Erythrocyte Morphology Research Articles

<b>Comparison of nuclear abnormalities in <i>Astyanax bifasciatus</i> Cuvier, 1819 (Teleostei: Characidae) of two sections of rivers from the middle Iguaçu

Most changes in water bodies are result of human activities that have the potential to undermine the environmental integrity of aquatic ecosystems. Changes in genetic material can be evidenced by the frequency of nuclear abnormalities in fish blood cells, in response to genotoxic agents even at low concentrations. Thus, we aimed at comparing the frequencies of nuclear abnormalities of fish kept in acclimation, with fish collected in Timbo river (Santa Cruz do Timbo, Santa Catarina State), under low anthropogenic interference, and fish collected in Iguacu river (Uniao da Vitoria, Parana State), a polluted river. The highest frequency of changes in nuclear morphology of fish erythrocytes was found in the urbanized area around the Iguacu river, while, the fish collected in areas with preserved riparian forest surrounding the Timbo river and acclimated fish showed no morphological changes. Our results suggest that genotoxic compounds in the Iguacu river are acting as stressors to aquatic communities, especially fish. In addition, the absence of nuclear abnormalities in fish from the Timbo river suggests that rivers without significant environmental changes can serve as reference point for comparative studies of genetic modifications for the species studied.

α1-and β-adrenergic antagonist labetalol induces morphological changes in human erythrocytes

Labetalol is one of the most used drugs for the treatment of hypertension. This molecule is able to bind to both alpha-1 (α1) and beta (β) adrenergic receptors present in vascular smooth muscle among other tissues. It has been determined that human erythrocytes possess both alpha receptors and beta-adrenergic receptors expressed on their surface. The objective of this work was to study the effect of labetalol on the morphology of human erythrocytes. To accomplish this goal, human erythrocytes and model membranes built of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) were used. These lipid species are present in the outer and inner monolayers of the red blood cell membrane, respectively. Our findings obtained by X-ray diffraction and differential scanning calorimetry (DSC) indicate that labetalol interacted with both lipids in a process dependent on concentration. In fact, at low concentrations labetalol preferentially interacted with DMPE. On the other hand, results obtained by scanning electron microscopy (SEM) showed that labetalol alters the normal biconcave form of erythrocytes to stomatocytes and knizocytes (cells with one or more cavities, respectively). According to the bilayers couple hypothesis, this result implied that the drug inserted in the inner monolayer of the human erythrocyte membrane.

THE MORPHOLOGICAL FEATURES OF ERYTHROCYTES IN STORED PACKED RED CELLs

Morfologi eritrosit Packed Red Cells (PRC) akan mengalami perubahan selama penyimpanan di suhu 2°–8°C. Eritrosit dalammempertahankan viabilitasnya membutuhkan adenosine triphosphate (ATP). Apabila kadar ATP intraseluler menurun, terjadi kerusakanlipid membran, penumpukan Natrium dan Kalsium intraseluler, penurunan kadar Kalium dan air intraseluler, dehidrasi sel, membranmenjadi kaku dan bentuknya berubah dari cakram menjadi sel krenasi, sferosit dan bite cell. Penelitian ini bertujuan untuk melihatpersentasi bentuk eritrosit crenated cell, sferosit dan bite cell PRC simpan pada hari ke-3 yang digunakan sebagai pembanding, hari ke-7,ke-14 dan ke-21 dari tanggal aktaf kantong darah. Penelitian observasional dengan pendekatan kajian kohort dilakukan pada bulanAgustus 2015 di BDRS RSUP Dr. Wahidin Sudirohusodo Makassar. Sampel sebanyak 30 selang kantong darah menjadi 120 hapusandarah slide. Dari 30 sampel kantong darah dengan golongan darah A 26,6%, B 13,3%, AB 16,6% dan O 43,3%, didapatkan peningkatanpersentase jumlah crenated cell, sferosit dan bite cell setelah penyimpanan hari ke-3, ke-7, ke-14 dan ke-21. Penyimpanan hari ke-3dijadikan pembanding. Data diolah dengan menggunakan uji Fiedman dan Wilcoxon dengan nilai kemaknaan p<0,001. Terdapatpeningkatan persentase perubahan morfologi eritrosit (crenated cell, sferosit dan bite cell) seiring dengan lamanya penyimpanan darahPRC. Pemakaian darah PRC dianjurkan tidak boleh melebihi 21 hari penyimpanan.

Effect on Morphology, Osmotic Fragility and Electro Kinetic Potential of Erythrocytes in Hypertension.

Hypertension is a common health problem concerning a large proportion of population and a leading global risk factor for the burden of cardiovascular disease (CVD). The purpose of this study was to assess the efficacy of the erythrocyte zeta potential as a potential additional indicator for cardiovascular disorder risk so that patients with this can be more rapidly identified and treated. In the present study, blood samples were collected in 5% dextrose solution from patients suffering from hypertension and healthy volunteers (Not taken any medication). The mobility of individual RBCs was tracked by equipped Zeta meter-ZM4DAQ software using microscopicallyacquired video images, data were recorded 10 times for each sample and average zeta-potential in mv was recorded. We found that mean erythrocytic ZP of control group was found to be 23.41 mv (± 1.87) whereas, erythrocytic ZP for Hypertensive patients was found to be 16.05 (±1.72) mV and Hypertensive patients with Diabetes is much lower from 6.96 mV to 22.76 (+3.88) mV along with structural deformities and increased osmotic fragility of erythrocytes. The data suggest that there are morphological changes in erythrocyte structure, increased osmotic fragility along with significantly lower ZP value as compared to that of healthy volunteers which may be the major cause for progression to the development of cardiovascular disease.

Addressing the impact of mercury estuarine contamination in the European eel (Anguilla anguilla L., 1758) – An early diagnosis in glass eel stage based on erythrocytic nuclear morphology

The decline of the European eel (Anguilla anguilla L., 1758) population throughout Europe has been partially attributed to pollution. As glass eel estuarine migration may represent a considerable threat, the impact of mercury (Hg) contamination at this stage was evaluated through an in situ experiment (7days). Total Hg (tHg) bioaccumulation was evaluated concomitantly with erythrocytic nuclear morphology alterations: erythrocytic nuclear abnormalities assay (ENA), frequency of immature erythrocytes (IE) and the erythrocytic maturity index (EMI). The ENA results suggested a genotoxic pressure at the most contaminated sites, in line with the tHg increase. The EMI data, together with IE frequency, showed that fish exposed to high levels of Hg exhibited alterations of haematological dynamics, translated into an erythropoiesis increment. Despite the presence of these compensatory mechanisms, the present findings suggest a harmful impact of Hg on genome integrity at this early development stage, potentially affecting eels' condition and ultimately the population sustainability.

The Effects of Cryoprotective Substances on the Mechanical Stability and Geometric Parameters of Human Erythrocytes

The aim of this study was to examine the effects of sucrose, dextran, polyethylene glycol, glycerol, and mannitol, which possess cryoprotective properties to various degrees, on the mechanical stability and the geometric parameters of human erythrocytes. All substances, except mannitol, contributed to a decrease in hemolysis, which was caused by the movement of small beads. Glycerol and polyethylene glycol, which provide the highest level of protection of erythrocytes during cryopreservation, also showed the maximum efficiency under mechanical stress. Changes in cell resistance may be associated with the transformation of their geometric parameters. According to the cytometry data, 5% solutions of all the substances, except mannitol, caused similar changes in the geometric parameters of the cell. The relationship between changes in the mechanical stability and the geometric parameters of erythrocytes under the influence of cryoprotective agents may be caused by the modification of the membrane−cytoskeleton protein complex, which controls the mechanoelastic properties and morphology of erythrocytes.

Evaluation of Erythrocyte Morphology and Antioxidant Enzymes in Patients of Systemic Lupus Erythematosus

Evaluation of Erythrocyte Morphology and Antioxidant Enzymes in Patients of Systemic Lupus Erythematosus

Surface property induced morphological alterations of human erythrocytes.

Microscopic investigations of any abnormality associated with erythrocyte/red blood cell morphology constitute an important segment of the age-old peripheral smear test. Though the test is conducted on a glass slide, the effect of glass and similar other solid substrates on erythrocyte morphology remained majorly unexplored. In the first of its kind investigation, we have outlined the effect of varying the substrate surface potential on erythrocyte morphology. Such a substrate induced phenomenon has been quantified for two distinctly different drying configurations (droplets and film) upon systematically varying the cell concentration. Experimental results and supporting theoretical analysis unambiguously show the surface potential of the solid substrate to be the most influential parameter in the process of morphological alteration. The findings of the present investigation may be utilized to formulate an error-free protocol for the baseline peripheral smear test of hematological diagnosis.

Inorganic mercury and cadmium induce rigidity in eukaryotic lipid extracts while mercury also ruptures red blood cells

Hg and Cd are non-essential toxic heavy metals that bioaccumulate in the tissues of living systems but less is known about their interactions with Eukaryotic lipid bilayers. Microscopy experiments showed that Hg and Cd changed the cell morphology of rabbit erythrocytes while Hg also induced cell rupture. As membranes are one of the first available targets, our study aimed to better understand metal-lipid interactions that could lead to toxic effects. Fluorescence spectroscopy (Laurdan Generalized Polarization) and dynamic light scattering were used to analyze metal-induced changes in membrane fluidity and the size of liposomes composed of Brain (Porcine), Liver (Bovine), Heart (Bovine) and Yeast (S. cerevisiae) lipid extracts. Under physiological chloride and pH levels, Hg irreversibly cleaves plasmalogens resulting in an increase in membrane rigidity. These lipids are enriched in Brain, Heart and Erythrocyte membranes and are important in signalling and the protection against oxidative stress. Interestingly, Hg had a heavily reduced effect on the plasmalogen-free Yeast extract membrane. In contrast, Cd induced rigidity by targeting negatively charged phosphatidic acid, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol and cardiolipin in these extracts. Metal-induced liposome aggregation depended on the proportion of negatively charged lipids/plasmalogen and even the order of metal addition. Our results show that data from model systems correlate with trends observed in complex biological extracts and red blood cells and serve as a predictive tool for analyzing metal-lipid interactions. The determination of the specific lipid targets for Hg and Cd provides new insights how these metals exert toxic effects on cell membranes.

Wide field of view common-path lateral-shearing digital holographic interference microscope.

Quantitative three-dimensional (3-D) imaging of living cells provides important information about the cell morphology and its time variation. Off-axis, digital holographic interference microscopy is an ideal tool for 3-D imaging, parameter extraction, and classification of living cells. Two-beam digital holographic microscopes, which are usually employed, provide high-quality 3-D images of micro-objects, albeit with lower temporal stability. Common-path digital holographic geometries, in which the reference beam is derived from the object beam, provide higher temporal stability along with high-quality 3-D images. Self-referencing geometry is the simplest of the common-path techniques, in which a portion of the object beam itself acts as the reference, leading to compact setups using fewer optical elements. However, it has reduced field of view, and the reference may contain object information. Here, we describe the development of a common-path digital holographic microscope, employing a shearing plate and converting one of the beams into a separate reference by employing a pin-hole. The setup is as compact as self-referencing geometry, while providing field of view as wide as that of a two-beam microscope. The microscope is tested by imaging and quantifying the morphology and dynamics of human erythrocytes.

Very Deep Convolutional Neural Networks for Morphologic Classification of Erythrocytes.

Morphologic profiling of the erythrocyte population is a widely used and clinically valuable diagnostic modality, but one that relies on a slow manual process associated with significant labor cost and limited reproducibility. Automated profiling of erythrocytes from digital images by capable machine learning approaches would augment the throughput and value of morphologic analysis. To this end, we sought to evaluate the performance of leading implementation strategies for convolutional neural networks (CNNs) when applied to classification of erythrocytes based on morphology. Erythrocytes were manually classified into 1 of 10 classes using a custom-developed Web application. Using recent literature to guide architectural considerations for neural network design, we implemented a "very deep" CNN, consisting of >150 layers, with dense shortcut connections. The final database comprised 3737 labeled cells. Ensemble model predictions on unseen data demonstrated a harmonic mean of recall and precision metrics of 92.70% and 89.39%, respectively. Of the 748 cells in the test set, 23 misclassification errors were made, with a correct classification frequency of 90.60%, represented as a harmonic mean across the 10 morphologic classes. These findings indicate that erythrocyte morphology profiles could be measured with a high degree of accuracy with "very deep" CNNs. Further, these data support future efforts to expand classes and optimize practical performance in a clinical environment as a prelude to full implementation as a clinical tool.

Analysis of various effects of abamectin on erythrocyte morphology in Japanese quails (Coturnix japonica).

Although previous studies have already confirmed the toxicological potential of abamectin (ABA) in different experimental models (from invertebrates to vertebrates), almost nothing is known about the impacts the exposure to this pesticide can cause on birds. Thus, the aim of our study is to investigate the cytotoxic effects on the erythrocytes of female Japanese quails (Coturnix japonica) exposed to low abamectin concentrations. In order to do so, three experimental groups were proposed: "control," composed of quails exposed to abamectin-free drinking water; "ABA 1% median lethal dose (LD50)," comprising birds exposed to water containing 15.5mg a.i./L of abamectin (via commercial formulation Kraft® 36EC), and "ABA 10% LD50," composed by birds exposed to water containing 155.0mg a.i./L of abamectin. The micronucleus test and the tests applied to other nuclear abnormalities in the peripheral blood of birds were conducted 40days after exposure. Our study revealed significant physical abnormalities in nuclear shapes (erythrocytes with asymmetric constriction nuclei, notched nuclei, indented and moved nucleus) of those birds exposed to higher abamectin levels. When all nuclear abnormalities were tallied, a significant dose-dependent trend was noted. Therefore, our study presents initial imprints on determination of abamectin-mediated cellular toxicity in avifauna which can be instrumental in checking polluted ecosystems.

Electrochemical activity and morphology of human erythrocytes at optically transparent ITO electrode

The electrochemical activity of a suspension of human erythrocytes was found at potentials above +0.4 V. The limiting current of electrooxidation was found to linearly depend on the cell concentration in the suspension. The electrochemical activity of an erythrocyte suspension is accompanied by reversible changes in the cell morphology only in non-buffered media.

Diagnosis of thalassemia and iron deficiency anemia using confocal and atomic force microscopy

Anemia is the most prevalent blood disorder, categorized into thalassemia and iron deficiency anemia. In anemia, the morphology of erythrocytes is disturbed, thus leading to abnormal functioning of the erythrocytes. Globally, thalassemia affects 1.3% of individuals and is one of the most widespread monogenic disorders in Pakistan. All over the World, women and children are most frequently affected by a type of nutritional deficiency known as iron deficiency anemia. The morphological changes that occur in erythrocytes due to these diseases are investigated in this study at the nano-scale level. Fifty samples of blood from individuals suffering from thalassemia or iron deficiency anemia were obtained from different hospitals in Rawalpindi and Islamabad. The blood samples were scanned using atomic force microscopy (AFM) and laser scanning confocal microscopy (LSCM) to check the morphological changes in both types of anemia. According to the present study, thalassemia is most prevalent in females in the age group between 5 and 15 years old, and iron deficiency is most prevalent in females in the age groups of 16–25 and 36–45 years old. Erythrocyte morphology is the significant determinant for diagnosing and discriminating between these two types of diseases. The study reports deformed erythrocytes in anemic patients, which were different from the ones that existed in the control. Thalassemia erythrocytes showed a crenated shape, iron deficiency anemia erythrocytes showed an elliptocyte shape and healthy erythrocytes showed a biconcave disk shape when using AFM and LSCM. These techniques seem to be very promising, cheap and less time consuming in determining the structure–function relationship of erythrocytes of thalassemic and iron deficiency anemic patients. The results of LSCM and AFM are quite useful in determining the morphological changes in erythrocytes and to study the disease at the molecular level within short period of time. Hence, we encourage employing these non-invasive techniques for the effective diagnosis of anemic patients.

Synthesis and pharmacological evaluation of pyridinyl-1,3,4-oxadiazolyl-ethanone derivatives as antimicrobial, antifungal and antitubercular agents

Nicotinic acid was converted into different substituted acetylated nicotinic acid derivatives by sequential transformation involving formation of hydrazide, Schiff’s base and finally acylated oxadiazole derivatives. The synthesized compounds were characterized by spectroscopic techniques and evaluated in vitro for the antimicrobial, antifungal, as well as antitubercular activity. Among all the synthesized derivatives, compounds 6b, 6d, 6e, 6g, and 6j demonstrated excellent antimicrobial activity on Bacillus subtilis. The compounds 6d, 6j and compounds 6b, 6f, 6h, and 6i exhibited maximum zone of inhibition against fungi Candida albicans as well as Aspergillus niger, respectively at the concentration of 500 µg/mL. The antitubercular activity exhibited by 6f, 6g, and 6d with minimum inhibitory concentration (MIC) values of 1.2, 3.1, and 7.8 µg/mL, respectively. The synthesized compounds were studied by molecular docking through Autodock Vina to evaluate their interaction at respective proteins. Further the effect of synthesized derivatives on surface morphology of human erythrocytes as well as hemolysis was also evaluated. The results demonstrated lesser extent of hemolytic toxicity.

One-year recombinant growth hormone therapy does not improve hemoglobin state and morphology of erythrocytes in growth hormone deficient children

An increase in growth rates of children suffering from growth hormone deficiency (GHD) subjected to recombinant growth hormone treatment (rGHT) was shown to be accompanied by acceleration of metabolic processes that may stimulate oxygen consumption in various organs and tissues. Therefore, oxygen-transporting properties of RBC should undergo considerable changes during the rGHT. The aim of this study was to examine the effects of rGHT on erythrocyte shape and hemoglobin state in GHD children. The level of oxyhemoglobin (Oxy-Hb) in RBC was analyzed by Raman spectroscopy. The RBC count, mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV) and other parameters were calculated. The blood of eleven treatment-naive prepubertal children with GHD (aged 3–9, median 5.7 years) was examined and compared with control group (aged 5–7; median 6.0 years) at three time points: 0, 3 and 12 months of rGHT. Before rGHT, the MI in GHD children was higher (median 0.48 vs 0.14 p=0.0018) and the RBC count was lower (median 4.20 vs 4.96 1012 cells/L p=0.0022) than in control group. After the treatment, cell count in GHD patients did not differ significantly from the control group, but Oxy-Hb level became higher (median 0.64 vs 0.41 p=0.0075). During rGHT, MCV decreased (median 80.3 vs 83.2μm3 p=0.0231). Morphological and functional characteristics of erythrocytes in GHD children were shown to differ significantly from the healthy control group. A twelve-month rGHT partially improved some of the studied parameters but Oxy-Hb level and echinocyte count remained high.

HAEMATURIA;

Introduction: Urine analysis was the first laboratory test performed in medicineand has been used for several thousand years. Today it continues to be a powerful tool inobtaining crucial information for diagnostic purposes in medicine.1 Aims and Objectives:To estimate the source of bleeding by erythrocyte morphology on PCM, in patients withhaematuria. Study Design: Descriptive. Setting: The Nephrology and Urology out Patients.Period: 2014 to 2015. Methods: Urinary samples were collected from 100 random patientswho presented with haematuria. Samples were examined by face contrast microscope.UrineRBCs were identified as isomorphic and dysmorphic, proportion was taken, >20% of eithercells were used for localizing the source of bleeding. Later on, finding was furtherconfirmed byultra sound or other investigations like renal Biopsies and cell cytology. Result: In our studywe found that not only the correct bleeding site can be located with a high specificity, but alsoadditional findings can be looked for with a little background knowledge of patient’s medicalhistory. Conclusion: Phase contrast microscopy should be used by the clinicians for gatheringthe primitive information in the patients with haematuria.

Exploring Erythrocytes as Blood Biomarkers for Alzheimer’s Disease

Peripheral biomarkers for dementia are few and far between. Despite research into blood plasma/serum biomarkers for dementia diagnostics, there is a lack of information on erythrocytes and their vast proteomes as potential biomarkers. This review identifies a number of relevant and potentially promising erythrocyte biomarkers for various subtypes of dementia. These include erythrocyte morphology, oxidative stress, and erythrocyte membrane proteins such as the glucose transporter (GLUT-1), amyloid-β, IgG, Hsp90, calpain-1, and band 3 protein. Of those proteins identified Hsp90, amyloid-β, calpain-1 and band 3 show the most promise as pre-clinical biomarkers. However, the most intriguing aspect of erythrocytes is their changed morphology in dementia. The altered morphology not only could be used as a diagnostic biomarker but may be crucial in early pathogenesis of the disease. Further work must be done to establish the pathological connection between the periphery and central disease processes.

Relationships of mean corpuscular volume with diameter and surface area of canine erythrocytes

Assessment of erythrocyte morphology is an important aid in diagnostic haematology. Anisocytosis which is changes in RBC size, correspond to changes in diameter and surface area (SA) of the cell, and are not always reflected in the Mean Corpuscular volume (MCV). This paper discusses the relationship of MCV with surface area and diameters (vertical and horizontal) in 2D views of erythrocytes of clinically healthy dogs using confocal microscopy. This information would be valuable for early detection of cellular changes in dogs in subclinical or clinical diseases. The average diameters, average SA, and MCV values of RBCs studied were in normal distribution and the mean values of 7.169 ± 0.648 μm for horizontal diameter (D1), 7.1245 ± 0.6646 μm for vertical diameter (D2), and 41.061 μm2 ± 6.866 for SA did not reveal a significant correlation or a strength of association with the MCV values, indicating that the MCV value has limitations as an objective measurement of detecting anisocytosis.

Novel oxolane derivative DMTD mitigates high glucose-induced erythrocyte apoptosis by regulating oxidative stress

Chronic hyperglycemia is one of the characteristic conditions associated with Diabetes Mellitus (DM), which often exerts deleterious effects on erythrocyte morphology and hemodynamic properties leading to anemia and diabetes-associated vascular complications. High glucose-induced over production of reactive oxygen species (ROS) can alter the blood cell metabolism and biochemical functions subsequently causing eryptosis (red blood cell death), yet another complication of concern in DM. Therefore, blocking high glucose-induced oxidative damage and subsequent eryptosis is of high importance in the better management of DM and associated vascular complications. In this study, we synthesized an oxolane derivative 1-(2,2-dimethyltetrahydrofuro[2,3][1,3]dioxol-5-yl)ethane-1,2-diol (DMTD), and demonstrated its efficacy to mitigate hyperglycemia-induced ROS generation and subsequent eryptosis. We showed that DMTD effectively inhibits high glucose-induced ROS generation, intracellular calcium levels, phosphaditylserine (PS) scrambling, calpain and band 3 activation, LDH leakage, protein glycation and lipid peroxidation, meanwhile enhances the antioxidant indices, osmotic fragility and Na+/K+-ATPase activity in erythrocytes. DMTD dose dependently decreased the glycated hemoglobin level and enhances the glucose utilization by erythrocytes in vitro. Further, DMTD alleviated the increase in ROS production, intracellular Ca2+ level and PS externalization in the erythrocytes of human diabetic subjects and enhanced the Na+/K+-ATPase activity. Taken together, the synthesized oxolane derivative DMTD could be a novel synthetic inhibitor of high glucose-induced oxidative stress and eryptosis. Considering the present results DMTD could be a potential therapeutic to treat DM and associated complications and open new avenues in developing synthetic therapeutic targeting of DM-associated complications.