Analytical Cellular Pathology - Volume 16, issue 1

Authors: Losa, Gabriele A. | Graber, Riccardo

Article Type: Research Article

Abstract: The proliferative capacity (%S‐phase fraction), DNA ploidy, apoptosis frequency (DNA fragmentation) and steroid hormone receptor status (estrogen receptor, ER; progesterone receptor, PR) of 110 samples of human breast tissues with ductal invasive carcinoma were measured using biochemical and cytofluorimetric procedures. The DNA fragmentation had a left‐skewed frequency distribution and an overall median value of 1.64%, whilst the median %S‐phase fraction was 8%. The median %DNA fragmentation and %S‐phase fraction were 1.96% and 16% in hyperdiploid tumours (n=29 ; DNA index > 1.1) higher than in hypodiploid tumors (n=10 ; DNA index < 0.96), 0.38% and 7.5%. …DNA diploid tumours (n=71 ) had median %DNA fragmentation and %S‐phase values of 1.68% and 6%, consistently lower than the median values of DNA hyperdiploid tumours. The ER content of hypodiploid tumours was about one half (median: 5.9 fmol/mg) the median values in hyperdiploid (10.6 fmol/mg) and diploid tumours (14.6 fmol/mg). This may correlate with the lowest frequency of apoptosis in hypodiploid tumours, at least when measured by biochemical methods which only detect cells in the late phases of apoptosis. In contrast, the median PR was lowest in hyperdiploid tumours than in hypo and/or diploid tumours. The %S‐phase/%fragmented DNA ratio for the hypodiploid tumours was 19.7, significantly higher than the ratios for hyperdiploid (8.2) and diploid tumours (3.6). These findings indicated that there is an imbalance between proliferative capacity and cell death or growth arrest in human breast tumours. This imbalance may well be linked to a loss of steroid hormone control. Show more

Keywords: Apoptosis, human breast cancer, DNA ploidy, DNA fragmentation, steroid hormone receptors, %S‐phase‐proliferative capacity

Citation: Analytical Cellular Pathology, vol. 16, no. 1, pp. 1-10, 1998

Authors: Hanselaar, Antonius G.J.M. | Poulin, Neal | Pahlplatz, Martin M.M. | Garner, David | MacAulay, Calum | Matisic, Jasenka | LeRiche, Jean | Palcic, Branko

Article Type: Research Article

Abstract: A retrospective analysis was performed on archival cervical smears from a group of 56 women with cervical intraepithelial neoplasia (CIN), who had received follow‐up by cytology only. Automated image cytometry of Feulgen‐stained DNA was used to determine the differences between progressive and regressive lesions. The first group of 30 smears was from women who had developed cancer after initial smears with dysplastic changes (progressive group). The second group of 26 smears with dysplastic changes had shown regression to normal (regressive group). The goal of the study was to determine if differences in cytometric features existed between the progressive and regressive …groups. CIN categories I, II and III were represented in both groups, and measurements were pooled across diagnostic categories. Images of up to 700 intermediate cells were obtained from each slide, and cells were scanned exhaustively for the detection of diagnostic cells. Discriminant function analysis was performed for both intermediate and diagnostic cells. The most significant differences between the groups were found for diagnostic cells, with a cell classification accuracy of 82%. Intermediate cells could be classified with 60% accuracy. Cytometric features which afforded the best discrimination were characteristic of the chromatin organization in diagnostic cells (nuclear texture). Slide classification was performed by thresholding the number of cells which exhibited progression associated changes (PAC) in chromatin configuration, with an accuracy of 93 and 73% for diagnostic and intermediate cells, respectively. These results indicate that regardless of the extent of nuclear atypia as reflected in the CIN category, features of chromatin organization can potentially be used to predict the malignant or progressive potential of CIN lesions. Show more

Keywords: Uterine cervix, dysplasia, image analysis, automation, DNA

Citation: Analytical Cellular Pathology, vol. 16, no. 1, pp. 11-27, 1998

Authors: Ranefall, Petter | Wester, Kenneth | Bengtsson, Ewert

Article Type: Research Article

Abstract: A method for quantification of images of immunohistochemically stained cell nuclei by computing area proportions is presented. The image is transformed by a principal component transform. The resulting first component image is used to segment the objects from the background using dynamic thresholding of the P^2 /A ‐histogram, where P^2 /A is a global roundness measure. Then the image is transformed into principal component hue, defined as the angle around the first principal component. This image is used to segment positive and negative objects. The method is fully automatic and the principal component approach makes …it robust with respect to illumination and focus settings. An independent test set consisting of images grabbed with different focus and illumination for each field of view was used to test the method, and the proposed method showed less variation than the intraoperator variation using supervised Maximum Likelihood classification. Show more

Keywords: Image analysis, principal components, automatic quantification, immunohistochemically stained cell nuclei

Citation: Analytical Cellular Pathology, vol. 16, no. 1, pp. 29-43, 1998

Authors: Lopes, José M. | Hannisdal, Einar | Bjerkehagen, Bodil | Bruland, Øyvind S. | Danielsen, Håvard E. | Pettersen, Erik O. | Sobrinho‐Simões, Manuel | Nesland, Jahn M.

Article Type: Research Article

Abstract: Controversy still exists regarding the validity of parameters commonly used in the evaluation of prognosis of patients with synovial sarcoma (SS). Forty‐nine cases of previously untreated primary SS (23 females and 26 males, ranging in age from 7 to 81, with 31 tumors located in the lower extremity, 8 at the upper extremity and 10 at the trunchus), without regional lymph‐node or distant metastases were studied. We investigated the relationship between (flow and image) DNA cytometry, proliferation activity, clinicopathologic parameters, and relapse‐free and overall survival of the patients. The prognostic value of gender, age, duration of symptoms, location, compartmentalization, size, …adequacy of surgical margins, residual tumor, adjuvant therapy, histologic subtype, extent of necrosis, glandular differentiation, calcification, and extent of hemangiopericytic areas, mitotic rate, amount of mast cells, blood vessel invasion, histologic (UICC and NCI) grades, DNA ploidy, percentage of cells in S and S+G2 phases, PCNA and Ki‐67 labeling indices (LI), and TNM (UICC) stage of the tumors, were evaluated by univariate and multivariate (Cox hazard model) analyses. Short duration of symptoms (<12 months), biphasic SS, scarcity of mast cells (< 10/10 HPF), high mitotic rate ({\scriptstyle\geq} 10/10 HPF), high histologic grade (grade 3), high PCNA‐LI ({\scriptstyle\geq} 20%), high Ki‐67‐LI ({\scriptstyle\geq} 10%), DNA aneuploidy, and advanced TNM stage (stage III) were features associated with significantly shorter relapse‐free and overall 5‐year survival rates in the univariate analyses. Scarcity of mast cells, high mitotic rate, or high PCNA‐LI were significant predictors of poor survival, in addition to TNM stage in the multivariate analyses. The amount of mast cells was inversely correlated with mitotic rate and PCNA‐LI. Scarcity of mast cells, high mitotic rate, or high PCNA‐LI are factors associated with poor prognosis, in addition to advanced TNM stage in patients with localized SS. Show more

Keywords: Synovial sarcoma, mast cells, mitotic rate, PCNA, staging, prognosis, Ki‐67, DNA flow cytometry, DNA image cytometry, soft tissue tumor

Citation: Analytical Cellular Pathology, vol. 16, no. 1, pp. 45-62, 1998