Analytical Cellular Pathology - Volume 16, issue 3

Authors: Yamaguchi, Taihei | Shindoh, Masanobu | Amemiya, Akira | Inoue, Nobuo | Kawamura, Masaaki | Sakaoka, Hiroshi | Inoue, Masakazu | Fujinaga, Kei

Article Type: Research Article

Abstract: Oral papilloma is a benign tumourous lesion. Part of this lesion is associated with human papillomavirus (HPV) infection. We analysed the genetical and histopathological evidence for HPV type 2 infection in three oral papillomas. Southern blot hybridization showed HPV 2a sequence in one lesion. Cells of the positive specimen appeared to contain high copy numbers of the viral DNA in an episomal state. In situ staining demonstrated virus capsid antigen in koilocytotic cells and surrounding cells in the hyperplastic epithelial layer. Two other specimens contained no HPV sequences by labeled probe of full length linear HPVs 2a, 6b, 11, 16, …18, 31 and 33 DNA under low stringency hybridization conditions. These results showed the possibility that HPV 2 plays a role in oral papilloma. Show more

Keywords: Human papillomavirus type 2, oral papilloma, in situ immunostaining, southern blot hybridization

Citation: Analytical Cellular Pathology, vol. 16, no. 3, pp. 125-130, 1998

Authors: Andersen, Lars | Kayser, Lars | Keiding, Niels | Thomsen, Jens

Article Type: Research Article

Abstract: Cells from 7 patients operated on for thyroid cancer were investigated. Samples of cells from the carcinoma and from the normal thyroid tissue were cultured with and without TSH stimulation. For light microscopy, serial sections of cells were cut and the size of nucleoli was measured and the number of nucleoli per cell counted. At the electron microscopic level the number and the volume of the fibrillar centres (FC) were estimated taking the Swiss cheese effect into account. The areal densities of FC, the fibrillar and granular component in nucleoli were determined by point counting. The results indicate that the …malignant transformation has no influence on the size of the FC, but the observed numbers as well as the total area of FC are larger in cancer cells than in the normal thyroid epithelial cells. The nucleolar density of the fibrillar component is larger and that of the granular component is smaller in thyroid carcinoma cells than in non‐malignant thyroid epithelial cells (p=0.0001 ). Thus simple morphometry at the electron microscopic level might be helpful to discriminate between thyroid epithelial cells and thyroid carcinoma cells in culture. Show more

Keywords: Thyroid gland, nucleoli, fibrillar centres, culture, cancer

Citation: Analytical Cellular Pathology, vol. 16, no. 3, pp. 131-140, 1998

Authors: Baldus, Stephan E. | Zirbes, Thomas K. | Weidner, Ion‐Corin | Flucke, Uta | Dittmar, Elke | Thiele, Juergen | Dienes, Hans P.

Article Type: Research Article

Abstract: Liver macrophages, which are involved in the different types of hepatitis, may indirectly induce hepatic fibrogenesis, since they have the possibility to activate hepatic stellate cells and fibroblasts by secretion of TGF‐\beta , TNF‐\alpha and IL‐1. To evaluate variations of the number of liver macrophages and their subpopulations, a quantification was carried out in normal human liver tissue, fatty liver, fatty liver hepatitis and hepatitis B. Identification was performed by the mab PG‐M1 (anti‐CD68) and, comparatively, four lectins, Griffonia simplicifolia agglutinin I (GSA‐I), Erythrina cristagalli agglutinin (ECA), peanut agglutinin (PNA) and soybean agglutinin (SBA). A …slight decrease in the frequency of macrophages in pericentral fields was observable in fatty liver and fatty liver hepatitis as compared to normal liver tissue. On the other hand, the number of CD68^{+} cells was significantly enhanced in hepatitis B with moderate and severe inflammatory activity. The highest incidence of macrophages was found in portal tracts of liver with fatty liver hepatitis and, particularly, hepatitis B. The fraction of cells stained by ECA, PNA or SBA did not increase significantly under pathological conditions. In contrast, the percentage of GSA‐I binding macrophages was higher in liver parenchyma of hepatitis B and in portal tract macrophages in fatty liver hepatitis and also hepatitis B. In conclusion, our results indicate that GSA‐I may aid in the detection of the subpopulation of activated macrophages which are assumed to play a pivotal role in liver pathology. Show more

Keywords: Kupffer cells, macrophages, CD68, lectins, Griffonia simplicifolia

Citation: Analytical Cellular Pathology, vol. 16, no. 3, pp. 141-150, 1998

Authors: Escribano, Luis | Orfao, Alberto | Villarrubia, Jesús | Díaz‐Agustín, Beatriz | Cerveró, Carlos | Rios, Agustín | Velasco, José L. | Ciudad, Juana | Navarro, José L. | San Miguel, Jesús F.

Article Type: Research Article

Abstract: The goal of the present paper was to define the immunophenotype of bone marrow mast cells (BMMC) from healthy controls and patients with hematologic malignancies (HM) based on the use of multiple stainings with monoclonal antibodies analyzed by flow cytometry. Our results show that BMMC from both groups of individuals display a similar but heterogenous immunophenotype. The overall numbers of BMMC are higher in the HM group of individuals (p=0.08 ). Three patterns of antigen expression were detected: (1) markers constantly positive in all cases analyzed (CD9, CD29, CD33, CD43, CD44, CD49d, CD49e, CD51, CD71, CD117, and Fc\varepsilon …RI), (2) antigens that were constantly negative (CD1a, CD2, CD3, CD5, CD6, CD11a, CD14, CD15, CD16, CD19, CD20, CD21, CD23, CD25, CD30, CD34, CD38, CD41a, CD42b, CD65, CD66b, HLA‐DR, and CD138), and (3) markers that were positive in a variable proportion of cases – CD11b (50%), CD11c (77%), CD13 (40%), CD18 (20%), CD22 (68%), CD35 (27%), CD40 (67%), CD54 (88%) and CD61 (40%). In addition, BMMC from all cases explored were CD45^+ , and this antigen was expressed at an intensity similar to that of mature granulocytes. In summary, our results show that BMMC from both healthy controls and HM patients display a relatively heterogenous immunophenotype. Interestingly, we have observed clear differences between the immunophenotype of BMMC and MC from other tissues. This could be due either to the heterogeneity of human MC according to their tissue localization or to the sensitivity of the method used for antigen detection. Show more

Keywords: Mast cells, immunophenotype, bone marrow, flow cytometry

Citation: Analytical Cellular Pathology, vol. 16, no. 3, pp. 151-159, 1998

Authors: François, Christine | Remmelink, Myriam | Petein, Michel | van Velthoven, Roland | Danguy, André | Wespes, Eric | Salmon, Isabelle | Kiss, Robert | Decaestecker, Christine

Article Type: Research Article

Abstract: Using a series of 105 renal cell carcinomas (RCCs) we investigated whether features quantitatively describing the appearance of Feulgen‐stained nuclei and, more particularly, of their chromatin (on the basis of computer‐assisted microscopy) can contribute any significant prognostic information. Thirty morphonuclear and 8 nuclear DNA content‐related variables were thus generated. The actual prognostic values of this set of cytometric variables was compared (by means of discriminant statistical analysis) to conventional diagnostic and/or prognostic markers including histopathological grades, tumour invasion levels and the presence or absence of metastases. We obtained complete clinical follow‐ups for 49 of the 105 RCC patients under study, …making it possible to define a subset of patients with a bad prognosis (i.e., who died in the 12 months following nephrectomy) and a subset of patients with a good prognosis (i.e., who survived at least 24 months following nephrectomy). An original method of data analysis related to artificial intelligence (decision tree induction) enabled a strong prognostic model to be set up. In the case of 10 new patients, this model identified all the dead patients as having a bad survival status, with a total of 8 correct predictions. Another prognostic model similarly generated enabled the correct predictions to be confirmed. Show more

Keywords: Renal cell carcinoma, prognosis, Feulgen staining, image cytometry, chromatin pattern, DNA ploidy, artificial intelligence

Citation: Analytical Cellular Pathology, vol. 16, no. 3, pp. 161-175, 1998

Authors: Hagedorn, Hjalmar G. | Tübel, Jutta | Wiest, Irmgard | Nerlich, Andreas G.

Article Type: Research Article

Abstract: The rate of cellular growth is mainly influenced by the balance between cell proliferation and cellular decay. Since to our knowledge, no study so far has analysed the rate of proliferation and apoptosis in the normal laryngeal mucosa and in invasive laryngeal carcinomas, we performed a morphological analysis on both parameters in biopsies from 30 patients with laryngeal carcinoma. We applied the TUNEL end labelling technique for the investigation of apoptosis and immunohistochemistry (Ki‐67 antigen) for the determination of the cell proliferation. In our study we demonstrated that invasive tumour growth of the larynx coincides with an increase of …both cellular proliferation and apoptosis. Both parameters, however, affected various tumour areas differently. While there was a preferential expression of the Ki‐67 antigen at the tumour–stroma interface, apoptotic figures could be found randomly distributed in the tumour. This indicates that the replication of tumour cells and tumour cell decay are differently distributed and possibly independently regulated. Since we observed a particularly strong increase of cell proliferation at the tumour–stroma interface which outnumbered the corresponding rate of apoptosis by far, the enhanced cell proliferation at the tumour border seems to be a main factor for tumour growth. A statistical evaluation revealed significant correlation between the apoptotic index and the degree of tumour cell differentiation, indicating that a high rate of apoptosis coincides with a high level of tumour cell differentiation. There was, however, no statistically significant correlation between prognostic clinical parameters and the rate of apoptosis or that of proliferation. Show more

Keywords: Larynx carcinoma, apoptosis, Ki‐67 antigen, prognostic parameters

Citation: Analytical Cellular Pathology, vol. 16, no. 3, pp. 177-184, 1998