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PubMed1998

[Fiber morphometry of the external intercostal muscle. Comparison of dominant and nondominant sides in patients with severe COPD].

The general morphometric characteristics of the external intercostal muscle (EIM) of patients with chronic respiratory disease have been well described. Because this muscle is highly accessible, it can provide an ideal model for longitudinal studies using consecutive biopsies of both sides. Whether or not the EIM fiber phenotype is homogeneous on dominant (D) and non dominant (ND) sides is unknown, however. To evaluate possible structural differences in right and left EIM in patients with COPD, eight patients (63 +/- 7 years of age) were enrolled. Lung function, respiratory muscle power, general muscle power and nutritional state were evaluated. Biopsies of the fifth EIM were taken from both sides. Specimens were processed in parallel manner to determine conventional morphometry (hematoxylin-eosin staining), including minimum diameter (Dm) and fiber area (Ar) in cross sections. Fibers were typed by ATPase (at pH 4.2, 4.6 and 9.4) and NADH-TR staining. Nutrition was normal in all patients. All patients had severe COPD (FEV1 27 +/- 7% of reference, limits 13 to 38% of reference) with air entrapment (RV 163 +/- 36% of reference, limits 181 to 276% of reference). None of the patients showed respiratory insufficiency at rest (PaO2 72 +/- 7 mmHg). Peripheral musculoskeletal power measured by manual dynamometer showed no significant right-left differences: D 29 +/- 2 and ND 28 +/- 3 dynes. Morphometric study of 16 muscle specimens showed no significant differences between fiber size on D and ND sides. DmD was 47 +/- 10 microns and ArD, was 2,595 +/- 1,249 microns2. DmD was 49 +/- 9 microns and ArD was 2,636 +/- 953 microns2. Likewise, no significant differences were found between D and ND fiber types: type ID 51 +/- 4% and type IID 49 +/- 5% versus type IND 52 +/- 4% and type IIND 48 +/- 4%. EIM on N and ND sides is homogeneous at the fifth intercostal space. This finding, along with the scarcely invasive nature of the technique for collecting specimens leads us to suggest that longitudinal studies might be performed on the structural effects of various pharmacological or physical treatments followed by COPD patients

Aged↗

PubMed1992

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.

Cell Nucleus↗

PubMed1992

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.

Diagnosis, Computer-Assisted↗

PubMed1992

Morphometry and prognosis in cancer of the pancreatic head.

The histological slides of 39 cases of cancer of the pancreatic head were analysed using an interactive image analyser system. Some 14 cases were classified as periampullary, 25 as ductal pancreatic cancer. All cases had undergone radical tumor resection according to Whipple's procedure. Morphometric data, tumor size and metastatic nodal involvement were correlated to prognosis. Univariate statistical analysis showed that the classical differentiation between ductal and periampullary cancer was a weaker prognosticator than morphometric variables. In fact, multivariant statistical analysis showed that the morphometric variable irregularity was the best prognosticator (p = 0.0001). No other variable added significant prognostic information. Irregularity is a newly developed variable describing the nuclear shape corrected for roundness. We conclude that morphometry can be of essential prognostic information for the clinician in cancer of the pancreatic head.

Carcinoma, Intraductal, Noninfiltrating↗

PubMed1992

Cytonuclear morphometry in the assessment of dysplasia in colorectal adenomatous polyps. A pilot study.

In order to objectify the degree of dysplasia, results of nuclear and cellular morphometry were compared with assessed grades of dysplasia in 32 colorectal adenomatous polyps. Of these adenomas 8 showed mild, 17 moderate and 7 severe dysplasia (according to blind duplicate assessments by two pathologists). Using a microscope video-overlay interactive digitizing measurement system, 100 nuclei were measured in each specimen according to a strict measurement protocol. In a stepwise discriminant analysis the best discriminating features appeared to be mean stratification index, N/C ratio, mean contour ratio and mean form AR. Overall with these features 65.6% correct jackknifed classification of the 32 cases could be achieved. On the other hand, a clear three-group distinction could not be obtained, even with the best set of discriminating features. Similarly to gastric dysplasia, the morphometric features might be used to design a two-grade (low, high) rather than a three-grade system to assess the degree of dysplasia. These findings make clear that objectifying the degree of dysplasia in adenomatous polyps is possible by means of interactive morphometric analysis.

Adenoma↗

PubMed1992

Prediction of response to adjuvant chemotherapy in premenopausal lymph node positive breast cancer patients with morphometry, DNA flow cytometry and HER-2/neu oncoprotein expression. Preliminary results.

The value of morphometry, DNA flow cytometry and HER-2/neu oncoprotein expression for prediction of response to adjuvant chemotherapy in premenopausal lymph node positive breast cancer patients was evaluated in a group of CMF treated patients and controls with long-term follow-up. In the treated group, the Morphometric Prognostic Index (cutpoint 1.1) was the best prognosticator (p less than 0.0001, MC = 16.9), followed by the Mitotic Activity Index, the volume percentage epithelium and the number of positive nodes. For the controls, only the % HER-2/neu oncoprotein expression revealed significant differences (p less than 0.0001, MC = 16.3). When directly comparing treated patients and controls stratified for a certain parameter, no significant differences were obtained, although a trend towards improved survival in the treated group was present for some of the subgroups for several parameters. These preliminary results indicate that morphometric features and quantitative HER-2/neu oncoprotein expression may be important factors for identifying cases that will or will not respond to adjuvant chemotherapy.

Adult↗

PubMed1992

Correlation of nucleolar organizer regions and nuclear morphometry assessed by automatic image analysis in breast cancer with aneuploidy, K167 immunostaining, histopathologic grade and lymph node involvement.

Silver-stained nucleolar organizer regions (AgNORs) in human breast carcinoma were studied using a computer-assisted system of image analysis. Standardized, automatic measurements of 7 morphometric parameters (area, perimeter, shape factor, bend energy, angle, and small and large diameters) performed on paraffin sections and cell imprint were compared and correlated with nuclear morphometry, histopathological grading, tumor growth fraction, (monoclonal Ki67-immunostaining), DNA nuclear content (stoechiometric Feulgen staining) and axillary lymph node invasion. The major findings were as follows: (i) variations in AgNORs and nuclear parameters were correlated, (ii) the ratio of AgNOR area/nuclear area was significantly different in low and high grade tumors, (iii) mean AgNOR parameter values increased significantly with the tumor growth fraction and tumor hyperploidy and were significantly higher in patients with axillary lymph node metastases and (iv) AgNOR evaluation was more accurate for cell preparations than for tissue sections.

Antibodies, Monoclonal↗

PubMed1978

Standardized fixation of human lung for radiology and morphometry; Description of a "two chamber"-system with formaldehyde vapor inflation.

In order to obtain optimal preservation of lungs for histology, morphometry and radiology a two-chamber system of lung fixation with hot formaldehyde vapor is introduced. Compared with other procedures this system offers small dead space and ensures quick and thorough fixation by imitating "inspiration" and "expiration". Volume- and pressure-regulated "breathing" enables fixation in a stage of the respiration cycle, which can be chosen. This procedure can be used routinely in the autopsy room. The only serious drawback seems to be air pollution, if the chambers are not absolutely insulated.

Air Pollution↗

PubMed1980

Ultrastructural morphometry of isolated cells: methods, models and applications.

Isolated cells constitute a favourable experimental model for the pathologist interested in ultrastructural morphometry. A particular advantage is that the same sampling regime may be appropriate for both normal and pathologically-altered material. Often this is not the case with other model systems. This report reviews alternative methodologies for analysing isolated cells, including free cells and dissociated cells in culture. We begin by attempting to define problems of more general concern: preparative techniques, methods of sampling, selection of stereological parameters and possible sources of error. A generalised morphometric model for isolated cells is followed by descriptions of models devised for specific cell types, notably mononuclear phagocytes and cells of the lymphocyte family. Typical results obtained by stereological analyses of these models are presented.

Animals↗

PubMed1986

Morphometry of breast cancer. I. Comparison of the primary tumours and the axillary lymph node metastases.

In 234 patients with primary breast cancer, morphometry was performed of three groups of tumour cells: 1. in the primary tumours without lymph node metastases (PRN); 2. in primary tumours with lymph node metastases (PRP) and; 3. in the axillary lymph node metastatic deposits (LMD). The morphometrical analysis included the cellularity index, the mitotic activity index and seven nuclear features. The highest mitotic activity indices (mean: 14.1) were found in the metastatic deposits (LMD), the lowest (mean: 12.5) in the lymph node negative primary tumours (PRN), whereas the lymph node positive primary tumours (PRP) fell in between (mean: 13.2). The cellularity index shows a similar, though reciprocal, trend. The largest standard deviations of the nuclear features were found in primary tumours with lymph node metastases (PRP). In contrast, the metastatic (LMD) nuclei had the smallest standard deviations. The nuclei of the latter are therefore more regular in size and shape. The differences found are not sufficiently large to distinguish PRN and PRP with an acceptable accuracy (the efficiency, sensitivity or specificity did not exceed 67% irrespective of the decision threshold used). Comparison of primary tumours with lymph node metastases and the metastatic deposits, from the same patients, revealed significant correlations of all morphometric features with the exception of the nuclear shape factor and the nuclear axes ratio. Thus, the nuclei of the metastatic cells in general are identical to the nuclei in the primary tumour from which they arise, but in a subset of patients the metastatic tumour cells have much more monomorphous ellipsoid nuclei. Further investigations are needed to see whether this phenomenon indicates a different malignant course.

Adult↗

PubMed1988

Pituitary pathology in Cushing's disease. Histology and morphometry of pituitary tissues removed through microsurgery.

58 cases of Cushing's disease were studied after selective transsphenoidal surgery, by means of histology, immunocytochemistry and morphometry. The qualitative and quantitative results, related to the clinical follow-up of the patients indicated that: 1) when a well-defined tumor was found (62% of the patients), successful outcomes occurred in 88% of the cases, 2) when a tumor could not be found, successful outcomes amounted to only 33% of the cases, a rate that might be related to a surgical exploration of the basophil cell zone "invading" the neural lobe, thus questioning a role of this zone, at least in some Cushing's diseases.

Adenoma↗

PubMed1989

Mapping intestinal metaplasia by histochemistry and morphometry.

A gastrectomy specimen housing an adenocarcinoma of intestinal type was divided into 60 blocks. Sections were stained with Alcian blue pH 2.5 (to detect foreign mucins). Alcian blue fields were quantitated with a MOP 30 connected to a computer. The mucosa analysed in the 60 sections measured 3788.3 mm2 and that occupied by Alcian blue stained goblet cells 300.9 mm2 (or 7.9% of the total mucosal area). The number of Alcian blue positive fields was significantly larger in the intermediate zone followed by the antral zone, the lowest value was found in the corpus. However, the gastric mucosa with Alcian blue positive cells was larger in the antrum than in the intermediate zone due to the fact that the area of the fields with Alcian blue positive cells was significantly larger in the antrum than in the intermediate zone. The size and number of Alcian blue positive fields were significantly larger along the greater than along the smaller curvature. All measurements were done by a trained technician. The method (using histochemistry, morphometry and skilled technical assistance) will be applied to study the distribution of intestinal metaplasia in gastrectomy specimens from populations with disparate incidences of gastric carcinoma.

Adult↗

PubMed1989

Potential of morphometry in grading transitional cell carcinoma of the urinary bladder.

The potential of morphometry in grading cases of transitional cell carcinoma of the urinary bladder was evaluated. Thirty cases of bladder cancer including all three WHO grades were studied. Three investigators measured the nuclear areas using the IBAS 1&2 image analyser system. The means, the standard deviations and the variances of the measurements were calculated. The nuclear areas covered values from 25.1 to 107.4 square micrometers, the mean +/- SD being 52.6 +/- 17.5. The total variance of measurements, including biological and methodological variation, was 304.6, and the methodological variance 10.4 (about 3.5% of the total variation). In a 3-grade grading system this would correspond to an efficiency of about 92%, and in a 2-grade system of about 96%, which are the percentages correctly graded by the grading system.

Carcinoma, Transitional Cell↗

PubMed1989

The value of morphometry to predict chemotherapy response in advanced ovarian cancer.

We evaluated the correlation of morphometric parameters to chemosensitivity. 63 patients with palliatively operated advanced epithelial ovarian cancer were investigated concerning their response to chemotherapy. Using multiple linear stepwise discriminant analysis of five morphometrical parameters 13 out of 17 responders and 37 out of 46 non-responders were correctly classified (76.5% sensitivity, 80.4% specificity, 79.4% efficiency). The five parameters were: nuclear area at the 10th percentile, standard deviation of the nuclear area, median value of the nuclear ovality, number of cells per area and mitotic activity index. To assess the performance of the discrimination formula when applied to new cases, the "leave one out" method was used. For our data the following corrected classification rates were obtained: 58.8% responders (10/17), 76.1% non-responders (35/46) (efficiency 71.4%). Morphometry is a fast and reproducible method to objectively record a tumor's morphology. Our results indicate that there is a correlation between morphometrical features, response to chemotherapy and survival, which should be tested in further studies.

Adult↗

PubMed1989

Morphometry of in vitro systems. An image analysis of two human prostate cancer cell lines (PC3 and DU-145).

Morphometric analysis on two human prostate cancer cell lines (PC3 and DU-145) was carried out to better characterize these cells and to approach some problems concerning their polymorphism. In our hands, these cells appear to differ either in their ability to oxidize testosterone or in their androgen receptor content. Morphometric evaluation was performed by means of an Interactive Image Analysis System (IBAS 1), which computes several parameters related to the whole cell and to the nucleus. Our results clearly indicate that nuclear parameters alone may discriminate PC3 from DU-145 cells. Particularly, the Nuclear Roundness Factor appeared important, since it quantifies changes in nuclear shape. Nevertheless, the value of this parameter for "in vitro" morphometry may be overshadowed by the polymorphism common to the cultured cells. However, morphometric evaluation of PC3 and DU-145 cells is relevant to a better understanding and definition of the biologic nature of these cells, even if polymorphism, heterogeneity and genetic instability, specially of PC3, require further investigations.

Cell Nucleus↗

PubMed1989

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.

Biopsy↗

PubMed1999

Benign, preinvasive and invasive ductal breast lesions. A comparative study with quantitative techniques: morphometry, image- and flow cytometry.

The histological distinction between ductal hyperplasia of the breast, atypical ductal hyperplasia and ductal carcinoma in situ is difficult and subjective. To gain a better understanding of these lesions, we performed a comparative study comprising 20 cases of ductal hyperplasia without atypia, 20 cases of ductal hyperplasia with atypia, and 30 cases of ductal carcinoma in situ (well-, moderately- and poorly-differentiated), using quantitative techniques: image cytometry analysis, morphometry and DNA analysis, and DNA flow cytometry. Our results confirm that the mean nuclear area and volume progressively decreased from ductal carcinoma in situ to ductal hyperplasia without atypia. The difference was significant (p < 0.05) when comparing hyperplasia without atypia with hyperplasia with atypia and hyperplasia with atypia with poorly differentiated ductal carcinoma in situ. Atypical ductal hyperplasia values were comparable to those of well-differentiated ductal carcinoma in situ. DNA image cytometry proved significant (p < 0.05) when comparing hyperplasia without atypia with hyperplasia with atypia and hyperplasia with atypia with moderately- and poorly-differentiated ductal carcinoma in situ. DNA flow cytometry revealed significant differences only in the distribution of the DNA ploidy patterns (p < 0.05) when comparing hyperplasia without atypia with moderately- and poorly-differentiated DCIS. A comparison of the results obtained by image and flow cytometry showed that in 90% of the cases the IC and FC values were coincident, whereas in the remaining cases the DNA index was aneuploid by IC and diploid by FC.

Breast Neoplasms↗

PubMed1998

Nuclear morphometry of the myocardial cells as a diagnostic tool in cases of sudden death due to coronary thrombosis.

Sudden cardiac death due to underlying coronary artery thrombosis is one of the leading causes of death. However, in a significant percentage of individuals who died suddenly, no indication of myocardial infarction is found during post-mortem examination, especially when the time interval between appearance of symptoms and death is short. In the present study, we have evaluated certain nuclear morphometric parameters, such as, minimum, maximum, mean and standard deviation of perimeter and area in 20 individuals who died of coronary artery thrombosis, within 1 h from symptoms onset. Furthermore, the above parameters were compared with those of a control population of 20 individuals whose sudden death was caused by traffic accidents. Statistical elaboration of the results by means of t-test, Mann-Whitney (U-test) and analysis of covariance (adjusting for age), showed a statistically significant difference for all variables except for the minimum area. With stepwise discriminant analysis method, the mean perimeter was selected as the best predictor of cardiac death. Mean perimeter achieved a correct reclassification percentage (based on Fisher's linear discriminant function) of 92.5% (85% and 100% for cases and controls, respectively). Moreover, by applying the cut-off of 172 microns, we could identify the individuals who died suddenly because of coronary artery thrombosis with a specificity of 100% (sensitivity 85%, P < 0.001). Our results show that nuclear morphometry of the myocardial cells is a reliable diagnostic tool for the diagnosis of coronary thrombosis based lesion in cases of sudden death, even when methods trying to verify the presence of infarction fail to do so.

Case-Control Studies↗