Assessment of inter-operator variability in peripheral monocyte subset gating strategy using flow cytometry in patients with suspected acute stroke.

BACKGROUND: Innovative tools to reliably identify patients with acute stroke are needed. Peripheral monocyte subsets, that is, classical-Mon1, intermediate-Mon2, and non-classical-Mon3, with their activation marker expression analyzed using flow-cytometry (FCM) could be interesting cell biomarker candidates. AIM: To assess the inter-operator variability in a new peripheral monocyte subset gating strategy using FCM in patients with suspected acute stroke. METHODS: In BOOST-study ("Biomarkers-algOrithm-for-strOke-diagnoSis-and Treatment-resistance-prediction," NCT04726839), patients ≥18 years with symptoms suggesting acute stroke within the last 24 h were included. Blood was collected upon admission to emergency unit. FCM analysis was performed using the FACS-CANTO-II® flow-cytometer and Flow-Jo™-software. Analyzed markers were CD45/CD91/CD14/CD16 (monocyte backbone) and CD62L/CD11b/HLA-DR/CD86/CCR2/ICAM-1/CX3CR1/TF (activation markers). Inter-operator agreement (starting from raw-data files) was quantified by the measure distribution and, for each patient, the coefficient of variation (CV). RESULTS: Three operators analyzed 20 patient blood samples. Median inter-operator CVs were below the pre-specified tolerance limits (10% [for Mon1 counts], 20% [Mon2, Mon3 counts], 15% [activation marker median-fluorescence-intensities]). We observed a slight, but systematic, inter-operator effect. Overall, absolute inter-operator differences in fractions of monocyte subsets were <0.03. CONCLUSION: Our gating strategy allowed monocyte subset gating with an acceptable inter-operator variability. Although low, the inter-operator effect should be considered in monocyte data analysis of BOOST-patients.

Effects of circadian variation, lifestyle and environment on hematological parameters: A narrative review.

The complete blood count (CBC) is the most widely prescribed laboratory test. It plays a key role in screening, diagnosing, and monitoring a variety of medical disorders. Preanalytical and analytical variables are responsible for more than 50% of laboratory errors that may lead to spurious CBC results. The effects of blood sampling, transport, storage, and analytical errors on hematological parameters have been well described. Circadian variation and changes in lifestyle and environment can also affect blood cells. It has been extensively studied in the past, but highly variable methodology and the presence of confounding factors have provided scattered and inconsistent results. We have investigated the literature to define the impact of circadian variation, modification of the sleep-wake cycle, acute and chronic exercise, eating habits, alcohol, tobacco, drugs of abuse, high-altitude, heat/cold exposure, and air pollution on CBC results. The affected cell type along with the intensity and duration of changes are detailed for each condition. We aim at providing a comprehensive overview of which situations may induce clinically significant changes and have to be taken into account by healthcare professionals before considering a hematological parameter as pathological and requesting complementary tests.

How I investigate basophilia in daily practice.

Basophilia is a rare disorder of the complete blood count (CBC) and its management in daily practice remains unclear. Two main factors explain this situation. On the one hand, the reliability of the basophil count is insufficient, whether it is performed by a microscopic slide examination or by a hematology analyser. On the other hand, our knowledge of conditions associated with basophilia is largely based on few case reports and on reviews that refer to older reviews. The association between basophilia and myeloid neoplasm, especially chronic myeloid neoplasm, is well established. Conversely, there are conflicting data on some benign medical conditions and it remains unclear where basophilia may be present. In this review, we have investigated the medical literature to define which medical conditions can lead to basophilia and which cannot, and we propose a practical approach to manage basophilia divided into 3 steps. First, we have to check the real existence of the basophilia by getting rid of spurious basophilia. Then, we have to look for symptoms that suggest reactive basophilia and for clue of a neoplastic cause. Finally, in case of suspicion of a myeloid neoplasm or persistence of the basophilia in the absence of a reactive cause, we have to decide which examinations need to be prescribed to confirm a neoplastic basophilia.

Clinically silent indolent T-cell leukemia.

The clinically silent, symptom-free T-cell prolymphocytic leukemia case that we report here confirms the major interest of the analysis of the blood smear as usual care of any emergent lymphocytosis. It also brings out the issue of the monitoring and follow-up of this uncommon presentation.

A composite lymphoma combining a Hodgkin lymphoma and a marginal zone lymphoma transformed into a diffuse large B-cell lymphoma.

Composite lymphoma is defined as the occurrence of two or more distinct lymphoma types in a single anatomic site. We report a case of Richter syndrome with both Hodgkin lymphoma and non-Hodgkin lymphoma in the bone marrow. This diagnostic was suspected because of discrepancies between histological and cytological results.

First successful use of eltrombopag before surgery in a child with MYH9-related thrombocytopenia.

MYH9-related disease (MYH9-RD) is one of the most frequent autosomal-dominant forms of inherited macrothrombocytopenias and is caused by mutations in MYH9 (nonmuscle myosin IIA), the gene coding for the heavy chain of the nonmuscle myosin IIA. Affected individuals can present with isolated thrombocytopenia, and whereas only some will have bleeding events requiring intervention, nearly all will require the use of prophylactic platelet transfusions before surgery. Here we report the first prophylactic use of eltrombopag before surgery in a child with MYH9-RD. Our patient was a 13-year-old girl with an MYH9 S96L missense mutation who required a tympanoplasty due to chronic otitis media. Pretreatment microscopic platelet count was 10 × 10(9)/L. The child was treated with eltrombopag starting 4 weeks before her planned surgery. On the day of surgery her platelet count was 70 × 10(9)/L. She required no platelet transfusions and no abnormal bleeding was reported either during surgery or postoperatively. Given these results, the first reported in a child, we suggest that the use of this thrombopoietic agent should be further evaluated as a useful presurgical prophylactic option in this hereditary thrombocytopenia, thus avoiding the use of platelet transfusions and their associated risks, which include alloimmunization and the transmission of infectious agents.