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.

Biological image analysis for microscopical diagnosis: the work-station S.A.M. (Shape Analytical Morphometry).

The S.A.M. (Shape Analytical Morphometry) software system and its related work-station give a generalized and easy-to-handle tool to face a classic and intriguing problem in biomedical morphological diagnosis. What is the shape of an object in a microscopic image? How can we understand the relationship between size and shape? According to Holloway: "Measurements such as length, width, height, whether in chords or arcs only describe space, ... and further run into the abyss of allometric correction ... if additional information (shape?) to size is expected, some method of allometric correction must be used." The S.A.M. software system assumes a logic architecture able to separate and to parametrize independently shape characteristics in terms of allometry and local pertubation by analytical procedures (polynomials, parabolic fitting, Fourier analysis) in addition to the classic evaluations of size and density carried out by image analyzers for microscopical diagnosis.

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.

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.

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.

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.

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)

Analytical morphies on mid-sagittal craniograms glabella-opisthocranion of Homo erectus and Homo sapiens neanderthalensis: Fourier parameters and synthesis of mean craniograms.

Among mathematic procedures used in morphological description, Fourier analysis was indicated as extremely effective in obtaining numerical representations of shape. In order to fully exploit its potentiality in morphology the worked data have to be referred exclusively to the shape of the investigated object and the application of suitable procedures of dimensional normalization are necessary, moreover the significance of the parameters obtained from the analysis must be referable univocally to the morphological datum. In these conditions the numerical characterization of shapes and the relations detectable from different parameters obtained from the description assume the significance of real "analytical morphies". These statements were verified by performing morphological description and comparison, by means of the Fourier harmonic analysis on two groups of mid-sagittal glabella-opisthocranIon craniograms to point out their possible distinctive analytical characteristics: the first group relates to classical neanderthalian group, the second one is made up of asiatic samples of Homo erectus. Some typical patterns of the obtained parameters were discussed and explained in terms of analytical morphies characterizing the given specimens, the traditional morphological classifications were verified and, above all, a numerical description of these samples was obtained.

Analytical morphies on mid-sagittal craniograms glabella-opisthocranion of Homo erectus and Homo sapiens neanderthalensis: Fourier parameters and multivariate discriminant analysis.

The analytical description of complicated morphologies offers the possibility to define patterns of parameters characterizing the investigated groups. These patterns must be considered as morphies useful in performing classification and comparison. Fourier parameters are extremely effective in describing and comparing complex irregular forms and since they are statistically independent we can use them in performing multivariate discriminant analysis. Two groups of mid-sagittal craniograms glabella-opisthocranion of asiatic samples of Homo erectus and of Homo sapiens neanderthalensis were described by means of Fourier harmonic analysis. The discriminatory power of all the obtained parameters (coefficients, amplitudes and phases) was tested. A discriminant function (error % = 0) was obtained using as parameters the first 4 sine/cosine coefficients, the 5th sine, the 6th and the 7th cosine components (11 parameters in all). When the information contained in the coefficient values is being subdivided into the two components of amplitude and phase, the amplitude component is not able to discriminate between the two groups (error % = 25), while the phase values of the first 7 harmonics are able to discriminate them (error % = 0 and distance between centroids = 47).

[Histochemical investigation on the mechanism of action of a synthetic mucolytic compound (Na 872 Cl): preliminary results]. / Indagine istochimica sul meccanismo d'azione di un mucolitico di sintesi (Na 872 Cl): risultati preliminarai.

A histochemical study about mucolytic capacity of Na872Cl compound revealed its action upon epitelial mucins of mouse colonic and tracheal mucosa but the results depend on the chemical type of fixative employed (formalin or acetone fixed sections); the mode of action especially regarding the influence of fixatives and the nature of sensitive substrates are discussed.

Analytical morphometry of sagittal fronto-facial profiles of 2nd-6th month human foetus.

A prominent problem in the morphological study of developing biological structures is shape parametrization with the aim to evaluate transformations in a non subjective way. We carried out an analytical morphometrical study on a series of human embryos, by means of the S.A.M. (Shape Analytical Morphometry) software package. After standardization and normalization of the fronto-facial sagittal profiles, the analytical procedures applied were: 1) Fourier analysis: each profile was considered as an irregular but periodic function obtained by the sum of sinusoids of increasing order. 2) Shape Asymmetry Evaluation: couples of profiles are compared by means of "Janus" procedure; by a parabolic fitting we obtained parameters able to evaluate allometric and isometric differences between the two profiles. The preliminary results of the applied procedures indicated that rate and direction of allometric and isometric growth are not constant during the time.

[Identification by superimposition of the skull on face photography using television: experimental application of a new method]. / Identificazione mediante sovrapposizione cranio-poto del viso a mezzo di circuito televisivo: applicazione sperimentale di una nuova metodica.

A new method of skull-face superimposition for forensic positive identification has been carried out. A single TV camera equipment with a special optical device was used. The tested specimens (22) were able to demonstrate the necessity to standardize the method in order to avoid errors.

[Vectorial description of the horizontal craniogram]. / Descrizione vettoriale della forma di craniogrammi orizzontali.

After the polynomial approximation we used Bezier's algorithm to obtain a vectorial description and to compare the horizontal craniogram morphology of hominid skulls. A minimum number of determinant points chosen on the original curves constitutes the vertices of a closed polygonal. By these points Bezier's interpolation gives new curves very similar to the polynomial curves obtained by the original ones. Each pair of consecutive segments was used to construct a parallelogram and so to have a vector. We applied the procedure on horizontal craniograms of Sts V. Pithecanthropus (erectus) VIII (Sartono), Circeo I (after S. Sergi) and a modern human female skull. Ten vectors for each skull were detected and compared. Some similarities in frontal and posterior regions of P VIII and Circeo I were explained in terms of common morphological solution to the problem of mechanical stability in the skulls with pronounced platycrany under chewing load.

[Evaluation of skull vault asymmetry using methods of analytical morphometry]. / Valutazione delle asimmetrie della volta cranica con metodi di morfometria analitica.

Some vertical frontal C.A.T. sections of 8 skull vaults were studied in evaluating shape asymmetry between left and right curves. Every profile was normalized by subdividing in 120 points and standardized by positioning the sagittal line of skull vault profile parallel to the ordinate axis of a Cartesian system. After these procedures, we tried to detect the asymmetry. In order to achieve these results we used a procedure that we named "Janus", a subset of S.A.M. (Shape Analytical Morphometry) software system, giving an artificial close curve by the mirror composition of left and right cranial vault profiles to match each other. This match was performed by a parabolic fitting for a standard positioning. The procedure was able to split the whole asymmetry information into the isometric and allometric components. We tried to detect allometry in terms of direction and versus with a numerical evaluator of its magnitude; this description was properly a vector. In this pilot study by using 24 sections, we found 20 left and 4 right predominance, expressed time by time by isometry, allometry or both.

[Definition of the analytical relation between the surface and internal perimeter of seminiferous tubules from subjects with normal spermatogenesis]. / Definizione della relazione analitica tra superficie e perimetro interno dei tubuli seminiferi del soggetto con normale spermatogenesi.

Luminal tubular measurements are executed in testis with normal spermatogenesis to individuate, by computerized morphometric method, the ratio between these two values. Statistical evaluations and calculation of the regression curves demonstrate that this ratio is well described by means of an exponential equation which coefficients are determined. This parameter results very useful in the diagnosis of Sertoli cell only syndrome and maturation arrest both characterized by tubular lumen modification.

[Analytical description and comparison of profiles and sections of the skull by polynomial equations. Illustration of the procedure]. / Descrizione analitica e comparazione di profili e sezioni del cranio a mezzo di equazioni polinomiali. Illustrazione della procedura.

Skull profiles and related sections may be worked in order to obtain the equation of the curve for best fitting. Polynomials of 6-7 nth degree give very high values of coefficient of determination. The technique is fully automatized and requires a TV-camera interfaced with a computer with specific hardware devices and software packages. Main routine gives coefficients of the equation, their standard deviation, standard errors, variance and covariance matrix. Accessory routines provide standardization of the position and apparent size of the image (skull or a photograph in a appropriate view) to work. General data on the fit of triplets of equations (total, frontal and facial of the anterior profile) are reported for Proconsul africanus, Australopithecus africanus (STS5, Plesianthropus transvaalensis), Australopithecus africanus (Taung I), Australopithecus Boisei (OH5, Zinjanthropus, rec. Tobias), Homo erectus (Pithecanthropus, rec. Weidenreich), Rhodesia man (Broken Hill I, E686), Homo S. neanderthalensis (Circeo I), Homo S. Sapiens (Grimaldi s.c. negroid, Grotta dei fanciulli VI, Predmost I, modern), Gorilla gorilla, Pan troglodytes, Macaca rhesus.

[Descriptive equations of the frontal-facial profile of Australopithecus africanus STS5 (Plesianthropus transvaalensis)]. / Equazioni descrittive del profilo fronto-facciale di Australopithecus africanus STS5 (Plesianthropus transvaalensis).

By means of the method previously carried-out by performing computerized image analysis we obtained three polinomial equations (7 degree) of best curve fitting for total (fronto-facial), frontal and facial profiles respectively of a standard lateral view of the skull Plesianthropus transvaalensis. The smooth function curves approximating the historical data are perfectly coincident with the scatter diagram for frontal (variance = 0,41) and facial (variance = 0,22) equations, while for the total contour variance raises to 90,8. The finding of the best fit equation is performed by analysis of variance simultaneously to the computation of the coefficients of progressively increasing order equations starting from the second. Then coefficient of determination, standard error of estimate. Standard deviations of coefficients and variance/covariance matrix are calculated with some dimensional parameters as length, eight and projections of profile, relate indexes and arc-chord ratio.

[Analytical morphometry of seminiferous tubules in maturation arrest: Fourier analysis of the internal contours of tubules]. / Morfometria analitica dei tubuli seminiferi nell'arresto maturativo: analisi di Fourier del contorno interno dei tubuli.

Shape of seminiferous tubuli in maturation arrest, is investigated by harmonic Fourier analysis and upper degree polynomials as attempt to find morphometric parameters useful in morphological diagnosis. Inner luminal contour is considered as a closed curve in a reference system and scattered in a series of points whose coordinates are automatically calculated. Then a software named S.A.M. (Shape Analytical Morphometry) is utilized to find the Kth-order equation and calculate the best-fit curve. Moreover an ellypse is drawn with the same barycenter and axes of original curve. These three curves (real curve, best-fit curve and ellypse) are considered as periodic functions and submitted to Fourier analysis to evaluate the coefficients of the series and the spectrum of harmonics. Among 20 contributors, the subsets of greatest amplitude are selected for comparison and classification in terms of fundamental shape and its perturbations.

[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.