Image cytometry determination of ploidy level, proliferative activity, and nuclear size in a series of 314 transitional bladder cell carcinomas.

Image cytometry was carried out on 281 superficial (Ta and T1) and 33 invasive (T2 to T4) bladder cancers. The parameters used to characterize these bladder tumors were: (1) histopathological grading, (2) clinical staging, (3) tumor size, (4) deoxyribonucleic acid (DNA) index (DI), (5) DNA histogram type (DHT), (6) percentage of euploid (diploid plus tetraploid) cells, (7) percentage of polyploid cells (> 5C DNA content), (8) proliferative activity (S phase fraction value), and (9) nuclear area (NA). The proliferative activity of the tumors was not related to either histopathological grade or to clinical stage, but it was related to the DHT parameter, which made it possible to identify diploid, hyperdiploid, triploid, hypertriploid, tetraploid, and polymorphic tumors. The hypertriploid tumors exhibited a significantly lower proliferative activity than the nonhypertriploid ones. Although both the DI and the NA values correlated significantly with histopathological grading, only the NA values correlated significantly with clinical staging. We further observed that some grade III bladder tumors were definitely diploid, whereas some grade I tumors were highly aneuploid. We thus hypothesize that the ploidy level of a given tumor reflects its age directly and its aggressiveness only very indirectly. In our opinion aneuploidy is only an indirect marker of aggressiveness because it reflects the fact that a malignant tumor is old, ie, has been present in a patient over a long period of time and has had ample time to express its malignancy at the clinical level. A significant relationship was accordingly obtained between tumor size and ploidy level with the highest proportion of aneuploid tumors and the highest percentage of polyploid cell nuclei being observed among the largest bladder tumors.

Digital cell image analysis of Feulgen-stained nuclei from human papillary, medullary, colloid, lobular and comedocarcinomas of the breast.

The morphonuclear characteristics (nuclear size and chromatin pattern), the proliferation index and the ploidy level were characterized in a series of 46 breast tumors including medullary (5 cases), papillary (6 cases), lobular (27 cases), colloid (4 cases) and comedo- (4 cases) carcinomas. The quantitative assessments were carried out by means of digital cell image analyses of Feulgen-stained nuclei from imprint smears. The results show that monovariate analyses (one-way variance analyses) were much less potent than multivariate analyses (principal components analyses followed by the canonical transformation of the data and discriminant analyses) in assessing the morphonuclear characteristics of these breast tumors. The multivariate analyses indicated that there might be a level of malignancy which increases according to the sequence papillary and medullary and colloid carcinomas-->comedocarcinomas-->lobular carcinomas. This assertion is corroborated by the ploidy-level-related results which revealed a higher proportion of highly aneuploid cases in the group of lobular carcinomas than in the group which included medullary, papillary and colloid carcinomas. However, since highly aneuploid cases were also encountered in this latter low malignancy level group, we expressed the hypothesis firstly that aneuploidy reflects two distinct biological properties, i.e. the aggressiveness of a tumor and its age, and secondly that a highly aneuploid but low malignancy tumor should correspond to old degenerating tumors.

Determination of DNA ploidy, nuclear size, and proliferative activity by means of the computer-assisted image analysis of Feulgen-stained nuclei in 68 soft tissue tumors of adults.

The diagnostic values of the ploidy level, the proliferative activity, and the nuclear size in a series of 68 soft tissue tumors of adults were determined by digital cell image analysis of Feulgen-stained nuclei from formalin-fixed, paraffin-embedded tissues. The DNA ploidy level was characterized by calculating the DNA index (DI) and the percentage of the diploid and polyploid cells, and by typing the DNA histogram. Proliferative activity assessments were a function of the determination of the proliferation index (PI), ie, the percentage of cells engaged in the S phase of the cell cycle (SPF value). The present series included 19 benign and 49 malignant soft tissue tumors. The results show that DNA aneuploidy, as assessed by both the DI and the DNA histogram type, cannot be used as a discriminatory parameter for distinguishing between benign and malignant soft tissue tumors. Indeed, some benign cases may be highly aneuploid, whereas some highly malignant soft tissue tumors may be definitely diploid. In contrast, the determination of the percentage of polyploid cell nuclei seems to be a useful parameter in distinguishing between benign and malignant cases. In fact, the benign soft tissue tumors showed a very significantly lower mean percentage value of polyploid cell nuclei than the malignant cases. The determination of the proliferative activity also discriminated significantly between the benign and the malignant cases, the former proliferating more slowly than the latter. Lastly, the determination of nuclear size made it possible to differentiate the primary malignant soft tissue tumors, whether recurrent or not, that were associated with metastasis from those free of metastasis.

Computer-assisted chromatin texture characterization of Feulgen-stained nuclei in a series of 331 transitional bladder cell carcinomas.

The chromatin patterns of Feulgen-stained nuclei in a series of six specimens of normal mucosa and 331 transitional bladder carcinomas, including 293 superficial (Ta and T1) and 38 invasive (T2-T4) cases, were quantitatively described by means of eight parameters relating to densitometric, run-length distribution, and co-occurrence matrix features. The results show that the chromatin texture of the superficial lesions was markedly different from that of the invasive tumours, which exhibited a distinctly more dense and heterogeneous chromatin pattern. The data also show that the increasing level of malignancy, as revealed by the increasing clinical stage, was accompanied by an increase in the overall chromatin condensation level. Only some areas of the nucleus actually increased in density; other pale areas appeared concomitantly with these increasingly denser chromatin areas. This chromatin density increase corresponded to a marked increase in the frequency of small dense chromatin clumps; these joined together into very large dense chromatin clumps, which were distributed more and more heterogeneously in the nucleus as the clinical stage of the tumour increased.