Comprehensive Morphological Assessment of Cord Blood: Normal Values and the Prevalence of Morphologically Aberrant Leukocytes.

INTRODUCTION: Cord blood (CB) is becoming a valuable source for stem cells utilized in a variety of cell therapy applications, as well as for newborn diagnostics. Some parameters of the CB cellular components can be provided by automated analyzers, while others, such as immature or aberrant cells, require blood film morphological assessment. The objectives of the study were to establish normal CB morphology and to determine the prevalence of morphologically aberrant leukocytes in CB. METHODS: We performed a comprehensive morphological analysis of 100 CB samples taken from healthy term and appropriate-for-gestational-age neonates born to healthy mothers, preterm neonates, neonates of diabetic mothers, and small-for-gestational-age neonates. Blood counts were assessed, and manual morphological analyses were performed by laboratory specialists. RESULTS: The manual differential count of normal CB samples established the following values: 47.8 ± 10.7% neutrophils, 31.2 ± 9.8% lymphocytes, 10.0 ± 4.0% monocytes, and 3.0 ± 2.5% eosinophils, with no significant sex-related differences. Blasts were observed in 44/100 samples with an average of 0.5 ± 0.7% per sample, and only a minor left shift was observed. There were significant populations of large granular lymphocytes (19.1 ± 10.6% of the total lymphocytes) and morphologically aberrant lymphocytes (12.4 ± 5.4% of the total lymphocytes) in the samples, irrespective of neonatal status. The differentials of preterm CB samples differ significantly from normal term CB samples, including the reverse of neutrophils/lymphocytes ratio, and the lack of basophils. CONCLUSIONS: Normal values and unique morphological features in the CB of neonates are described. The abundant morphologically aberrant lymphocytes in CB may represent an immature state of the immune system at birth.

Low FXIII activity levels in intensive care unit hospitalized COVID-19 patients.

BACKGROUND: COVID-19 infection is associated with a hypercoagulable state. Severe COVID-19 patients present with high plasma fibrinogen levels, continuous deposition of fibrin and the presence of microthrombi in their lungs, accompanied by significant fibrinolysis, resulting in high D-dimer levels. Due to the role of FXIII in fibrin crosslinking and clot stabilization, we analyzed its activity levels and dynamics in COVID-19 patients hospitalized in the intensive care unit (ICU). METHODS: FXIII levels were measured in thirty four COVID-19 patients hospitalized in the ICU and in fourteen non-severe COVID-19 patients. FVIII levels were measured for comparison. Laboratory data and clinical variables were recorded. RESULTS: The average FXIII activity level in 34 ICU hospitalized COVID-19 patients was 69.9±33 %, significantly lower compared to an average of 120±20.9 % FXIII activity in 14 non-severe COVID-19 patients. FXIII activity levels were below the low normal value (< 79 % FXIII activity) in 74 % of the ICU hospitalized COVID-19 patients. In contrast, high FVIII activity was measured among all severe COVID-19 patients. Consecutive measurements, performed in fourteen ICU hospitalized COVID-19 patients, pointed to a significant decrease in FXIII activity from the average of 85.7±28.2 %, (which is in the normal range), to an average of 68.0±20.4 %, below the low normal range, within 6.4±3.4 days of ICU hospitalization. Liver functions did not differentiate between patients with low and normal FXIII activity. No inhibitor to FXIII activity was found in the plasma of severe COVID-19 patients. Levels of FXIII-A antigen correlated with FXIII activity, and were low in severe COVID-19 patients. CONCLUSIONS: Low FXIII activity levels were found in COVID-19 patients hospitalized in the ICU, with gradual decline during their hospitalization. A mechanism of consumption may account for the low FXIII activity in these patients.

The Clinical Significance of Circulating Lymphocytes Morphology in Diffuse Large B-Cell Lymphoma As Determined by a Novel, Highly Sensitive Microscopy.

Chimeric Antigen Receptor T-cell (CAR T) therapy has become the preferable treatment in relapsed/refractory diffuse large B-cell lymphomas (DLBCL) patients. Detection of CAR Ts in peripheral blood smear (PBS) is challenging due to insufficient data regarding their morphology and low sensitivity. The morphological evolution of CAR Ts along their production process, and in patients, was established by Full-Field Morphology (FFM), a novel digital microscopy approach that provides highly sensitive PBS analysis. At day 8 of production, 42.7 ± 10.8% of the CAR T transduced cells exhibited activated morphology compared with 9.3 ± 3.8% in untransduced cells. Moreover, engagement of transduced CAR Ts with target cells resulted in further morphological transformation into activated morphology (83 ± 5.6% of the cells). In patients, the average number of day 5 CAR Ts, and their sustained presence, were significantly higher in patients obtaining complete response. A high number of activated morphology CAR Ts at day 14 was associated with prolonged cytokine release storm. Overall, CAR Ts exhibited heterogeneous morphology, with the activated morphology attributed predominantly to transduced cells following engagement with target cells. Post-transfusion CAR T detection was associated with increased complete responses. FFM CAR T surveillance in PBS may serve as a simple inexpensive method to provide clinically relevant insights into this treatment modality.

Remote Digital Microscopy Improves Hematology Laboratory Workflow by Reducing Peripheral Blood Smear Analysis Turnaround Time.

BACKGROUND: The demand for morphological diagnosis by peripheral blood smear (PBS) analysis with clearly defined turnaround times (TAT), coupled with a shortage of morphologists and increasing cost containment, is driving digitalization to the forefront of laboratory workflow. Labor-intensive manual PBS review affects weekend workflow with limited staff availability. The impact of remote analysis of PBS on the performance of hematology laboratories has not yet been assessed. OBJECTIVES: Following implementation of fully remote digital microscopy within our laboratory, we measured its impact on morphology workflow efficiency, TAT, and hours saved per month. METHODS: A retrospective study of the effects of remote PBS analysis on the morphology workflow in a tertiary medical center using the Scopio Labs X100 Full-Field PBS system was conducted. 10,704 PBS samples were analyzed pre-and post -implementation, over a 5-month period. Overall PBS workload, and average TAT of PBS samples over weekends and the first two weekdays were collected and evaluated. RESULTS: Remote weekend viewing resulted in a 15.8% reduction in the overall morphology TAT of the laboratory (p <0.03) over a 5-month period, despite similar overall workload. PBS analysis TAT on Fridays was reduced by 41.4% (p <0. 006), and by 59.1% on the first weekday (p <0.02). The additional hours incurred over the weekend were offset against a reduced need for double weekday shifts resulting in approximately 12.76 work hours saved per month. Internet links to clinically relevant cases are provided. CONCLUSION: The Scopio Labs Full-Field X100 PBS system with remote analysis capacity significantly reduced PBS TAT and improved the morphology workflow of the hematology laboratory. PBSs with significant clinical findings are now available for remote viewing by on-call clinicians located outside the medical center perimeter. Remote PBS viewing, coupled with the overall monthly cost savings, merit consideration for the implementation of full digitalization for remote PBS review.

Evaluation of Scopio Labs X100 Full Field PBS: The first high-resolution full field viewing of peripheral blood specimens combined with artificial intelligence-based morphological analysis.

BACKGROUND: Current digital cell imaging systems perform peripheral blood smear (PBS) analysis in limited regions of the PBS and require the support of manual microscopy without achieving full digital microscopy. We report a multicenter study that validated the Scopio Labs X100 Full Field PBS, a novel digital imaging system that utilizes a full field view approach for cell recognition and classification, in a decision support system mode. METHODS: We analyzed 335 normal and 310 abnormal PBS from patients with various clinical conditions and compared the performance of Scopio's Full Field PBS as the test method, with manual PBS analysis as the reference method. Deming regression analysis was utilized for comparisons of WBC and platelet estimates. Measurements of WBC and platelet estimation accuracy along with the agreement on RBC morphology evaluation were performed. Reproducibility and repeatability (R&R) of the system were also evaluated. RESULTS: Scopio's Full Field PBS WBC accuracy was evaluated with an efficiency of 96.29%, sensitivity of 87.86%, and specificity of 97.62%. The agreement between the test and reference method for RBC morphology reached 99.77%, and the accuracy for platelet estimation resulted in an efficiency of 94.89%, sensitivity of 90.00%, and specificity of 96.28%, with successful R&R tests. The system enabled a comprehensive review of full field PBS as shown in representative samples. CONCLUSIONS: Scopio's Full Field PBS showed a high degree of correlation of all tested parameters with manual microscopy. The novel full field view of specimens facilitates the long-expected disengagement between the digital application and the manual microscope.