J. A. M. van Unnik

Cyvadic in advanced soft tissue sarcoma: A randomized study comparing two schedules: A study of the EORTC soft tissue and bone sarcoma group

Two hundred forty‐six adults with advanced progressive soft tissue sarcoma received combination chemotherapy with cyclophosphamide, vincristine, Adriamycin (doxorubicin), and DTIC. They were randomly allocated to receive the four drugs simultaneously every 4 weeks (S1: CYVADIC), or pairs of drugs (S2: ADIC‐CYV) alternating at 4 weekly intervals. One hundred sixty‐two patients completed 8 weeks of chemotherapy, and were considered to be evaluable for response. There were 18 complete remissions and 25 partial remissions, an overall response rate of 26%, with a highly significant difference between the two arms in favor of S1 (38% versus 14%, P = 0.001). There were no significant differences between S1 and S2 in terms of median duration of remissions (62 versus 39 weeks), and median survival of responders (85 versus 80 weeks) and of all evaluable patients (43 versus 45 weeks). Karnofsky index (KI) was the single most important prognostic factor. Patients with KI 90–100 showed a remission rate of 41% (56% on the S1 regimen) in contrast with 14% in those with KI 50–80. No patient with a KI of 50 responded to chemotherapy. The main toxicities were nausea, vomiting, anorexia, alopecia and myelosuppression, but did not differ significantly between the two regimens. Our findings suggest that stratification according to KI is essential for studies on chemotherapy for advanced soft tissue sarcomas in order to make a valuable comparison of treatment results.

Grading of soft tissue sarcomas : experience of the EORTC soft tissue and bone sarcoma group

A practical grading system for soft tissue sarcomas was developed, based on 282 eligible patients entered in an EORTC adjuvant clinical trial. The primary tumours in this trial had to be adequately treated. Histopathological parameters, which appeared significant in two preceding studies, were tested. These parameters were differentiation of the tumour, presence and amount of necrosis, the presence and amount of myxoid areas and the number of mitoses. In addition, the size of the tumour was also analysed. The quantitative data (mitotic count and size of the tumour) were not a priori grouped, but were divided into categories based on the results of the statistical analysis. Based on a multivariate analysis only mitotic count, the presence or absence of necrosis and the size of the tumour were significantly correlated with the duration of survival or the time to distant metastases. Of these parameters, the mitotic count was the most important.

A study to analyze the origin of tumor cells in malignant fibrous histiocytomas a multiparametric characterization

To study the derivation of tumor cells of malignant fibrous histiocytomas (MFH), their phenotypical marker profile was investigated and compared with those of malignant histiocytosis (MH) and of different types of soft tissue tumors (STT). The presence of the following markers was investigated: on paraffin sections, alpha‐1‐antichymotrypsin (ACT); on frozen sections antigens associated with lymphocytes, macrophages and fibroblasts, the enzymes acid phosphatase, nonspecific esterase, and beta‐glucuronidase; and, on isolated and cultured cells, the receptors for EA‐gamma and complement. Furthermore, the capacity to phagocytose sensitized erythrocytes and carbon particles was studied in vitro. MFH tumor cells and a part of other types of STT shared the expression of ACT and lysosomal enzymes with MH. They differed, however, from MH by the absence of monocyte/macrophage‐associated antigens and by the expression of fibroblast‐associated antigens, which property they had in common with other STT. MFH tumor cells were not able to form rosettes or to phagocytose Ig‐sensitized erythrocytes, but they showed phagocytosis of carbon particles. The results strongly indicate that MFH tumor cells originate from (primitive) fixed mesenchymal cells and are not related to monocyte‐derived histiocytes.

Radiation therapy in primary non-Hodgkin's lymphomas of the CNS

Twelve patients with primary non-Hodgkins lymphomas of the CNS are described. Out of 5 CSF cytologies performed, 4 were positive. Radiotherapy was given to the tumour area in 3 patients, or to the whole brain in 5 patients. Four cases received radiotherapy to the spinal cord as well. Patients receiving whole CNS irradiation, including the spinal cord, seem to have a longer survival than patients with brain irradiation only. Out of the 5 patients with total brain irradiation, 2 showed a relapse in the spinal cord. It is suggested that therapy should be given not only to the tumour bearing areas, but should comprise the entire CNS.

Detection of calcitonin-encoding mRNA by radioactive and non-radioactive in situ hybridization: improved colorimetric detection and cellular localization of mRNA in thyroid sections.

The localization of mRNA encoding calcitonin was studied by in situ hybridization using 35S-labeled RNA probes and biotin-labeled DNA probes. Radiolabeled probes were detected by autoradiography and biotin-labeled probes by streptavidin-biotin-peroxidase. To intensify the colorimetric signal, the indirect avidin-biotin complex (ABC) method was performed. However, the results were often variable. To improve the sensitivity, the peroxidase reaction signal was enhanced with a gold-silver deposit intensification reaction. To shorten the incubation times and to enhance the colorimetric reaction, several reaction steps were performed in a microwave oven. The localization of calcitonin mRNA in thyroid tissue, as detected with in situ hybridization, was confirmed by immunohistochemical localization of the calcitonin polypeptide. The results of in situ hybridization using biotinylated probes were compared to in situ hybridization using radioactive probes. Our data show that the results of in situ hybridization applied on frozen and paraffin-embedded sections using biotinylated DNA probes, detected with an indirect streptavidin-biotin-peroxidase reaction and intensified by silver-gold enhancement, were comparable to those obtained with radioactive probes. The localization of calcitonin encoding mRNA was in agreement with the localization of the calcitonin polypeptide.

Two cell lines with epithelial cell-like characteristics established from malignant fibrous histiocytomas

Two malignant fibrous histiocytoma (MFH) cell lines were established: one from a storiform‐pleomorph subtype and the other from a myxoid one (codes, MFH‐3 and MFH‐4). Light microscopic examination revealed large rounded cells, growing mostly separately, in both cell lines. Their ultrastructure was different in various aspects. The MFH‐3 cells showed abundant lysosomal activity, a well‐developed Golgi apparatus, and a few desmosome‐like cell contacts. The MFH‐4 cells had a well‐developed rough endoplasmic reticulum, delicate bundles of tonofilaments, the formation of pseudoacini, and the presence of small completely developed desmosomes. Based on immunostaining and immunoblotting assays of cultured cells, both cell lines expressed immunoreactivity for vimentin; cytokeratins 7, 8, and 18; desmin; and laminin, but they lacked reactivity for cytokeratins 10 and 19, neurofilament, α‐smooth muscle actin, S‐100 protein, collagen type IV, carcinoembryonic antigen, and antigens specific for macrophages. Fibronectin and, to a variable extent, glial fibrillary acid protein and epithelial membrane antigen (EMA) were detectable in MFH‐3 cells only. Furthermore, a 60‐kilodalton band was present in both cell lines which was reactive for cytokeratins 8 and 18. The MFH‐3 cells had the capacity to grow as xenografts with a carcinoma‐like pattern. The cells retained their immunoreactivity for vimentin and cytokeratin 8 and showed the presence of desmosomes. Several of these immunophenotypic features also were noticed in established sarcoma cell lines and in short‐term cultures of fibroblasts, smooth muscle cells, and endothelial cells. However, experimental data on the two MFH cell lines show that the MFH cell line may express some immunophenotypic and ultrastructural features considered to be specific for epithelial cells. The MFH cells may originate from multipotential mesenchymal cells with a capacity to differentiate to fibroblast‐like cells, and less frequently, to epithelial cells, smooth muscle cells, and Schwannian cells. Such a differentiation became evident when these cells were adapted to culture conditions or grew in nude mice.

European Experience of Adjuvant Chemotherapy for Soft Tissue Sarcoma: Interim Report of a Randomized Trial of Cyvadic Versus Control

A Multidisciplinary approach to the management of soft tissue sarcomas has led to improvements in the rates of local control, despite increasing use of limb salvage procedures. However, metastasis, which seems to correlate with higher histological grade, remains a distressingly frequent problem, occurring in approximately 30% of individuals, often in the absence of local failure.

Non-Hodgkin's Lymphoma. Immunohistochemical and electron microscopical findings in relation to lightmicroscopy A study of 74 cases

From 74 patients with non‐Hodgkins lymphoma (NHL), fresh biopsy material from involved tissue was investigated histochemically and by light and electron microscopy. The results were compared with the light microscopical conclusions based on three currently utilized classifications, namely of Rappaport, Lukes, and Lennert. The separate groups of low‐grade NHL appeared to consist of homogeneous cell populations, both in immunohistological as well as in enzymehistochemical and electron microscopical sense. On the contrary, high‐grade NHL constituted a heterogeneous group in which supplementary (immunohistochemical, enzymehistochemical, and electron microscopical) investigation is very useful. All NHLs reacted with anti‐human lymphocyte serum, including the so‐called histiocytic NHLs according to the Rappaport classification, which proves their lymphocytic origin. Consequently the prefix „histiocytic”︁ should be altered in a morphogenetically correct way. Without exception the B‐cell NHLs were characterized by the presence of monoclonal immunoglobulin on the cell membrane and/or intracytoplasmically. In a part of the immunologically non‐T‐/non‐B‐/(„null cell”︁) NHLs, a B‐cell origin was indicated by the presence of ATPase and/or 5′ nucleotidase and a T‐cell origin by the presence of dot‐like acid phosphatase.

Histiocytic and dendritic reticulum cells in follicular structures of follicular lymphoma and reactive hyperplasia. A quantitative electron microscopical analysis.

SummaryHistiocytic reticulum cells (HRC) and dendritic reticulum cells (DRC) are integral parts of germinal centres. These cell types are also present in follicles of follicular lymphomas, the neoplastic analogues of physiological germinal centres. In this study the distribution and ultrastructural appearances of HRC and DRC present in normal germinal centres and in neoplastic follicles were established by means of morphometric methods.The number of HRC was significantly lower in malignant follicles than in their reactive counterparts. Quantitative analysis of the cytoplasm and phagolysosomes suggest that HRC are smaller and that their activity is lower in malignant follicles. DRC were present in smaller numbers in these structures, as measured by nuclear counts and their relative volume within the follicles. The ultrastructural features indicate that DRC in follicular lymphoma are functionally less active than in reactive lymph nodes.The possibility that differences between the reticulum cells from reactive and neoplastic follicles may be related to the absence of an immune reaction in malignant follicular lymphoma is discussed. The frequency and appearance of HRC and DRC are suitable as additional parameters to differentiate reactive secondary germinal centres from their malignant analogues.

Malignant lymphoma of follicle centre cells with marked nuclear lobation

SummaryFour cases of malignant B-cell lymphoma characterized by a conspicuous component of tumour cells with markedly lobtated nuclei are described. Two exhibited a follicular and two a diffuse growth pattern. The tumour cell population formed a continuous spectrum comprising both cells resembling normal follicle centre cells and multilobated lymphoma cells. Cytomorphological analysis of the multilobated cell group indicated a differentiation series from centroblast-like cells with moderately lobated nuclei to large and medium-sized cells with marked nuclear lobation which revealed features of centrocytes. In three cases (1, 3, and 4) the majority of these multilobated cells showed plasmacytoid differentiation in their cytoplasm in conjunction with the synthesis of monotypical cytoplasmic immunoglobulin. No plasmacytoid features were present in a fourth case (2). In only one case (4) monotypical surface immunoglobulin was detectable on the tumour cells.A close relationship between the multilobated tumour cells and follicle centre cells was further substantiated by the finding of a similar cell variant in the follicle centres of a control group of non-neoplastic lymph nodes. It included cells with plasmacytoid differentiation which synthesized polytypical immunoglobulin.We consider this type of B-cell lymphoma with a conspicuous component of cells with lobated nuclei as a variant of malignant lymphoma, centroblastic/centrocytic.