Nuclear areas in breast cancer: relationship with estrogen and progesterone receptor expression.

The relationship between nuclear area and the expression of estrogen (ER) and progesterone receptors (PR) was evaluated in a series of 66 breast carcinomas (50 primaries, 16 metachronous recurrences). Nuclear measurements were directly performed on touch imprints previously reacted for ER and PR by the immunocytochemical method. Nuclei expressing ER or PR were found to be smaller than negative ones. Moreover, area values of PR+ nuclei overlapped with those of ER+ ones. The pattern of receptor expression co-existence was observed to be closely related to nuclear dimensions, since a significant increase in mean nuclear areas was detected in cells from ER-/PR-tumours, as compared with cells from ER+/PR+, ER+/PR- and ER-/PR+ sub-groups. Nuclear area values were also determined in both steroid receptor-positive and steroid receptor-negative neoplastic cells co-existing in the same breast carcinoma. No significant differences in nuclear area values were found between receptor-positive and receptor-negative neoplastic cells in tumours featuring a positive status for at least one of the steroid receptors (ER+/PR+, ER+/PR- or ER-/PR+). On the contrary, in breast carcinomas characterized by a negative steroid receptor status (ER-/PR-), receptor-negative neoplastic nuclei were significantly larger than receptor-positive ones. It is suggested that heterogeneous steroid receptor expression may be caused by a completely different pathogenetic mechanism in receptor-positive and receptor-negative breast carcinomas.

Metastatic capacity and differentiation in murine melanoma cell lines. A morphometric study.

A morphometric analysis was carried out on electron micrographs of cells of the F1 (low metastatic capacity) and F10 (high metastatic capacity) variant sublines of the murine B16 melanoma, both in in-vitro cultures and in lung-metastatic nodules developed after the intravenous injection of neoplastic cells in syngeneic C57 black male mice. A group of 28 morphometric parameters was derived to describe quantitatively each neoplastic cell profile. No qualitative difference was observed between the two cell lines. The quantitative expression of subcellular organelles was dissimilar in the two sublines, being consistently characterized, both in in-vitro cultured cells and in lung-metastatic colonies, by a significant decrease in the mean values of parameters related to melanosomes in the high metastatic capacity cell line (B16-F10). Moreover, in in-vitro cultured cells, indices describing heterochromatin masses and cytoplasmic membranous compartments displayed statistically significant differences between the two sublines. In this experimental system, an inverse relationship between metastatic capacity and differentiation is detected, since cells with a more aggressive metastatic behavior exhibit a decreased degree of differentiation.

A morphometric comparison of subcellular structures between primary breast tumors and their nodal metastases.

A morphometric study of subcellular structures in human primary breast tumors (of the ductal infiltrating type) and their synchronous lymph node metastases was carried out using planimetry and stereology. Each primary neoplasm was compared separately with its axillary metastatic growth in order to detect differences in the quantitative estimates of their subcellular components. A set of 24 morphometric parameters was used to quantitatively describe each neoplastic cell profile. Although some subcellular parameters showed significant differences between the cells of a primary tumor and its metastases, no parameter was found to be consistently altered in all cases. No consistent increase or decrease in the values of the affected parameters was observed in the metastases. More than being due to the expression of a biologically variant (more-or-less differentiated) metastatic phenotype, the differences in stereologic parameters detected in metastatic cells appear to reflect the existence of different functional states of the same cell type.

The morphometric analysis of cell nuclei from fine needle aspirates of thyroid follicular lesions does not improve the diagnostic accuracy of traditional cytologic examination.

In order to evaluate the usefulness of the morphometric analysis in improving the accuracy of the traditional cytologic examination of follicular thyroid lesions, fine needle aspirates from 17 adenomatous goiters, 19 follicular adenomas and 18 follicular carcinomas were examined. In each case 20-50 follicles, accounting from 100 to 200 thyroid cells, were studied by operators unaware of the histologic diagnosis. Nuclear areas (mean, maximum and minimum area) were derived by means of a semiautomatic system and follicular thyroid lesions were classified into three groups (goiter, adenoma and follicular carcinoma) utilizing cut-off values. The results were then compared to the cytologic diagnosis carried out according to traditional (optic microscopy) criteria. Mean values of nuclear areas showed significant differences among the three groups, but a considerable overlap occurred in the size distribution of cell nuclei. At present, therefore, planimetric measurements of cell nuclei in fine needle aspirates do not improve the accuracy of standard cytologic examination in the differential diagnosis of follicular thyroid lesions.

Proliferative lesions of the breast lobules. A morphometric analysis.

A morphometric study was carried out on proliferative lesions of the breast lobules, including cases of atypical lobular hyperplasia (ALH), lobular carcinoma in situ (LCIS) and infiltrating lobular carcinoma (ILC) with axillary metastases (META). Seven nuclear parameters were assessed, such as perimeter, area, shape factor, diameter of circle of equivalent area, longest axis, shortest axis and axes ratio. In comparison with nuclei of normal lobules, a statistically significant increase in the mean values of 5 nuclear parameters was observed in all pathologic lesions examined, whereas shape factor and axes ratio exhibited no significant difference. Nuclear area was found to be the most valuable parameter. No difference was observed between nuclear estimates of ALH and LCIS. Mean nuclear values were higher in ILC cases than in in-situ lesions, but no difference was detected between ILC and axillary metastases.

Computer-assisted morphometry of the peripheral nervous system. A diagnostic tool.

A formal systematization of the set of procedures required in peripheral nerve morphometry has been implemented in a computer program, in an attempt to provide an easy-to-handle diagnostic tool in current neuropathological practice. The hardware is composed of an IBM-PC compatible computer and a graphic tablet. Programs are written in BASIC and run in computers compatible with the MS-DOS operating system. The flow of operative steps involved in the morphometric study is controlled by the software. Sequential measurements are also tested for congruence. Nerve fibre measurements are stored in pluri-dimensional matrices for subsequent statistical calculations, i.e. evaluation of size-frequency distribution of nerve fibres and correlation analysis between nerve fibre subcomponents. Stereological estimates of the parenchymal components are also derived. The increase in accuracy achieved using area instead of diameter measurements is stressed. Compared to "dedicated" machines such personal computer-assisted systems exhibit substantial advantages in terms of low cost and flexibility.

Morphometric studies on fine-needle aspirates from follicular proliferative lesions of the thyroid.

A morphometric analysis was carried out on smears of fine needle aspirates obtained from proliferative follicular lesions of the thyroid. Mean values, range as well as histograms of nuclear areas were calculated in histologically-diagnosed cases of non-toxic goiters, follicular adenomas and well-differentiated follicular carcinomas. Mean nuclear areas were found to be significantly higher in carcinomas than in adenomas and in adenomas than in non-toxic goiters. In size-frequency distribution histograms, differences among corresponding classes of the three groups were not statistically significant. Linear discriminant analysis was ineffective in differentiating not only benign from malignant follicular tumors, but also non-toxic goiters from adenomas. Morphometric studies on nuclear areas of thyreocytes in fine needle aspirates do not appear to be able to differentiate between proliferative follicular lesions of the thyroid.

Morphometric changes in sympathetic nerve fibres in diabetes.

A morphometric study was carried out on electron micrographs of lumbar sympathetic nerve fibres surgically removed from 12 patients with arteriosclerosis obliterans: 4 diabetics, 4 non-diabetics and 4 subjects with glucose intolerance. In patients with diabetes and glucose intolerance a significant decrease in the percent area fraction of unmyelinated axons and an increase in the percent area fraction of the endoneurial space were demonstrated.

Personal computer software for the stereologic evaluation of sympathetic ganglia.

Software was developed on a personal computer for a quantitative study of some ultrastructural aspects of sympathetic ganglia (the neuronal cell body, the surrounding satellite cell layer and the nearby capillaries). A low-cost digitizer, originally designed for teaching, semiprofessional graphics applications and games, was utilized to sample areas and distances on electron micrographs. The processing program derives stereologic parameters from the crude data stored in the memory. Further algorithms then assess the mean thickness and the volume densities of the satellite cell layer, endothelial basal lamina and endothelial layer.

Algorithms for morphometric measurements on cancer cells in electron microscopy. Pilot tests.

Automated procedures were designed for handling quantitative data derived from a morphometric study of neoplastic cells on electron micrographs. Pilot studies were carried out on 3 breast carcinomas. Each tumor cell was coded by the sequence of 30 stereological parameters that described the general organization of subcellular constituents. Wide standard deviations, as large as 90% of the mean, were observed in some parameters. The practical application of correlation analysis between stereological parameters turned out to be of limited value, since only parameters related to the same subcellular structure showed correlation.