Plasmablastic transformation of a double hit follicular lymphoma: An emerging entity.

ABSTRACT: Plasmablastic transformation of follicular lymphoma is very rare and has been reported in only 5 cases till date. We report a case of simultaneous identification of extranodal, soft tissue plasmablastic lymphoma in the ankle and bone marrow involvement by follicular lymphoma. This unusual case presentation is a challenge for the treating physician with the patient becoming resistant to chemotherapy and succumbing to the disease within a few months of diagnosis. These cases are known to have an aggressive clinical course with very poor prognosis and survival rate of less than 6 months. This report broadens the spectrum of morphological transformation of follicular lymphoma and it may represent a new category of high-grade transformation of follicular lymphoma.

Rapid Detection of Plasmodium vivax by the Hematology Analyzer for Population Screening.

In India, where malaria is endemic, the prompt and accurate detection of infections is crucial for disease management and vector control. Our study aimed to evaluate the "iRBC" flag, a novel parameter developed for routine hematology analyzers, for its sensitivity and specificity in detecting Plasmodium vivax (P. vivax) infections. We used residual blood samples from patients with suspected malaria and compared the iRBC flag results with microscopy, which serves as the gold standard. Additionally, we compared the results with rapid immuno-chromatographic tests (RDTs) commonly used in the field. Our study included 575 samples, of which 187 were positive for P. vivax. The iRBC flag demonstrated a high sensitivity of 88.7% and 86.1% on the XN and XN-L hematology analyzers, respectively, and a clinical specificity of 100% on both analyzers. Furthermore, the scattergram derived from each positive dataset exhibited distinct patterns, which facilitated rapid confirmation by laboratory specialists. Notably, the iRBC flag remained effective even in the presence of interfering conditions. Overall, our results indicate that the iRBC flag is a reliable and rapid screening tool for identifying P. vivax in routine blood testing. Our findings have significant implications for malaria detection and control in endemic regions like India.

Performance Evaluation of Fully Automated 5-Part Differential Hematology Analyzer ELite 580 (ERBA).

The ELite 580 (Erba Lachema) Hematology Analyzer is a newly developed automated hematology analyzer that uses impedance and flow cytometry technologies. The analyzer provides 25 reportable parameters including a 5-part differential and additionally 4 research parameters with a throughput of 80 samples per hour. To evaluate the performance of the fully automated 5-part differential hematology analyzer ELite 580 as per the international standards and correlate the results with a reference analyser, LH 780 from Beckman Coulter. We evaluated the analyzer's performance by determining background noise, carryover, precision, linearity and correlation of complete blood cell count parameters between the ELite 580 from Erba Lachema and the LH 780 from Beckman Coulter at a tertiary care hospital, according to the Clinical and Laboratory Standards Institute guidelines. The ELite 580 showed minimal background noise and carryover, high precision, accuracy and linearity over a wide analytical range for white blood cells, hemoglobin, red blood cells, hematocrit, and platelet parameters. Correlation between the ELite 580 and the LH 780 was good for all parameters (R2 > 0.90) except for mean corpuscular hemoglobin concentration (MCHC) and the basophil count. ELite 580 Hematology Analyzer with its precise and accurate results and ease of operation is suitable for clinical use in medium to large sized hematology laboratories. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12288-021-01423-y.

Immune Modulation and Cytomegalovirus Reactivation in Sepsis-induced Immunosuppression: A Pilot Study.

INTRODUCTION: Sepsis is a life-threatening condition caused due to dysregulated immune response to an infection and progressive immunosuppression. Reactivation of cytomegalovirus (CMV) occurs frequently in sepsis and is found associated with adverse outcomes. The study objective was to evaluate the association between incidence of CMV reactivation and immune alteration in sepsis-induced immunosuppression in patients with prolonged sepsis. PATIENTS AND METHODS: Patients admitted to intensive care unit (ICU), with severe sepsis and CMV immunoglobulin G (IgG) seropositivity, were prospectively enrolled. Other manifest immune suppression causes were excluded. Samples were collected on enrolment and further once a week until day 21 or death/discharge. CMV viral load was quantified using qPCR. Lymphocyte subset analysis (CD3+, CD4+, CD8+, CD19+, CD16+/CD56+, and CD25+CD127- regulatory T cells), human leukocyte antigen-DR isotype (HLA-DR) expression on monocytes, programmed death-1 (PD-1) expression on T lymphocytes, and proinflammatory (interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ)), anti-inflammatory cytokines levels (IL-2, IL-4, and IL-10) were analyzed by flow cytometry as markers for immunosuppression. RESULTS: A total of 25 CMV IgG-positive patients and 11 healthy controls were included. CMV reactivation occurred in 20 patients. Patients with CMV reactivation had T-cell lymphopenia. PD-1 expression on CD4+ and CD8+ T cells was markedly elevated (p <0.02) in CMV-reactivated patients compared to nonreactivated patients. HLA-DR expression was significantly low on monocytes in all septic patients (p <0.01) compared to healthy controls. IL-6 levels showed elevation at day 7, whereas IL-10 was found to be significantly higher from day 0 in CMV-reactivated group. CONCLUSION: Our study concluded that immune suppression markers and cytokine levels in patients with severe sepsis were found to be significantly associated with the incidence of CMV reactivation. HOW TO CITE THIS ARTICLE: Lambe G, Mansukhani D, Khodaiji S, Shetty A, Rodrigues C, Kapadia F. Immune Modulation and Cytomegalovirus Reactivation in Sepsis-induced Immunosuppression: A Pilot Study. Indian J Crit Care Med 2022;26(1):53-61.

COVID-19-Associated Bone Marrow Necrosis-A Case Report.

We report, herein, a rare case of vertebral bone marrow necrosis in a patient at 1-month post-novel coronavirus disease 2019 (COVID-19) pneumonia complicated with disseminated intravascular coagulation (DIC). The commonly observed radiological features on the imaging modalities like computed tomography (CT), magnetic resonance imaging (MRI), and 18-F fluorodeoxyglucose positron emission tomography (FDG PET) have been discussed here followed by a brief discussion on the role of in-phase and opposed-phase imaging in differentiating the disease from malignant infiltrative pathologies. Histopathological findings on bone marrow smear that confirm the diagnosis have also been illustrated.

Evaluation of automated malaria diagnosis using the Sysmex XN-30 analyser in a clinical setting.

BACKGROUND: Early and accurate diagnosis of malaria is a critical aspect of efforts to control the disease, and several diagnostic tools are available. Microscopic assessment of a peripheral blood smear enables direct visualization of parasites in infected red blood cells and is the clinical diagnostic gold standard. However, it is subjective and requires a high level of skill. Numerous indirect detection methods are in use, but are not ideal since surrogate markers of infection are measured. This study describes the first clinical performance evaluation of the automated Sysmex XN-30 analyser, which utilizes fluorescence flow cytometry to directly detect and quantitate parasite-infected red blood cells. RESULTS: Residual EDTA blood samples from suspected malaria cases referred for routine diagnosis were analysed on the XN-30. Parasitaemia was reported as a percentage, as well as absolute numbers of infected red blood cells, and scattergrams provided a visual image of the parasitized red blood cell clusters. The results reported by the XN-30 correlated with microscopy and the analyser demonstrated 100% sensitivity and specificity. Measurements were reproducible and storage of samples at room temperature did not affect the parameters. Several Plasmodium species were detected, including Plasmodium falciparum, Plasmodium vivax and Plasmodium ovale. The XN-30 also identified the transmissible gametocytes as a separate cluster on the scattergrams. Abnormal red blood cell indices (low haemoglobin and raised reticulocyte counts), haemoglobinopathies and thrombocytopenia did not interfere with the detection of parasites. The XN-30 also generated a concurrent full blood count for each sample. CONCLUSIONS: The novel technology of the Sysmex XN-30 provides a robust, rapid, automated and accurate platform for diagnosing malaria in a clinical setting. The objective enumeration of red blood cells infected with Plasmodium species makes it suitable for global use and allows monitoring of the parasite load once therapy has been initiated, thereby providing an early marker of drug resistance. The automated generation of a full blood count for each sample provides an opportunity for detecting unsuspected cases. Asymptomatic carriers can also be identified, which will be useful in blood transfusion centres, and will enable treatment of these individuals to prevent the spread of the disease.

Age-related reference intervals for immunoglobulin levels and lymphocyte subsets in Indian children.

BACKGROUND: In children, innate and adaptive immunity varies with age, disease status, and ethnicity, reflected by lymphocyte subsets and serum immunoglobulin (Ig) levels. The paucity of such data from the Indian subcontinent necessitated this study. AIMS: This study aims to determine reference ranges of Ig and lymphocyte subsets in Indian children from birth to 5 years. SETTINGS AND DESIGN: Neonates, infants, and children from a tertiary care hospital were selected and categorized into 5 groups from cord blood/newborn to 5 years. MATERIALS AND METHODS: Samples were taken from cord blood and healthy children up to 5 years of age. Complete blood counts, serum Ig levels (by turbidimetry), and lymphocyte subsets (by flow cytometry) were studied, and reference ranges calculated. RESULTS: Four hundred and three samples were analyzed; 53 from cord blood and 350 from children 1 month to 5 years. High IgG levels were noted at birth, which decreased in the first 6 months followed by a rise thereafter. IgM remained low in infancy and peaked at 13-36 months. IgA levels were very low at birth but increased with age. CD4 counts were high in cord blood till 3 years of age and then declined. CD8 and CD19 counts remained steady till 5 years of age. CD56 increased after the age of 2 years. CONCLUSIONS: While our data correlated well with published literature, notable differences were higher IgM levels seen in 1-3 years' age group and higher natural killer cells through all age groups in our study. Our results provide the largest database of its kind from our country.

Sehgal index: A new index and its comparison with other complete blood count-based indices for screening of beta thalassemia trait in a tertiary care hospital.

INTRODUCTION: Beta thalassemia trait (BTT) must be differentiated from iron deficiency anemia to avoid unnecessary iron therapy and for the prevention of thalassemia major by genetic counseling. In a tertiary care hospital, it is vital that the screening tool is not only sensitive but also specific so as to be cost effective and save time. AIM: The aim of this study was to evaluate the new Sehgal index and compare it to existing complete blood count-based indices for the best combination of sensitivity and specificity to predict BTT. MATERIALS AND METHODS: Study was done in 2 phases - Phase 1: A retrospective analysis of 1022 consecutive high-performance liquid chromatography (HPLC) cases from July 2008 to June 2011. Phase 2: A prospective analysis of 973 consecutive HPLC cases from July 1, 2011 to June 10, 2013 was done to confirm the results of Phase 1 and the applicability of the new Sehgal index. RESULTS: Prevalence of BTT was 28.8% (294/1022) and 25.39% (247/973) in Phase 1 and Phase 2, respectively. Receiver operating characteristic-area under the curve and Youden index was highest for new Sehgal index, followed by Mentzers index <14. The prospective study shows results similar to those in Phase 1 confirming the superiority of the above two indices. CONCLUSION: Sehgal index and Mentzers index <14 showed the best combination of sensitivity and specificity in predicting BTT. The best indices or combination can be used as a "validated flag rule" in the analyzer middleware program in a hospital for identifying suspected cases of BTT.

Unusual immunophenotype of T-cell large granular lymphocytic leukemia: report of two cases.

Large granular lymphocytes (LGL) leukemias are commonly of the T-cell or NK-cell type. T-cell LGL leukemia is typically a disorder of mature CD3, CD8 and T-cell receptor TCR (TCR - T cell receptor)-αß positive cytotoxic T-cells. Rare variants include TCRγδ+ variants and CD4 + TCRαß+ cases. We report a case of each of these rare variants. An 83-year-old female presented with anemia and lymphocytosis with LGLs on peripheral smear. Six-color multiparametric flowcytometric analysis showed expression of CD3, heterogeneous CD7, dim CD2 and TCRγδ and lacked expression of CD5, TCRαß, CD56, CD4 and CD8. A final diagnosis of TCRγδ+ T-cell LGL leukemia was made. Differentiation between TCRγδ+ T-cell LGL leukemia and other γδ+ T-cell malignancies is of utmost importance due to the indolent nature of the former as compared to the highly aggressive behavior of the latter. An 85-year-old male diagnosed with liposarcoma was identified to have lymphocytosis during preoperative evaluation. Peripheral smear showed presence of LGLs. Flowcytometric immunophenotyping showed expression of TCRαß, CD3, CD2, CD5, CD4, dim CD8, CD56 with aberrant loss of CD7 expression. Vß repertoire analysis by flowcytometry showed 97% cells with Vß14 clonality. A final diagnosis of TCRαß+ CD4 + T-cell LGL leukemia was made. CD4 + T-cell large granular lymphocytic leukemias have an indolent, less aggressive course when compared to their CD8 + counterparts and are not necessarily associated with cytopenias. However, their association with secondary neoplasia (29% of the cases) warrants a high degree of suspicion in the diagnosis as also noted in the index case. Use of a wide panel of antibodies and newer modalities such as Vß repertoire analysis helps in accurate subtyping of LGL leukemia.

Comparison of platelet counts by sysmex XE 2100 and LH-750 with the international flow reference method in thrombocytopenic patients.

BACKGROUND: There are several methods for counting platelets, of which the international flow reference method (IRM) is considered to be the gold standard. We compared the platelet count given by this method to the count given by automated analyzers using other methods, such as optical fluorescence and impedance. AIMS: The aim of this study is to compare the platelet counts obtained by Sysmex XE 2100 by Impedance (Sysmex-I), optical florescence (Sysmex-O) and reported (Sysmex-R) based on the switching algorithm and LH-750 by Impedance (LH-750) with the IRM in thrombocytopenic blood samples. To calculate the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of various technologies at the clinically relevant transfusion thresholds of 10 × 10 9 /l and 20 × 10 9 /l. MATERIALS AND METHODS: A total of 118 blood samples with platelet count of <50 × 10 9 /l were selected for the study. Platelet counts of all samples were analyzed by all methods using the Sysmex analyzer, LH-750 and IRM in parallel within 6 h of collection. STATISTICAL ANALYSIS USED: Pearson correlation, bland Altman analysis, sensitivity and specificity, PPV and NPV. RESULTS AND CONCLUSIONS: Sysmex-R had the least Bias and 95% limits of agreement (95%LA) range and thus correlated best with IRM values. LH-750 had a higher Bias compared to Sysmex-O and Sysmex-R, but a strikingly similar 95% LA ensures similar results in all three methods. In fact, in the oncology subset, it had the narrowest 95% LA, which made it the best performer in this subgroup. Of the three Sysmex results, Sysmex-I had the highest bias, widest 95% LA and highest potential risk of over transfusion. Hence, Sysmex-R and LH-750 were found to be reliable tools for estimation of platelet count in thrombocytopenic patients.

Spontaneously developing autoantibody to factor VIII in an elderly woman: diagnostic and therapeutic challenges.

An elderly woman with a continuously bleeding small wound was investigated for the presence of antibodies to FVIII using activated partial time-based screening and confirmatory tests. A late acting coagulation factor inhibitor was detected. The same was characterised to be a low titre antibody against FVIII (5.2 Bethesda units). Cryoprecipitate infusions, corticosteroids and topical desmopressin were unsuccessful in controlling the bleeding. Addition of cyclophosphamide brought about stoppage of bleeding and disappearance of the autoantibody.