The possibility that computerized image analysis could increase the reproducibility of grading of bladder carcinoma as compared to conventional subjective grading made by pathologists was investigated. Object, texture and graph based analysis were carried out from Feulgen stained histological tissue sections. The object based features were extracted from gray scale images, binary images obtained by thresholding the nuclei and several other images derived through image processing operations. The textural features were based on the spatial gray‐tone co‐occurrence probability matrices and the graph based features were extracted from the minimum spanning trees connecting all nuclei. The large numbers of extracted features were evaluated in relation to subjective grading and to factors related to prognosis using multivariate statistical methods and multilayer backpropagation neural networks. All the methods were originally developed and tested on material from one patient and then tested for reproducibility on entirely different patient material. The results indicate reasonably good reproducibility for the best sets of features. In addition, image analysis based grading showed almost identical correlation to mitotic density and expression of p53 protein as subjective grading. It should thus be possible to use this kind of image analysis as a prognostic tool for bladder carcinoma.

The prognostic value of 435 cytochemical, cytometrical, morphological, epidemiological, and clinical variables was analyzed in a prospective study of 179 breast cancer patients followed for five years after mastectomy. A variable reduction was obtained by first selecting variables correlated with recurrence rate in direct (Student's t test) or correlation analysis with consideration of the type of variable analyzed (nominal, interval, ordinal). The 20 variables most strongly correlated with recurrence were analyzed by logistic stepwise regression analysis in order to find out what combination of variables had most discriminatory power in predicting recurrence. It was found that axillary metastization as such was correlated with a combination of variables describing mitotic frequency, size of primary tumor and differentiation of the primary tumor (average cluster size in fine‐needle biopsies). It was also found that there was a strong time dependency in the predictive power of the variables, so that different variable combinations predicted the recurrence rate during the first 2.5 year period (size of axillary metastases and primary tumor, number of lymphocytes around the tumor, mitotic frequency, and degree of differentiation) compared with the second 2.5 year period (variance of DNA content among tumor cell nuclei, number of lymphocytes around the tumor, occurrence of multiple tumors in the operated breast and occurrence of breast cancer among relatives). While other factors previously shown to be correlated with risk of recurrence were also found to be positively correlated here, they were neither as highly predictive as, nor did they increase the predictive value of the above mentioned combined variables. The current study strongly emphasizes that, at the present time, studies of recurrence prediction in human breast cancer should be based on an optimal combination of a number of variables which, independently, influence the prognosis. Further, the current study indicates that prerequisite methods for predicting breast cancer recurrence exist today.

SUMMARY The inter-and intraobserver reproducibilities of the histopathological systems of breast cancer classification suggested by the World Health Organisation (WHO), the Armed Forces Institute of Pathology (AFIP) and Ackerman have been analysed. The reproducibilities of the three classification systems were only "fair" to "moderate" and no correlation with the five-year recurrence rate was found. Our results indicate that these classification systems are without biological significance and are useless for prognosis in the individual patient.When the tumours were classified according to degree of differentiation (high, moderate, low) or graded according to WHO (which includes both differentiation and nuclear atypia), however, there was a significant correlation with the five-year recurrence rate. Yet even such "reduced" subdivisions are of no value in judging prognosis for the individual patient at the time of diagnosis; rather, they are useful only in the follow-up analysis of groups of patients. The cancers were unilateral and none had been given preoperative radiotherapy. The methods of operation varied. Either a simple mastectomy, radical mastectomy or mastectomy with exploration or exaeresis of the axilla was performed. After the operation for mammary carcinoma the patients were regularly checked at their local hospitals. Information concerning the occurrence of metastases or death was obtained from hospital records. The patients were followed up for five years. TYPES OF SPECIMENSMaterial for histopathological examination was obtained from all the patients subjected to surgical intervention-that is, exploratory biopsy or mastectomy. On removal all specimens were marked by the surgeon at the 12 o'clock position. Immediately after removal, the tumour was cut through and the two longest, perpendicular diameters were measured.A slice (approx 3 mm thick) of the tumour and its 392 on 31 March 2019 by guest. Protected by copyright.

An evaluation of cytometric features of 142 human breast cancers was performed. Twenty-two cytometric features describing density, structure and shape were correlated with mitotic frequency and labeling index of some of the tumors and with the observed recurrence rate of the tumors within 3 years after removal of the breast. A new cytometric measure was also created by combining the four features most strongly correlated with mitotic frequency of the tumors. It was found that the variance of the nuclear area was the feature that was most strongly correlated with mitotic frequency and recurrence rate. No new cytometric measure created by combining the four features most strongly correlated with mitotic frequency, correlated as well as variance of nuclear area even though the four features were optimized by multiple regression analysis for their correlation with mitotic frequency.Key terms: Breast cancer, mitotic frequency, cytometric features One of the important components of cytometric work in cancer diagnosis is the evaluation of features (13) and their correlation to other biologically important factors. Surprisingly little has been accomplished in the field of feature evaluation although more than 20 years ago several groups using different modes of spectrophotometry in visible light began to pursue DNA determinations in tumor materials. Although abnormal DNA distribution patterns were noted in most human tumors few or no attempts have been made to "grade" these abnormal patterns or to relate them to other observations such as mitotic frequency or patient survival rather than conventional morphology. In many tumors, including breast carcinoma, the mitotic frequency and/or labeling index after incubation with tritiated thymidine are expressions of DNA-synthesis activity.In a study of the possible correlations between morphologic and epidemiologic characteristics of breast cancer we have developed a method of cytometric characterization of tumor cell nuclei populations (14). This method had a comparatively high reproducibility and consistency.In the present study the various morphometric features of I Supported by NIH/NCI Contract NOlCB53968.the nuclei populations of breast carcinoma are correlated with mitotic frequency of the tumors and in one sample of the populations also with the labeling index.Materials and Methods Determination of Cytometric Features: The cytometric features were obtained from the tumors of I42 breast cancer patients. After removal each specimen was taken directly to the laboratory. Cells were aspirated from the tumor with a 22-gauge needle and a 20-ml syringe in a syringe holder. This method produces a proportion of single dispersed cells that are well-suited to cytometric studies. The cytologic specimens were fixed immediately in fresh Carnoy's solution and stained with chrome-alum gallocyanin (3, 4). One hundred nuclei from each specimen were recorded. Randomly selected undamaged epithelial cells were chosen for recording.A diode array scanner coupled with a light microscope was used...