Whole genome sequencing of hematologically stained cells catapulted from Cell smears

Analyzing archived peripheral blood smears is a potential route towards gaining cell morphology and genome information of blood cell types from various diseases. Yet, acquiring whole genome information from morphologically targeted cells was difficult, especially for rare cell types. The main causes for such difficulty were the inevitable usage of cell stains leading to whole genome amplification inhibition, and insufficient cell isolation performance of previously introduced laser microdissection (LMD) techniques. Here, we introduce a new laser-based cell isolation technique and a whole genome amplification (WGA) protocol optimized for whole genome analysis from minute input of hematologically stained cells. We were able to perform whole genome copy number profiling and SNP analysis from as little as 5 cells.

Weakly Supervised Ternary Stream Data Augmentation Fine-Grained Classification Network for Identifying Acute Lymphoblastic Leukemia

Due to the high incidence of acute lymphoblastic leukemia (ALL) worldwide as well as its rapid and fatal progression, timely microscopy screening of peripheral blood smears is essential for the rapid diagnosis of ALL. However, screening manually is time-consuming and tedious and may lead to missed or misdiagnosis due to subjective bias; on the other hand, artificially intelligent diagnostic algorithms are constrained by the limited sample size of the data and are prone to overfitting, resulting in limited applications. Conventional data augmentation is commonly adopted to expand the amount of training data, avoid overfitting, and improve the performance of deep models. However, in practical applications, random data augmentation, such as random image cropping or erasing, is difficult to realistically occur in specific tasks and may instead introduce tremendous background noises that modify actual distribution of data, thereby degrading model performance. In this paper, to assist in the early and accurate diagnosis of acute lymphoblastic leukemia, we present a ternary stream-driven weakly supervised data augmentation classification network (WT-DFN) to identify lymphoblasts in a fine-grained scale using microscopic images of peripheral blood smears. Concretely, for each training image, we first generate attention maps to represent the distinguishable part of the target by weakly supervised learning. Then, guided by these attention maps, we produce the other two streams via attention cropping and attention erasing to obtain the fine-grained distinctive features. The proposed WT-DFN improves the classification accuracy of the model from two aspects: (1) in the images can be seen details since cropping attention regions provide the accurate location of the object, which ensures our model looks at the object closer and discovers certain detailed features; (2) images can be seen more since erasing attention mechanism forces the model to extract more discriminative parts’ features. Validation suggests that the proposed method is capable of addressing the high intraclass variances located in lymphocyte classes, as well as the low interclass variances between lymphoblasts and other normal or reactive lymphocytes. The proposed method yields the best performance on the public dataset and the real clinical dataset among competitive methods.

ARTIFACTS OF FORMED ELEMENTS AND PLASMA OF BIRD BLOOD, THEIR GENESIS AND DIAGNOSTIC SIGNIFICANCE

The work is devoted to the study of literature data (humane and veterinary medicine) and the practical analysis of artifacts formed elements and plasma in peripheral blood smears of birds in a model of broiler chickens Gallus gallus L. of early postnatal ontogenesis. The age of the studied clinically healthy chicks and young hens was: day 1, day 7, day 23 and day 42 (n = 40). We studied 158 (n = 158) high-resolution color micrographs of the fields of view, in blood smears stained according to Pappenheim. As a result, single artifacts of avian erythrocytes were identified: cytoplasmic vacuoles of various pattern character, scalloped «bitten» edges of cells. Artifacts of blood plasma were found: pericellular and adhesioned on the cell surface of colored coagulated granularity. In some cases, this granularity imitated the toxic forms of granulocytes and agranulocytes in the peripheral blood of birds. It is necessary to distinguish artifacts of cells and plasma in peripheral blood smears from adaptive changes and symptoms of infectious, invasive and non-infectious diseases.

How Reproducible Is the Data from Sysmex DI-60 in Leukopenic Samples?

Digital morphology (DM) analyzers are widely applied in clinical practice. It is necessary to evaluate performances of DM analyzers by focusing on leukopenic samples. We evaluated the analytical performance, including precision, of a Sysmex DI-60 system (Sysmex, Kobe, Japan) on white blood cell (WBC) differentials in leukopenic samples. In a total of 40 peripheral blood smears divided into four groups according to WBC count (normal, mild, moderate, and severe leukopenia; each group n = 10), we evaluated precision of WBC preclassificaiton by DI-60. %coefficients of variation (%CVs) of precision varied for each sample and for each cell class; the fewer cells per slide, the higher %CV. The overall specificity and efficiency were high for all cell classes except plasma cells (95.9–99.9% and 90.0–99.4%, respectively). The largest absolute value of mean difference between DI-60 and manual count in each group was: 10.77, normal; 10.22, mild leukopenia; 19.09, moderate leukopenia; 47.74, severe leukopenia. This is the first study that evaluated the analytical performance of DI-60 on WBC differentials in leukopenic samples as the main subject. DI-60 showed significantly different performance depending on WBC count. DM analyzers should be evaluated separately in leukopenic samples, even if the overall performance was acceptable.

Educational cost-effective intervention to reduce pathologist’s peripheral blood smears reviews with non-contributory findings: an academic institution experience

Peripheral blood smear (PBS) review by a pathologist is a necessary and invaluable diagnostic tool. However, innovative highly sophisticated haematology analysers that flag peripheral blood abnormalities have decreased the need for a PBS review. Ordering practices including PBS reviews lumped as part of an ‘order set’ or with complete blood count (CBC) constituted most PBS requests at our institution. A retrospective review of all PBS review orders from 1 April 2016 to 31 January 2017 was performed to investigate the ordering practices at our institution. A total of 2864 PBS were ordered during the above study period. In many cases, the PBS report did not add any significant clinical information beyond that acquired by the CBC and differential count. These findings inspired policy changes within our institution for pathologist PBS reviews. Within the electronic order system, all PBS orders for inpatients were linked to a pop-up window with criteria for peripheral smear review and instructions on the approval policy. Outpatient orders required clinicians to request pathology approval. This implementation reduced total number of PBS orders by 42.5% with no adverse effect on patient management. Empowering pathologists and clinicians with guidelines on PBS review orders is a beneficial educational exercise of resource utilisation. Discussion with physicians regarding clinical indications reduces non-contributory PBS reviews, provides guidance to appropriate testing, and aptly allocates pathologist and laboratory staff time and resources.

Cytotoxic and Genotoxic Evaluation of the Aqueous and Hydroalcoholic Leaf and Bark Extracts of Crataegus oxyacantha in Murine Model

Crataegus oxyacantha has been mainly used in traditional medicine for the treatment of cardiovascular diseases. However, its safety profile has not been fully established, since only the genotoxic effects of C. oxyacantha fruit have been described. Therefore, the objective of this work was evaluating the cytotoxicity and genotoxicity of the aqueous and hydroalcoholic leaf and bark extracts of C. oxyacantha by means of the micronucleus test in a murine model. Doses of 2000, 1000, and 500 mg/kg of both extracts were administered orally for 5 days in mice of the Balb-C strain. Peripheral blood smears were performed at 0, 24, 48, 72, and 96 h after each administration. The number of polychromatic erythrocytes (PCEs), micronucleated polychromatic erythrocytes (MNPCEs), and micronucleated erythrocytes (MNEs) was determined at the different sampling times. Our results showed that the leaf and bark of C. oxyacantha increase the number of MNEs at the 2000 mg/kg dose, and only the aqueous leaf extract decreases the number of PCEs at the same dose. Therefore, the aqueous and hydroalcoholic leaf and bark extracts of C. oxyacantha showed genotoxic effects, and only the aqueous leaf extract exhibited cytotoxic effects.

Clinical and immunohistochemical findings of splenic mast cell tumour in a cat: A case report

A 6-year-old, spayed, female, domestic shorthair cat presented with a 4-month history of chronic intermittent vomiting and anorexia. The haematologic results indicated moderate anaemia and a circulating mast cell population. The abdominal radiography revealed a markedly enlarged spleen. The cytological analysis of the spleen showed a uniform population of mast cells, and a diagnosis of systemic mastocytosis (splenic mast cell tumour with mastocytaemia) was made. This diagnosis was subsequently confirmed by the histopathological examination of the spleen. The immunohistochemistry for KIT showed KIT pattern II (focal cytoplasmic expression). A splenectomy and chemotherapy with vinblastine and prednisolone resulted in remission of the anaemia and other clinical signs. This case report highlights the importance of cytological evaluations of peripheral blood smears and/or aspirates of enlarged spleens for diagnosing splenic mast cell tumours and for quickly initiating the appropriate treatment.

Performance of the CellaVision DC-1 digital cell imaging analyser for differential counting and morphological classification of blood cell

AimsRecently, a new automated digital cell imaging analyser (Sysmex CellaVision DC-1), intended for use in low-volume and small satellite laboratories, has become available. The purpose of this study was to compare the performance of the DC-1 with the Sysmex DI-60 system and the gold standard, manual microscopy.MethodsWhite blood cell (WBC) differential counts in 100 normal and 100 abnormal peripheral blood smears were compared between the DC-1, the DI-60 and manual microscopy to establish accuracy, within-run imprecision, clinical sensitivity and specificity. Moreover, the agreement between precharacterisation and postcharacterisation of red blood cell (RBC) morphological abnormalities was determined for the DC-1.ResultsWBC preclassification and postclassification results of the DC-1 showed good correlation compared with DI-60 results and manual microscopy. In addition, the within-run SD of the DC-1 was below 1 for all five major WBC classes, indicating good reproducibility. Clinical sensitivity and specificity were, respectively, 96.7%/95.9% compared with the DI-60% and 96.6%/95.3% compared with manual microscopy. The overall agreement on RBC morphology between the precharacterisation and postcharacterisation results ranged from 49% (poikilocytosis) to 100% (hypochromasia, microcytosis and macrocytosis).ConclusionsThe DC-1 has proven to be an accurate digital cell imaging system for differential counting and morphological classification of WBCs and RBCs in peripheral blood smears. It is a compact and easily operated instrument that can offer low-volume and small satellite laboratories the possibilities of readily available blood cell analysis that can be stored and retrieved for consultation with remote locations.

High altitude epidemic malaria in Bamian province, central Afghanistan

We report an epidemic of Plasmodium falciparum malaria in the remote valley of Bamian [altitude 2250 m-2400 m] in the central highl and s of Afghanistan. A team of malaria experts from the World Health Organization and HealthNet International carried out the investigation. A total of 215 peripheral blood smears were obtained and 63 cases of malaria [90.5% P. falciparum, the remainder P. vivax] were confirmed. The study revealed that areas vulnerable to malaria in Afghanistan are more widespread than previously recognized. The area had been malaria-free until recently, when the disease appears to have been introduced as a consequence of protracted conflict and resultant population movement, and transmitted locally during the short summer months. The outbreak led to severe morbidity and high mortality in a province having only a few poorly-provisioned health care facilities

Deep Optical Blood Analysis: COVID-19 Detection as a Case Study in Next Generation Blood Screening

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.