Analytical Cellular Pathology - Volume 23, issue 1

Authors: Castelli, Christian | Losa, Gabriele A.

Article Type: Research Article

Abstract: Fractal morphometry was used to investigate the ultrastructural features of the plasma membrane, perinuclear membrane and nuclear chromatin in SK‐BR‐3 human breast cancer cells undergoing apoptosis. Cells were incubated with 1 μM calcimycin (A23187) for 24 h. Cells in the early stage of apoptosis had fractal dimension (FD) values indicating that their plasma membranes were less rough (lower FD) than those of control cells, while their perinuclear membranes were unaffected. Changes of the chromatin texture within the entire nucleus and in selected nuclear domains were more pronounced in treated cells. This confirms that the morphological reorganization imputable to a loss …of structural complexity (reduced FD) occurs in the early stage of apoptosis, is accompanied by the inhibition of distinct enzymatic events and precedes the onset of conventional cellular markers, which can only be detected during the active phases of the apoptotic process. Show more

Keywords: Apoptosis, breast cancer cells, electron microscopy, fractal dimension, gray level tresholding segmentation, nuclear chromatin, flow cytometry, acidic sphingomyelinase, γ‐glutamyltransferase

Citation: Analytical Cellular Pathology, vol. 23, no. 1, pp. 1-9, 2001

Authors: Wang, Naining | He, Qimin | Skog, Sven | Eriksson, Staffan | Tribukait, Bernhard

Article Type: Research Article

Abstract: The cytosolic thymidine kinase 1 (TK1) is one of the enzymes involved in DNA replication. Based on biochemical studies, TK1 is activated at late G1 of cell cycle, and its activity correlates with the cell proliferation. We have developed a polyclonal anti‐TK1 antibody against a synthetic peptide from the C‐terminus of human TK1. Using this antibody, here we demonstrate the exclusive location of TK1 in the cytoplasm of cells. Cell cycle dependent TK1 expression was studied by simultaneous fluorescence staining for TK1 and bromodeoxyuridine, by using elutriated cells, and by quantitation of the amount TK1 in relation to the cellular …DNA content. TK1, which was strongly expressed in the cells in S+G2 period, raised at late G1 and decreased during mitosis. The amount of TK1 increased three folds from late G1 to G2. TK1 positive cells were demonstrated in areas of proliferation activity of various normal and malignant tissues. The new anti‐TK1 antibody works in archival specimens and is a specific marker of cell proliferation. Show more

Keywords: Thymidine kinase 1, polyclonal antibody, cell proliferation, cell cycle, immunochemistry

Citation: Analytical Cellular Pathology, vol. 23, no. 1, pp. 11-19, 2001

Authors: Arnerlöv, Conny | Emdin, Stefan O. | Cajander, Stefan | Bengtsson, Nils‐Olof | Tavelin, Björn | Roos, Göran

Article Type: Research Article

Abstract: To study intratumoral DNA ploidy heterogeneity and S‐phase fraction (SPF) variability, we prospectively collected five different samples from 48 breast carcinomas and each sample was analysed separately by flow cytometry. Aneuploidy rate was 89.6% after analysis of four or five samples. DNA ploidy heterogeneity, i.e., different samples classified as either DNA euploid or DNA aneuploid in the same tumor was seen in 17%, and DNA index heterogeneity, i.e., tumor populations with different DNA indices (DIs) seen in different samples was 44%. A statistical model defining SPF heterogeneity is proposed. SPF heterogeneity as defined by us was 71%, and as expected …the SPF heterogeneity rate increased significantly with increasing number of analysed samples. Four or more samples are needed to detect the most deviant (highest) SPF values. An unrecognized intratumor heterogeneity of DNA ploidy and SPF may partly explain the conflicting results reported in the literature on the above prognostic indicators. Show more

Keywords: Breast cancer, DNA ploidy, heterogeneity, s‐phase fraction, SPF

Citation: Analytical Cellular Pathology, vol. 23, no. 1, pp. 21-28, 2001

Authors: Mattfeldt, Torsten | Wolter, Hubertus | Kemmerling, Ralf | Gottfried, Hans‐Werner | Kestler, Hans A.

Article Type: Research Article

Abstract: Comparative genomic hybridization (CGH) is a modern genetic method which enables a genome‐wide survey of chromosomal imbalances. For each chromosome region, one obtains the information whether there is a loss or gain of genetic material, or whether there is no change at that region. Usually it is not possible to evaluate all 46 chromosomes of a metaphase, therefore several (up to 20 or more) metaphases are analyzed per individual, and expressed as average. Mostly one does not study one individual alone but groups of 20–30 individuals. Therefore, large amounts of data quickly accumulate which must be put into a logical …order. In this paper we present the application of a self‐organizing map (Genecluster) as a tool for cluster analysis of data from pT2N0 prostate cancer cases studied by CGH. Self‐organizing maps are artificial neural networks with the capability to form clusters on the basis of an unsupervised learning rule, i.e., in our examples it gets the CGH data as only information (no clinical data). We studied a group of 40 recent cases without follow‐up, an older group of 20 cases with follow‐up, and the data set obtained by pooling both groups. In all groups good clusterings were found in the sense that clinically similar cases were placed into the same clusters on the basis of the genetic information only. The data indicate that losses on chromosome arms 6q, 8p and 13q are all frequent in pT2N0 prostatic cancer, but the loss on 8p has probably the largest prognostic importance. Show more

Keywords: Artificial neural networks, chromosome aberrations, cluster analysis, comparative genomic hybridization, multivariate analysis, prognostic factors, prostatic cancer, self‐organizing maps, tumor suppressor genes

Citation: Analytical Cellular Pathology, vol. 23, no. 1, pp. 29-37, 2001

Authors: Saretzki, G. | Fischer, H. | Kaufmann, I.‐G. | Schewe, C. | Nadjari, B. | Blohmer, J. | Hauptmann, St.

Article Type: Research Article

Abstract: It is well known that almost all carcinoma cells including those of the uterine cervix have re‐established their telomerase activity. However, until now there is no conclusive picture on the telomerase activity in cervical dysplasias and about their relationship to HPV infection. To investigate this question, material from 34 patients (15 with normal epithelium, 11 with LGSIL, 8 with HGSIL) obtained by conventional cervical brushing was used and subjected to non‐radioactive TRAP‐ELISA (Boehringer Mannheim). The HPV analysis was performed by PCR on formalin‐fixed, paraffin‐embedded biopsy material obtained after cytological investigation. We could show that telomerase activity is detectable in normal …cervical epithelium, and that an gradual increase exists for both telomerase activity and HPV positivity from normal epithelium to HGSIL. However, HPV infection and telomerase activity appear to be independent of each other. The high frequency of telomerase positivity in patients with normal cervical epithelium indicates that telomerase activity is not a useful differential diagnostic aid. Whether patients with telomerase‐positive dysplasias have a higher probability to progress into an invasive carcinoma remains to be clarified by follow‐up studies. Show more

Keywords: Cervix uteri, dysplasia, telomerase activity, HPV infection, gynecological cytology

Citation: Analytical Cellular Pathology, vol. 23, no. 1, pp. 39-43, 2001