Nuclear morphometry in squamous cell carcinoma (SCC) of the tongue.

In the histological grading of oral squamous cell carcinoma (OSCC) nuclear features are very important. Nevertheless evaluation is usually performed in a subjective and not highly reproducible way. The aim of this work was to investigate the relationship between nuclear shape and survival in 30 cases of carcinoma of the tongue. All the patients were divided into two groups: short-term survival and long-term survival. Twenty nuclei for each tumour were submitted to a morphometrical study by the shape analytical morphometry (SAM) software system. It was thus possible to evaluate not only nuclear dimensions but also nuclear contour irregularities and nuclear shape asymmetries. Multivariate discriminant analysis (MDA) of the quantitative parameters obtained by the morphometrical study distinguished the patients of the two groups with only a 10% error; moreover successful cluster analysis was performed by using Fourier parameters. Both these sets of results were achieved mainly owing to the parameters for contour irregularities.

Nuclear morphology and histological grading of oral squamous cell carcinoma (OSCC). A morphometric study.

Histological grading of oral squamous cell carcinoma (OSCC) is usually evaluated in a semiquantitative way and this causes a low grade of reproducibility of this prognostic evaluation. The aim in this preliminary phase is to investigate the relation between nuclear shape (studied by an analytical procedure) and histological grading to evaluate the weight of nuclear shape and dimension in grading formulation. Nine cases of OSCC were studied (3 cases of G1, G2 and G3 respectively) with 20 nuclei for each case. All the tumors were localized on the lip. Histologic grading according to the Bryne system was performed. Each nucleus was submitted to a morphometrical study by the SAM (Shape Analytical Morphometry) system which, apart from dimensions allows us to evaluate nuclear contour irregularities and nuclear shape asymmetry. Nuclei of squamous cell carcinoma show a progressive increase in dimensions (area, perimeter and diameter) from G1 to G3. Nuclei of G3 carcinoma are characterized by the most irregular shape. Size differences (mean value) are significant to differentiate G1 nuclei from G2 and G3 nuclei while shape parameters are useful in distinguishing G3 nuclei from G1 and G2. Multivariate discriminant analysis showed that by using only nuclear parameters it is possible to correctly classify about 80% of nuclei. This study, using quantitative methods, confirmed nuclear pleomorphism to be the most important feature in grading evaluation.

A new approach to the histologic study of ovarian tumors by analytical morphometry.

In recent years, borderline epithelial tumors of the ovary have been investigated by morphometric techniques to allow for a differential diagnosis from benign and malignant neoplasms. In order to enhance this discriminant power, we have applied a new analytical procedure to the evaluation of the nuclear shape in epithelial ovarian tumors. Sixty nuclei of benign ovarian serous neoplasms, 60 nuclei of serous borderline tumors and 60 nuclei of serous carcinomas (18 cases in all) were examined using the software system SAM (Shape Analytical Morphometry). The morphometric procedure consisted of three different phases: (i) extraction of nuclear fundamental curve: this is a function curve giving the smoothing of the original contour by two parametric equations (separately for x and y values as dependent variables); (ii) evaluation of nuclear contour irregularities by Fourier analysis; (iii) evaluation of nuclear shape asymmetry by SAE (Shape Asymmetry Evaluator): this is the ratio between the length of a segment of a parabola interpolating the original curve points and a straight line joining its extremities for a 180 degrees barycentric rotation according 10 degrees steps. All the obtained independent parameters were submitted to statistical analysis. Nuclei of borderline tumors showed dimensional parameters which were intermediate between those of benign and malignant tumors. Both the asymmetry and the irregularities of nuclear contour were greatest in carcinomas.

Laryngeal intraepithelial neoplasia (LIN). An analytical morphometric approach.

Laryngeal Intraepithelial Neoplasia (LIN) is graded in 3 levels (LIN Grade I-II-III), corresponding to the classic aspects of mild, moderate and severe dysplasia-in situ carcinoma, on the basis of the number and position of mitoses and of the undifferentiated or atypical cells limited to the basal or extended to the intermediate or the superficial layers of epithelium. In order to reduce the subjective imprecision of these parameters we have applied not only traditional dimensional evaluators but also procedures of analytical morphometry to the nuclear shape. By using the software system S.A.M. (Shape Analytical Morphometry) we have examined fifty nuclei of the basal layer in LIN grade I, II and III, fifty nuclei in normal laryngeal mucosa and fifty nuclei in invasive carcinoma of the larynx (twenty-five cases in all). Normal and dysplastic nuclei did not show any dimensional differences, while the carcinomatous nuclei were significantly larger. An asymmetric distortion of the nuclear contour was noted in the moderate and severe dysplasia, but not in carcinomatous cells. Also the Fourier parameters, increased in severe dysplasia, decreased dramatically in carcinomatous cells which showed nuclei with minor contour irregularities than the normal cells. These findings outline the discriminative power of the analytical morphometry and suggest a possible correlation between nuclear shape and cell biology.

Morphometrical investigation of medulloblastoma nuclei by S.A.M. (Shape Analytical Morphometry) software system.

In order to characterize medulloblastomas and to get over the difficulties sometimes encountered in differential diagnosis, a double morphometric procedure has been applied to its nuclei. The first consisted of size measurements (maximum diameter, area and perimeter), the latter is represented by S.A.M. (Shape Analytical Morphometry) software-system specifically implemented to describe shape of biological structure by analytical parameters. Analytical and dimensional parameters submitted to Hotelling's multivariate discriminant analysis gave the best results when used together in convenient discriminant subsets, thereby allowing a good distinction between medulloblastoma in comparison with neuroblastoma, Ewing's tumor, lymphoblastic and lymphocytic lymphoma. These results underline the usefulness of morphometric characterization also for practical diagnostic purposes.

Multiparametric discrimination of serous ovarian tumors by analytical morphometry.

In order to enhance the discrimination power in the field of serous ovarian tumors, we applied the software system SAM (Shape Analytical Morphometry) to the analytic studies of biological forms. Besides the usual dimensional evaluations (perimeter, area, maximum diameter and shape index), this procedure permits the description of the nuclear form using analytical parameters: 1) extraction of nucleus fundamental curve; that is a functional curve giving the "smoothing" of the original contour by two parametric equations (separately for x and y values as independent variables); 2) evaluation of nuclei contour irregularities by Fourier analysis; 3) evaluation of shape asymmetry by SAE (Shape Asymmetry Evaluator); that is the ratio between the length of a segment of a parabola interpolating the original curve points, and a straight line joining its extremities for a 180 degrees barycentric rotation according 10 degrees steps. All parameters resulted to be independent and were submitted to multivariate discriminant analysis. We studied 180 nuclei from 18 cases of serous ovarian tumors, (6 benign, 6 borderline and 6 malignant tumors). With respect to the dimensional parameters, the application of analytical morphometry permitted us to reduce the minimum percentage error in the discrimination of the different classes. In fact, in the distinctions of benign and malignant nuclei, the minimum percentage error was 13.30%, against the 18.3% error when using dimensional morphometry. Furthermore, in the comparison of malignant and borderline nuclei there was a reduction of error from 23.3% to 22.5%, and in the comparison of benign and borderline nuclei, the error was reduced from 37.5% to 30%.(ABSTRACT TRUNCATED AT 250 WORDS)

Using analytical morphometry to distinguish severe dysplasia and large bowel carcinoma.

The aim of the present paper is to find analytical parameters to distinguish severe dysplasia from cancer. A population of 150 nuclei was selected randomly from: 5 cases of large bowel carcinoma, 5 cases of adenomatous polyps with severe dysplasia--according to Kozuka's criteria--(without any reference to the architecture: tubular, tubulo-villous and villous) and 5 cases of normal mucosa as control. The files of the coordinates of the nuclear contours were submitted to the S.A.M. (Shape Analytical Morphometry) work-station, which produces quantitative parameters (area, perimeter, maximum diameter and Roundness Factor) as well as analytical shape-descriptors. The latter were extracted by the S.A.E. (Shape Asymmetry Evaluator) procedure, which evaluates the symmetry of each contour studied, and by Fourier harmonic analysis which breaks up each curve into a defined set of harmonics which are then used to perform statistical tests. In the comparison between normal and severe dysplastic nuclei, normal and carcinomatous nuclei and severe dysplastic and carcinomatous nuclei univariate statistical evaluation (Student t test) proved to be highly significant for the most of the analytical parameters.

[Nuclear morphometry and estrogen receptors in infiltrating ductal carcinoma of the breast]. / Morfometria nucleare e recettori estrogenici (RE) nel carcinoma duttale infiltrante della mammella.

In this study ten cases of breast infiltrating ductal carcinoma have been considered. In all of them the content of ER has been evaluated by using monoclonal antibodies. Five of them were ER positive and five were ER negative. For the morphometric study ten nuclei of each case have been considered. By using the S.A.M. (Shape Analytical Morphometry) work-station an analytical study of the nuclear shape was performed. The first step was the extraction of fundamental shape which describes the basic shape of original contour without its irregularities. It was obtained by using two parametric equations. The second step was the evaluation of shape asymmetry by S.A.E. (Shape Asymmetry Evaluator). Finally the contour irregularities were evaluated by Fourier analysis. Along with analytical parameters, dimensions (area, perimeter and maximum diameter) were considered too. All obtained data were submitted to univariate statistical analysis (Student's T test) to compare the two groups (ER positive and ER negative tumors). Area, perimeter and maximum diameter were significatively greater in ER negative cases while analytical parameters were not discriminant between the two groups.

Hand-mirror (HM) cells in acute lymphoblastic leukaemia (A.L.L.). An attempt at cellular classification by morphometric analytical parameters using the S.A.M. (Shape Analytical Morphometry) system.

A morphometric study of the nuclear and cytoplasmic shape of a blastic population in a case of HM-A.L.L. was performed by comparing the two differently shaped populations with and without HM configuration. The results obtained using analytical size-independent parameters created by the S.A.M. work-station enable us to characterize the shape of both blastic cell populations quantitatively, and strongly suggest the existence of shape modulation from one cellular type toward the other. Thus a possible sequence from blastic cells (having regular, rather round nuclei) to HM cells (characterized by high distortion of both nucleus and cytoplasm) was proposed.

Non-Hodgkin's lymphoma diagnosis aided by the S.A.M. system.

In the attempt to discriminate between centroblasts and immunoblasts, an analytical morphometrical procedure was adopted, considering the nuclei of a randomly selected centroblastic and immunoblastic population from non-Hodgkin's lymphomas and the nucleoli of a more restricted number of the same neoplastic population. All the size-independent shape-descriptor parameters extracted for each step of the mathematical analyses used were submitted to Hotelling's multivariate discriminant analysis that enabled us to achieve a good distinction (3% error) between centroblasts and immunoblasts when nuclear and nucleolar parameters were used together.

Differential diagnosis between thyroid follicular adenoma and carcinoma. Analytic morphometric approach.

In this study some nuclear dimensional and analytical parameters were evaluated in order to distinguish follicular atypical adenoma from follicular carcinoma of the thyroid. Eighty nuclei from carcinomas, 80 from adenomas and 80 from normal thyroid were studied. Analytical parameters obtained by the nuclear shape study (by S.A.M. system) as well as dimensional parameters were submitted to univariate statistical analysis. On the ground of our results atypical adenoma could be considered as an intermediate aspect of a progressive change from benign to malignant even if they are closer to normal thyroid than to carcinoma.

Relation between primary tumor shape and biological behavior in breast cancer.

The aim of this study was to investigate the biological significance of tumor shape in breast cancer by considering the shape not as a casual event but as an expression of the behavior and natural history of the tumor. The shape was studied by an analytical morphometry procedure and was related to axillary metastases, which up to now are the most meaningful prognostic factors in this disease. Fifty cases of infiltrating breast carcinoma (25 N+ and 25 N-) were investigated. The shape, studied on subgross sections of the tumor, was considered as the result of two components: the subtle contour irregularities and the main distortions of the figure. The procedures used allowed us to distinguish and to parametrize these two components in order to submit them to univariate analysis (Student's t test), a principal components analysis and, finally, a multivariate discriminant analysis (Hotelling test). The utilized analytical procedures by work-station S.A.M. (S.A.M. work station is a product of "Metamorphosis") consisted of three steps: 1) Extraction of tumor function curve obtained by Kth order polynominals which gives a smoothing effect to the original curve; 2) Evaluation of contour irregularities by Fourier harmonic analysis; 3) Evaluation of shape asymmetry by SAE (shape asymmetry evaluator). We considered also the roundness factors of the original and function curves and finally the maximum tumor diameter. Three parameters relating to contour irregularities (sum and mean value of Fourier harmonic amplitude and percentage of superimposed points) and parameters relating to main distortions of the figure (mean value of SAE) were highly significant (p less than 0.001). The roundness factor of the original curve was more significant (p less than 0.001) than that of the function curve (p less than 0.01) whereas maximum tumor diameter was not significant when tested by Student's t test. Multivariate discriminant analysis allowed 20% of error to be reached by using 3 parameters relating to the shape analysis and the two roundness factors. By using 8 parameters, including maximum tumor diameter, the percentage error was 16%. The results obtained, while they stress the usefulness of the employed procedure, reveal that shape of the tumor, together with its dimensions, is an important expression of the biological behavior relating to metastatic spread.

Morphometric analytical evaluators of lymphoid populations in nonneoplastic lymph nodes.

A morphometric analysis using an Apple II microcomputer and shape analytical morphometry (SAM) software was performed of the nuclear shape in the lymphoid population of normal (nonneoplastic) lymph nodes to provide baseline parameters useful for distinguishing between different subtypes of lymphomas. Fourier harmonic analysis and two indices, a contour asymmetric evaluator (CAE) and a shape asymmetric evaluator (SAE), are proposed as non-dimensional shape parameters. Results obtained with the CAE and SAE indices indicate that they represent sensitive shape descriptors able to reveal even subtle irregularities of nuclear outline. With these parameters, centrocyte nuclei were easily differentiated from those of all other lymphoid cells while an interesting close relationship was found between lymphocytes and centroblasts. Lymphocyte nuclear shape was shown to be less regular than generally reported, suggesting the existence of subtle differences between stimulated and unstimulated nuclei. Immunoblast nuclei were shown to have the most regular shapes.

[Description of the shape of carcinomatous nodules of the breast by means of high degree polynomial equations and Fourier harmonic analysis]. / Descrizione della forma dei noduli carcinomatosi della mammella mediante equazioni polinomiali di grado superiore e analisi armonica di fourier.

Breast cancer nodule shape has been investigated by two analytical procedures: upper degree polynomials and harmonic Fourier analysis. An automized method (TV camera interfaced with a computer) has been employed to find the coefficients of kth order equation giving the best fit. Then the difference between real node contour and function curve has been calculated and submitted to harmonic analysis. This gives the sine/cosine coefficients of the two series and the amplitude of the harmonics. The results point out that no difference exists between metastatic and non metastatic tumors and suggest that parametrized descriptors calculated may represent a peculiar behaviour in the growth of the tumor.

[Simulation of the metastatic spread of mammary carcinoma by evaluation of the shortest routes in a computerized model of the lymphatic system represented in a three-dimensional map]. / Simulazione della diffusione metastatica del carcinoma mammario mediante valutazione dei percorsi minimi in un modello computerizzato del sistema linfatico rappresentato da un grafo tridimensionale orientato.

A simulation model of mammary lymphatics has been realized by a computerized system to draw a tridimensional map of all lymphnode stations (axillary, supraclavear and internal mammary lymphnodes) draining from the breast. The purpose was to identify the minimal routes between tumor and lymphnodes and among different nodal points that represent lymphnodes of the lymphatic net situated at various distance from tumor. A simulation of node metastatization has been performed giving to every route of the model a different weight represented by the distance between the nodes.