Hartmut Arps

Prognostic factors in medullary thyroid carcinomas. Survival in relation to age, sex, stage, histology, immunocytochemistry, and DNA content

Patients with medullary thyroid carcinomas (MTC) were analyzed according to age, sex, and tumor stage. In addition, the MTC were screened for the predominant histologic pattern, immunocytochemical spectrum (60 tumors), and DNA content (DNA cytophotometry and DNA flow cytometry, 25 tumors). These findings were correlated with follow‐up data available for 45 of these patients. Forty‐eight percent of the tumors revealed a polygonal cell pattern, whereas 22% showed spindle‐cell predominance. All tumors contained cytokeratin, chromogranin A, and calcitonin (CT). Calcitonin gene‐related peptide (CGRP) was present in 92%, carcinoembryonic antigen (CEA) in 77%, neuron‐specific enolase (NSE) in 75%, and vimentin in 53% of cases. Positivity for neurotensin, somatostatin, neurofilaments, bombesin, and alpha human chorionic gonadotropin (a‐hCG) and serotonin ranged between 3% and 27%. All MTC were negative for substance P, adrenocorticotropic hormone (ACTH), thyroglobulin (TG), or S‐100 protein. Local recurrences and regional lymph node metastases revealed identical staining patterns as the primaries. Prognosis of MTC was found not to be related to histologic features (dominant architectural pattern, cellular shape, presence of amyloid deposits) or immunocytochemical pattern. Instead, survival was significantly correlated to age, sex, and stage of disease. The best prognosis was seen in women younger than 40 years and revealing an early stage of disease. DNA measurements added valuable information in assessing the prognosis of MTC.

Morphological detection and quantification of lipoprotein(a) deposition in atheromatous lesions of human aorta and coronary arteries

Lipoprotein(a), as an atherogenic particle, represents an independent risk factor for coronary heart disease. In the present study the morphological distribution of apoprotein (a) and apoprotein B within the arterial wall is described. Apoprotein B, a constituent of very low-density lipoprotein, low-density lipoprotein and lipoprotein(a) has previously been demonstrated in atheromatous lesions. Lipoprotein(a) possesses an additional protein, designated apoprotein (a). Autopsy material (n = 74) from the left coronary artery and from the thoracic aorta has been examined by means of immunohistochemistry and both apoprotein (a) and apoprotein B were detected, primarily associated with the extracellular matrix and accumulating in lesions in the arterial wall. The staining pattern for both antigens was almost always found to be congruent, suggesting that the detection of (a)-antigen has to be attributed at least in part to the presence of lipoprotein(a). It is concluded that both low-density lipoprotein and lipoprotein(a) have an important role in the pathogenesis of atherosclerosis.

Impact of overall treatment time of fractionated irradiation on local control of human FaDu squamous cell carcinoma in nude mice

A series of experiments were performed to determine the impact of overall treatment time on local control of human FaDu squamous cell carcinoma irradiated with 30 fractions under ambient conditions in nude mice. The TCD50 increased with treatment time between 15 days and 10 weeks from 43 Gy to 102 Gy. The data can be well described by a single linear function. The dose recovered per day is 1.0 Gy. However, the data can also be adequately fitted by two components with an initial delay of about 30 days followed by a steep increase at a rate of 1.5 Gy per day. Assuming that the increase of TCD50 is solely caused by repopulation of clonogenic tumor cells, and that the cellular radiation sensitivity in vitro reflects the radiation sensitivity of FaDu cells in vivo, the doubling time of clonogenic tumor cells during treatment is estimated to be approximately 1.8 days for the one-component model and, after an initial delay, approximately 1.2 days for the two-component model. Both values are shorter than the doubling time of clonogenic cells in untreated FaDu tumors and similar to the potential doubling time determined by flow cytometry after BrdUrd labelling. It is concluded that the dose necessary to control FaDu squamous cell carcinoma increases considerably with increasing time of a fractionated radiation treatment. It appears most likely that this increase is caused by repopulation of clonogenic tumor cells; however, other mechanisms such as an increasing fraction of hypoxic tumor cells can not be ruled out at present.

Mucoepidermoid tumors of the salivary glands. Correlation of cytophotometrical data and prognosis.

The mucoepidermoid tumors of the Salivary Gland Registry, Institute of Pathology, University of Hamburg, Western Germany, were evaluated retrospectively with regard to epidemiologic data, clinical follow‐up, and cytophotometric data. Clinical data were obtained in 71 cases. Tissue from 46 cases was studied by single cell scanning cytophotometric analysis. Two thirds of the tumors were located in the major salivary glands, the parotid being the most common site, one third occurred in the minor salivary glands. The age range was from 6 to 81 years; peaks were observed in the fourth and seventh decades; the sex distribution was almost equal. By means of a single cell scanning cytophotometric device, a division into “diploid” and “atypical” patterns was possible. The clinical course was well correlated with these two groups, the atypical group showing generally an unfavorable course. Especially in poorly differentiated tumors, selection of clinically aggressive tumors was possible by their atypical DNA distribution pattern. Consequently, single cell DNA assessment can be a useful supplementary tool in the clinicopathologic and prognostic evaluation of mucoepidermoid tumors of the salivary glands.

In vitro Prediction of Cytostatic Drug Resistance in Primary Cell Cultures of Solid Malignant Tumours

The in vitro monolayer proliferation assay (MP-assay) described here enables predictive determination of the efficacy of anticancer drugs considered for clinical application. The assay was designed (1) to achieve a high plating efficiency, (2) to adapt in vitro growth as close as possible to in vivo conditions, and (3) to prove that the cells in vitro correspond with the in vivo tumour cells they were derived from. From 452 freshly explanted or biopsied tumours, 321 (71%) proliferating cultures could be established. To prove malignant origin of the incubated cells each strain was characterised by DNA-cytophotometry for aneuploidy and by immunocytochemistry for marker proteins. Drug potency was determined by comparing the number of living cells in drug-treated cultures with non-treated controls. Drug concentrations in vitro corresponded with those achievable in tumour tissue and thus represented clinically relevant levels. Growth inhibition in vitro was correlated with in vivo tumour response. Two hundred in vitro/in vivo correlations were performed (50 retrospective, 150 prospective). Overall predictive accuracy of the MP-assay was 86%, with correct indication of resistance in 94.5% and of sensitivity in 75.8% (P < 0.001). The results show that the proposed assay is capable of estimating the response probability of cytostatic drugs in individual tumours and thus can contribute to reducing the applications of non-effective drugs and, within limitations, to improving the basis of drug selection.

DNA cytophotometry and prognosis in ovarian tumors of borderline malignancy. A clinicomorphologic study of 80 cases

Surgical specimens of 80 ovarian tumors of borderline malignancy (OTBM) were investigated by scanning DNA cytophotometry. Diploid or euploid DNA histograms were found for 21 tumors, whereas 59 OTBM showed noneuploid or aneuploid DNA patterns. All patients were followed‐up after surgery for at least 3 years (mean observation period, 6.7 years). Follow‐up showed 11 cases of recurrent disease and 6 deaths. DNA findings and several other morphologic and clinical details (including patient age, histologic type and stage of disease, and extent of therapy) were correlated to the postoperative course. Statistical analyses disclosed that, of these parameters, only DNA content significantly affected prognosis. Recurrences and deaths resulting from tumor exclusively were observed among patients with noneuploid or aneuploid OTBM, whereas none of the diploid or euploid tumors recurred (P < 0.05). DNA cytophotometry thus might be regarded as an effective complementary means to assess the prognosis of individual OTBM cases.

The encapsulated follicular carcinoma of the thyroid

In a retrospective study of 86 follicular carcinomas of the thyroid gland, 35 lesions were classified as encapsulated carcinomas (40.7%). In two of these, lymph node metastases were detected initially. Another patient presented with distant metastases. The biological behaviour of these 35 tumours was studied over a long-term follow-up period (0.4–19.1 years, mean 10.3 years) which featured three cases of death from thyroid carcinoma 0.4–5.0 years after thyroidectomy. Another patient suffered from local recurrence of a follicular carcinoma 13.9 years later. The morphological and clinical findings of those five patients who initially presented with metastases and/or whose follow-up registered the local recurrence of thyroid cancer or death as a result of it, were compared with the remaining 30 cases which were of a benign clinical course. Statistical analysis showed that the prognosis of encapsulated follicular carcinoma is more serious when tumours occur in patients older than 65 years of age and when the tumour diameter is 5.0 cm or more. There was a tendency towards poorer prognosis in those tumours exclusively composed of oxyphilic epithelium.

Nuclear DNA content of borderline tumors of the ovary: correlation with histology and significance for prognosis

Scanning-DNA cytophotometry was applied to Feulgen stained sections of 22 borderline tumors of the ovary (BOT). The DNA content was related to conventional histology. In 11 cases clinical follow up for more than 5 years was available. The DNA measurements disclosed two subgroups in the group of BOT. One showed a nuclear DNA content not exceeding tetraploidy (4c) indicating proliferative activity without malignant change and a second one exhibited DNA values higher than 4c indicating malignant transformation. Correlation of histological evaluation with the DNA content revealed a good agreement in 15 cases. However, a discrepancy was found in 7 cases: either the histological evaluation aroused suspicion for malignant potential but the histogram showed DNA values not higher than 4c (n=4), or histology showed well differentiated lesions with an atypical histogram (n=3). Clinical monitoring revealed no recurrence or tumor spread in all but one case of the group of lesions with DNA values up to 4c, whereas in the group with atypical DNA histograms (DNA values>4c) relapse appeared in 6 out of 7 cases. The results suggest that DNA analysis has prognostic significance for BOT.

Effect of continuous vs intermittent application of 3-OH-tamoxifen or tamoxifen on the proliferation of the human breast cancer cell line MCF-7 M1

SummaryThe antiproliferative potency of 5x10-7M tamoxifen (TAM) and 3-hydroxytamoxifen (3-OH-TAM) was investigated during continuous (8 days) or intermittent (2 h every 2nd or 3rd day, respectively) application to the oestrogen-receptor-positive, estradiol-sensitive human mammary carcinoma cell line MCF-7 M1, a variant of MCF-7 wild type. Growth modulation was evaluated in parallel by counting cells and by measuring DNA content. Continuous incubation resulted in a growth inhibition to 21.8±3.2% by 3-OH-TAM and to 39.5±4.8% by TAM when compared with control cultures defined as 100%. Intermittent addition induced a growth reduction to 23.0±2.1% by 3-OH-TAM and to 41.2±2.4% by TAM in relation to 100% controls. Addition of 3-OH-TAM for 2 h only at day 1 resulted in an inhibition to 70.3±3.2%, again in relation to 100% controls. When TAM was administered once for 2 h at day 1 it induced an inhibition to 79.0±4.9% at day 8. The in vitro results indicate that (a) at 5x10-7M3-OH-TAM has a better antiproliferative effectiveness than TAM, (b) the intermittent application is as effective as continuous application (no significant difference), and (c) the addition once a week reveals only a slight growth reduction after 8 days of culture.Application of the long-living TAM results in continuously high serum concentrations, which have been shown to create resistant cell clones. Compared to TAM the 3-OH metabolite has a considerably shorter half-life and its application in vivo reveals rise and fall of its serum concentrations. Since the presented data demonstrate that 3-OH-TAM is more potent than TAM and that the intermittent application is as effective as the continuous form, interval therapy with 3-OH-TAM may slow down the process of acquiring resistance to antioestrogens.

Pancreatic endocrine carcinoma with ectopic PTH-production and paraneoplastic hypercalcaemia

In a case of pancreatic endocrine carcinoma hypercalcaemia without bone metastases and normal parathyroid glands prompted our suspicion that there was paraneoplastic production of an osteoclast activating substance by the tumour tissue. This view was further confirmed by bone histology. Immunohistology post mortem revealed the production of PTH in the primary tumour and a liver metastasis. The usefulness of immunohistology in detecting paraneoplastic secretion of hormonal substances is discussed.