Analytical Cellular Pathology - Volume 17, issue 3

Authors: Fležar, Margareta | Doudkine, Alexei | Us‐Krašovec, Marija

Article Type: Research Article

Abstract: The effect of three primary fixation procedures, used in the preparation of routine cytological samples: air‐drying, Delaunay, and Saccomanno fixation, with postfixation in modified Böhm–Sprenger fixative, on nuclear features as a function of hydrolysis time is reported. Three different cell types: lymphatic cells (tonsil), epithelial cells (buccal mucosa) and mesenchymal cells (uterine myometrium) were used for the study. Our findings show, that generally not all features have the same plateau times as the IOD (integrated optical density), and that many features show different values depending on cell type and fixation method. It is therefore recommended that for any primary fixative …used in routine clinical work and for each cell type, the hydrolysis curve for all nuclear features to be used in sample analysis should be established. Show more

Keywords: Image cytometry, fixation, Feulgen reaction, hydrolysis, nuclear texture features

Citation: Analytical Cellular Pathology, vol. 17, no. 3, pp. 131-144, 1998

Authors: Forestier, Erik | Holmgren, Gösta | Roos, Göran

Article Type: Research Article

Abstract: Flow cytometric DNA‐index (DI^{\rm FCM} ) and karyotype were analysed in 82 consecutive children with acute lymphoblastic leukemia (ALL) during a 10 year period. A statistically significant correlation existed between modal chromosome number and DI^{\rm FCM} (p=0.009 ). DI^{\rm FCM} could reliably identify leukemias with > 51 chromosomes, whereas only three out of 12 cases with modal chromosome numbers between 47–51 were classified as aneuploid by DI^{\rm FCM} . In the pseudodiploid group only one out of 20 leukemias had a DI^{\rm FCM}>1.0 . Five leukemias with a diploid karyotype showed …an aneuploid DI^{\rm FCM} and in three patients the flow cytometric measurement revealed biclonality undetected by karyotyping. During treatment aneuploid clones could be detected by DI^{\rm FCM} in a substantial number of cases where the cytogenetic analysis was normal, and the opposite was also demonstrated in one case. DI^{\rm FCM} gave prognostic information, showing that cases with a DI >1.12 (corresponding to 51 chromosomes) had a superior outcome with treatment protocols today in use. Show more

Keywords: Lymphoblastic leukemia, childhood, flow cytometry, DNA‐index, karyotype, event free survival

Citation: Analytical Cellular Pathology, vol. 17, no. 3, pp. 145-156, 1998

Authors: van Rossum, M.M. | Mommers, J.M. | van de Kerkhof, P.C.M. | de Jongh, G.J. | van Hooijdonk, C.A.E.M. | van Erp, P.E.J.

Article Type: Research Article

Abstract: Multi parameter flow cytometrical assays permit simultaneous assessment of proliferation, differentiation, and inflammation parameters. In this study, the validation of TO‐PRO‐3 iodide (TP3) compared to propidium iodide (PI) and DE‐K10 compared to RKSE60 were evaluated in tape stripping induced hyperproliferation. No occlusion, Duoderm (intermediate occlusion) and Blenderm (maximal occlusion) were used as a model to evaluate the effect of occlusion on epidermal regeneration. Proliferation in the keratin 10‐negative compartment measured with TP3 proved to be a good approximation of proliferation measured with PI. Other epidermal subpopulations (keratin 10‐dim and ‐bright cells) did not make a relevant contribution to hyperproliferation. DE‐K10 …is probable more sensitive than RKSE60 to distinguish populations that differ in degree of differentiation. Occlusion of tape stripped skin resulted in decreased proliferation and increased differentiation. This effect was most pronounced with maximal occlusion. This study showed that occlusion is a therapy, which realises normalisation of hyperproliferative skin disorders. Show more

Citation: Analytical Cellular Pathology, vol. 17, no. 3, pp. 157-165, 1998

Authors: Visscher, Daniel W. | Gingrich, Debra S. | Leon‐Armin, Carmen | Tabaczka, Pamela | Crissman, John D.

Article Type: Research Article

Abstract: Two‐color, multiparametric synthesis phase fraction (SPF) analysis of cytokeratin‐labeled epithelial cells was flow cytometrically performed on both benign (SPFb) and malignant tissue samples (if available, SPFt) from 132 mastectomy/lumpectomy specimens. These data were then correlated with clinicopathologic features, including (1) tumor differentiation, (2) the proportion of tumor comprised of duct carcinoma‐in situ (DCIS), and (3) the histology of accompanying benign breast tissue, classified by predominant microscopic pattern as intact, normal terminal duct lobular units (NTDLU, 34% of cases), atrophic (AT, 33% of cases), proliferative fibrocystic (PFC, 26% of cases), and non‐proliferative fibrocystic (NPFC, 7% of cases). SPFt was inversely correlated …with extent of DCIS (DCIS =0{-}20 % tumor volume – 12.7% mean SPFt, vs. DCIS > 20% tumor volume – 6.4% mean SPFt, p=0.001 ). SPFt also correlated with the histology of background benign breast tissue (NTDLU – 14.8% mean SPFt vs. AT – 6.9% mean SPFt vs. PFC – 12.7% mean SPFt, p=0.05 ) but it did not correlate with patient age or SPFb (overall mean =0.73 %). SPFb was correlated with patient age ({>}56 yr – 0.59% mean SPFb vs. {<}56 yr – 0.84% mean SPFb, p=0.02 ), with background histology (NTDLU – 1.1% mean SPFb vs. AT – 0.43% mean SPFb vs. PFC – 0.70% mean SPFb, p<0.02 ) and with the grade of the neoplasm (well/moderate – 0.58% mean vs. poorly differentiated – 0.85% mean, p=0.04 ). Patients having a background of PFC were significantly older than patients with a background of NTDLU (45.2 yr vs. 60.2 yr, p=0.01 ). We conclude: (1) breast carcinomas arising from a background of more actively cycling pre‐involutional or proliferative fibrocystic epithelium have a greater proliferative fraction than tumors arising from atrophic epithelium, implying that the differentiation status of target cells may impact the effect(s) of tumorigenic events; (2) PFC may represent delayed, abnormal or interrupted involution rather than a hyperproliferative state relative to NTDLU, suggesting that it facilitates neoplasia by extending the period of exposure to promoter agents such as endogenous hormones, and (3) lower SPFt in breast neoplasia with more abundant “residual” DCIS may reflect a lengthier pre‐invasive disease interval due to intrinsically less aggressive phenotype. Show more

Keywords: Synthesis phase fraction, fibrocystic disease, flow cytometry, breast carcinoma

Citation: Analytical Cellular Pathology, vol. 17, no. 3, pp. 167-175, 1998

Authors: Nielsen, Niels H. | Arnerlöv, Conny | Cajander, Stefan | Landberg, Göran

Article Type: Research Article

Abstract: Cyclin E is a part of the cell cycle machinery and aberrantly expressed in several malignancies including breast cancer. Since cyclin E is cell cycle specifically expressed, we wanted to examine the relation between proliferation and expression of cyclin E with special attention to tumours with overexpression of the protein. Seventy‐four breast tumours were analysed for the expression of cyclin E by immunohistochemistry and Western blotting and related to the growth fraction determined by Ki‐67. Significant correlations were obtained between the growth fraction, the percentage of cyclin E positive cells, the intensity of cyclin E and total amount of cyclin …E determined by Western blotting. The majority of the tumours had less cyclin E than Ki‐67 positive cells indicating a conserved cell cycle specific expression of the protein which further was supported by flow cytometric analysis of breast cancer cell lines. The cell cycle specificity of cyclin E was found even in tumours with inactivated retinoblastoma protein (pRB) demonstrating the existence of a pRB independent regulation of cyclin E. A fraction of the tumours had considerably elevated cyclin E levels that were not in relation to the proliferative activity as observed for the other tumours. These tumours were in general highly proliferative and considered to overexpress cyclin E. Patients with tumours of high proliferative activity, high total cyclin E levels or disproportionally elevated cyclin E expressions in relation to proliferation had significantly increased risk of death in breast cancer, whereas the intensity of the immunohistochemical cyclin E staining did not affect the survival. Show more

Keywords: Cyclin E, proliferation, cell cycle, breast cancer, Ki‐67, prognosis

Citation: Analytical Cellular Pathology, vol. 17, no. 3, pp. 177-188, 1998