Identification of hyperdiploidy in fixed cells from pediatric acute lymphoblastic leukemia cases using flow cytometry and cytogenetic analysis.

Cytogenetic analysis is of value in predicting clinical outcome of pediatric cases of acute lymphoblastic leukemia (ALL). Hyperdiploidy in these patients has significant impact on therapeutic outcome. Because of the technologic limitations of cytogenetic analysis in determining hyperdiploidy in some of these cases, we devised a method to analyze cytogenetically fixed material for cellular DNA content by standard flow cytometric methods. This technique greatly enhances out ability to interpret the observance of single cell anomalies and assess whether these cells are predictive of clonal stemlines or are a result of random events.

Lymphocytic lymphoma with associated gamma heavy chain and IgM-lambda paraproteins. An unusual biclonal gammopathy.

Evaluation of proteinuria in a 66-year-old white female detected the presence of two distinct anomalous proteins: a gamma heavy chain devoid of light chains and an intact IgM-lambda paraprotein. The gamma heavy chain protein was incomplete, antigenically corresponding to the Fc portion of the molecule. Subsequent diagnostic studies of the patient revealed the simultaneous presence of an adenocarcinoma of the colon and an extranodal well-differentiated lymphocytic lymphoma involving the lungs, pulmonary hilum, and upper gastrointestinal tract. Levels of the two paraproteins were unaltered by surgical excision of the colonic carcinoma but were reduced by chemotherapy directed at the lymphoma.

Pseudoangiomatous hyperplasia of mammary stroma. Some observations regarding its clinicopathologic spectrum.

Pseudoangiomatous hyperplasia of mammary stroma (PHMS) is a benign proliferation of keloid-like fibrosis, containing slit-like pseudovascular spaces. Its main importance is its distinction from angiosarcoma; however, the clinicopathologic spectrum of PHMS remains incompletely described. We report two new cases and describe our findings in 200 consecutive breast specimens evaluated for the presence of PHMS. The first patient presented with peau-de-orange change in the overlying breast skin, thus mimicking inflammatory breast carcinoma. Furthermore, this patient's PHMS lesion had been diagnosed and treated inappropriately as a low-grade angiosarcoma. The second case showed the more typical, fibroadenoma-like presentation of PHMS. In addition, PHMS changes occur commonly in routine breast biopsy specimens. In fact, our review of 200 consecutive breast specimens showed PHMS in at least one microscopic focus in 23% of cases. The PHMS changes occurred in younger patients than the control population and were associated with fibrocystic changes, in fibroadenomas, in gynecomastia, in normal breast tissue, and in sclerosing lobular hyperplasia. Ultrastructural and immunohistochemical studies of one case showed that the capillary-like spaces were either acellular or lined by fibroblasts. Pseudoangiomatous hyperplasia of mammary stroma represents a clinicopathologic spectrum, extending from focal, insignificant microscopic changes to cases where PHMS produces a breast mass. Increased awareness of PHMS and its clinicopathologic spectrum will allow its differentiation from other vascular tumors of the breast, especially low-grade angiosarcoma.

T-cell lymphomas containing Epstein-Barr viral DNA in patients with chronic Epstein-Barr virus infections.

Fatal T-cell lymphomas developed in three patients with a chronic illness manifested by fever, pneumonia, dysgammaglobulinemia, hematologic abnormalities, and extraordinarily high titers of antibody to the Epstein-Barr virus (EBV) capsid antigen (greater than 10,000) and early antigen (greater than 640) but low titers to the EBV nuclear antigen (less than or equal to 40). To understand the pathogenesis of these tumors better, we determined the immunophenotype of the tumor cells and analyzed tumor-cell DNA for EBV genomes and for lymphoid-cell gene rearrangements. More than 80 percent of the cells in tumors had an activated helper T-cell phenotype (T4, T11, la positive). The EBV genome was found by in situ hybridization in tumor tissue from each patient. Southern blot assay of DNA digests from one patient showed the same pattern as that of the EBV-infected marmoset line, B95-8. DNA digests from two patients showed a monoclonal proliferation of T cells determined on the basis of uniform T-cell-receptor gene rearrangements and a single band for the joined termini of the EBV genome. We conclude that EBV may infect T cells and contribute to lymphomas in selected patients with severe EBV infections.

Use of monoclonal antibodies for analyzing the distribution of the intermediate filament protein vimentin in human non-Hodgkin's lymphomas.

A series of human non-Hodgkin's lymphomas was examined for immunoreactivity with monoclonal antibodies to the intermediate filament protein vimentin with the use of an avidin-biotin immunoperoxidase method. The lymphoid cell nature of each tumor was established with the use of a panel of monoclonal antibodies to lymphoid cell differentiation antigens. There were 28 B-cell and 2 T-cell lymphomas in the series; of the 30 tumors, 11 (37%) were immunoreactive for vimentin. There was no correlation between vimentin immunoreactivity and the histopathologic type of lymphoma. In some tumors, there was nonspecific stromal immunoreactivity for vimentin, but the neoplastic lymphocytes were not immunoreactive. The selective expression of vimentin in non-Hodgkin's lymphomas may be due to masking of the appropriate epitopes or to selective expression of the vimentin gene in certain tumors. On the basis of these results, monoclonal antibodies to vimentin appear to be of limited usefulness in establishing the diagnosis of non-Hodgkin's lymphoma.

Detection of Borrelia in acridine orange-stained blood smears by fluorescence microscopy.

Tick-borne borreliosis (relapsing fever) can be an important, unsuspected cause of febrile illness. The diagnosis is generally made by identifying Borrelia spirochetes in stained peripheral blood smears. Since Borrelia may be difficult to detect with Romanowsky stains, an alternative method, using acridine orange (AO), was used to screen blood smears. Duplicate blood smears of seven patients were examined with the AO technique and Romanowsky stains. In all seven cases spirochetes were easily identified with the AO-stained smears compared with only five cases with Romanowsky stains. In a double-blind laboratory experiment, six of ten duplicate smears from a single patient with mild spirochetemia were positive by AO, whereas only two of ten were positive by Romanowsky stain. We concluded that the AO stain is simple, rapid and more sensitive than Romanowsky methods for detecting cases of low-level spirochetemia.

Primary malignant lymphoma of the cauda equina.

A case report of paraplegia secondary to a malignant B-cell lymphoma primary in the cauda equina is presented. Initial diagnosis was suggested on cytocentrifuge preparations of cerebrospinal fluid with subsequent tissue confirmation following bilateral laminectomy (T12-L3). Histologically, the tumor was a diffuse "histiocytic" lymphoma by Rappaport's or large noncleaved FCC lymphoma by Lukes and Collins classification. Immunologic studies typed the tumor as a B-cell neoplasm with lambda light chains. Following an extensive evaluation of the patient, the lymphoma was found to be limited to the lower spinal cord. Although radiotherapy was initiated there was no improvement of her neurological symptoms, and CSF cytology remains positive for rare malignant cells 2 months after diagnosis. A complete reevaluation at 3 months after laminectomy was negative for lymphoma involvement of other sites.

The role of carboxylic acids in EDTA-dependent panagglutination.

The investigation of a patient blood sample showing a discrepancy between cell grouping and serum confirmation demonstrated a serum agglutinin which reacted with all red blood cells tested when exposed to EDTA. This reaction was 4+ macroscopic at room temperature, 2+ macroscopic with hemolysis at 37 degrees C in albumin, and 1+ macroscopic in the anti-human globulin phase. Agglutination was abolished following dithiothreitol treatment of the patient's serum or following saline washing of the EDTA-exposed test cells. The agglutination reaction was not limited to EDTA, but could be produced with polycarboxylic acids (citrate, L-tartrate, succinate) and monocarboxylic acids (acetate, lactate, propionate, valerate, butyrate). Non-carboxylic acids and low molecular weight ketones or alcohols failed in the agglutination reaction. This study reports an additional example of an IgM "EDTA dependent agglutinin" and demonstrates the dependence on carboxyl groups for its agglutinating activity.