Reversal of Glanzmann thrombasthenia platelet phenotype after imatinib treatment in a pediatric chronic myeloid leukemia patient.

Chronic Myelogenous Leukemia (CML) is a myeloproliferative neoplasm characterized by proliferation of Philadelphia positive clonal pluripotent hematopoietic cells. Bleeding is a rare presentation of CML that can occur due to platelet dysfunction. Both pre-treatment and post-treatment platelet function abnormalities in CML have been described in the literature. We describe a rare case of childhood CML who presented with mucocutateous bleeding manifestations. On laboratory workup, a Glanzmann Thrombasthenia (GT) like platelet phenotype was demonstrated along with confirmation of diagnosis of CML in chronic phase. The acquired nature of platelet function defect was confirmed by demonstrating recovery of platelet antigens glycoprotein IIb/IIIa after achieving complete hematological response with Imatinib. Due to presenting complaint of bleeding diathesis and absence of hepatosplenomegaly, the case was undiagnosed for CML until the patient reported to us. Careful evaluation of complete blood counts, peripheral blood picture and detailed laboratory workup was the window to proper diagnosis and treatment in this case.

Study of Bone Marrow Lymphocyte Subset in Acute Myeloid Leukemia.

Introduction Acute myeloid leukemia (AML) is a heterogenous disorder consisting of clonal expansion of myeloblasts. Tumor immunity plays an important part in the pathobiology of AML. Understanding the components of tumor immunity is important for understanding tumor pathogenesis and the principles of immunotherapy. Methods We studied 41 patients with AML, for total lymphocyte, CD4 positive helper T cells, CD8 positive cytotoxic T cells, and CD16/56 positive natural killer (NK) cells proportion. Quantification was done on bone marrow aspirate sample by flowcytometry. Whenever available, post induction bone marrow was also analyzed for the lymphocyte subset. Results No significant difference was noted in the percentage of blasts among the three risk categories: favorable, intermediate, and adverse. However, there was significant difference in the total lymphocyte among the risk stratification groups, being highest in the favorable group and lowest in the adverse group. CD8 positive cytotoxic T cells were significantly less in Acute Promyelocytic Leukemia (APML) cases ( p = 0.001). Total lymphocytes were, however, more numerous in APML ( p = 0.005). NK cell proportion was not significantly different between APML and non-APML patients. On completion of induction chemotherapy, bone marrow samples for 12 patients could be processed for lymphocyte subset. On comparing the baseline against the post induction bone marrow, it was observed that there was significant increment in the proportion of CD4 positive T lymphocytes ( p = 0.046). Conclusion There is a difference in lymphocyte subset amongst patients with AML. Larger studies including functional aspects are needed to better define the role of lymphocytes in disease pathogenesis.

Evaluation of utility of immunohistochemistry markers as a tool for objective diagnosis of low-grade myelodysplastic syndrome in routine reporting: Prospective observational study.

Purpose: Diagnosis of myelodysplastic syndrome (MDS) primarily relies on the detection of morphological dysplasia in bone marrow. It is subjective and many studies have reported lack of interobserver agreement in reporting. Biopsy is preferred specimen for megakaryocyte assessment. We studied 43 bone marrow biopsies from 40 suspected MDS patient having persistent undiagnosed cytopenia. Utility of immunohistochemistry (IHC) with CD61 and p53 in detecting low-grade MDS was analyzed over routine morphology. Method and Results: Total number of megakaryocytes and number of dysplastic megakaryocytes seen on CD61 IHC was significantly higher than that on H and E stain (P value < 0.05) Out of total 43 biopsies, 13 [30.2%] cases showed dysplastic megakaryocytes that were confirmed by interobserver agreement after IHC. From 30 cases with no significant dysplasia on morphology, 21/43 [48.8%] cases showed >10% dysplastic megakaryocytes on CD61 (P value 0.0001). Nine cases showed no significant dysmegakaryopoiesis with either H and E or CD61 IHC. Fourteen cases could meet higher cut off (30%) of dysmegakaryopoiesis with CD 61 IHC. Out of total 34 cases showing significant dysplasia 7 cases (20.6%) showed positivity for p53 on IHC, which is little less than that reported in low-grade MDS. Conclusion: CD61 IHC is helpful in making correct diagnosis of MDS in cases with minimal dysplasia and should be performed before excluding possibility of MDS on morphology in a patient with undiagnosed cytopenia. IHC is cost effective tool for MDS diagnosis in developing world where access to extensive flow cytometery and molecular testing is limited.

A case of splenomegaly with RK-39 positivity: A diagnostic pitfall.

Long-standing moderate to marked splenomegaly suggests several differential diagnoses, both haematological and infectious, particularly leishmaniasis and malaria in endemic areas. Non-infectious causes may be missed in these regions, especially if pitfalls of serological testing are not considered. Careful patient evaluation is necessary to arrive at the correct diagnosis. We report a case of a young male whose hereditary spherocytosis was initially missed because of RK-39 positivity, splenomegaly and the fact that he hailed from an endemic region.

The utility of a single tube 10-color flow cytometry for quantitative and qualitative analysis in myelodysplastic syndrome- a pilot study.

INTRODUCTION: Assessment of myelodysplasia (MDS) by flow cytometry (FCM) includes elaborate panels, and interpretation is observer-dependent. This study evaluates single tube 10-color FCM in a test cohort of clinically suspected MDS patients. METHODS: We analyzed fifty-six bone marrow (BM) samples from clinically suspected MDS patients in a morphology-blinded manner along with controls using a 10-color single tube flow cytometry. We analyzed the reproducibility of Ogata score and modified FCM scores, additionally incorporating the proportion of CD15, CD11b, CD56, and CD38MFI on CD34+CD19-cluster for each patient. Patients were grouped as proven-MDS, suspected-MDS, and non-MDS groups based on morphology and cytogenetics. Optimized multi-axial radar-plots were also used to analyze maturation patterns in the granulocytic, monocytic, and blast progenitor compartments of proven-MDS cases and controls. RESULTS: Flow cytometric abnormalities ≥3 were present in proven-MDS (n = 23) with a sensitivity and specificity of 78 % and 94 %, respectively, as per Ogata score. The addition of CD38 MFI to the score yielded sensitivity and specificity of 82 % and 88 %, respectively. Additional analysis of aberrant expression of CD15, CD11b, and CD56 increased the diagnostic power of the FCM score. A qualitative analysis of data also showed differences in maturation patterns in proven-MDS compared to the control group. CONCLUSION: Single tube 10-color FCM scoring, including Ogata score, modified-FCM scores, and radar plots pattern analysis, showed significant abnormalities in proven-MDS cases in this pilot study. Large databases, including FCM-scoring and pattern-based analysis for normal BM maturation, could be further validated and standardized for screening MDS.

Hypercholesterolemic arthritis: A case report with emphasis on synovial fluid cytology.

Hyperlipidemia is a risk factor of osteoarthritis. In the present case report, a case of hypercholesterolemic arthritis secondary to diabetes mellitus is described in a 40-year-old male along with the cytological features of synovial fluid.

Cytodiagnosis of Extraskeletal Ewing's Sarcoma and its Confirmation by Fluorescence in situ Hybridization.

Extraskeletal Ewing's sarcoma is an aggressive malignant small round cell tumour usually occuring in children and adolescents. It needs to be differentiated from other malignant small round cell tumours and immunohistochemistry plays a pivotal role in establishing the diagnosis. Fluorescence in situ hybridization or real time-polymerase chain reaction helps in confirming the diagnosis by demonstration of EWS-FLI1 translocation, which is found in approximately 85% of the cases. We report a case of extraskeletal Ewing's sarcoma in a10-year-old male, who presented with a right gluteal region mass. Fine needle aspiration and cell block preparation followed by a panel of immunohistochemical markers were performed. Immunohistochemistry for CD99 and FLI1 was positive. EWS-FLI1 translocation was confirmed by fluorescence in situ hybridization.

An Affordable, Indigenous Polarizer-Analyser System with Inbuilt Retardation Plate Function to Detect Birefringence using 3D Glasses: An Experience.

INTRODUCTION: Polarizing microscope plays a vital role in few but unique situations. A pair of cross polarizers is used to confirm the presence of birefringent substances. Also, a red retardation plate is needed to evaluate the sign of birefringence. However, a polarizing microscope especially with retardation plate is very expensive. Thus, an affordable yet effective substitute using the 3D Polaroid glasses used for '3D movies' would enable widespread use of the polarizing system. AIM: To study the use of 3D polaroid glasses procured from cinema halls in detecting birefringence substances and to study the red retardation plate function in them. MATERIALS AND METHODS: Passive 3D Polaroid glasses were procured from cinema halls. They were arranged in aspecific manner to obtain polarized light. Red retardation plate function can be obtained by changing the arrangement of the glasses. These glasses were used with various available models of different light microscope manufacturers. Various specimens observed included amyloid deposits, woven and lamellar bone, skeletal muscle striations, urate crystals, cholesterol crystals, suture material and glove powder. The comparison was based on subjective interpretation of intensity and quality of birefringence. Sign of birefringence was also determined whenever relevant. RESULTS: The birefringence observed by our system was comparable to the commercially available polarizing system with respect to intensity and quality. Also, there were no false positive /negative results when compared with the commercial Polarizing microscope. Moreover, the system had an inbuilt red retardation plate to determine sign of birefringence. CONCLUSION: The system is efficient, cheap, easily accessible, portable and compatible with all models of light microscopes.

Paratesticular Spindle Cell Rhabdomyosarcoma.

Spindle cell rhabdomyosarcoma is a rare variant of embryonal rhabdomyosarcoma that affects young males and most commonly involves the paratesticular region. We report a case of paratesticular spindle cell rhabdomyosarcoma in a 14-year-old boy, who presented with a painless scrotal mass. Left inguinal orchidectomy was performed. Histopathological and immunohistochemical examination of the mass revealed spindle cell rhabdomyosarcoma of the paratesticular region.