Analytical Cellular Pathology - Volume 23, issue 3-4

Authors: Grote, Hans Jürgen | Friedrichs, Nicolaus | Pomjanski, Natalia | Guhde, Helen Friderike | Reich, Olaf | Böcking, Alfred

Article Type: Research Article

Abstract: Objective: To assess the prognostic value of DNA‐image cytometry in cervical carcinoma of the uterus and its relation to other established prognostic factors. Study design: The study included 116 cases of cervical carcinoma FIGO stages IB and II which were treated with radical abdominal hysterectomy. The median follow‐up was 55 months (range 1–162 months). DNA image cytometry was performed on cytologic specimens prepared by enzymatic cell separation from formalin‐fixed, paraffin‐embedded tissues. DNA stemline ploidy, DNA stemline aneuploidy, 5c exceeding rate, 9c exceeding rate, 2c deviation index, and DNA malignancy grade were computed. DNA‐variables as well as various clinical and …histological variables were related to survival rates. Results: In multivariate statistical analysis DNA stemline ploidy using 2.2c as a cut‐off value and FIGO stage showed to be statistically significant available presurgery predictors of survival, whereas the postsurgical parameters lymphonodal status, tumor size and parametrial involvement were significantly correlated with survival. The synopsis of all parameters in a multivariate Cox model indicated that – with declining relevance – the number of positive pelvic lymph nodes, DNA stemline ploidy using a cut‐off level at a modal value of 2.2c, largest pelvic lymph node, 5c exceeding rate, and ratio of carcinoma area to cervix area, were of predictive value for survival. Conclusions: Our results suggest that prognostic information deducted from classical staging parameters is successfully complemented by DNA image cytometry which can be applied pretherapeutically. Show more

Keywords: Cervical cancer, staging, DNA image cytometry, aneuploidy, prognosis, grading

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 97-105, 2001

Authors: Davidson, Ben

Article Type: Research Article

Abstract: Carcinoma of the ovary is the leading cause of death from gynecological cancer in western countries. Ovarian carcinoma is commonly associated with the accumulation of fluid containing malignant cells in the peritoneal, and not infrequently in the pleural cavity. The differentiation of these cells from reactive mesothelial cells is at times difficult. In addition, tumor progression in ovarian carcinoma and the biological characteristics of carcinoma cells in effusions compared to their counterparts in solid tumors are poorly understood. This review details the current knowledge regarding diagnostic and biologic aspects of effusion cytology, with emphasis on ovarian carcinoma. Results from our …first studies of effusions are subsequently presented. These attempt to address several issues. First, to improve the diagnostic ability to detect cancer cells in effusions using antibodies designed for the differentiation of epithelial cells from mesothelial cells. Secondly, to study genotypic and phenotypic differences between ovarian carcinoma cells in effusions, solid primary tumors and metastatic lesions, as well as to compare malignant cells in peritoneal and pleural effusions. These studies of carbohydrate antigens, E‐cadherin complex and matrix metalloproteinases (MMP) attempted to evaluate whether ovarian carcinoma cells in effusions possess true metastatic properties, or are similar to the cells in primary tumors, thereby merely representing the result of a shedding process. Finally, the prognostic role of these molecules was studied in solid tumors from a patient cohort consisting of long‐ and short‐term survivors, followed for up to 20 years. Figure 1 on http://www.esacp.org/acp/2001/23‐3,4/davidson.htm. Show more

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 107-128, 2001

Authors: Okoń, Krzysztof | Tomaszewska, Romana | Nowak, Krystyna | Stachura, Jerzy

Article Type: Research Article

Abstract: The aim of the study was to test applycability of neural networks to classification of pancreatic intraductal proliferative lesions basing on nuclear features, especially chromatin texture. Material for the study was obtained from patients operated on for pancreatic cancer, chronic pancreatitis and other tumours requiring pancreatic resection. Intraductal lesions were classified as low and high grade as previously described. The image analysis system consisted of a microscope, CCD camera combined with a PC and AnalySIS v. 2.11 software. The following texture characteristics were measured: variance of grey levels, features extracted from the grey levels correlation matrix and mean values, variance …and standard deviation of the energy obtained from Laws matrices. Furthermore we used moments derived invariants and basic geometric data such as surface area, the minimum and maximum diameter and shape factor. The sets of data were randomly divided into training and testing groups. The training of the network using the back‐propagation algorithm, and the final classification of data was carried out with a neural network simulator SNNS v. 4.1. We studied the efficacy of networks containing from one to three hidden layers. Using the best network, containing three hidden layers, the rate of correct classification of nuclei was 73%, and the rate of misdiagnosis was 3%; in 24% the network response was ambiguous. The present findings may serve as a starting point in search for methods facilitating early diagnosis of ductal pancreatic carcinoma. Show more

Keywords: Pancreatic duct, hyperplasia, classification, chromatin, image processing, neural networks

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 129-136, 2001

Authors: Mello, Maria Luiza S. | Lareef, Mohamed H. | Vidal, Benedicto C. | Russo, Jose

Article Type: Research Article

Abstract: RNA relocation and the incidence of nucleolus‐like bodies accumulated during mitosis were studied cytochemically in benzo[a]pyrene (BP)‐transformed human breast epithelial MCF‐10F cells after microcell‐mediated transfer of normal chromosomes 11 and 17. The changes resulting from the transfer of these two chromosomes in tumorigenic MCF‐10F cells (BP1‐E cell line) were examined, since alterations in these chromosomes are involved in the expression of the transformed and tumorigenic phenotypes in the MCF‐10F cell series. In addition, the frequency of nucleolus‐like bodies decreases drastically with transformation and tumorigenicity in MCF‐10F cells, thus being conceivable that it would be affected in presence of normal chromosomes …11 or 17. The pattern of RNA relocation associated with the mitotic spindle did not vary in the cell lines analyzed. The introduction of chromosome 17 in BP1‐E cells either decreased or did not affect the frequency of persistent nucleolus‐like bodies. In contrast, in cells which received a normal chromosome 11, the frequency of nucleolus‐like bodies was closer to that of non‐transformed MCF‐10F cells. These results suggest that a normal chromosome 11 but not chromosome 17 contributes to the maintenance of an RNA surplus which accumulates in nucleolus‐like bodies during cell division of the human breast epithelial cells, at least in vitro. Some loci which were retained in the BP1‐E cells which received a normal chromosome 11 are probably involved with the control of RNA transcript production. Figure 1 on http://www.esacp.org/acp/2001/23‐3,4/mello.htm Show more

Keywords: RNA, nucleolus‐like bodies, human breast epithelial cells, microcell‐mediated chromosome transfer, chromosome 11, chromosome 17

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 137-141, 2001

Authors: Gustafsson, Ulf | Einarsson, Curt | Eriksson, Lennart C. | Gadaleanu, Virgil | Sahlin, Staffan | Tribukait, Bernhard

Article Type: Research Article

Abstract: Gallstones are a risk factor for the development of gallbladder cancer. We studied DNA ploidy and cell cycle composition by flow cytometry in archival specimens from 52 gall bladder carcinomas in relation to histopathological grade, tumour stage, gallstone number and survival. 69% of the gallbladder carcinomas showed aneuploidy. All tumours with single stones (N=11) were aneuploid while only 61% of tumours with multiple stones (N=41) were aneuploid (p=0.002). DNA aneuploidy was related to increase in T‐category (p=0.01), grade (p=0.02), and nuclear pleomorphism (p=0.0005). The distribution of DNA ploidy shifted from tetraploid in low stage towards triploid positions in high stage …tumours (p=0.02) combined with higher S‐phase values in triploid tumours (p=0.05). S‐phase fraction increased during development from normal tissue to dysplasia, cancer in situ and cancer in diploid cases (p=0.0002), and further at the change from diploid to aneuploid (p=0.004). At a median cancer specific survival time of four months patients with diploid tumours had a better survival than those with aneuploid tumours (p=0.02). In multivariate analysis of the tumour characteristic, only T‐category and tumour grade were independent prognostic factors. The shift from diploid to aneuploid and the further shift of ploidy within aneuploid tumours are in agreement with the concept of a clonal development of gallbladder cancer. These changes are combined with a stepwise increase in the fraction of S‐phase cells. Low frequency of symptoms in single stone patients may be the reason for detection of malignancy at a late stage of tumour development. Show more

Keywords: Gallbladder cancer, flow cytometry, DNA ploidy, S phase fraction, cell cycle, gallstone number, survival

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 143-152, 2001

Authors: Smolle, Josef | Kahofer, Peter

Article Type: Research Article

Abstract: Objective: To evaluate the feasibility of the CART (Classification and Regression Tree) procedure for the recognition of microscopic structures in tissue counter analysis. Methods: Digital microscopic images of H&E stained slides of normal human skin and of primary malignant melanoma were overlayed with regularly distributed square measuring masks (elements) and grey value, texture and colour features within each mask were recorded. In the learning set, elements were interactively labeled as representing either connective tissue of the reticular dermis, other tissue components or background. Subsequently, CART models were based on these data sets. Results: Implementation of the CART classification rules into …the image analysis program showed that in an independent test set 94.1% of elements classified as connective tissue of the reticular dermis were correctly labeled. Automated measurements of the total amount of tissue and of the amount of connective tissue within a slide showed high reproducibility (r=0.97 and r=0.94, respectively; p<0.001). Conclusions: CART procedure in tissue counter analysis yields simple and reproducible classification rules for tissue elements. Show more

Keywords: Image analysis, malignant melanoma, connective tissue, classification and regression tree, tissue counter analysis

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 153-158, 2001

Authors: Frimmel, Hans | Egevad, Lars | Busch, Christer | Bengtsson, Ewert

Article Type: Research Article

Abstract: Objectives. When analysing the 3D structure of tissue, serial sectioning and staining of the resulting slices is sometimes the preferred option. This leads to severe registration problems. In this paper, a method for automatic registration and error detection of slices using landmark needles has been developed. A cost function takes some parameters from the current state of the problem to be solved as input and gives a quality of the current solution as output. The cost function used in this paper, is based on a model of the slices and the landmark needles. The method has been used to register …slices of prostates in order to create 3D computer models. Manual registration of the same prostates has been undertaken and compared with the results from the algorithm. Methods. Prostates from sixteen men who underwent radical prostatectomy were formalin fixed with landmark needles, sliced and the slices were computer reconstructed. The cost function takes rotation and translation for each prostate slice, as well as slope and offset for each landmark needle as input. The current quality of fit of the model, using the input parameters given, is returned. The function takes the built‐in instability of the model into account. The method uses a standard algorithm to optimize the prostate slice positions. To verify the result, s standard method in statistics was used. Results. The methods were evaluated for 16 prostates. When testing blindly, a physician could not determine whether the registration shown to him were created by the automated method described in this paper, or manually by an expert, except in one out of 16 cases. Visual inspection and analysis of the outlier confirmed that the input data had been deformed. The automatic detection of erroneous slices marked a few slices, including the outlier, as suspicious. Conclusions. The model based registration performs better than traditional simple slice‐wise registration. In the case of prostate slice registration, other aspects, such as the physical slicing method used, may be more important to the final result than the selection of registration method to use. Show more

Keywords: Registration, landmarks, 3D model, prostate

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 159-165, 2001

Article Type: Other

Citation: Analytical Cellular Pathology, vol. 23, no. 3-4, pp. 167-169, 2001