Blood Cell Morphology Research Articles

A Machine-Learning Derived Red Blood Cell Morphology Tool Enables Differential Diagnosis and Novel Single-Cell Analyses

Microscopic examination of red blood cell (RBC) morphology in a peripheral blood smear is a standard clinical test with broad utility even in resource-limited settings. Yet this analysis remains subjective, semi-quantitative, and low throughput, limiting its potential in clinical practice and translational research. Prior attempts to develop quantitative RBC morphology tools have been hampered by limited scope, reproducibility challenges, and sparse clinical validation. Here, we present a novel, open-source machine-learning approach leveraging durable geometric features to quantify clinically relevant RBC morphologies-elliptocytes, microcytes, macrocytes, schistocytes, sickle cells, spiculated cells, and teardrop cells-and generate an RBC differential ('RBC-diff'), analogous to the widely used white cell differential. Analysis of >300 thousand smear images across two medical centers shows algorithm concordance with expert human morphology estimates at both the individual cell and full-field smear level, and when compared to standard clinical grading of smears. This algorithm illustrates high intra-sample reproducibility while providing greatly improved speed and resolution comparative to current standards. To test the utility of this new tool we examined: (i) diagnostic signatures in the RBC differential, (ii) large-scale cohort prognostics, (iii) single-cell mechanistic studies, and (iv) an animal model of hematologic disease. Diagnostic RBC differential signatures were discovered by applying the RBC-diff to a cohort of patients with thrombotic microangiopathy (TMA). In particular, the RBC-diff facilitated rapid identification and differential diagnosis of patients with thrombotic thrombocytopenic purpura (TTP) or hemolytic uremic syndrome (HUS), providing greater specificity than manual review (72% vs. 41%, p < 0.001), while maintaining high sensitivity (94-100%). Because TTP is a life-threatening disorder requiring immediate therapy, this new rapid diagnostic tool may provide significant benefit, as existing gold-standard diagnostic tests for TTP are slow (hours to days). Large-scale cohort prognostic value was uncovered through an analysis of smears from a general cohort of 58,950 inpatients that found that elevated schistocyte counts are associated with increased all-cause mortality (9.5% six-month mortality for >1% schistocytes vs. 4.7% for schistocytes < 0.5%, p < 0.001), after adjusting for patient demographics, comorbidities, and manual smear review flags. This tool also allowed for analysis of morphologically annotated single-cell volume distributions, providing insights into how dysmorphology can affect mean corpuscular volume (MCV) and the red cell distribution width (RDW); note that current technology provides only whole-population measures for these variables. For example, in iron-deficient patients who underwent intravenous iron transfusions, this technique highlighted that successful hematologic resolution involved volume increases for all cell types, rather than an isolated reduction in microcyte levels. Finally, without any retraining the model was successfully applied to animal-derived blood smears, highlighting expected differences between a wild-type (WT) and sickle cell disease mouse model (SCD) (0% sickle cells for WT, 1.2% for SCD), demonstrating potential value for basic research investigations. Usage of the red cell differential has the potential to greatly benefit hematologic research and clinical practice. Here we present the RBC-diff, an extensively validated method for efficient calculation of this differential from commonly collected blood smear images. As a resource to the community, we provide open-source code for calculating the RBC-diff. We also provide a repository of 5000 individual RBC images, and 50 full-field smears, each manually labelled by a team of expert pathologists, to facilitate further developments of improved computational methods of RBC classification. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Red blood cells morphology and morphometry in adult, senior, and geriatricians dogs by optical and scanning electron microscopy.

Red blood cells (RBC) morphologic evaluation through microscopy optical (OM) and SEM, provides information to forecast, evaluate, and monitor the functioning of many organs. Factors, such aging and diseases affect RBC morphology in both, human and animals. SEM is useful to evaluate RBC morphology, although its use in diagnosis and evaluation in dogs is limited, due to the availability and cost. The aim of this research was to assess the normal RBC morphology in adult, senior and geriatrician dogs, clinically healthy by OM and SEM. In addition to evaluating the age effect, sex, body size, and their interaction on erythrocyte morphometry. To carry out the research 152 blood samples were evaluated from dogs of different sexes and body sizes (small, medium, and large). Three groups were made based on dogs age: group I adults (1-7.9 years old), group II senior (8-11.9 years old), and group III geriatricians (>12 years old). Erythrocyte parameters were evaluated by OM (diameter, height, and axial ratio). Per each dog, the parameters of 20 erythrocytes were measured. A total of 2,600 cells were scanned with the AmScope™ Software scale. In addition, the RBC morphology was evaluated by SEM. Statistical analyses used analysis of variance and a general linear model, which allows the comparison of multiple factors at two or more levels (p < 0.05). The results of this study showed that diameter and height were lower in adult dogs than in senior and geriatrician dogs (p < 0.05). Whereas, sex, body size, and the interaction did not show a significant effect (p > 0.05). Additionally, some images of anisocytosis, polychromasia, and poikilocytosis (echinocytes, acanthocytes, codocytes, spherocytes, stomatocytes, dacryocytes quatrefoil, and elliptocytes) were obtained by OM and SEM. Our study provides information about the morphological and morphometry alterations of adult, senior, and geriatrician dogs RBC. This work contributes to future investigations and the diagnosing diseases, where it is necessary to evaluate the morphology of RBC.

Numerical investigation on red blood cell flow based on unstructured grid.

Prediction of blood cell flow is known as the difficult research by reason of the complexity of blood vessel. In this study, considering the complex structure of blood vessels, a mechanical model for red blood cell (RBC) based on unstructured grid has been established to study the flow characteristics of RBCs in complex blood vessels. In the model, the strain-energy function by Skalak is employed to model the shear elasticity and surface-area conservation of the membrane, and the hinge spring is used to describe the forces originating from local bending of the membrane. The immersed boundary method is utilized to couple the interphase force. Using the model, the stretching test of RBC is compared with the experiment data, and the good agreement verified the validation of the present model. The morphology of red blood cell and the blood viscosity in micro-vessel are studied. RBCs move with a symmetric shape (parachute shape) in small blood vessels, and the buckling instability is observed when the RBC flow slowly through a micro-vessel or a converging-diverging capillary. When the vessel diameter is around 10μm, the reverse Fahraeus-Lindqvist effect is presented. The blood apparent viscosity shows linear increase with the blood hematocrit. In addition, Malaria infection can make the RBC deformability decreased and the blood apparent viscosity increased.

New formulation and activity of rhenium-platinum antitumor system

Two-component Rhenium-Platinum system (Re-Pt system) is based on administration of a cluster dirhenium(III) compound and cisplatin to tumor bearing animals followed by a significant antitumor effect and decreased toxic effect of cisplatin on normal cells. The aim of this work was to obtain solid lipid nanoparticles (SLN) from surface lipids (waxes) of Chelidonium majus L. (Papaveraceae) leaves and to estimate whether capsulation of dirhenium(III) as a component of the Re-Pt system into SLN will affect its antitumor activity and red blood cells (RBC) morphology in a rat model of Guerin’s carcinoma growth. Fourier-transform infrared spectroscopy, gas-liquid chromatography, microscopy, light scattering were used in the research. Solid lipid nanoparticles were obtained, characterized, loaded with cluster dirhenium(III) and being introduced together with cisplatin to rats with Guerin’s carcinoma resulted in RBC morphology preservation and a significant decrease in tumor weight. It was concluded that the lipid coating of the rhenium cluster compound did not reduce the antitumor effect of the Re-Pt system and protected RBC from toxic cisplatin influence­. A new formulation of the Re-Pt system is proposed. Keywords: carcinoma, rhenium cluster compound, rhenium-platinum antitumor system, solid lipid nanoparticles, surface lipids

ALECTINIB-RELATED ERYTHROCYTE MEMBRANE CHANGES, NON-IMMUNE HEMOLYSIS AND ERYPTOSIS

Alectinib is an anaplastic lymphoma kinase (ALK) inhibitor which standard initial treatment for patients with advanced ALK rearranged non-small-cell lung cancer (NSCLC). The current study was planned by the incidental observation of non-immun hemolysis signs in several alectinib-treated patients, and its aim was to comprehensively characterize eritrocyte changes under this drug. We analyzed retrospectively 13 patients treated with alectinib for ALK+ NSCLC at the Bakirkoy Sadi Konuk and Kartal Dr Lutfi Kırdar Hospital Medical Onkology Depertment.Almost all patients were consulted with the hematology clinic because of anemia and elevated lactate dehydrogenase during alectinib.Laboratory tests requested for characterization of anemia included reticulocyte count, indirect bilirubin,haptoglobin, direct antiglobulin test, and LDH.All patients were examined by peripheral smear The analyzed patients, hematological tests results are showed that: Anemia was present in approximately all of patients and was mostly mild, (lowest hgb 8.5gr/dl).Reticulocytes were increased and the direct antiglobulin (Coombs) test was negative in all patients. Peripheral blood smears showed signs of eryptosis, abnormal red blood cell morphology in all patients, with anisocytosis, a predominance of acanthocytes, as well as occasional echinocytes, spherocytes, dacrocytes and rare fragmentocytes We have reported 13 cases of significant alterations in erythrocyte morphology secondary to alectinib. Patients predominantly showed mild anemia, but one patient developed significant Coombs-negative hemolysis. In this case, hemolytic markers improved after alectinib was discontinued. This study highlights the need for both clinicians and haematologists to be observant for unrecognised off-target effects of novel agents.

Comparative analysis of different water samples as Buffer in Leishman stain.

Staining of blood films remains one of the most essential part of haematology. The different blood cells have different intracellular structure that take up stains according to chemical nature. Leishman stain is one of the Romanowsky stains used in the haematology laboratory for the staining of blood films. It contains methylene blue and eosin dye prepared in an alcohol medium and is diluted with a buffer. Phosphate buffer is used during Leishman stain as it acts as a mordant enhancing staining reactions and it is the recommended and best buffer for staining. Aim This comparative study was aimed at determining if different water samples used as buffer show significant effects on the morphology of blood cells during Leishman staining. Methods pH analyses of different water sources such as water from borehole, waterboard, rainwater, fridge water, distilled water and air conditioned water were carried out and used as buffers to carry out Leishmann staining on blood films. The stained slides were examined macroscopically and microscopically using 100x objective. Results The study has shown that deionized and tap water used as buffer could produce moderately good film with well stained red cells, white cells and platelets. Rainwater, distilled, fridge and air conditioned water showed a significant change in the morphology of blood cells both in an apparently healthy film and a diseased film Conclusion This study demonstrates that deionized water and tap water devoid of chlorine can be used as substitutes in the right pH of 6.8 in a situation where phosphate buffer is not available. It is therefore recommended that quality control should be carried out to check the quality of stains and buffer used for staining films in order to produce quality results.

Analysis of ridges and grooves shape in grating coupler for optimization of integrated optics sensor structures.

The paper presents a theoretical analysis of a sensor structure based on a planar waveguide and grating coupler designed to determine selected physical properties of blood – hemoglobin concentration and oxidation level. In particular analysis were focused on optimization of selected geometrical properties of grating coupler (shape of ridges and grooves) to obtain maximum efficiency of uncoupling of light from the sensor structure. The analysis were carried out for three type of ridges and grooves shape in grating coupler: rectangular, triangular and sinusoidal. Full Text: PDF ReferencesI. . Singh, A.Weston, A. Kundur, G. Dobie, Haematology Case Studies with Blood Cell Morphology and Pathophysiology; Elsevier: Amsterdam, The Netherlands, (2017). DirectLink P. Jarolim, M. Lahav, SC. Liu, J. Palek, "Effect of hemoglobin oxidation products on the stability of red cell membrane skeletons and the associations of skeletal proteins: correlation with a release of hemin", Blood 76, 10 (1990). CrossRef E. Beutler, J. Waalen, "The definition of anemia: what is the lower limit of normal of the blood hemoglobin concentration?", Blood 107, 5 (2006). CrossRef M. Kiroriwal, P. Singal M. Sharma, A. Singal, "Hemoglobin sensor based on external gold-coated photonic crystal fiber", Optics &amp; Laser Technology 149, 107817 (2022). CrossRef A. A. Boiarski, J. R. Busch, B. S. Bhullar, R. W. Ridgway, V. E. Wood, "Integrated optic sensor with macro-flow cell", Proc. SPIE Integrated Optics and Microstructures 1793 (1993). CrossRef L. Cheng, S. Mao, Z. Li, Y. Han and H. Y. Fu, "Grating Couplers on Silicon Photonics: Design Principles, Emerging Trends and Practical Issues", Micromachines 11, 666 (2020). CrossRef P. Struk, "Design of an Integrated Optics Sensor Structure Based on Diamond Waveguide for Hemoglobin Property Detection", Materials 12, 175 (2019). CrossRef P. Struk, "Numerical analysis of integrated photonics structures for hemoglobin sensor application", Phot. Lett. Poland 12, 2 (2020). CrossRef P.V. Lambeck, "Integrated optical sensors for the chemical domain", Meas. Sci. Technol. 17, (2006). CrossRef W. Lukosz, "Integrated optical chemical and direct biochemical sensors", Sens. Actuators B Chem 29 (1995). CrossRef P. Struk, T. Pustelny, K. Gołaszewska,E. Kaminska, M.A. Borysiewicz, M. Ekielski, A. Piotrowska, "Hybrid photonics structures with grating and prism couplers based on ZnO waveguides", Opto-Electron. Rev. 21, (2013). CrossRef P. Struk, "Design of an integrated optics sensor structure for hemoglobin property detection", Proc. SPIE 11204, (2019). CrossRef OptiFDTD Technical Background and Tutorials - Finite Difference Time Domain Photonics Simulation Software, Optiwave Systems Inc. (2008). DirectLink K. Yee, "Cutoff Frequencies of Eccentric Waveguides", IEEE Transactions 14, 3 (1966). CrossRef

Utility of Differential White Cell Count and Cell Population Data for Ruling Out COVID-19 Infection in Patients With Community-Acquired Pneumonia

IntroductionThe main aim of this study was to assess the utility of differential white cell count and cell population data (CPD) for the detection of COVID-19 in patients admitted for community-acquired pneumonia (CAP) of different etiologies. MethodsThis was a multicenter, observational, prospective study of adults aged ≥18 years admitted to three teaching hospitals in Spain from November 2019 to November 2021 with a diagnosis of CAP. At baseline, a Sysmex XN-20 analyzer was used to obtain detailed information related to the activation status and functional activity of white cells. ResultsThe sample was split into derivation and validation cohorts of 1065 and 717 patients, respectively. In the derivation cohort, COVID-19 was confirmed in 791 patients and ruled out in 274 patients, with mean ages of 62.13 (14.37) and 65.42 (16.62) years, respectively (p<0.001). There were significant differences in all CPD parameters except MO-Y. The multivariate prediction model showed that lower NE-X, NE-WY, LY-Z, LY-WY, MO-WX, MO-WY, and MO-Z values and neutrophil-to-lymphocyte ratio were related to COVID-19 etiology with an AUC of 0.819 (0.790, 0.846). No significant differences were found comparing this model to another including biomarkers (p=0.18). ConclusionsAbnormalities in white blood cell morphology based on a few cell population data values as well as NLR were able to accurately identify COVID-19 etiology. Moreover, systemic inflammation biomarkers currently used were unable to improve the predictive ability. We conclude that new peripheral blood biomarkers can help determine the etiology of CAP fast and inexpensively.

Coagulopathy during COVID-19 infection: a brief review.

The COVID-19 pandemic caused by SARS-CoV-2 continues to spread rapidly due to its virulence and ability to be transmitted by asymptomatic infected persons. If they are present, the symptoms of COVID-19 may include rhinorrhea (runny nose), headache, cough, and fever. Up to 5% of affected persons may experience more severe COVID-19 illness, including severe coagulopathy, acute respiratory distress syndrome (ARDS) characterized by respiratory failure that requires supplementary oxygen and mechanical ventilation, and multi-organ failure. Interestingly, clinical evidence has highlighted the distinction between COVID-19-associated coagulopathy (CAC) and disseminated intravascular coagulation (DIC). Patients with CAC exhibit different laboratory values than DIC patients for activated partial thromboplastin time (aPTT) and prothrombin time (PT) which may be normal or shortened, varying platelet counts, altered red blood cell morphology, unique bleeding complications, a lack of schistocytes in the peripheral blood, and no decrease in fibrinogen levels. In this review, we consider the search for 1) laboratory results that can diagnose or predict development of CAC, including serum levels of D-dimers, fibrinogen, interleukin-6 (IL-6) and the growth factor angiopoietin-2 (Ang-2), 2) mechanisms of CAC induction, and 3) novel therapeutic regimens that will successfully treat COVID-19 before development of CAC.

WBC Image Segmentation Based on Residual Networks and Attentional Mechanisms.

White blood cell (WBC) morphology examination plays a crucial role in diagnosing many diseases. One of the most important steps in WBC morphology analysis is WBC image segmentation, which remains a challenging task. To address the problems of low segmentation accuracy caused by color similarity, uneven brightness, and irregular boundary between WBC regions and the background, a WBC image segmentation network based on U-Net combining residual networks and attention mechanism was proposed. Firstly, the ResNet50 residual block is used to form the main unit of the encoder structure, which helps to overcome the overfitting problem caused by a small number of training samples by improving the network's feature extraction capacity and loading the pretraining weight. Secondly, the SE module is added to the decoder structure to make the model pay more attention to useful features while suppressing useless ones. In addition, atrous convolution is utilized to recover full-resolution feature maps in the decoder structure to increase the receptive field of the convolution layer. Finally, network parameters are optimized using the Adam optimization technique in conjunction with the binary cross-entropy loss function. Experimental results on BCISC and LISC datasets show that the proposed approach has higher segmentation accuracy and robustness.

Morphology and cytochemical patterns of peripheral blood cells of tiger frog (Rana rugulosa).

Tiger frog (Rana rugulosa) is a national second-class protected amphibian species in China with an important ecological and economic value. In recent years, due to excessive human hunting, pollution and habitat loss, the wild population of tiger frog has declined sharply. To protect wildlife resources, the artificial breeding of tiger frogs has rapidly developed in China. Diseases are increasing and spreading among tiger frogs due to the increasing scale of artificial farming. The blood examination is the most straightforward and less invasive technique to evaluate the animal health condition. Thus, it is essential to obtain the normal hematological indicators of tiger frogs. The objective of this study was to investigate the morphometry, microstructure and cytochemical patterns of peripheral blood cells in tiger frogs. The number of blood cells in tiger frogs was counted on a blood count board, and the cell sizes were measured by a micrometer under light microscope. The morphology and classification of blood cells were studied by Wright-Giemsa staining, and the cytochemical pateerns was investigated by various cytochemical staining including periodic acid-Schiff (PAS), Sudan black B (SBB), peroxidase (POX), alkaline phosphatase (AKP), acid phosphatase (ACP), chloroacetic acid AS-D naphthol esterase (CAE) and α-naphthol acetate esterase (ANAE) staining. Besides erythrocytes and thrombocytes, five types of leukocytes were identified in tiger frogs: neutrophils, eosinophils, basophils, lymphocytes and monocytes. The mean erythrocyte, leukocyte and thrombocyte counts were 1.33 ± 0.15 million/mm3, 3.73 ± 0.04 × 104/mm3 and 1.7 ± 0.01 × 104/mm3, respectively. Small lymphocytes were the most abundant leukocytes, followed by large lymphocytes, Neutrophils, eosinophils and monocytes, basophils were the fewest. Eosinophils were strongly positive for PAS, positive for SBB, POX, ACP, CAE, ANAE, while weakly positive for AKP staining; basophils were strongly positive for PAS, ACP, positive for SBB, CAE, weakly positive for ANAE, negative for AKP, POX staining; neutrophils were strongly positive for ACP, SBB, positive for PAS, POX, weakly positive for AKP, CAE and ANAE staining; monocytes were positive for PAS, SBB, ANAE, weakly positive for ACP, AKP, POX, CAE staining; large lymphocytes and thrombocytes were positive for PAS, ACP, weakly positive for ANAE, while negative for SBB, POX, AKP, CAE; small lymphocytes were similar to large lymphocytes, except for strongly positive for PAS and ACP staining. The blood cell types and morphology of tiger frogs were generally similar to those of other amphibians, while their cytochemical patterns had some notable species specificity.Our study could enrich the knowledge of peripheral blood cell morphology and cytochemistry in amphibians, and provide baseline data for health condition evaluation and disease diagnosis of tiger frogs.

A multiple instance learning approach for detecting COVID-19 in peripheral blood smears.

A wide variety of diseases are commonly diagnosed via the visual examination of cell morphology within a peripheral blood smear. For certain diseases, such as COVID-19, morphological impact across the multitude of blood cell types is still poorly understood. In this paper, we present a multiple instance learning-based approach to aggregate high-resolution morphological information across many blood cells and cell types to automatically diagnose disease at a per-patient level. We integrated image and diagnostic information from across 236 patients to demonstrate not only that there is a significant link between blood and a patient's COVID-19 infection status, but also that novel machine learning approaches offer a powerful and scalable means to analyze peripheral blood smears. Our results both backup and enhance hematological findings relating blood cell morphology to COVID-19, and offer a high diagnostic efficacy; with a 79% accuracy and a ROC-AUC of 0.90.

Development of a non-viral gene vector for enhancing gene transfection efficiency

Construction of a safe and high transfection efficiency non-viral gene delivery vector to improve the effectiveness of gene delivery is vital for cancer treatment. Herein, we used 1, 2-Dioleoyl-3-Trimethylammonium-Propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol to prepare cationic liposomes in combination with poly(aspartate-graft-PEI423) (PAE) and plasmid DNA to prepare Liposomes-poly(aspartate-graft-PEI423)/DNA (LP-PAE/DNA) to improve gene transfection efficiency. LP-PAE/DNA particle size was about 220 nm, the zeta potential was about 20 mV and the morphology was spherical. The LP-PAE/DNA lipopolyplexes showed higher transfection efficiency at a weight ratio of 10:1:1 in HeLa, A549 and SPC-A1 cell line. LP-PAE/DNA lipopolyplexes toxicity in different cell lines and the main endocytic pathways through which LP and LP-PAE mediate DNA into cells was examined. In addition, the results of hemolysis test, red blood cell morphology observation and blood routine experiments showed that LP-PAE/DNA would not cause systemic hematological toxicity. The histological analysis including hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay confirmed lipopolyplexes/liposomes mediate the entry of therapeutic genes into the body not causing damage to normal organs and enhancing tumor tissue apoptosis relative to more traditional approaches. Therefore, it could be hoped the feasibility of LP-PAE as an efficient and promising platform for gene delivery.

Hematology, Ultrastructure and Morphology of Blood Cells in Rufous-Winged Buzzards (Butastur liventer) from Thailand.

Simple SummaryComplete blood counts (CBCs) are a manual tool used for the screening of raptor health. These tests require knowledge of blood cell morphology. This study aims to describe the quantitative and qualitative information of blood cells in rufous-winged buzzards (RWB) admitted into the rehabilitation center. The CBCs of 12 out of 17 RWBs were included for descriptive hematologic values. Heterophils were the most prevalent white blood cells in RWBs. One non-parasitized RWB showed hypochromic erythrocytes with PCV 0.18 L/L, which indicated that anemia in RWBs resulted from non-parasitic causes. The blood cells in RWBs are similar to those in other diurnal raptors, except for the lymphocytes. The electron micrographs confirmed the photomicrograph. This report can be used for the baseline information for rehabilitation of the RWBs in Thailand.In attempt to treat injured raptors and promote conservation awareness, the Kasetsart University Raptor Rehabilitation Unit (KURRU) was established in 2007. The complete blood counts (CBCs) are a manual tool used for the screening of raptor health. These tests require knowledge of blood cell morphology. This study aimed to describe the preliminary information of the hematology, ultrastructure, and morphology of blood cells in rufous-winged buzzards (RWB). There were 17 RWBs admitted into the KURRU. CBCs were manually performed by veterinary technicians. The morphology and morphometry of blood cells were observed from Wright-stained blood smears. Ultrastructure was observed from uranyl acetate and lead citrate-stained sections. The hematologic values were analyzed and described from individual RWBs that were clinically healthy, negative for blood parasites, and had PCV > 0.30 L/L. Consequently, CBCs of 12 out of 17 RWBs were included for descriptive hematologic values. Heterophils were the most prevalent white blood cells in RWBs. Of these 17 RWBs, 1 non-parasitized RWB showed hypochromic erythrocytes with PCV 0.18 L/L, which indicated that anemia in RWBs resulted from non-parasitic causes. The morphology of blood cells in RWBs was similar to those in other diurnal raptors, except that the lymphocytes showed pale or colorless cytoplasm. The electron micrographs highlighted that the basophil contained two types of granules: homogeneous electron-dense granules and reticulated electron-dense granules. The photomicrographs in this report are the scientific reference for identification of blood cells in RWBs. The CBCs from non-parasitized RWBs (clinically healthy) can be used as a cage mate reference in the KURRU. Additionally, we found evidence that evaluations of blood smears together with CBC examination were important in raptors.

Haematological profile, blood cell characteristic and serum biochemical composition of cultured brown trout, Salmo trutta fario with respect to sex

The purpose of this research is to see how sex affects several haematological, biochemical and blood cell morphology in cultured brown trout, Salmo trutta fario. Different haematological parameters, for instance haemoglobin (Hb) concentration, haematocrit (Hct), red blood cell (RBC) and white blood cell (WBC) counts, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) and serum biochemical parameters such as glucose, cholesterol, total protein, albumin, globulin, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), sodium, potassium and phosphorus were analysed. Throughout the study period, male fish had considerably greater Hb content, RBC count and Hct content than female fish, but insignificant (P > 0.05) difference in WBC count, differential leucocyte count, MCV, MCH and MCHC was seen between the two sexes of Salmo trutta fario. The majority of biochemical markers showed no significant (P > 0.05) difference, with the exception of AST, ALP, sodium, potassium, and phosphorus, which showed a significant (P < 0.05) difference. Under light microscopy, RBC and differential leukocyte count (neutrophils, lymphocytes, and monocytes) as well as thrombocytes were characterised and discriminated, revealing that the majority of the cells were normal and intact in shape. As a result of the foregoing findings, it is suggested that the data generated be used to identify the health state of fish and to determine the occurrence of various clinical and subclinical disorders, which might assist in enhancing the overall fish population production.

Characteristics of haematological and morphology in pulmonary tuberculosis patients using drugs with ADR at the National Lung Hospital from January 2021 to June 2021

Background: ADR (Adverse drug reactions) is also known as an adverse drug effect. ADRs of anti-tuberculosis drugs are quite common, causing changes in haematological tests. Objective: to study the characteristics of some haematological index and peripheral blood cell morphology in pulmonary tuberculosis patients with tuberculosis drugs with ADR who were treated at the National Lung Hospital from January 2021 to June 2021. Subjects and methods: 134 patients with pulmonary tuberculosis were diagnosed with tuberculosis drugs with ADR at the National Lung Hospital using a cross-sectional descriptive study. Results showed that the mean erythrocyte index was 3.69±0.92x1012/l in men and 3.81±0.74x1012/l in women, mean haemoglobin was 102.42±26.44 g/l in men and 96±22.81 g/l in women. The rate of anaemia was 73.47% in men and 61.11% in women. Average white blood cell count was 11.88±6.38x109/l, average platelet index was 371.45±198.16x109/l. Common morphology: roulaux cells, target cells and hypochromic RBCs. Neutrophils increase segmentation and granulocytosis. Giant platelets. Conclusion: pulmonary tuberculosis patients with tuberculosis drugs with ADR have anaemia, appearance of granulocytosis, giant platelets, rouleaux cells, target cells and hypochromic RBCs.

Effects of biodegradable biomedical porous MnO2 nanoparticles on blood components and functions

In recent years, manganese dioxide (MnO2) nanoparticles with unique physicochemical properties have been widely used in many biomedical fields, such as biosensors, contrast agents, tumor therapy, and drug delivery. From these applications, MnO2 nanoparticles have great clinical translation potential. However, by contrast, the in vitro and in vivo biosafety of MnO2 nanoparticles have been deeply and thoroughly clarified for the clinical translation, which hinders their clinical applications. In this work, we deeply investigated the blood safety of MnO2 nanoparticles by conducting a series of in vitro and in vivo experiments. These included the effects of MnO2 nanoparticles on morphology of red blood cells, activation of platelets, coagulation functions, and toxicity of key organs. The obtained results show that these effects displayed a concentration-dependent manner of MnO2 nanoparticles. Different safe concentration ranges could be found in the different experimental index. This study provides important guidance for the specific biomedical applications of MnO2 nanoparticles, greatly accelerating their laboratory development and clinical translation.

E3 ubiquitin ligase RNF123-deficient mice exhibit reduced parasitemia and mortality in rodent malaria (Plasmodium yoelii 17XL) infection.

Increased levels of several human ubiquitin ligases, including ring finger protein 123 (RNF123), in red blood cells with Plasmodium falciparum infection, have been reported. RNF123 is an E3 ubiquitin ligase that is highly expressed in erythroid cells. However, the function of the RNF123 gene and the relationship between the RNF123 gene and malarial parasite has not been clarified in vivo. In this study, we generated RNF123-deficient mice using the CRISPR/Cas9 system, and analyzed malaria susceptibility and erythrocyte morphology. The levels of parasitemia 5days post-infection and mortality 21days post-infection with the lethal type of rodent malaria (Plasmodium yoelii 17XL) in RNF123-deficient mice was significantly lower than that in wild-type mice. In contrast, red blood cell morphology in RNF123-deficient mice was almost normal. These results suggest that erythrocytic RNF123 plays a role in susceptibility to rodent malaria infection, but does not play a role in erythrocyte morphology.

A Novel Animal Model in Diabetes: Regeneration of β cells in the Axolotl Salamander?

Diabetes is an ever‐growing health issue affecting an estimate of 463 million people worldwide. Type 1 diabetes is characterized by hyperglycemia and loss of pancreatic β cell mass. Currently, there is no curative treatment of β cell loss. Humans have a poor natural ability to regenerate β cells, empathizing the importance of conducting research in regenerative medicine. However, this field of study is limited by the animal models available due to their inadequate intrinsic ability to regenerate, size and/or remoteness from humans. As the axolotl salamander (Ambystoma mexicanum) has an intrinsic ability to regenerate limbs and organs throughout life, it is an advantageous animal model for studying the physiological and molecular mechanisms of regeneration. Compared to zebrafish, a well‐established regenerative animal model of diabetes, the size of the axolotl (up to 150 g) is fit for pre‐clinical imaging systems and serial blood samples throughout an experiment without causing any detrimental effects.We hypothesize that the axolotl has the ability to regenerate β cells, giving it the potential of becoming a new regenerative animal model of diabetes. To generate a diabetes‐like axolotl, streptozotocin, a cytotoxic glucose analogue targeting β cells specifically, was injected intraperitoneally. Initially, a protocol inspired by studies of zebrafish were used, in which 0.35 mg/g body mass was given at day 0, 1, 2, 11, and 18 to group 1 (n=12). However, this dosage caused high mortality, and the experiment was terminated prematurely. Therefore, three new dosage regimes inspired by rodent studies were investigated. For group 2 (n=3), animals were treated with 0.05 mg/g body mass at 5 consecutive days, group 3 (n=3) was given 0.05 mg/g body mass at day 0, 2, 4, 12, and 19, and group 4 (n=3) was given a single dose of 0.2 mg/g body mass at day 0. Disease development was detected by glucose tolerance tests and will further be investigated with immunofluorescence. Potential off target effects of streptozotocin will be characterized by examining changes in red blood cell morphology and the histology of liver and kidney.We found that the treatment given to group 2 and 3 resulted in diabetes‐like symptoms when compared to a control group. Group 3 resulted in low‐grade hyperglycemia, whereas the 5 consecutive injections given to group 2 caused moderate hyperglycemia.Our results demonstrate that the axolotl is able to develop diabetes‐like symptoms by streptozotocin treatment. We anticipate that further adjustments to the dosage regime given to group 2 will result in severe hyperglycemia and thus model a diabetic axolotl, and that this animal model will be able to regenerate the lost β cells. Thus, this animal model will be useful for studying the cellular source of the regenerated cells and the molecular mechanisms of regeneration.

Human HMGB1 does not induce eryptosis &lt;i&gt;in vitro&lt;/i&gt;

Aim: To study the ability of human high mobility group box protein 1 (HMGB1) to induce eryptosis in vitro. Material and methods: Blood collected from six healthy volunteers was incubated with HMGB1 (0-0.2-1-5 ng per ml). Eryptosis of red blood cells was assessed by Annexin V staining and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) staining by flow cytometry. The forward scatter (FSC) fluorescence was used to evaluate the morphology of red blood cells. Results: Exposure of erythrocytes to HMGB1 did not affect the morphology of erythrocytes, evidenced by no changes in the percentage of cells with small volume, i.e. shrunken cells, and erythrocytes with large volume, i.e. enlarged cells. HMGB1 had no impact on phosphatidylserine externalization, which is confirmed by the absence of statistically significant changes in the amount of phosphatidylserine-displaying cells and the mean fluorescence intensity (MFI) values of Annexin V-FITC in cells exposed to different concentrations of HMGB1. Furthermore, H2DCFDA staining revealed that the HMGB1 did not induce oxidative stress. Conclusion: HMGB1 does not promote eryptosis of human erythrocytes at concentrations of up to 5 ng/ml. Key words: high mobility group box protein 1, erythrocytes, inflammation, cell death