Kappa and lambda determination by single colored immunohistochemistry and chromogenic in situ hybridization techniques in B-cell malignancies

Background: kappa and lambda light chains detection in bone marrow trephine sections help in the determination of B-cell clonality through evaluation of light chain restriction

Aim of the Work: was to compare the efficacy of single color detection-based immunohistochemistry [IHC] and chromogenic in situ hybridization [CISH] in evaluating kappa/lambda expression in tissues harboring B-lymphoid lesions

Patients and Methods: Forty patients were enrolled in this study. They were divided into three groups chronic lymphocytic leukemia [CLL/SLL] group I [n=13], non-Hodgkin lymphoma [NHL] group II [n=24] and hairy cell leukemia [HCL] group III [n=3]. The 24 NHL cases comprised of [11 diffuse large B-cell lymphoma, 6 mantle cell lymphomas, 3 marginal zone lymphoma, 2 lymphoplasmacytic lymphoma, 1 follicular lymphomas and 1 Burkitt's lymphoma]. Kappa and lambda light chains were detected in their bone marrow trephine sections using single colored immunohistochemistry, chromogenic in situ hybridization and the results were compared to the flowcytometry as reference method

Results: Light chain restriction [LCR] was detected by FCM in 100% of the cases followed by CISH [52.1%; 12/23] of the cases and finally IHC [43%; 18/40]

Conclusion: Both conventional CISH and IHC are effective in determining monoclonality in cases of mature B- cell neoplasm that has plasmacytic differentiation and with high amount of cytoplasmic Ig light chains such as MZL and LP. However, they are not effective in determining monoclonality in cases with low amount of Ig light chain such as cases of pregerminal and germinal center lymphoma. Yet, CISH is more informative than IHC due to the lack of background staining which allowed for greater discrimination between absence and presence of monoclonality

Accuracy of dose planning for prostate radiotherapy in the presence of metallic implants evaluated by electron spin resonance dosimetry

Radiotherapy is one of the main approaches to cure prostate cancer, and its success depends on the accuracy of dose planning. A complicating factor is the presence of a metallic prosthesis in the femur and pelvis, which is becoming more common in elderly populations. The goal of this work was to perform dose measurements to check the accuracy of radiotherapy treatment planning under these complicated conditions. To accomplish this, a scale phantom of an adult pelvic region was used with alanine dosimeters inserted in the prostate region. This phantom was irradiated according to the planned treatment under the following three conditions: with two metallic prostheses in the region of the femur head, with only one prosthesis, and without any prostheses. The combined relative standard uncertainty of dose measurement by electron spin resonance (ESR)/alanine was 5.05%, whereas the combined relative standard uncertainty of the applied dose was 3.35%, resulting in a combined relative standard uncertainty of the whole process of 6.06%. The ESR dosimetry indicated that there was no difference (P>0.05, ANOVA) in dosage between the planned dose and treatments. The results are in the range of the planned dose, within the combined relative uncertainty, demonstrating that the treatment-planning system compensates for the effects caused by the presence of femur and hip metal prostheses.

Local anesthesia with epinephrine is safe and effective for oral surgery in patients with type 2 diabetes mellitus and coronary disease: a prospective randomized study

OBJECTIVE: To investigate the variations in blood glucose levels, hemodynamic effects and patient anxiety scores during tooth extraction in patients with type 2 diabetes mellitus T2DM and coronary disease under local anesthesia with 2% lidocaine with or without epinephrine. STUDY DESIGN: This is a prospective randomized study of 70 patients with T2DM with coronary disease who underwent oral surgery. The study was double blind with respect to the glycemia measurements. Blood glucose levels were continuously monitored for 24 hours using the MiniMed Continuous Glucose Monitoring System. Patients were randomized into two groups: 35 patients received 5.4 mL of 2% lidocaine, and 35 patients received 5.4 mL of 2% lidocaine with 1:100,000 epinephrine. Hemodynamic parameters (blood pressure and heart rate) and anxiety levels were also evaluated. RESULTS: There was no difference in blood glucose levels between the groups at each time point evaluated. Surprisingly, both groups demonstrated a significant decrease in blood glucose levels over time. The groups showed no significant differences in hemodynamic and anxiety status parameters. CONCLUSION: The administration of 5.4 mL of 2% lidocaine with epinephrine neither caused hyperglycemia nor had any significant impact on hemodynamic or anxiety parameters. However, lower blood glucose levels were observed. This is the first report using continuous blood glucose monitoring to show the benefits and lack of side effects of local anesthesia with epinephrine in patients with type 2 diabetes mellitus and coronary disease. .

Inmunodeficiencia común variable: Epidemiología y manifestaciones clínicas en 69 pacientes / Common variable immunodeficiency: Epidemiology and clinical manifestations in 69 patients

La inmunodeficiencia común variable (IDCV) se caracteriza por una alteración en la producción de anticuerpos y una mayor susceptibilidad a infecciones por bacterias extracelulares capsuladas, principalmente del tracto respiratorio. Analizamos las características clínicas de 69 pacientes, evaluados en un período de 10 años en tres centros de la ciudad de Buenos Aires. Al inicio del estudio se encontraban en seguimiento 14 pacientes y al finalizar 60; la mayoría fueron derivados por infecciones o hipogammaglobulinemia, y casi la mitad con diagnóstico establecido de inmunodeficiencia. Sesenta y cinco (94.2%) pacientes presentaron infecciones por gérmenes capsulados, cuatro (6.1%) sepsis y dos tuberculosis. La edad promedio de comienzo de los síntomas infecciosos fue de 18.1 años, la edad promedio al momento del diagnóstico fue de 29.6 años y el retraso diagnóstico de 11.9 años. En 41 (59.4%) pacientes se registró el antecedente de diarreas recurrentes o crónicas. En 22 (31.9%) se diagnosticaron 13 enfermedades autoinmunes, siendo las más frecuentes las hematológicas y el hipotiroidismo. Ocho pacientes tuvieron linfoproliferación policlonal histológica, cuatro (5.8%) como enfermedad granulomatosa de hígado, laringe y piel, y cuatro como neumonía intersticial linfoidea (NIL). Diecinueve (27.5%) pacientes presentaron esplenomegalia y 23/57 (40.3%) imágenes sugestivas de procesos granulomatosos o linfocíticos en la TAC de tórax (incluidos los 4 con NIL). Tres (4.3%) pacientes desarrollaron linfoma B, cuatro (5.8%) adenocarcinoma de estómago y uno cáncer de mama. El estudio tuvo una mediana de seguimiento de 54 meses, rango 1-353 y durante el período del mismo murieron cuatro pacientes (5.8%).

Common variable immunodeficiency (CVID) is characterized by an impaired antibody production and an increased susceptibility to recurrent infections of the respiratory tract, mainly by extracellular encapsulated bacteria. We analyzed the clinical characteristics of 69 patients evaluated over a period of 10 years at three centers in the city of Buenos Aires. At the onset of the study 14 patients were on follow up, and at its end the number of patients reached to 60. Most of them consulted for infection or hypogammaglobulinemia and nearly half had an established diagnosis of immunodeficiency. Sixty-five (94.2%) patients had infections by encapsulated bacteria, four (6.1%) sepsis and two tuberculosis. The average age of onset of infectious symptoms was 18.1 years; the average age at diagnosis was 29.6 years and the delay to diagnosis 11.9 years. Forty one (59.4%) patients reported a history of recurrent or chronic diarrhea. In 22 (31.9%) 13 autoimmune diseases were diagnosed, being the most frequent the hematological disorders and hypothyroidism. Eight patients had histological polyclonal lymphoproliferation, four (5.8%) with granulomatous disease affecting the liver, the larynx and/or the skin; and four as lymphoid interstitial pneumonitis (LIP). Nineteen (27.5%) patients had splenomegaly and 23/57 (40.3%) images suggestive of lymphocytic or granulomatous processes (including the 4 with LIP) in the chest CT. Three (4.3%) patients developed B cell lymphoma, four (5.8%) stomach adenocarcinoma and one breast cancer. The study had a median follow-up of 54 months, range 1-353 and four patients (5.8%) died during the follow up.

Morphologic, immunphenotypic and clinical discriminators between T-cell / histiocyte-rich large B-cell lymphoma and lymphocyte-predominant Hodgkin lymphoma

Features of T-cell/histiocyte rich large B-cell lymphoma [THRLBCL] overlap with those of lymphocyte predominant Hodgkin lymphoma [LPHL]. The two lymphomas may represent a spectrum of the same disease, and differentiation between the two can sometimes be difficult. We looked at histomorphologic, immunophenotypic and clinical information that may help differentiate the two entities. Cases of THRLBCL and LPHL were blindly reviewed and studied for histological pattern [nodular vs. diffuse], nuclear features and pattern of expression of CD20, CD30, CD57, epithelial membrane antigen [EMA] and Epstein-Barr virus [EBV]. A score encompassing diffuse histology, high nuclear grade, CD20 single-cell pattern, CD30+, CD57-, EMA-, and EBV+ was estimated for the diagnosis of TCHRLBCL. There were 58 cases, including 30 cases of TCHRLBL and 28 cases of LPHL. The median age was 36 years for TCHRLBCL and 21 years for LPHL [P=0.0001]. Three types of nuclei were identified [lymphocytic/histocytic, Reed-Sternberg and centroblast-like]. The latter two high-grade nuclei were suggestive of TCHRLBCL. TCHRLBCL and LPHL, respectively, showed diffuse histology, 90% vs. 4% [P=0.001], single CD20+ cells, 93% vs. 3.5% [P=0.00004], CD30+ cells, 30% vs. 0% [P=0.01], CD57+ cells, 41% vs. 93% [P=0.008], EMA+ cells, 27% vs. 60% [P=0.113], EBV+ cells, 24% vs. 0% [P=0.117], high nuclear grade, 70% vs. 0% [P=0.001], total score 2-7 [mean 4.68] vs. 0-2 [mean 0.72] [P=0.001], high stage, 86% vs. 7% [P=0.0001]. Our findings indicate that a combination of multiple parameters can help differentiate between the two diseases. Two cases originally diagnosed as LPHL were re-assigned the diagnosis of THRLBCL

Proliferaciones linforreticulares del pulmón / Pulmonary lymphoproliferative lesions

The present review describes the current classification of the pulmonary lymphoproliferative lesions as proposed by the WHO in 2004 with emphasis in the clinical picture and histopathological features. The definition of these entities includes the clinical picture, histopathology, immunohistochemistry and molecular features. The differential diagnosis of the most important entities is also briefly discussed

En el presente trabajo de revisión se describe la clasificación actual de las lesiones linfoproliferativas del pulmón propuesta por la OMS el año 2004 con énfasis en el cuadro clínico y los aspectos histopatológicos. La definición de estas entidades incluye cuadro clínico, histopatología, inmunohistoquímica y características moleculares. Se discute brevemente el diagnóstico diferencial de las formas más importantes

Epstein-barr nuclear antigen-1 (EBNA-1) in diffuse large B-cell lymphoma and its relationship to the bcl-2 protein.

AIM: To determine EBNA-1 expression in the tissue of patients with diffuse large B-cell lymphoma and its relationship to bcl-2 expression. METHODS: Paraffin-embedded tissue from 24 cases of diffuse large B-cell lymphoma were stained immunohisto chemically with monoclonal antibody anti-EBNA-1 and anti-bcl-2 using Streptavidin Biotin Complex method. The bcl-2 expression was measured as negative (no staining), positive 1 (weak staining), and positive 2 (moderate to strong staining). The relationship between the immunohisto chemistry results was examined. RESULTS: From the 24 samples, 12 (50%) were EBNA-1 positive and 14 (58.3%) showed expression of bcl-2. Eleven of the 14 samples (90%) showed expression of bcl-2 were EBNA-1 positive, while only 3 of these samples (10%) were EBNA-1 negative. The relationship between bcl-2 expression and EBNA-1 was statistically significant (Chi square-uncorrected 11.2571, p= 0.0036). CONCLUSION: Our results showed that 50% of diffuse large B-cell lymphoma were EBNA-1 positive. Furthermore, EBNA-1 seemed to be correlated with bcl-2 expression.

Rosette formation in non Hodgkin's lymphoma in the bone marrow--a case report.

This is a case report of rosette formation in non-Hodgkin's lymphoma in the bone marrow. A 66 year old female on treatment for metastatic papillary carcinoma thyroid and non-Hodgkin's lymphoma of the nodular small cleaved cell type was found to have bone marrow infiltration by a low grade B cell lymphoma. Bone marrow aspirate, imprint and trephine biopsy showed rosettes of the abnormal lymphoid cells. The associations of papillary carcinoma thyroid, hyperglobulinemia and retroperitoneal fibrosis in this case which explain the histogenesis of rosette formation are discussed.

Clinical relevance of soluble CD44 in serum as indicator of tumor burden in patients with B-lymphoid malignancies

In this study, the serum level of sCD44 was studied in a group of B- lymphoid malignancies at presentation trying to define any association with tumor burden and clinical staging and to correlate the soluble marker with other clinical, laboratory and biologic parameters. It was conducted on 100 adult patients with different B-lymphoid malignancies divided into two groups. ELISA determinations revealed a significant elevation of serum sCD44 values in the entire patients group and the three subgroups of lymphoma/leukemia as well as HD group in comparison with the controls. On comparing the three subgroups as regards sCD44 mean values, a significant difference was observed with the least mean value expressed by the highly aggressive subgroup. 68% of the studied patients showed serum sCD44 above the cut-off value and 16% showed levels above 1000 ng/ml

Linfoma cutâneo de células B / Cutaneous B-cell lymphomas

Linfomas cutâneos de células B fazem parte de um grupo heterogêneo de alteraçöes do sistema linforreticular que que podem acometer a pele. Nesta apresentaçäo relatamos o caso de um paciente que apresentava dados clínicos, histopatológicos e imunoistoquímicos compatíveis com o diagnóstico de linfoma cutâneo de células B. Durante o estadiamento se encontrou comprometimento da medula óssea. O paciente foi tratado com quimioterapia sistêmica e radioterapia localizada, havendo a regressäo das lesöes.

A clinicopathologic study on the diffuse malignant lymphoma: a morphologic and immunophenotypic analysis in 62 patients at Harbor-UCLA Medical Center

In order to compare the prognoses of patients with diffuse malignant lymphomas on the basis of histology and immunophenotypes, we retrospectively studied 62 cases of diffuse lymphoma arising in lymph nodes. We also evaluated the reactivity patterns of monoclonal antibodies (MoAb) LN1, LN2 and LN3 to determine the criteria for making a differential diagnosis in B cell lymphomas. The immunologic phenotypes were determined by the avidin biotin peroxidase complex method, using frozen or paraffin fixed tissues. The majority (66.3%) were B cell with the remaining 20.9% being T cell and 12.9% were non-B, non-T cell lineage. Immunological heterogeneity was found especially in the mixed small and large cell and the immunoblastic lymphomas. There was no significant difference between B- and T-cell lymphomas with respect to survival and death (P > 0.05). Histologically 79% (49/62) of the lymphoma was large cell and 21% (13/62), small cell lymphoma. There was a difference in prognosis between low, intermediate and high-grade of lymphomas. However there were no significant differences among the subtypes of the diffuse aggressive lymphomas. Factors associated with poor prognosis were advanced stages (P < 0.025) and histology of the malignant lymphomas. MoAb LN1, LN2 and LN3 gave positive staining in 83.3%, 91.7% and 60% of B cell lymphomas, respectively. The most common phenotypic pattern in B cell lymphomas was LN1+, LN2+, LN3+/-, suggestive of follicular center cell origin. As a panel, phenotypic patterns of MoAb LN1, LN2 and LN3 may be useful in differentiation of follicular center cell lymphoma from others.