealed that the size of red blood cells is associated with body size in some lizard taxa, and this finding was interpreted in the context of the metabolic theory. In the present study, we examined the numbers and the size of blood cells in 2 species of monitor lizards, the mangrove-dwelling monitor (Varanus indicus) and the savannah monitor (V. exanthematicus), and we compared these traits in individuals of different body size. The results revealed that during the course of ontogeny, the size of red blood cells increases with body mass. Because the mass-specific metabolic rate decreases with body size and the cell volume-to-surface ratio decreases with the cell size, changes in the erythrocyte size might be the result of oxygen transport adjustment.

The effect of Malathion was studied orally on Japanese quail males (Cotrunix Cotrunix Japonica). The Median lethal Dose LD50 of malathion through 24 hours was found to be 163.6 mg / kg. Malathion was provided in doses of 75% 122.7 mg / kg b.wt., 50% 81.8 mg / kg b.wt., 25%40.9 mg/kg b.wt. LD50 of body weight plus cyclophosphamide 20 mg/kg b.wt as positive control and Corn oil as a negative control, The current study was carried out to detect the effects of malathion on cell cytotoxicity based on genetic cytotoxicity tests such as Micronuclei Test (MN for mature red blood cells in peripheral blood and immature blood cells in bone marrow for 18, 20 and 22 hours per treatment and Chromosomal Aberration (CA) test for immature red blood cells in bone marrow, And 24 hours for each treatment with subcutaneous injection of colchicine 0.5gm prior to 3-hour incubation. The results of the statistical analysis Suggested that a significant increase in P≤0.05of micronucleated mature red blood cells in peripheral blood and micronucleated immature blood cells in bone marrow for 18, 20 and 22 hours per treatment in the formation of micronuclei compared to negative control. The results of the study also Suggested that a significant increase in P≤ 0.05 on the induction of chromosomal Aberration of immature red blood cells in bone marrow.

The effect of Malathion was studied orally on Japanese quail males (Cotrunix Cotrunix Japonica). The Median lethal Dose LD50 of malathion through 24 hours was found to be 163.6 mg / kg. Malathion was provided in doses of 75% 122.7 mg / kg b.wt., 50% 81.8 mg / kg b.wt., 25%40.9 mg/kg b.wt. LD50 of body weight plus cyclophosphamide 20 mg/kg b.wt as positive control and Corn oil as a negative control, The current study was carried out to detect the effects of malathion on cell cytotoxicity based on genetic cytotoxicity tests such as Micronuclei Test (MN for mature red blood cells in peripheral blood and immature blood cells in bone marrow for 18, 20 and 22 hours per treatment and Chromosomal Aberration (CA) test for immature red blood cells in bone marrow, And 24 hours for each treatment with subcutaneous injection of colchicine 0.5gm prior to 3-hour incubation. The results of the statistical analysis Suggested that a significant increase in P≤0.05of micronucleated mature red blood cells in peripheral blood and micronucleated immature blood cells in bone marrow for 18, 20 and 22 hours per treatment in the formation of micronuclei compared to negative control. The results of the study also Suggested that a significant increase in P≤ 0.05 on the induction of chromosomal Aberration of immature red blood cells in bone marrow. http://dx.doi.org/10.25130/tjps.23.2018.164

ISSUE HIGHLIGHTS ‐ March 2020

Many passerine species lay eggs that are speckled with dark protoporphyrin pigmentation. Because protoporphyrin is mainly derived from the blood, we here formulate and test a new hypothesis that links an increase in anaemia along the laying sequence to within-clutch variation in egg pigmentation. More intense pigmentation is expected if pigments accumulate during enhanced red blood cell production in response to anaemia. Reduced pigmentation is expected if pigments are derived from the degradation of red blood cells that circulate in smaller numbers due to blood loss. To test this hypothesis, we manipulated anaemia in great tit (Parus major) females by infesting the nests with hen fleas (Ceratophyllus gallinae) prior to egg laying. Polychromatophil (i.e., immature red blood cells) percentage, as a measure of blood cell production, was positively correlated with parasite load confirming that female great tits experienced stronger anaemia when infested with haematophagous parasites during egg laying. We found a positive relationship between spot darkness and laying order that weakened under high parasite load. This result suggests that anaemia in females due to blood-sucking parasites led to diminished protoporphyrin from disintegrated red blood cells and hence a decreased deposition of protoporphyrin. However, the overall increase in pigment darkness along the laying sequence suggests that pigments also accumulate by enhanced red blood cell production caused by anaemia due to egg production itself.