Blood Smear Analysis Research Articles

Hematological and molecular finding of Trypanosoma evansi in buffaloes of North Gujarat

Trypanosoma evansi is definitely one of the most common pathogenic trypanosomes in all over the world, obviously infecting domestic and wild animals. Before it contributes to even more economic losses, a fast and confirmatory early diagnosis is a must to curb the spread and development of the disease. The current study was aimed to identify Trypanosoma spp. in buffalo blood by blood smear analysis and polymerase chain reaction (PCR) as well as comparison of these two methods. This investigation was performed on 475 buffaloes with anaemia, recurrent fever, muscular weakness and loss of appetite. Out of 475 blood samples, 57 were found anaemic (Hb

Virtual Staining, Segmentation, and Classification of Blood Smears for Label-Free Hematology Analysis.

Objective and Impact Statement. We present a fully automated hematological analysis framework based on single-channel (single-wavelength), label-free deep-ultraviolet (UV) microscopy that serves as a fast, cost-effective alternative to conventional hematology analyzers. Introduction. Hematological analysis is essential for the diagnosis and monitoring of several diseases but requires complex systems operated by trained personnel, costly chemical reagents, and lengthy protocols. Label-free techniques eliminate the need for staining or additional preprocessing and can lead to faster analysis and a simpler workflow. In this work, we leverage the unique capabilities of deep-UV microscopy as a label-free, molecular imaging technique to develop a deep learning-based pipeline that enables virtual staining, segmentation, classification, and counting of white blood cells (WBCs) in single-channel images of peripheral blood smears. Methods. We train independent deep networks to virtually stain and segment grayscale images of smears. The segmented images are then used to train a classifier to yield a quantitative five-part WBC differential. Results. Our virtual staining scheme accurately recapitulates the appearance of cells under conventional Giemsa staining, the gold standard in hematology. The trained cellular and nuclear segmentation networks achieve high accuracy, and the classifier can achieve a quantitative five-part differential on unseen test data. Conclusion. This proposed automated hematology analysis framework could greatly simplify and improve current complete blood count and blood smear analysis and lead to the development of a simple, fast, and low-cost, point-of-care hematology analyzer.

Aetiological Profiles of Pancytopenia in Children between 2 months to 12 years of AgeA Retrospective Study from a Tertiary Care Centre, Chennai, India

Introduction: Pancytopenia is a medical condition that generally exists among children in which the peripheral blood cell lineages are found to reduce in blood. The condition is diagnosed as a common haematological problem with an extensive differential diagnosis and vary according to the geographical distribution and genetic mutation. The condition is reversible and easily treatable if identified at the earliest. Aim: To determine the frequency of aetiological spectrum of pancytopenia in children between the age of 2 months to 12 years from Chennai, Tamil Nadu, India. Materials and Methods: A retrospective descriptive study was carried out at the Institute of Child Health, Madras Medical College, Chennai, Tamil Nadu, India, from January 2016 to December 2016. Out of 12,869 patients admitted, 91 children were identified with pancytopenia based on peripheral blood smear analysis. Complete blood count, peripheral smear, red blood cell indices, bone marrow examination, and serum vitamin B12 levels were analysed. The complete data was tabulated in Microsoft excel sheet and frequency(n) and percentage (%) analysis was performed. Results: Total of 91 children of pancytopenia (45 males and 46 females , aged from 6 months to 12 years) were included in the study and analysed. Majority of the pancytopenia children belonged to the age group of 1 to 6 years (40, 43.9%). The identified common clinical presentations for admission include the unexplained fever (82.92%), pallor (10.8%), and bleeding (9.79%). The common major aetiologies were megaloblastic anaemia (23, 25.2%), acute lymphoblastic leukaemia (17, 18.6%), and aplastic anaemia (12, 13.1%). Infections due to microorganisms in children also cause pancytopenia and there was one child each with the infectious symptoms due to Human Immunodeficiency Virus (HIV), tuberculosis, and Epstein-Barr Virus (HBV). Conclusion: Acute leukaemia and bone marrow failure were identified as the most common causes of pancytopenia in children. It is highly advisable to record the symptoms related with iron deficiency anaemia along with the symptomatic issues of certain viral and bacterial infections among children from the developing nations while diagnosing pancytopenia.

Early Diagnosis and Results of Enzyme Replacement Therapy in the Patient with Mucopolysaccharidosis Type VI: Clinical Case

Background. Mucopolysaccharidosis type VI (MPS VI, Maroteaux–Lamy syndrome) is rare autosomal-recessive multisystem disease, one of the group of lysosomal storage diseases. The MPS VI pathogenesis is determined by arylsulfatase B enzyme deficiency caused by mutations in the ARSB gene. There are only few published clinical examples of this disease that covers the results of early enzyme replacement therapy (ERT) onset.Clinical case description. The child was suspected to have lysosomal storage disease at the age of 1.5 months, it was based on microscopic analysis of blood smears: Alder abnormality was revealed (granulations and red-violet inclusions in neutrophils, monocytes, lymphocytes cytoplasm). The diagnosis was confirmed at the age of 3 months: increased glycosaminoglycans (GAGs) concentration in the urine, arylsulfatase B activity decrease in dried blood spots, and pathogenic variant c.943C>T (p. R315X) in the ARSB gene in homozygous state were revealed. ERT with galsulfase was started at the age of 7 months. There was decrease in excretion of GAGs in urine to normal level after 9 and 15 months of therapy. Normal growth and body proportions for the patient’s age were determined 3 years after continuous ERT. However, there was progression of multiple dysostosis and joint stiffness, as well as eyes lesion.Conclusion. Early ERT onset cannot completely stop MPS VI progression but it allows to reduce the severity of several symptoms and improves patient’s quality of life.

New distributional data on <em>Haemogregarina stepanowi</em> (Apicomplexa) and <em>Placobdella costata</em> (Hirudinea) parasitising the Sicilian pond turtle <em>Emys trinacris</em> (Testudines)

The host-parasite system “Emys trinacris – Placobdella costata – Haemogregarina stepanowi” is known for Sicily, but scarce information is available to date about the distribution of the two parasites on the island. Therefore, an extensive sampling effort through visual census and collection and analysis of blood smears of the endemic Sicilian pond turtle E. trinacris was carried out in 46 water bodies scattered throughout mainland Sicily. Our findings revealed that the distribution of both parasites is limited to the Nebrodi area, where the infection of H. stepanowi has shown a high incidence on the local turtle populations. Our data suggest no correlation between the current distribution of the two parasite species and environmental features. The current distribution of H. stepanowi and P. costata seems not to be relictual, but rather the outcome of a recent colonisation process. Considering the possible negative impact of both H. stepanowi and P. costata on their turtle host, their long-term effect on E. trinacris should be investigated.

A Study on Prevalence of Anemia among Pregnant Women Attending Antenatal Clinic at Rural Health Centre (RHC) and Hospital, Jabalpur City, M.P.

Weakness is a wholesome lack issue, and 56 percent of the women living in developing countries are pallid (World Health Organization). Iron deficiency is the second leading cause of maternal death in India, accounting for 20% of all maternal deaths. To focus on the incidence of sickliness in pastoral expecting women and investigate the factors coupled with paleness. The review focused on pregnant women, who visited the Maternity Clinic of the Rural Health Preparation Center and saw a clinic in India. The study will last one year, beginning on September 1, 2021, and ending on September 7, 2021. According to World Health Organization guidelines, iron insufficiency was designated. The total number of participants was 269, as determined by a pre-planned and pre-tested survey. The review subjects ranged in age from 20 to 30 and above 30. We've also found iron deficiency in the first, second, and third trimesters. Continuous renal sickness and postpartum drain were restricted as of late blood bonded. The conclusion of iron deficiency was attempted using a fringe blood smear analysis and a conventional haemoglobin analysis using Shale's approach. The prevalence of iron insufficiency is 58.36 percent across 269 individuals, including mild, moderate, and severe weakness. The mother's age, education, socioeconomic standing, equality, and food preferences were discovered to be profoundly important determinants. Family structure, size, and menarche completion were not found to be significant predictors of illness. The significant incidence of iron deficiency (58.36 percent) indicates that the National Nutrition Anemia Prophylaxis Program is not being implemented properly.

Automated Digital Morphometry of Peripheral Blood Smears Detects Both Infrequent Events and Cellular Population Patterns in Myelodysplastic Syndrome

Background: Complete Blood Count (CBC) analytical capacity is falling short of recognizing informative RBC morphology or WBC dysplastic morphological changes. Current morphologic peripheral blood smear (PBS) analysis is performed manually using a semi-quantitative scale on a limited number of cells introducing high degree of subjectivity and low sensitivity. The novel Full-Field Morphology (FFM) technology developed by Scopio Labs performs PBS analysis on a significantly larger scale of 1000 fields of 100X view in a routine manner, allowing a precise and highly sensitive automated quantification of cellular and sub-cellular morphological parameters. Current diagnosis of myelodysplastic syndrome (MDS) is based on invasive bone marrow aspirate, followed by subjective morphological analysis. In this study, we applied this digital morphometric approach to compare PBS morphology of MDS patients with age-matched controls.Methods: 32 MDS (average age 80+10, [range 41-97] y, F:M ratio 14:18) and 30 age-matched control (average age 79+9, [range 65-100] y, F:M ratio 13:17) PBS were scanned by the Scopio Labs system, and evaluated according to three distinct morphological features with known significance in MDS: blast percentage per 100 or 1000 WBC; neutrophil cytoplasmic granulation per 1000 neutrophils; RBC morphology of at least 150,000 RBC. Quantitative determination of neutrophils granulation, was measured by Granulation Index (GI, between 0-1) and GI Distribution Width (GIDW, between 0-1). RBC measurements included the quantitative measurements of RBC size, namely macro- and microcytosis, and RBC contour changes (deformation), i.e. the percent of RBC that deviate from normal RBC shape.Results: The mean GI of MDS samples was 0.36+0.15, [range 0.14-0.63] (Fig. 1A middle, Fig. 1E), significantly (p<10 -4) lower compared with the mean GI of age-matched control samples 0.53+0.10, [range 0.24-0.64] (Fig. 1A top, Fig. 1E). Mean GI were highly diverse among MDS samples compared with age-matched controls (Fig. 1E), but with no significant differences in GIDW (not shown). Interestingly, two sub-populations of neutrophils were detected in some of the MDS samples, differ in their mean GI (0.26 for one sub-population, 0.52 for the second one, Fig. 1A bottom). Such fingerprint, suggesting the presence of an abnormal and normal clones, was not detected in the control samples. Percentage of blasts was determined per 100 or 1000 WBC counts/sample (Fig. 1B). Blasts were detected in 13/32 (41%) of MDS samples compared with 1/30 (3%) of age-matched controls, when counts were performed per 100 WBC. However, when 1000 WBC were analysed, blasts were detected in 27/32 (84%) of MDS samples compared with 6/30 (20%) of age matched controls, a highly significant difference (p<10 -6). The percentage of blasts per 1000 WBC counts/sample of MDS samples was 0.92+1.35, [range 0-5] %, significantly (P<0.0008) higher compared with the percentage of blasts per 1000 WBC counts/sample of age-matched control samples 0.02+0.05, [range 0-0.2] % (Fig. 1E). RBC analysis revealed significant differences between MDS and age-matched samples (Fig. 1C). As expected, mean RBC size (49+4, [range 41-59] mm 2 MDS; 45+3, [range 40-51] mm 2 age matched) and % of macrocytosis (17+15, [range 2-60] % MDS; 3+4, [range 0-15] % age matched) were significantly (p<10 -5) higher in the MDS samples compared with the age matched controls (Fig. 1E). We found that MDS PBS contained significantly (p<10 -6) higher number of abnormally-shaped RBC (8+1, [range 5-12] %), compared with age-matched controls (6+1, [range 5-8] %) (Fig. 1E). Representative summaries of morphometric analyses of MDS and age-matched control are shown in figure 1D. Representative PBS scans of MDS and control samples are available in https://demo.scopiolabs.com/?_org=VCdaE756rjYwZW3Z#/scans.Conclusion:Our study demonstrates that FFM-based digital PBS analysis enables the detection and quantification of unique WBC and RBC morphologic alterations associated with MDS. The expanding therapeutic options for MDS, including for patients at early disease stages, makes the establishment of an accurate diagnosis of MDS, even at early stages, to be highly important. The proposed novel digital imaging technology opens the opportunity to screen patients, diagnose them early, based on peripheral blood morphology, and potentially, monitor their responsiveness to therapy. [Display omitted] DisclosuresKatz: Scopio Labs: Consultancy. Karni: Scopio Labs: Current Employment. Shimoni: Scopio Labs: Current Employment. Natan: Scopio Labs: Current Employment. Shaham: Scopio Labs: Current Employment. Pozdnyakova: Scopio Labs: Consultancy. Mittelman: Janssen · Roche · Novartis · Takeda · Medison / Amgen · Neopharm / Celgene / BMS · Abbvie · Gilead: Research Funding; Novartis · Takeda · Fibrogen · Celgene / BMS · Onconova · Geron: Other: Clini; Onconova · Novartis · Takeda · Silence: Membership on an entity's Board of Directors or advisory committees; MDS HUB: Consultancy; Celgene / BMS · Novartis: Speakers Bureau. Avivi: Novartis: Speakers Bureau; Kite, a Gilead Company: Speakers Bureau.

Automated Peripheral Blood Cell Differentiation Using Artificial Intelligence - a Study with More Than 10,000 Routine Samples in a Specialized Leukemia Laboratory

Background: Cytomorphology is the gold standard for quick assessment of peripheral blood (PB) and bone marrow samples in hematological neoplasms and is used to orchestrate specific diagnostics. Artificial Intelligence (AI) promises to provide an unbiased way of interrogating blood smear data as reproducibility varies across labs. This is a prospective clinical study (ClinicalTrials.gov Identifier: NCT04466059) conducted on our approach outlined at ASH 2020.Aim: Use an AI model to classify cell images to produce differential counts of PB smears side-by-side to routine diagnostics.Methods: We enrolled 10,082 patient samples which were sent to our lab between 01/2021 and 07/2021 for cytomorphology with a suspected hematologic neoplasm. Blood smears were differentiated by highly skilled technicians (median 5y in lab) and all were reviewed by hematologists. In parallel, all samples were scanned on a MetaSystems (Altlussheim, Germany) Metafer Scanning System (Zeiss (Oberkochen, Germany) Axio Imager.Z2 microscope, automatic slide feeder). Areas of interest were defined and leukocyte positions were flagged by pre-scan in 10x magnification followed by high resolution scan in 40x to generate cell images for analysis.We set up a supervised Machine Learning model based on ImageNet-pretrained Xception using Amazon Sagemaker (AS) and trained it on 8,425 carefully annotated color images to identify 21 predefined classes (including 1 garbage class). Overall accuracy of this model against hold-out-set (10%) was 96%. The algorithm consumes 144x144pixel cell images and produces probability scores (PS) for each class in every image.Results: For routine diagnostics in median 100 cells/sample (range 82 - 103) were differentiated manually, overall 988,130. The automated process gathered 500 cell images/sample (range 101 - 500), overall 4,937,389. Average capture times for 500 cells: 4:37 min. Cropped images were uploaded to a cloud storage and exposed to an AS endpoint to initiate classification and the computation of a PS for each of the predefined 21 classes in the model.For the study we only considered images with a probability of at least 90% (n=3,781,670/4,937,389) and excluded normoblasts, smudge cells and images identified as garbage (together n=2,120,258). Final diagnosis included: no lymphoma detectable (2,186), MDS (1,152), AML (369), in these 11 APL, MPN (658), CLL (558), other mature B-cell neoplasms (377), CML (326), multiple myeloma (155), but also rare entities such as hairy cell leukemia variant (2) or PPBL (3).Comparing the benign normal cells in peripheral blood we identified (all values normalized) segmented neutrophils (manual (M): 516,648=52% vs AI: 882,538=53%), eosinophils (M: 24,860=2.52% vs. AI: 55,699=3.36%), basophils (M: 7159=0,72% vs. AI: 11,957=0,72%), monocytes (M: 74,113=7.5% vs. AI: 110,126=6.64%), lymphocytes (M: 313,518=31.7% vs. AI: 399,249=24%).Pathogenic blasts were detected in 16,048 (0.97%) images by AI (M: 16,290=1.65%). In routine diagnostics 536 cases with blast cells, including “questionable blasts” were identified. The AI identified 493 (91%) of these cases. At least one atypical/malignant lymphocyte was found in 2,323 samples manually, out of which the AI identified 2,279 (98%). In few cases manual differentiation relies on the number of pathogenic cells from an immunophenotyping analysis, which the AI does not had.During the course of the study by chance we identified at least 3 instances, were the AI detected pathogenic cells (blasts, atypical promyelocytes (APL) or bilobulated promyelocytes (APL-v)) which were initially missed manually (in some case WBC below .5 G/l) or flagged during subsequent immunophenotyping/molecular genetic analysis. Upon manually revisiting the smear, we could verify the presence of the AI-anticipated cells, revealing the higher sensitivity of the 5 time increase in cells/sample investigated by AI and power of algorithms.Conclusion: We present data of a prospective, blinded clinical study comparing blood smear analysis between humans and AI head-to-head. The concordance is extremely high with 95% for pathogenic cases. Misclassified cells are used for retraining to continuously improve the model and benefit from large datasets even for rare cell types. The model's cloud based implementation makes it easy to connect scanning devices for automated, unbiased classification. DisclosuresHaferlach: MLL Munich Leukemia Laboratory: Other: Part ownership. Kern: MLL Munich Leukemia Laboratory: Other: Part ownership. Haferlach: MLL Munich Leukemia Laboratory: Other: Part ownership.

Utilisation of Deep Learning-Enabled Image Analysis for Detection and Enumeration of Plasmodial Forms in Red Cells

Abstract Introduction/Objective Objective: This work investigates utilisation of deep learning enabled peripheral blood smear image analysis for automated detection and enumeration of red cell parasites. Methods/Case Report Methods: Peripheral blood smear red cell images from 30 individuals identified as positive and/or negative for plasmodilum falciparum forms were used. Blood submitted to hematology laboratory for complete blood count was evaluated on Sysmex XN 3100 analyzer with routine peripheral blood slides performed on instrument associated SP50 stainer. Images of red cells obtained with Xfinity DX40 microscope mounted camera were subjected to classification using deep learning software (Cognex, ViDi Suite 4.1). The training of the classification tool was performed on 200 peripheral blood smear images divided in two dataset classes: normal/negative and abnormal/positive for plasmodia, 50% of training images represented positive data set. Performance of the developed model was tested on 300 images including 66% positive for plasmodia obtained from 20 patients. Enumeration of parasitic forms was performed for each case. Model performance was compared to expert hematopathology reviewer which was used as gold standard. Results (if a Case Study enter NA) Results: Overall, Cognex ViDi Suite 4.1 demonstrates the effectiveness in discriminating between images positive and negative for red cell plasmodial forms as well as enables parasite quantification. Following performance specifications were determined for parasite detection: sensitivity (0.969230769), specificity (0.99383217). High correlation coefficients (0.9961) between automatically detected parasites and ground truth, on both image level and patient level, demonstrate the practicality of our method. Conclusion Deep learning enabled image analysis of peripheral blood smears is a promising alternative to manual identification and enumeration of red cell plasmodial forms with performance comparable to expert hematopathology reviewer.

Effects of Pucuk Merah (Syzygium myrtifolium (Roxb.) Walp.) Leaves Extract on Lymphocytes Count and Spleen Index of Male Balb/C Strain Mice (Mus musculus L.)

Antibiotics are applied to directly kill pathogenic microbes as a causal agent of infection. The excessive use of antibiotics as therapeutical agents on pathogen infections could trigger microbial resistance inside human body and even cause new diseases. As an alternative to reduce the use of antibiotics, therapy with natural ingredients contained in medicinal plants can help body by stimulating immune cells to eliminate invading pathogens. These medicinal plants are utilized as immunostimulant. As a tropical country, Indonesia has a variety of plants that can be used as medicinal plants, which one of them is the ornamental plant Pucuk Merah (Syzygium myrtifolium (Roxb.) Walp.). In this study, we tested the effect of Pucuk Merah (Syzygium myrtifolium (Roxb.) Walp.) leaves extract on lymphocytes count and spleen index of Balb/C mice (Mus musculus L.) as test animals. Pucuk Merah leaves extract was obtained by maceration method using 96% ethanol as solvent. Phytochemical compounds were analyzed using qualitative biochemical test and Gas Chromatography-Mass Spectrophotometry (GC-MS). Three doses used in the administration of the extract are 0.127, 0.510 and 0.765 mg/g of body weight. The extract was administered orally using a cannula. Lymphocytes count of treated mice were mainly based on blood smear analysis. Spleen of tested animal were weighed to determine its organ index. Our results showed that Pucuk merah leaves extract contained flavonoid, steroid, and tannin as detected by biochemical assays. Phytochemical quantitative test using GC-MS instrument detected 15 compounds with 1,2,3-Benzenetriol (37,80 %) and caryophyllene (27,49 %) as dominant compounds. Lymphocyte count of mice was tended to be decreased on the treatment of low dose (0.510 mg/g of body weight or less) of Pucuk Merah extract, but high dose (0.765 mg/g of body weight) was observed to increase mice lymphocyte count. Spleen index of Pucuk Merah extract-treated mice was not correlated with its ......

Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.

Manual microscopic inspection of fixed and stained blood smears has remained the gold standard for Plasmodium parasitemia analysis for over a century. Unfortunately, smear preparation consumes time and reagents, while manual microscopy is skill-dependent and labor-intensive. Here, we demonstrate that deep learning enables both life stage classification and accurate parasitemia quantification of ordinary brightfield microscopy images of live, unstained red blood cells. We tested our method using both a standard light microscope equipped with visible and near-ultraviolet (UV) illumination, and a custom-built microscope employing deep-UV illumination. While using deep-UV light achieved an overall four-category classification of Plasmodium falciparum blood stages of greater than 99% and a recall of 89.8% for ring-stage parasites, imaging with near-UV light on a standard microscope resulted in 96.8% overall accuracy and over 90% recall for ring-stage parasites. Both imaging systems were tested extrinsically by parasitemia titration, revealing superior performance over manually-scored Giemsa-stained smears, and a limit of detection below 0.1%. Our results establish that label-free parasitemia analysis of live cells is possible in a biomedical laboratory setting without the need for complex optical instrumentation. We anticipate future extensions of this work could enable label-free clinical diagnostic measurements, one day eliminating the need for conventional blood smear analysis.

3095 – IMMUNE THROMBOCYTOPENIA AND BONE MARROW HEMOPHAGOCYTOSIS ASSOCIATED WITH SARS COV-2

The clinical presentation of patients infected with SARS-CoV-2 is remarkably diverse. Likewise, the underlying pathophysiological mechanisms are proving complex. Disturbances in the blood coagulation system and cytokine storm, such as seen in hemophagocytic syndrome, are among the most serious ones. We present the case of a female 79-year-old patient with marked thrombocytopenia of 4 (150-450) x10^9/L occurring in the context of a confirmed SARS-CoV-2 infection. Clinically, one episode of epistaxis and petechiae was observed, otherwise no signs of bleeding occurred. Diagnostic workup included microscopic blood smear analysis, bone marrow cytologic evaluation and flow cytometric immunophenotyping. Hematological malignancies, thrombotic microangiopathies and common infections were excluded as cause of the low platelet count. In the bone marrow, cytology, the megakaryocytic lineage presented normocellular. However, several large hemophagocytes with engulfed hematopoietic cells were detected. A further evaluation of markers frequently associated with hemophagocytic syndrome was performed. Ferritin was 2888 (0-150) ng/mL, CRP and GOT were slightly elevated. The white blood count was normal with a marked decrease of lymphocytes to 0.13 (1.10-3.60) × 10^9/L. There was no fever or organomegaly and the patient was in good clinical constitution. Thus, we did not diagnose hemophagocytic syndrome. Due to no other explanation for the clinical and laboratory findings, the patient was diagnosed with immune thrombocytopenia and concomitant bone marrow hemophagocytosis associated to SARS-CoV-2. The first-line treatment consisting of prednisolone and intravenous immunoglobulins failed to induce an increase in the platelet count. As second-line treatment therapy with Eltrombopag, a TPO-agonist, was started and a sustainable response with platelets in the normal range was achieved.

First report of Trypanosoma vivax infection in sheep from Nicaragua.

In Central America, outbreaks of trypanosomiasis by Trypanosoma vivax have been recorded only in cattle. This is the first report of an outbreak of trypanosomiasis by T. vivax in 30 Pelibuey sheep (2 to 7years old, male and female) from Nicaragua, which occurred in 2009. Clinical signs included fever, apathy, pale mucous membranes, weakness, progressive weight loss, and sudden death. Infection by T. vivax was detected in 22 (73.3%) sheep by blood smear analysis and/or PCR. Trypanosomes were morphologically identified in 11 (36.7%) blood smear samples, whereas 17 (85%) of the 20 samples subjected to PCR were positive for T. vivax. Eighteen (81.8%) of the 22 infected sheep presented a packed red cell volume (PCV) lower than 25%. Upon diagnosis, the treated animals were clinically recovered and no parasites could be observed in subsequent examinations. Tabanids were potential mechanical vectors of T. vivax in the farm. This is the first report of T. vivax in Nicaragua and for the first time this haemoparasite is recorded in sheep in Central America.

Determination the Effect of Ionizing Radiation in Blood Cells Parameters among Sudanese Radiology Workers

Background: Radiology work involves risks that affect most blood components, leading to chronic diseases Design: This analytical study was conducted in Khartoum State from August to December 2020 Objective: The goal is to determine the effect of ionizing radiation on blood values in healthy radiologists using the analysis of complete blood count and blood smear And to determine the effect of the number of years of work, the number of working hours per day, the years of work: are they continuous or discontinued, the intensity of radiation, wearing a protective shield made of lead on the values of the complete blood count

Impaired Iron Homeostasis and Haematopoiesis Impacts Inflammation in the Ageing Process in Down Syndrome Dementia.

Down syndrome (DS) subjects are more likely to develop the clinical features of Alzheimer’s disease (AD) very early in the disease process due to the additional impact of neuroinflammation and because of activation of innate immunity. Many factors involved in the neuropathology of AD in DS, including epigenetic factors, innate immunity and impaired haematopoiesis, contribute significantly towards the pathophysiology and the enhanced ageing processes seen in DS and as a consequence of the triplication of genes RUNX1, S100β and OLIG2, together with the influence of proteins that collectively protect from cellular defects and inflammation, which include hepcidin, ferritin, IL-6 and TREM2. This study is aimed at determining whether genetic variants and inflammatory proteins are involved in haematopoiesis and cellular processes in DS compared with age-matched control participants, particularly with respect to neuroinflammation and accelerated ageing. Serum protein levels from DS, AD and control participants were measured by enzyme-linked immunosorbent assay (ELISA). Blood smears and post-mortem brain samples from AD and DS subjects were analysed by immunohistochemistry. RUNX1 mRNA expression was analysed by RT-PCR and in situ hybridisation in mouse tissues. Our results suggest that hepcidin, S100β and TREM2 play a critical role in survival and proliferation of glial cells through a common shared pathway. Blood smear analysis showed the presence of RUNX1 in megakaryocytes and platelets, implying participation in myeloid cell development. In contrast, hepcidin was expressed in erythrocytes and in platelets, suggesting a means of possible entry into the brain parenchyma via the choroid plexus (CP). The gene product of RUNX1 and hepcidin both play a critical role in haematopoiesis in DS. We propose that soluble TREM2, S100β and hepcidin can migrate from the periphery via the CP, modulate the blood–brain immune axis in DS and could form an important and hitherto neglected avenue for possible therapeutic interventions to reduce plaque formation.

Gain of Function Mutant p53 Drives Clonal Hematopoiesis and the Pathogenesis of Myelodysplastic Syndromes

Clonal hematopoiesis increases with age, where a single mutant hematopoietic stem or progenitor cell (HSPC) contributes to a significant, measurable clonal proportion of mature blood lineages. Evolution of mutant clonal hematopoiesis with age predisposes the elderly to myelodysplastic syndromes (MDS) and other aging-associated diseases, suggesting that mutations identified in HSPCs contribute to disease development. Previous studies using whole exome sequencing of blood DNAs identified a high frequency of TP53 mutations in aged healthy individuals . TP53 mutations are also present in 10% of MDS cases and correlate with worse clinical outcomes. However, the role of TP53 mutations in clonal hematopoiesis and the pathogenesis of MDS is largely unknown. Ample data indicate that mutant p53 proteins not only lose their tumor suppressive functions, but also gain new abilities that promote tumorigenesis. Here, we report that specific gain-of-function (GOF) mutant p53 proteins drive the development of pre-leukemic hematopoietic stem cells (HSCs), predisposing aged mutant p53 mice to MDS development.Pre-leukemic HSCs have been shown to contribute to normal blood development and harbor a selective growth advantage compared to normal HSCs. We found that GOF mutant p53 (p53R248W) enhances the competitive advantage of HSCs without affecting terminal differentiation in steady state. Further, we observed that p53R248W mutant promotes clonal expansion of HSCs following genotoxic stresses. RNA-seq analysis revealed that HSC and leukemia stem cell signatures were significantly enriched in mutantHSCs compared to wild-type HSCs. Thus, HSCs expressing GOF mutant p53 recapitulate several key features of pre-leukemic HSCs. GOF p53 proteins are capable of dramatically altering tumor cell behavior, primarily through their interactions with other cellular proteins and regulation of cancer cell transcriptional programs. We discovered that GOF mutant p53 proteins, including p53R248W, p53R273H, and p53R175H, but not wild-type p53, interact with histone methyltransferase EZH2 and enhance H3K27me3 in HSCs. EZH2, a catalytic component of the Polycomb repressive complex 2 (PRC2), trimethylates histone H3 at lysine 27 (H3K27me3) to repress the transcription of target genes. Indeed, we found that the expression of HSC regulators, including Elf4 and Gadd45g, was significantly downregulated in mutant HSCs compared to wild-type HSCs. ChIP-seq analysis show enrichment of H3K27me3 in the promoter regions of Elf4 and Gadd45g in mutant HSCs. Importantly, we discovered that EZH2 inhibitor treatment decreased the replating potential of mutantbone marrow cells to wild-type cell level. Thus, we demonstrated that GOF mutant p53 drives the development of pre-leukemic HSCs through enhancing EZH2 activity.Given that most homozygous p53-/- and p53R248W/R248Wmice develop spontaneous tumors, including lymphoma, thymoma, and sarcoma, and die within 3 to 6 months after birth, it was not clear whether heterozygous mutant p53 mice (p53R248W/+) would develop myeloid malignancies during aging. To evaluate the role of GOF mutant p53 in the pathogenesis of myeloid malignancies, we maintained p53+/+ and p53R248W/+ mice for more than a year and monitored their survival and tumor development. While p53R248W/+ mice show extended life span compared to p53-/- and p53R248W/R248W mice, most of these mice died within 16-18 months after birth. During the observation period, approximately 30 % of heterozygous mutant p53mice developed MDS based upon pathological analysis of BM, spleen, and peripheral blood smear. Histological assessment revealed hypercellular BM in some aged p53R248W/+ mice. BM of some aged p53R248W/+ mice show neutrophil, erythroid precursor and megakaryocyte dysplasia. We further demonstrated that mutant p53 driven-MDS is transplantable. Thus, we provided evidence that GOF mutant p53 drives MDS development during aging.In summary, we have identified a critical role for GOF mutant p53 in clonal hematopoiesis and the pathogenesis of MDS. DisclosuresNo relevant conflicts of interest to declare.

British Society for Haematology guidelines for the diagnosis and evaluation of prognosis of Adult Myelodysplastic Syndromes.

British Society for Haematology guidelines for the diagnosis and evaluation of prognosis of Adult Myelodysplastic Syndromes.

First detection of Trypanosoma vivax in cattle in the state of Bahia, Brazil, based on parasitological and molecular analyses

We report the first detection of Trypanosoma vivax in Bahia state based on blood smear and PCR analyses. A total of 623 bovine blood samples were collected over two years. Parasitological analysis by smear technique detected the presence of T. vivax in 0.3%, while molecular analysis by PCR showed a prevalence in 18.9% of the samples. This study demonstrated the higher sensitivity of molecular analysis in the diagnosis of hemoparasitosis caused by T. vivax in dairy cattle herds.

Hydrogel-Based Stamping Technology for Solution-Free Blood Cell Staining.

An accurate microscopical analysis of blood smears requires a reproducible and convenient method of staining. Solution-based staining procedures can be cumbersome. Especially in low- and middle-income countries, the lack of skilled technicians and adequate laboratory facilities, as well as insufficient water and reagent quality, often become confounding factors. To overcome these obstacles, we developed a new cell staining method based on sequential stamping of agarose gel patches that contain eosin, methylene blue/oxidized methylene blue, Azure B, and buffer, respectively. Our method, termed "hydrogel staining", provides a simple, reproducible, solution-free, and inexpensive approach to stain blood cells. We have optimized incubation times to achieve the optimal transfer of dyes to fixed blood cells on a glass slide, with outcomes comparable to conventional solution-based methods for white blood cells and malaria-infected red blood cells. This hydrogel staining method does not require special skills to produce excellent quality stained blood film slides. The new method could enhance the accuracy of microscopical examination of blood smears, especially in resource-limited settings.

Detection and Classification of Leukocytes in Blood Smear Images

Manual analysis of microscopic blood smears by highly expert pathologists is labor-intensive, time-consuming, and is subject to inter-observer variations. Recent innovations in image processing and computer vision techniques have improvised digital pathology in terms of objectivity and reproducibility. Traditional computer vision-based methods of recognition of white blood cell (WBC) from a pathological blood smear image includes the process of detection, segmentation, and classification. This paper presents a review of state-of-the-art detection, segmentation, and classification techniques for white blood cell analysis. The goal of this work is to present an introduction to the field, provide enough information about the analysis methods developed so far, and to be an appropriate reference for the researchers looking forward in this field. The methods under review are classified into intensity and feature based. The crucial steps involved in these techniques, mathematical foresights, performance evaluation techniques, issues, and future directions are discussed.