Gelatinous transformation of bone marrow associated with ring sideroblasts: A diagnostic pitfall.

Gelatinous bone marrow transformation (GBMT) is a rare condition characterized by adipocyte atrophy, deposition of extracellular gelatinous substance in the bone marrow and associated hypoplastic hematopoiesis. The underlying pathogenic mechanisms of GBMT remain poorly understood. Here we describe 3 cases of GBMT associated with ring sideroblasts. An electronic search of institutional archives was conducted via the laboratory information system to identify patients with a body mass index (BMI) of <18.5 who underwent bone marrow evaluation. The slides and reports for these bone marrow specimens were reviewed. Bone marrow specimens of 10 patients were identified and reviewed. Three (30 %) were found to have GBMT and ring sideroblasts, ranging from 2 to 20 %. Blasts were not increased and there was no other morphologic evidence of dysplasia. Every patient had one or more peripheral blood cytopenias. In one patient, copper deficiency was proven providing an explanation for the ring sideroblasts. To the best of our knowledge, ring sideroblasts have not been well documented in GBMT and aims to contribute to a better understanding of disease recognition and pathogenesis and also to prevent potential misdiagnosis as a myelodysplastic syndrome.

Clinical, immunological, and molecular characterization of hyper-IgM syndrome due to CD40 deficiency in eleven patients.

PURPOSE: Hyper-IgM syndrome due to CD40 deficiency (HIGM3) is a rare form of primary immunodeficiency with few reported cases. In this study, we further characterize the clinical, immunological, and molecular profiles of the disease in a cohort of 11 patients. METHODS: Molecular genetic analysis and a comprehensive clinical review of patients diagnosed with HIGM3 at our tertiary care center from 1994 to 2011 were undertaken. RESULTS: Eleven patients from seven families were enrolled. The patients had a median age of 9 years [ranging from 2 to 22 years old]. All 11 patients had recurrent chest infections at presentation. Pneumocystis jiroveci pneumonia was confirmed in three patients. Five patients had sclerosing cholangitis, and five patients had Cryptosporidium isolated from their stool. Six patients had nasal and sinus infections, and two of these patients had destructive nasal fungal infections. Eight patients had neutropenia. All of the patients had low IgG and normal or high IgM levels. IgA was undetectable in all but three patients. Two novel mutations were found: a splice site for intron 3 and a missense mutation located in the coding region of exon 3. Two patients underwent successful stem cell transplantation from a matched donor. Four patients are doing well on prophylaxis; two are very sick, one with protracted diarrhea and persistent Cryptosporidium and the other with neurological complications. Three patients died early in life as a result of severe sepsis. CONCLUSIONS: To our knowledge, this report provides the largest cohort of patients with this disease with a very long follow-up period. Our cohort showed variable disease severity

A Rare Case of CD4 Positive Cytotoxic PTCL, NOS With Leukemic Presentation.

Peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS) is a relatively rare mature T-cell lymphoma that cannot be categorized under any of the well-defined category. This type of aggressive lymphoma mostly involves the lymph nodes, though any other organ can be affected. Leukemic presentation is extremely rare. No case report of isolated leukemic presentation was found after detail literature search. Herein we present a case of PTCL, NOS with leukemic presentation.

Flow cytometry improves the diagnostic value of fine needle aspiration cytology in a spleen with neuroendocrine tumor: A case report.

Fine needle aspirations are infrequently performed on the spleen due to concerns for hemorrhagic complications. As a result, splenic lesions can be challenging to diagnose given the limited amount of available specimen. Metastasis to the spleen is rare and metastatic neuroendocrine tumors to the spleen are scarce in literature. The diagnosis of splenic lesions from fine needle aspirate entails processing which prolongs the turnaround time, particularly if the cytomorphology is non-typical and a limited sample can further complicate this process. We describe a case in which flow cytometry performed on fine needle aspiration of a splenic lesion suggested a diagnosis of neuroendocrine neoplasm involving the spleen. Further workup confirmed this diagnosis. Flow cytometry can recognize neuroendocrine tumors involving the spleen in a timely manner so that appropriate immunohistochemistry tests on limited specimens can be performed to aid in their accurate diagnosis.

Proposal of Diagnostic Approach of Periodontal Primary Non Hodgkin Lymphoma of Bone with Flow Cytometry as an Essential Diagnostic Component.

BACKGROUND: Primary bone lymphoma is a rare type of lymphoid neoplasm with favorable prognosis, where Primary Non Hodgkin Lymphoma of bone (PB-NHL) is most common with the subtype. Amongst PB-NHL, diffuse large Bcell lymphoma represents the majority of cases. The mandible is a very uncommon site of involvement, presenting as a painful bone mass with high suspicion of osteomyelitis. METHODS: We report the case of a 45-year-old male with no significant past medical history who was admitted to the hospital with a large right jaw mass and pain after recent tooth removal. The original tissue biopsy was not diagnostic, and cultures were found to be negative for microorganisms. Due to enlargement of the mass, a fine needle aspiration (FNA) was done. At the time of rapid onsite evaluation of the FNA, atypical lymphoid cells were seen, and material was obtained for flow cytometry (FC) evaluation. This revealed an aberrant clonal B-cell population. The consequent immunohistochemical evaluation of original material supported the diagnosis of PB-NHL. After chemotherapy patient improved. RESULTS: After an extensive English language literature review, we identified and summarized the clinical presentations, diagnostic procedures, histopathologic features, treatment methods, and outcomes of forty-two cases of periodontal PB-NHL. Based on our findings, we propose a set of clinical features at initial presentation to increase the clinical suspicion of periodontal PB-NHL for practitioners. CONCLUSION: Based on our institution's experience and the literature review conclusions, we propose the University of Texas Medical Branch diagnostic approach for PB-NHL and suggest that FNA and FC should be utilized as the essential diagnostic component. The fast and efficient diagnosis of PB-NHL can facilitate the correct treatment and sufficiently improve patient care.

Grading of follicular lymphoma using flow cytometry.

BACKGROUND: The treatment and prognosis of follicular lymphoma (FL) is dependant on the grade of the disease. In the World Health Organization classification of lymphoma, grading of FL into low grade (1 and 2) and high grade (3) is recommended. Grading of FL is possible in excision biopsy; histological grading is subjective and inconsistent. Grading is extremely difficult in needle core biopsies and fine needle aspirates. We attempted to grade FL using flow cytometry (FCM) and CD19/ forward scatter. MATERIALS AND METHODS: Cases of FL seen in our institution and submitted for FCM were evaluated for the percentage of cells detected beyond the 500-channel mark (on a 1024 scale) on a CD19/forward scatter dot plot. We hypothesized that these cells most likely represent centroblasts and their percentage would reflect the grade of the disease. Histological grading of the lymphoma on the open biopsies constituted the reference for FL grade. RESULTS: Thirty-six cases of FL, including 22 males and 14 females, ranging in age from 19 to 92 years (median, 42 years), were studied. There were 17 cases of low grade (grade 1; n=10 and grade 2; n=7) and 19 cases of high grade (grade 3) FL. The percentage of cells identified beyond the 500-channel mark on CD19/forward scatter dot plot ranged from 0.12% to 12.55% (median, 4.9%) in low grade (grade 1 and 2) whereas the percentage of those cells in high grade FL ranged from 6.22% to 51.95% (median, 21%; P=0.00001). CONCLUSION: Our findings suggest that using a CD19/forward scatter dot plot can help identify centroblasts in FL making grading possible on FCM, especially in small biopsies and fine needle aspirates.

Validation of a modified cryopreservation method for leukemic blasts for flow cytometry assessment.

BACKGROUND: Cryopreservation, a common method for storing human cells, has advantages when cells are used in retrospective studies of selected cell populations. Frozen lymphocytes can be used for tissue typing, for monitoring cell-mediated immunity, and for various immunological tests. Our report describes an efficient, simple and inexpensive method for cryopreservation of human acute leukemia cells. METHODS: Leukemia cells from 20 newly diagnosed cases were frozen at -80 degrees C after cryopreservation with 5% dimethysulfoxide and then assayed by flow cytometry for antigen expression determined by monoclonal antibodies at different time intervals. RESULTS: All cases had viability above 75% at presentation. After 4 weeks, 91% of pre-B ALL, 88% of T-ALL, 100% of AML, and 100% of biphenotypic aliquots had viability over 75%. Viability continued to be reliably above 75% at 6 weeks from cryopreservation. CONCLUSION: We confirm that the method does not significantly alter the viability of cells and it preserved the antigenic expression of leukemia cells.

Expression of CD66c and CD25 in acute lymphoblastic leukemia as a predictor of the presence of BCR/ABL rearrangement.

BACKGROUND: Expression of myeloid or T cell lymphoid in precursor B cell acute lymphoblastic leukemia (pre-B cell ALL), which is referred to as aberrant expression, is quite a common phenomenon. CD66c is a myeloid marker which has aberrant expression in pre-B cell ALL, with strong correlation with non-random genetic changes (BCR/ABL rearrangement). Another leukemia associated marker (CD25) is frequently expressed in pre-B cell ALL. The frequency of CD25-expressing lymphoblasts has been found to be significantly higher in BCR/ABL-positive vs. BCR/ABL-negative patients. METHODS: In a cohort of 103 patients diagnosed with pre-B cell ALL or biphenotypic leukemia and studied for expression of CD66c and CD25 at presentation, we evaluated the frequency of expression of either or both in BCR/ABL positive cases. RESULTS: Surface CD66c was expressed by 70 cases (68%) and CD25 was expressed by 33 cases (32%) while both were expressed together on 29 cases (28%). BCR/ABL was positive in 18/103 patients. All BCR/ABL positive cases were positive for surface CD66c and CD25. CONCLUSION: Positivity for both leukemia-associated antigens CD66c and CD25 in combination can predict the presence of BCR/ABL rearrangement in pre-B cell ALL. While this finding does not replace the detection of BCR/ABL abnormality by cytogenetic or molecular techniques, it does provide an early and handy tool for prediction and management of high-risk cases of pre-B cell ALL, especially in centers with limited laboratory facilities.