E. Grundmann

Differential pattern of DNA-Aneuploidy in human malignancies

The differential pattern of DNA-aneuploidy, detected by flow cytometry (FCM) regarding its frequency, grade and multiclonality, was investigated and correlated to tumor type, malignancy grade, tumor stage and prognosis in a multi-institutional study at the University of Münster. High resolution measurements using admixed normal blood reference cells were undertaken in 2413 cases of 13 different malignant diseases and in 776 benign lesions or samples. The incidence of DNA-aneuploidy was highest in melanomas, carcinomas, testicular tumors, sarcomas (75%-95%) and myelomas (65%). Acute leukemias showed an intermediate DNA-aneuploidy rate of 40% with special subgroups represented by common ALL (44%), p less than 0.05) and myelomonocytic/monocytic AML (47%, p less than 0.01). The lowest DNA-aneuploidy-rate was found in basal cell skin carcinomas (19%) and congenital melanocytic nevi (9%). No case of DNA-aneuploidy was observed in the 776 benign lesions or samples.--DNA-indices giving the grade of DNA-aneuploidy with 1.0 for normal diploid G1/0 cells were found distributed predominantly between 1.0 and 2.0 in the solid tumors, except testicular tumors, clustering around a triploid maximum at 1.5. DNA-indices of myelomas and acute leukemias generally ranged below 1.25 with lower DNA-aneuploidy grades in AML than in ALL (p less than 0.01).--In melanomas the aneuploidy rate was higher (86%) in metastases than in the primary tumors (54%, p = 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Tumor heterogeneity in osteosarcoma as identified by flow cytometry

Measurements of the cellular DNA content by flow cytometry were carried out in 25 untreated osteosarcomas to identify the frequency of DNA aneuploidies and heterogeneous DNA stemlines in relation to histopathology. Analyzing multiple specimens from each single tumor (2–16; median, 4), highly malignant osteosarcomas were found to express DNA aneuploidies in 18 of 21 cases (86%) with multiple aneuploid DNA stemlines in 10 cases (48%). In three paraosteal osteosarcomas, no DNA aneuploidy was detected and a significantly lower proportion of cells in S‐phase was observed as compared to the highly malignant osteosarcomas (mean 8.6% vs. 18.8%; P <0.05). Like in the paraosteal osteosarcomas, no DNA aneuploidy and a low fraction of cells in S‐phase was found in the predominant cell population of one of the very rare sclerosing small cell osteosarcomas, which also revealed a second DNA stemline with a DNA index of 2.0. These results demonstrate a high degree of DNA stemline heterogeneity in highly malignant osteosarcomas. The data emphasize the usefulness of DNA measurements for the characterization of bone tumors and indicate the possibility of discriminating highly malignant from low‐grade osteosarcomas. Cancer 59:324–328, 1987.

Malignant Lymphomas with Primary Bone Manifestation

18 malignant lymphomas with initial manifestation in bone were selected from the Bone Tumor Registry of Westfalia at the Münster Institute of Pathology where they had been documented between 1975 and 1985, and evaluated under clinical, radiological and histological aspects. Non-Hodgkin lymphomas were reclassified according to the Kiel nomenclature. Paraffin-embedded material was subjected to immunohistochemical analysis in order to assess the features that could add to the correct characterization of these lymphomas, and to their differentiation from other round cell tumors of bone. Non-Hodgkin lymphomas (NHL) were more common in the collective than Hodgkins lymphomas (HL). Of 13 NHL, 2 were of low, 11 of high-grade malignancy. The former comprised one centrocytic and one centroblastic/centrocytic, the latter 7 centroblastic, 2 immunoblastic, and 2 lymphoblastic lymphomas. Seven NHL patients with localized tumors survived up to 11 years (mean survival span: 6 yrs) after local therapy (radiation and/or resection). Another NHL patient, however, had multiple bone lesions, and died within a year. Analysis for leukocyte common antigen was positive in 9/12 NHL cases, reaction with Ki-B-3, a marker of B-lymphocytes, was positive in 7/12 cases. No lymphoma cells were found to react positively with lysozyme, alpha 1-antitrypsin, or alpha 1-antichymotrypsin. It is concluded that localized lymphomas of bone respond well to appropriate local therapy; immunohistochemical investigation may be useful to characterize the true cellular origin of these tumors, and may help to differentiate them from other round cell tumors of bone.

Tumor cell types in osteosarcoma as revealed by electron microscopy. Implications for histogenesis and subclassification.

SummaryUltrastructural studies in 26 osteosarcomas of high grade malignancy which were in diverse locations and of varied histological types revealed seven different tumor cell types. They were characterized by their features as follows: 1) anaplastic cells of malignant blast structure: 2) osteoblastic cells —some of them with dot-like intranuclear bodies ; 3) osteocytelike cells surrounded by mineralized matrix; 4) fibroblast-like cells; 5) cells of myofibroblastic differentiation ; 6) chondroblast-like cells in chondroblastic areas, and even 7) angioblastic cells that may be differentiated from the angioblastic and endothelial structures of capillaries. Histogenetically, osteosarcoma may be derived from stromal mesenchymal cells with a potential for differentiation into these seven tumor cell types, any tumor including, however, the osteoblastic and the osteocyte-like cells with tumor osteoid. This matrix serves as the specific criterion for identifying a tumor as „osteosarcoma“, but almost every osteosarcoma of high grade malignancy will show these seven tumor cell types. The predominance of one or the other cell in the population may provide the basic information for achieving a cytologic subclassification of osteosarcoma in order to obtain relevant morphologic criteria in terms of prognosis.

Biologic characterization of human bone tumors. III. Giant cell tumor of bone. A combined electron microscopical, histochemical, and autoradiographical study.

The cytogenesis of giant cell tumors of bone was studied in 6 cases by combined electron microscopical, histochemical and autoradiographical investigations. Electron microscopy identified two different types of mononuclear stromal cells: fibroblast-like cells with spindly shape and numerous membranes of the granular ER occur together with macrophages bearing many large lysosomes and a prominent Golgi apparatus. Enzyme histochemical results reflect the same diversity: One portion of mononuclear cells exhibits strong alpha-naphthyl acetate esterase (ANAE) activity, known as a marker for cells of the mononuclear phagocyte system, while the other, fibroblast-like cell type is ANAE negative. Tumor giant cells contain numerous membranes of granular ER, mitochondria, and a few isolated lysosomes. They lack the typical brush border of osteoclasts. Moderate to strong ANAE activity of these giant cells reflects their belonging to the mononuclear phagocyte system. Consequently, the giant cell tumor of bone consists of two different cell types, i.e. fibroblast-like cells and cells of the mononuclear phagocyte system, and so is appraised as a fibrohistiocytic tumor. A new inference from our autoradiographic findings is that tritiated thymidine is incorporated only by mononuclear cells, but not by giant cells. Electron microscopical autoradiography demonstrated that among the mononuclear cells, only fibroblasts are found to proliferate, but not macrophages. Thus, the giant cell tumor of bone is seen as a neoplasm of fibroblastic cells with a strong reactive infiltration of cells from the mononuclear phagocyte system.

Malignant fibrous histiocytoma of bone and osteosarcoma. A comparative light and electron microscopic study.

Malignant fibrous histiocytomas are well-described tumors of the soft tissues. Recent investigations have shown that malignant histiocytoma may also occur as a primary bone tumor. However, difficulties may arise to distinguish malignant histiocytoma of bone from other malignant bone tumors, such as osteosarcoma. In the present study, the ultrastructure of five cases of malignant fibrous histiocytoma of bone is compared with that of osteosarcoma. The results show that malignant fibrous histiocytoma is composed mainly of histiocytic cells and fibroblastic cells. In addition, xanthomatous cells, undifferentiated cells, and giant cells may be observed. By contrast, the predominant cell type in osteosarcoma is the neoplastic osteoblast, characterized by abundant rough endoplasmic reticulum. Signs of matrix calcification in the intercellular matrix between the collagen fibrils are regularly observed in osteosarcoma, but not in malignant histiocytoma. From these results it is concluded that the ultrastructure of malignant fibrous histiocytoma arising in bone is morphologically identical with the soft tissue counterpart of this tumor. The components of the tumor are derived from neoplastic histiocytes. This cytogenesis differs from that of osteosarcoma, which is derived from neoplastic osteoblasts. Therefore, from the ultrastructural point of view, malignant fibrous histiocytoma of bone should be accepted as a distinct histologic entity among bone tumors.

Histochemical Identification of T and B Areas in Paraffin-Embedded Lymphoid Tissues by Demonstration of α-Naphthyl Acetate Esterase (ANAE) Activity

A strong activity of acid alpha-naphthyl acetate esterase (ANAE) in T lymphocytes and macrophages within mouse lymphoid organs is demonstrable even after paraffin-embedding, if a rapid embedding procedure with acetone as dehydrating agent is used. The lymphoid tissue structure is better preserved in paraffin sections procured after this rapid embedding procedure than in frozen sections commonly used for histochemical demonstration of ANAE. In the paraffin sections the pattern of the ANAE reaction allows a clear delineation of the T and B areas in the lymphoid organs, and the intracellular location of the focal ANAE activity in T lymphocytes is readily discernible.

Improved grading of bone tumors with the monoclonal antibody Ki-67

SummaryA total of 60 bone tumors and tumor-like lesions presenting various grades of malignancy were investigated immunohistologically with the monoclonal antibody Ki-67 directed against a cell proliferation-associated nuclear antigen. The results obtained agree well with those of flow cytometric and autoradiographic studies on similar tumor entities. The monoclonal antibody Ki-67 was found to be a handy and reliable tool for improved grading of bone tumors.

Metastasizing Eccrine Porocarcinoma Report of Two Cases with Fatal Outcome

The clinical, histopathological and ultrastructural features of a metastasizing eccrine porocarcinoma are described in a 75-year-old woman and a 82-year-old woman. The previously not recognized tumors were identified by its distinct pattern of metastasis. Metastatic spreading was restricted for years to a circumscribed region of the skin. Histology and electron microscopy disclosed pronounced epidermotropism of the PAS-positive tumor cells. The characteristic pattern of dissemination obviously represents a homing phenomenon of the tumor cells derived from the intraepidermal sweat gland duct.

DNA Aneuploidy — A Common Cell Marker in Human Malignancies and Its Correlation to Grade, Stage and Prognosis

Numerous cytogenetic [1, 14, 30] as well as single cell cytophotometric [31] studies have revealed karyotype or cellular DNA content abnormalities as a typical finding in various malignant diseases. Reliable detection of clonal DNA aneuploidy, however, was made possible only by modern flow cytometers (FCM) [9, 10] combining high accuracy DNA fluorometry with a rapid flow through system, thus providing representative DNA distribution of cell populations. On the basis of FCM measurements in various malignancies by several investigators (overview see [3]) DNA aneuploidy may be regarded the most common specific cell marker in cancer and leukemia. By means of FCM, DNA aneuploidies were first observed in acute leukemias [6], and some were later described in a series of different solid tumors by the MD Anderson Hospital group in Houston and by our group [2, 32].