Erythrocyte Morphology Research Articles

Numerical Simulations and Experiments on Changes in Erythrocyte Morphology under Continue Flow Ventricular Assisted Devices

Ventricular assist devices (VADs) implanted in human body produce different levels of shear stress due to mechanical structure, causing damage for erythrocytes. The study is focusing on the morphology changes of erythrocytes causing by continue flow VADs. This study used a Hemodynamic Shearing Device (Thermo Electron Corporation) to expose erythrocytes to shear stress produced by VADs. Then free hemoglobin of plasma is measured and blood smears are respectively made to count the number of abnormal erythrocytes. The results show that the correlation coefficient of the percent of abnormal erythrocytes (PAE) and shear stress is 0.725 (p=0.027, <0.05). After blood shearing experiments the number of abnormal erythrocytes has increased with shear stress under constant exposure time, and both shear stress and exposure time contribute to morphology changes in erythrocytes. The conclusion indicates that low shear stress (lower than 450 Dynes/cm2) damage of erythrocyte is an important factor in the application of VADs.

The haematology of six species of native catfish from northern Australia

Haematology has the potential to be a valuable tool in determining the health status of wild fish populations and wider ecosystem health. However, limited haematological data are available for wild Australian fish species, and the interpretation and nomenclature of leukocytes is variable and inconsistent in fish. The morphology and cytochemical reactions of erythrocytes, thrombocytes and leukocytes of 189 wild eel-tailed catfish (Tandanus tandanus), Wet Tropics tandan (Tandanus tropicanus), Hyrtyl’s tandan (Neosilurus hyrtlii), black catfish (Neosilurus ater), lesser salmon catfish (Neoarius graeffei), and silver cobbler (Neoarius midgleyi) are described. Erythrocytes, thrombocytes, lymphocytes, monocytes and heterophils in all six species are morphologically similar to those reported in other Siluriformes. Basophils and eosinophils are rarely reported in fish; however basophils were observed in peripheral blood smears of T. tropicanus and N. ater, and eosinophils were observed in N. graeffei. Periodic acid Schiff positive granular leukocytes were observed in N. graeffei, N. midgleyi, N. ater and T. tandanus. This is the first report on the leukocyte morphology and cytochemistry of any native Australian catfish species, and provides useful baseline data for future assessments of fish health and ecosystem integrity in Australia.

Effects of iron overload and exenatide on erythrocyte deformability in a rat model.

Iron overload is known to affect erythrocyte membrane properties and erythrocyte shape. We hypothesized that iron overload which directly affects the erythrocyte morphology may also interfere with erythrocyte deformability (ED). Exenatide, a glucagon like peptide -1 (GLP-1) analogue used in the treatment of diabetes, is known to have beneficial pleiotropic effects on endothelial function and blood flow which are different from its glucose-lowering effects. In our study we aimed to test the effect of iron overload on ED in a rat model (1) and to evaluate the effect of exenatide on ED in the same model (2). For this purpose, the animals were randomly divided into three groups, each containing 6 rats. Rats in the control group (Group C) were given intraperitoneal injections of saline as placebo. The second group (Group Fe) was given intraperitoneal iron dextran (60 mg/kg/day) five days a week for 4 weeks to develop iron overload. The third group (Group Fe +E) received subcutaneous injections of 10 mcg exenatide (Byetta® Lilly Pharma) in two divided doses for 4 weeks in addition to iron dextran. We observed that ED index was significantly higher in Group Fe when compared to Group C and Group Fe+E (p Keywords: erythrocyte deformability, iron, exenatide.

Misdiagnosis of two cases of hereditary spherocytosis in a family and review of published reports

BackgroundHereditary spherocytosis is usually diagnosed based on a combination of clinical and family histories, physical examination, and laboratory data. Milder or atypical cases can be difficult to diagnose. MethodsWe report one case each of mild and moderate hereditary spherocytosis whose diagnoses were made by the First Affiliated Hospital, Guangxi Medical University in China. ResultsIn case 1, laboratory test results were as follows: hemoglobin 81.40g/l, mean corpuscular hemoglobin concentration 339.00g/l, mean corpuscular volume 82.88fl, reticulocyte count 19%, mean sphered corpuscular volume 71.66fl, total bilirubin 57.60μmol/l, and blood smears showed increased spherocytosis. In case 2, laboratory test results were as follows: hemoglobin 133.10g/l, mean corpuscular hemoglobin concentration 358.60g/l, mean corpuscular volume 96.75fl, reticulocyte count 17%, mean sphered corpuscular volume 77.78fl, total bilirubin 62.50μmol/l, and blood smears showed increased spherocytosis. ConclusionsMild and moderate hereditary spherocytosis can be easily misdiagnosed. Assessment of total, direct, and indirect serum bilirubin, erythrocyte morphology and red cell characteristics (particularly mean corpuscular volume and mean sphered corpuscular volume) clearly distinguishes hereditary spherocytosis from autoimmune hemolytic anemia, glucose-6-phosphate dehydrogenase deficiency, thalassemia, and autoimmune hepatitis.

Optical assay of erythrocyte function in banked blood.

Stored red blood cells undergo numerous biochemical, structural, and functional changes, commonly referred to as storage lesion. How much these changes impede the ability of erythrocytes to perform their function and, as result, impact clinical outcomes in transfusion patients is unknown. In this study we investigate the effect of the storage on the erythrocyte membrane deformability and morphology. Using optical interferometry we imaged red blood cell (RBC) topography with nanometer sensitivity. Our time-lapse imaging quantifies membrane fluctuations at the nanometer scale, which in turn report on cell stiffness. This property directly impacts the cell's ability to transport oxygen in microvasculature. Interestingly, we found that cells which apparently maintain their normal shape (discocyte) throughout the storage period, stiffen progressively with storage time. By contrast, static parameters, such as mean cell hemoglobin content and morphology do not change during the same period. We propose that our method can be used as an effective assay for monitoring erythrocyte functionality during storage time.

Ultrastructural changes of erythrocytes in whole blood after exposure to prospective in silico-designed anticancer agents: a qualitative case study

BackgroundNovel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16). These compounds were designed to have improved bioavailability when compared to their source compound, 2-methoxyestradiol. This theoretically would be due to their increased binding affinity to carbonic anhydrase II, present in erythrocytes. Since the novel compounds under investigation are proposed to be transported within erythrocytes bound to carbonic anhydrase II, the morphological effect which they may exert on whole blood and erythrocytes is of great significance. A secondary outcome included revision of previously reported procedures for the handling of the whole blood sample.The purpose of this study was twofold. Firstly, the ultrastructural morphology of a healthy female’s erythrocytes was examined via scanning electron microscopy (SEM) after exposure to the newly in silico-designed compounds. Morphology of erythrocytes following exposure to ESE-15-ol and ESE-16 for 3 minutes and 24 hours at 22°C were described with the use of SEM. The haemolytic activity of the compounds after 24 hours exposure were also determined with the ex vivo haemolysis assay. Secondly, storage conditions of the whole blood sample were investigated by determining morphological changes after a 24 hour storage period at 22°C and 37°C.ResultsNo significant morphological changes were observed in the erythrocyte morphology after exposure to the novel anticancer compounds. Storage of the whole blood samples at 37°C for 24 hours resulted in visible morphological stress in the erythrocytes. Erythrocytes incubated at 22°C for 24 hours showed no structural deformity or distress.ConclusionsFrom this research the optimal temperature for ex vivo exposure of whole blood samples to ESE-15-ol and ESE-16 for 24 hours was determined to be 22°C. Data from this study revealed the potential of these compounds to be applied to ex vivo study techniques, since no damage occurred to erythrocytes ultrastructure under these conditions. As no structural changes were observed in erythrocytes exposed to ESE-15-ol and ESE-16, further ex vivo experiments will be conducted into the potential effects of these compounds on whole blood. Optimal incubation conditions up to 24 hours for whole blood were established as a secondary outcome.

Japanese guidelines of the management of hematuria 2013.

Japanese guidelines of the management of hematuria 2013.

Postartesunate delayed hemolysis is a predictable event related to the lifesaving effect of artemisinins

AbstractPatients with severe malaria treated with artesunate sometimes experience a delayed hemolytic episode. Artesunate (AS) induces pitting, a splenic process whereby dead parasites are expelled from their host erythrocytes. These once-infected erythrocytes then return to the circulation. We analyzed hematologic parameters in 123 travelers treated with AS for severe malaria. Among 60 nontransfused patients observed for more than 8 days, 13 (22%) had delayed hemolysis. The peak concentration of circulating once-infected erythrocytes was measured during the first week in 21 patients and was significantly higher in 9 patients with delayed hemolysis than in 12 with other patterns of anemia (0.30 vs 0.07; P = .0001). The threshold of 180 million once-infected erythrocytes per liter discriminated patients with delayed hemolysis with 89% sensitivity and 83% specificity. Once-infected erythrocyte morphology analyzed by using ImageStream in 4 patients showed an 8.9% reduction in their projected area, an alteration likely contributing to their shorter lifespan. Delayed clearance of infected erythrocytes spared by pitting during AS treatment is an original mechanism of hemolytic anemia. Our findings consolidate a disease framework for posttreatment anemia in malaria in which delayed hemolysis is a new entity. The early concentration of once-infected erythrocytes is a solid candidate marker to predict post-AS delayed hemolysis

Mice Deficient in the Putative Phospholipid Flippase ATP11C Exhibit Altered Erythrocyte Shape, Anemia, and Reduced Erythrocyte Life Span*♦

Transmembrane lipid transporters are believed to establish and maintain phospholipid asymmetry in biological membranes; however, little is known about the in vivo function of the specific transporters involved. Here, we report that developing erythrocytes from mice lacking the putative phosphatidylserine flippase ATP11C showed a lower rate of PS translocation in vitro compared with erythrocytes from wild-type littermates. Furthermore, the mutant mice had an elevated percentage of phosphatidylserine-exposing mature erythrocytes in the periphery. Although erythrocyte development in ATP11C-deficient mice was normal, the mature erythrocytes had an abnormal shape (stomatocytosis), and the life span of mature erythrocytes was shortened relative to that in control littermates, resulting in anemia in the mutant mice. Thus, our findings uncover an essential role for ATP11C in erythrocyte morphology and survival and provide a new candidate for the rare inherited blood disorder stomatocytosis with uncompensated anemia.

Study of the surface architectonics of human erythrocytes by the method of scanning electron microscopy in the presence of doxycycline.

The effects of doxycycline on the morphology and function of donor erythrocytes were studied by scanning electron microscopy. The antibiotic in concentrations of 7.8 × 10(-5) and 7.8 × 10(-6) mol/liter induced heterogeneous changes in the surface architectonics in the erythrocyte population. These data indicated stimulation of erythrocyte "aging" under the effect of doxycycline.

Modified activin receptor IIB ligand trap mitigates ineffective erythropoiesis and disease complications in murine β-thalassemia

AbstractIn β-thalassemia, unequal production of α- and β-globin chains in erythroid precursors causes apoptosis and inhibition of late-stage erythroid differentiation, leading to anemia, ineffective erythropoiesis (IE), and dysregulated iron homeostasis. Here we used a murine model of β-thalassemia intermedia (Hbbth1/th1 mice) to investigate effects of a modified activin receptor type IIB (ActRIIB) ligand trap (RAP-536) that inhibits Smad2/3 signaling. In Hbbth1/th1 mice, treatment with RAP-536 reduced overactivation of Smad2/3 in splenic erythroid precursors. In addition, treatment of Hbbth1/th1 mice with RAP-536 reduced α-globin aggregates in peripheral red cells, decreased the elevated reactive oxygen species present in erythroid precursors and peripheral red cells, and alleviated anemia by promoting differentiation of late-stage erythroid precursors and reducing hemolysis. Notably, RAP-536 treatment mitigated disease complications of IE, including iron overload, splenomegaly, and bone pathology, while reducing erythropoietin levels, improving erythrocyte morphology, and extending erythrocyte life span. These results implicate signaling by the transforming growth factor-β superfamily in late-stage erythropoiesis and reveal potential of a modified ActRIIB ligand trap as a novel therapeutic agent for thalassemia syndrome and other red cell disorders characterized by IE.

Lysophosphatidic acid and human erythrocyte aggregation

The effects of lysophosphatidic acid on the morphology and aggregation of human erythrocytes has been studied. Morphology of erythrocytes and their aggregates were studied by light microscopy. It has been shown that lysophosphatidic acid changes the shape of red blood cells: diskocyte become echinocytes. Aggregation of red blood cells (rouleaux) was significantly reduced in autoplasma. At the same time there is a strong aggregation of echinocytes. This was accompanied by the formation of microvesicles. Adding normal plasma to echinocytes restores shape and aggregation of red blood cells consisting of "rouleaux". A possible mechanism of action of lysophosphatidic acid on erythrocytes is discussed.

Effect of adenosine on the shape, aggregates morphology and aggregability of ATP-depleted erythrocytes

The effect of adenosine on the shape, aggregate morphology and aggregability of ATP-depleted erythrocytes was studied. It is shown that the ATP-depletion of erythrocyte leads to the change in their shape: diskocytes transform to echinocytes. It is found that the aggregability of such cells in autoplasma significantly decreased. Incubation of echinocytes with adenosine largely restored discoid shape and erythrocyte aggregability.

Detection of erythrocytes in patients with Waldenström macroglobulinemia using atomic force microscopy

The pathological changes of erythrocytes are detected at the nanometer scale, which is important for revealing the onset of diseases and diagnosis. The aim of this study is to examine the ultrastructural changes of erythrocytes in Waldenström macroglobulinemia (WM) at a nanometer scale. Blood samples were collected from two healthy volunteers, two WM patients, and three multiple myeloma (MM) patients when they were first diagnosed. The changes of morphology in the erythrocytes were studied at the nanometer level by high-resolution atomic force microscopy imaging (AFM). Compared with the healthy controls and the MM patients, there were dramatic deformations in the overall shape and surface membrane of the erythrocytes in WM patients. Healthy, pathological WM, and MM erythrocytes could be distinguished by several morphological parameters, including the width, length, length-to-width ratio, valley, peak, peak-to-valley, and Ra. AFM is able to detect the morphological differences in the red blood cells from WM patients, healthy controls, and MM patients. Therefore, the erythrocyte morphology is an important parameter for the diagnosis of WM, which can be used to distinguish WM from MM. The changes of ultrastructure in red blood cells may provide a clue to reveal the mechanism of WM.

Imaging flow cytometry for automated detection of hypoxia‐induced erythrocyte shape change in sickle cell disease

In preclinical and early phase pharmacologic trials in sickle cell disease, the percentage of sickled erythrocytes after deoxygenation, an ex vivo functional sickling assay, has been used as a measure of a patient's disease outcome. We developed a new sickle imaging flow cytometry assay (SIFCA) and investigated its application. To perform the SIFCA, peripheral blood was diluted, deoxygenated (2% oxygen) for 2 hr, fixed, and analyzed using imaging flow cytometry. We developed a software algorithm that correctly classified investigator tagged "sickled" and "normal" erythrocyte morphology with a sensitivity of 100% and a specificity of 99.1%. The percentage of sickled cells as measured by SIFCA correlated strongly with the percentage of sickle cell anemia blood in experimentally admixed samples (R = 0.98, P ≤ 0.001), negatively with fetal hemoglobin (HbF) levels (R = -0.558, P = 0.027), negatively with pH (R = -0.688, P = 0.026), negatively with pretreatment with the antisickling agent, Aes-103 (5-hydroxymethyl-2-furfural) (R = -0.766, P = 0.002), and positively with the presence of long intracellular fibers as visualized by transmission electron microscopy (R = 0.799, P = 0.002). This study shows proof of principle that the automated, operator-independent SIFCA is associated with predictable physiologic and clinical parameters and is altered by the putative antisickling agent, Aes-103. SIFCA is a new method that may be useful in sickle cell drug development.

Gambaran Anemia pada Lanjut Usia di Panti Sosial Tresna Werdha Abiyoso Yogyakarta Tahun 2013

Background: Anemia is a main health problem of elderly. The aging process causes many psychological changes in nearly all body systems of elderly that causes many kinds of processes of nutrient absorptions in the body disturbed. There is an opinion says that generally elderly has lower hemoglobin levels compared to the younger people. Other research showed that elderly often suffered from calorie and protein malnutrition and micronutrient. It happened because the HCL and pepsin secretion was reduced. Consequently, the absorption of Vitamin B12 and iron decreased that causes Anemia.
 Method: It was a descriptive analysis with laboratory tests. The sample of this research was 60 year-old and older patients in TresnaWerdhaAbiyoso Nursing Home Yogyakarta. There were 96 patients. The data of research results were analyzed descriptively and presented in the forms of tables and graphs.
 Results: Of the 96 old patients, there were 31 patients suffered from anemia that consisted of 27 patients with mild anemia and 10 patients with moderate anemia. Based on the morphology of erythrocytes in elderly, there were two kinds of anemia. They were normocytic-normochromic anemia with 84 patients and microcytic-hypochromic anemia with 12 patients.
 Conclusion: The description of anemia in elderly who stayed in TresnaWerdhaAbiyoso Nursing Home Yogyakarta showed that 32.29% of the elderly suffered from anemia. 72.97% suffered from mild anemia. 27.03% suffered from moderate anemia. The anemia classifications based on morphology in elderly in Tresna Werdha Abiyoso Nursing Home Yogyakarta were 87.5% normocytic-normochromic anemia and 12.5% microcytic-hypochromic anemia.

The effect of metabolites and impurities of glyphosate on human erythrocytes (in vitro)

The toxicity of herbicides to animals and human is an issue of worldwide concern. The present study was undertaken to evaluate toxic potential of widely used pesticide – glyphosate, its metabolites: aminomethylphosphonic acid (AMPA); methylphosphonic acid and its impurities: N-(phosphonomethyl)iminodiacetic acid (PMIDA), N-methylglyphosate, hydroxymethylphosphonic acid and bis-(phosphonomethyl)amine.We evaluated the effect of those compounds on hemolysis, hemoglobin oxidation, reactive oxygen species (ROS) formation and changes in morphology of human erythrocytes. The erythrocytes were exposed to different concentrations of glyphosate and its metabolites and impurities (0.01–5mM) for 1, 4 and 24h.Glyphosate, its metabolites and impurities induced a little hemolysis and hemoglobin oxidation. All changes were very low, even after 24h incubation. Most of the investigated compounds induced reactive oxygen species formation from 0.25mM, except the N-methylglyphosate which caused an increase in ROS formation from 0.5mM. Moreover, the investigated xenobiotics did not change the size and shape (except bis-(phosphonomethyl)amine) of the human erythrocytes.Changes in human erythrocytes were observed only when high concentrations of the compounds were applied. Some investigated metabolites and impurities caused a slight stronger damage to human erythrocytes than a glyphosate.The results clearly show that the changes induced in the erythrocytes can occur only as a result of poisoning with these compounds.

Determining The Diameter Of Erythrocyte By Means Of Manual Laser

The article gives instructions on carrying out the school experiment with laser light. Numerical value of the diameter of erythrocyte has been determined. By applying this technique various erythrocyte morphology can be noticed. Size and shape of a blood cell are usually determined by means of a microscope. This method is quite boring, time-consuming and nonapplicable to a large number of specimens and cells. Relative to this method there is a simple and quick way of determining an average size and shape of a blood cell by means of laser light diffraction. Blood specimens are taken from healthy people and patients suffering from various types of disease (cancer). The laser light diffraction method is very quick and easy to estimate an average size and shape blood cells. The new and sophisticated method that uses the laser tecnique has been developed for determining erythrocyte size, and the apparatus consists of a specimen (microscope slide), a laser and a screen where the radius/diameter of human erythrocytes is measured. The specimen, a drop of blood smeared on the microscope slide (a thin layer) , is placed between the laser and the screen. He-Ne laser whose power is P=2MW is used for the diffraction. When passing through the blood specimen the laser beam forms an excellent diffraction image on the screen (picture 1). The slide with erythrocytes in a single layer has been provided by a hospital laboratory. When being placed on the optical bench between the laser and the screen, the slide ought to be turned towards the light source. The slide should be moved until a sharp and clear image of diffraction rings is formed, then the radius/diameter, r, of the first (and the second) diffraction circle(s) is measured, as well as the distance from the laser to the screen L. The diffraction angle is determined by tanθ =r/L. The mean diameter of blood cells is determined by using the wave length of laser light λ = 632,2nm. PICTURE 1: Diffraction on erythrocyte done in “Foto Sretenovic“ photo shop; photographed by Vladimir Sretenovic According to Rayleigh’s expression (William Strutt) cell diameter d can be written as d = 1.22 λ/ tanθ. By replacing tanθ the equation for erythrocyte diameter d is obtained: d = n·λ·L/r 1. r – radius of the black circle, L – distance between the microscope slide and the screen (milimeter paper), λthe laser light wavelength (632,2nm) ; n = 1.22;2.23;3.24 is used for the rings. We used a manual laser manufactured by MITT INFIZ, the power of which is 5 mW and the wavelength : λ= 67о nm. (One must not look in the direction of the laser light.) Applying the equation 1 the first diameter of an erythrocyte is calculated when the expression d1 = 1.22 λ·L/r1 = 9.31μm is used, for the second diameter of an erythrocyte d2 = 2.23 λ·L/r2 = 8. 81 μm, and the third diameter of an erythrocyte d3 = 3. 24 λ·L/r3 = 9.2μm. The obtained results imply that the patient is ill. Determining The Diameter Of Erythrocyte By Means Of Manual Laser www.iosrjournals.org 21 | Page Reference [1]. R. Krsnik. Suvremene ideje u metodici nastave fizike. Skolska knjiga. Zagreb,2008 (R. Krsnik, Modern Ideas in Methodics of Teaching Physics, Skolska knjiga, Zagreb,2008) [2]. I. Janjic. Fizika II deo Novi Sad 1971 Janjic, Physics, Part 2, Novi Sad, 1971) [3]. Kvant. Naucno-popularni fizicko-matematicki žurnal. 1989 (Kvant, Popular Science Journal of Physics and Mathematics,1989) [4]. J. Simovic, D. Koledin. Praktikum iz biofizike, Medicinska knjiga. Beograd-Zagreb, 1991 [5]. (J. Simovic, D. Koledin, Book of Exercises in Biophysics, Medicinska knjiga, Belgrade-Zagreb, 1991) [6]. C. Ramakrishna Rao, Kaleemed Jaleeli , B.S. Bellubbi1 &Adeel Ahmad

Hematology of wild caught Dubois's tree frog Polypedates teraiensis, Dubois, 1986 (Anura: Rhacophoridae).

Blood was analyzed from eighty (forty males and forty females) adult individuals of Polypedates teraiensis to establish reference ranges for its hematological and serum biochemical parameters. The peripheral blood cells were differentiated as erythrocytes, lymphocytes, eosinophils, neutrophils, monocytes, basophils, and thrombocytes, with similar morphology to other anurans. Morphology of blood cells did not vary according to sex. The hematological investigations included morphology and morphometry of erythrocytes, morphometry of leucocytes, packed cell volume (PCV), hemoglobin content (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), erythrocyte or red blood cell (RBC) count, leukocyte or white blood cell (WBC) count, differential leukocyte count, and neutrophil to lymphocyte (N/L) ratio. Besides, protein, cholesterol, glucose, urea, uric acid, and creatinine content of blood serum were assayed. Hematological parameters that differed significantly between sexes were RBC count, length and breadth of RBC, neutrophil %, N/L ratio, area occupied by basophils, and diameter of large lymphocyte and eosinophils. The level of glucose, urea, and creatinine in blood serum also significantly differed between sexes.

Effect of radiographic contrast media (Iodixanol, Iopromide) on hemolysis.

Radiographic contrast media can lead to drastic changes of the morphology of erythrocytes. The change of the erythrocyte morphology is associated with a decreased deformability possibly resulting from distinctions in the loss of constituents of the membrane cytoskeleton. However, it is unclear whether there is an intravascular hemolysis as a consequence of the disintegration of the erythrocyte membrane. The results of this study showed, that free haemoglobin increased from 16.8 ± 10.0 mg/dl to 21.6 ± 12.6 mg/dl after Iopromide application (p = 0.240), while it slightly decreased from 20.5 ± 10.3 mg/dl to 19.5 ± 12.2 mg/dl after Iodixanol application (p = 0.547). The slight decrease of free haemoglobin after application of Iodixanol differed significantly compared to the increase of free haemoglobin after Iopromide application (p < 0.05). This different response is thought to give evidence to the assumption that the erythrocyte membrane integrity was compromised leading to the release of free haemoglobin as an indicator of hemolysis as well.