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G Haroske

Publications and source records attributed to G Haroske.

At least 37 records · Page 2Linked to original sources
PubMed1998

Loss of caveolin expression in type I pneumocytes as an indicator of subcellular alterations during lung fibrogenesis.

Caveolin is a major structural protein of caveolae, also known as plasmalemmal vesicles, which are particularly abundant in type I pneumocytes and capillary endothelial cells of lung parenchyma. Here we demonstrate that caveolin expression in the alveolar epithelium of rats and mini pigs is strikingly downregulated after irradiation-induced lung injury. Indirect immunoperoxidase staining with polyclonal anti-caveolin antibodies, confirmed by double fluorescence studies with type I cell-specific monoclonal anti-cytokeratin antibodies or lectins, revealed a dramatic loss of caveolin immunoreactivity in type I pneumocytes. In contrast, caveolin expression increased in endothelial cells. Immunoblotting of lung homogenates from normal and irradiated rats using specific anti-caveolin antibodies confirmed the presence of caveolin in normal tissue and its marked decrease of expression in fibrotic tissue. The loss of caveolin as an important structural protein of caveolae in alveolar epithelial cells may be an early indicator of serious type I cell injury during fibrogenesis. The increase of caveolin immunoreactivity in endothelia of blood vessels may indicate that different types of caveolae and/or different regulatory mechanisms of caveolin expression exist.

Animals↗
PubMed1998

1997 ESACP consensus report on diagnostic DNA image cytometry. Part I: basic considerations and recommendations for preparation, measurement and interpretation. European Society for Analytical Cellular Pathology.

A task force of invited experts in the field of diagnostic DNA image cytometry, especially consisting of participants from the PRESS (Prototype Reference Standard Slides) and EUROPATH (European Pathology Assisted by Telematics for Healthcare) projects, but open to any other scientist or physician revealing experience in that new diagnostic procedure (names are given in the Annex A) agreed upon the following updated consensus report during the 5th International Congress of the ESACP 1997 in Oslo. This report is based on the preceeding one [9] and on results of the above mentioned European research projects. It deals with the following items: Biological background and aims of DNA image cytometry, Principles of the method, Basic performance standards, Diagnostic interpretation of DNA measurements, Recommendations for practical use.

Aneuploidy↗
PubMed1998

Prognostic relevance of DNA image cytometry in oral cavity carcinomas.

OBJECTIVE: To evaluate the relation of DNA content of squamous cell carcinomas of the oral cavity to clinicopathologic features by image cytometry. STUDY DESIGN: A series of 52 patients with operable oral cavity carcinomas at stage T1-4 and NO-3 were studied. The tumors were classified according to the TNM classification and graded histopathologically. RESULTS: A positive correlation between tumor size and ploidy status was observed. There was a significant correlation between ploidy status and histologically proven existence of cervical lymph node metastases (61% nondiploid versus 29% diploid, P < .03). A relationship between histopathologic grade and ploidy status could not be proven. The cumulative five-year survival rate among patients with diploid DNA findings as opposed to those with nondiploid findings was significantly increased (P < .03). CONCLUSION: When assessing the prognosis of squamous cell carcinomas, DNA cytometry analysis can be used as an additional method. The significantly higher N stage, a higher frequency of metastases and a significantly lower survival rate with nondiploid tumors underline the importance of this examination method.

Adult↗
PubMed1998

Remote quantitation server for quality assurance in DNA ploidy analysis.

OBJECTIVE: To remotely test a stepwise framework of quality control measures, developed according to the 1997 European Society of Analytical Cellular Pathology consensus on diagnostic DNA image cytometry, in two series of measurements by means of the quantitation server EUROQUANT. STUDY DESIGN: In each of these series, 104 fine needle aspiration biopsies, imprints from breast cancer specimens and 28 rat liver imprints were measured twice at two different cytometry workstations. Further measurements on special rat liver specimens for evaluation of the stability of the machinery and of the preparation process were done at both workstations. Afterwards the measurement data from both machines were transferred to the quantitation server and analyzed automatically. Beside the classical DNA histograms, a set of further evaluations was performed to detect optical errors as well as inhomogeneities in the measurements. Running values for mean integrated optical density values, mean corrective factors and mean coefficients of variations of corrective factors were computed to control the stability of the entire methodology over time. RESULTS: The study demonstrated the preconditions and outcome of server-based quality control in DNA ploidy analysis. The results show that such a remote analysis is feasible and comparable to local DNA ploidy analysis. It is also demonstrated how quality control tests reflect the process performance of ploidy analysis at its different levels. CONCLUSION: Quality control must be an inherent part of the daily routine to allow a reliable diagnostic interpretation and ensure steadily high product quality.

Animals↗
PubMed1997

DNA analysis of squamous cell carcinoma in the oral cavity by image cytometry and its rating in the assessment of tumor prognosis.

Image cytometric measurements on the DNA content of squamous cell carcinoma of the oral cavity were carried out to evaluate its relationship with clinicopathological features. In the prospective study 52 patients with operable oral cavity carcinoma in stage T1-4 and N0-3 were studied. The tumors were classified according to the TNM classification (UICC 1987) and graded histopathologically. A positive correlation between tumor size and ploidy status was observed. There was a significant correlation between ploidy status and histologically confirmed cervical lymph node metastases (P < 0.03). A relationship between histopathological grading and ploidy status was not found. The cumulative 5-year survival rate was significantly higher (P < 0.03) among patients with diploid DNA findings than among those with nondiploid findings. When assessing the prognosis of squamous cell carcinoma, DNA cytometry analysis should be used as an additional method. Particularly the primarily existing, significantly higher N stage, a higher frequency of metastases, and a significantly lower survival rate with nondiploid tumors underline the importance of this examination method.

Adult↗
PubMed1997

Morphological heterogeneity of p53 positive and p53 negative nuclei in breast cancers stratified by clinicopathological variables.

The study was aimed to detect differences in nuclear morphology between nuclear populations as well as between tumours with different p53 expression in breast cancers with different clinicopathological features, which also reflect the stage of tumour progression. The p53 immunohistochemistry was performed on paraffin sections from 88 tumour samples. After the cells had been localised by means of an image cytometry workstation and their immunostaining had been categorised visually, the sections were destained and stained by the Feulgen protocol. The nuclei were relocated and measured cytometrically by the workstation. There were significant differences in the nuclear features between tumours as well as between nuclear populations with different p53 expression in the most subgroups. The variability of nuclear shape in tumour groups, classified by the tumour size or the lymph node status, increase with the p53 immunoreactive score, whereas in tumours grouped by the Bloom-Richardson grade features of the chromatin distribution were different between the p53 staining categories. The nuclear subpopulations showed differences in the amount and distribution of chromatin in most subgroups. The results demonstrate the relationship between the nuclear morphology and the p53 expression in different stages of breast cancers. The p53 status is an important factor of the biological behaviour but not the only one.

Breast Neoplasms↗
PubMed1997

Correlation between p53 status, DNA ploidy, proliferation rate and nuclear morphology in breast cancer. An image cytometric study.

The study was designed to detect differences in the nuclear morphology of tumours and tumour cell populations with different p53 expression in correlation with DNA ploidy and proliferation rate. The paraffin sections from routinely processed samples of 88 breast cancers were immunostained with the monoclonal p53-antibody DO-1. After localization and evaluation with a scoring system the sections were destained and stained by the Feulgen method. The nuclei were relocated automatically and measured by means of the image cytometry workstation. Significant differences between the tumours and tumour cell populations with different p53 expression were found in the euploid tumours as well as in the aneuploid tumours and in the breast cancers with a high proliferation rate. The breast cancers with a low immunoreactive score (IRS 1-4) differ from the negative cancers as well as from the cancers with a higher immunoreactive score (IRS 5-12). Evaluating the nuclear populations of the p53 positive cancers, there were differences in the features of the chromatin amount and distribution in the groups of the euploid breast cancers and in cancer with a high proliferation rate. In contrast, the nuclear populations of the aneuploid cancers did not show any differences in their nuclear morphology. The results showed the different impacts of the p53 expression, DNA ploidy and the proliferation rate on the nuclear morphology in breast cancer.

Adult↗
PubMed1997

DNA histogram interpretation based on statistical approaches.

Image cytometric DNA measurements provide data which are most often interpreted as equivalent to the chromosomal ploidy although the chromosomal and the DNA ploidy are not identical. The common link between them is the cell cycle. Therefore, if destined for DNA ploidy interpretations, the DNA cytometry should be performed on a population-oriented stochastic basis. Using stochastic sampling the data can be interpreted by applying the rules of stochastic processes. A set of statistical methods is given that enables a DNA histogram to be interpreted objectively and without human interaction. These statistics analyse the precision and accuracy of the entire measurement process. They give in error probabilities for accepting a measurement as reliable, for recognition of stemlines, stemline aneuploidy, and for evaluating so-called rare events. Nearly 300 image cytometric DNA measurements from breast cancers and rat liver imprints examples have been selected to demonstrate the efficiency of the statistics in each step of interpreting DNA histograms.

Aneuploidy↗
PubMed1997

Nuclear image analysis of p53-positive and -negative cells in breast carcinoma.

OBJECTIVE: To detect differences in nuclear morphology in breast carcinoma cells with different p53 expression. STUDY DESIGN: Immunohistochemical reactions were performed on paraffin sections from 88 breast cancers with the monoclonal antibody DO-1. By means of an image cytometry workstation, 500 nuclei were localized and categorized in an immunohistochemical staining scoring system in each case. After destaining and Feulgen staining, the nuclei were relocated and measured by means of the workstation. RESULTS: P53-positive nuclei showed greater irregularity of the chromatin pattern and stronger granularity of the chromatin. There were also differences in the chromatin texture and nuclear shape between tumors different in their p53 expression. The p53-negative nuclear population from the p53-positive tumors and the whole nuclear population of the p53-negative tumors were morphologically different, also. CONCLUSION: Nuclear image analysis of immunohistochemically characterized cells allows further insight into the relationship of structure and function in malignant tumor cells.

Adult↗
PubMed1997

Diagnostic and prognostic relevance of malignancy-associated changes in cervical smears.

OBJECTIVE: For approximately 15 years, malignancy-associated changes (MACs) have been consistently found by means of high-resolution cytometry in different tissues, especially in visually normal appearing cervical cells. Their biologic nature is not yet fully understood. The aim of this investigation was to assess the expression of MACs in cervical smears and to evaluate their prognostic relevance. STUDY DESIGN: This study was performed on normal intermediate cells obtained from 53 cytologically positive and 78 cytologically negative cervical smears. From a second sample, 31 cases showing negative cytology were selected for a prospective longitudinal study. Densitometric and texture features were generated, and MACs were described on the basis of multivariate discriminant analysis. RESULTS: Discrimination between positive and negative cases was possible, with a correct classification rate of approximately 80%. After a mean period of 29.5 months, we noted no statistically significant increase in the incidence of cervical intraepithelial neoplasia in the group of healthy but MAC-positive women as compared to those who were MAC negative. CONCLUSION: MACs were constantly expressed in the epithelium of the cervix. Although their prognostic relevance remains unclear, MACs play an important role in the effort to automate cervical cytology.

Adult↗
PubMed1996

Nuclear image analysis of immunohistochemically stained cells in breast carcinomas.

Hitherto, the relationship between malignancy-associated morphological features in single tumour cells and the expression of markers indicating functional properties of these cells remained widely unknown. This study was aimed at describing differences in the size, shape and chromatin structure between tumour cells with different marker expression for progesterone receptors (PgR) and p53. Two series of breast cancers, consisting of 50 PgR-positive, and 39 p53-negative and 49 p53-positive mammary carcinomas, were investigated. The immunohistochemical staining was performed on paraffin sections using 3-amino-9-ethylcarbazole as the chromogenic substrate. By means of a cytometry workstation equipped with a computer-controlled motorised scanning stage, about 500 positive and negative tumour cells in each case were localised in the microscope and categorised by a scoring system for their staining intensity. After destaining, the tissue sections were Feulgen-stained. Then, all the tumour cells were relocated automatically and analysed by high resolution image cytometry. Among the numerous size, shape, and texture features used in the system, several variables of the nuclear contour and chromatin structure were found to be significantly different between the positive and negative tumour cell populations. Nuclei without PgR had more malignancy-associated morphological features than PgR-positive cells. Whereas p53-negative nuclei had a higher degree of regularity, their positive counterparts exhibited higher DNA ploidy values.

Biomarkers, Tumor↗
PubMed1996

Distribution of von Willebrand factor in capillary endothelial cells of rat lungs with pulmonary fibrosis.

To determine the value of von Willebrand factor (vWF) antigen as a marker of endothelial injury in radiation-induced fibrosis of rat lungs, we studied endothelial immunoreactivity to antibodies against vWF using the indirect immunoperoxidase technique combined with morphometric analysis. Using immunoelectron microscopy of LR White embedded lung samples to detect vWF, immunogold-labelled Weibel Palade bodies were found in endothelial cells of capillary endothelium. The irradiated lungs showed a statistically significant elevation of vWF expression, ie, vWF positive endothelia per unit area, and a significant increase of vWF expression per unit of parenchyma as well. The results suggest that vWF antigen expression and the number of vWF positive structures is modulated under condition of injury in radiation-induced fibrosis.

Animals↗
PubMed1996

'Blind' multicenter evaluation of the prognostic value of DNA image cytometric and morphometric features in invasive breast cancer.

The prognostic value of classical prognosticators in 92 invasive breast cancers with long follow-up has been compared with a number of DNA-related image cytometric features (2c deviation index (2cDI)), Auer (histogram) type, 5c exceeding rate (5cER), DNA malignancy grade (MG), and the morphometrically assessed mitotic activity index (MAI) and mean nuclear area (MNA). The quantitative analyses were carried out without knowledge of the outcome nor of clinicopathological features. The cytometric analyses and assessments of MNA were performed on Feulgen stained, 4-microns thick tissue sections in Dresden. The MAI was assessed in Amsterdam. Thereafter, the survival data were matched with the classical variables, the cytometric features, the MAI and the MNA in univariate and multivariate analyses. Lymph node status (LN) and tumour stage were significantly different, tumour stage being the strongest discriminator between survivors and non-survivors (Mantel-Cox value = MC = 30.7, P < 0.0001). Grade was (just) significant (P = 0.05). The cytometric features, the MAI and the MNA were all highly significant prognosticators. The strongest cytometric feature was the Auer type followed by the 5cER, and the 2cDI. The MNA was also significant. The MAI was the strongest single prognostic factor in this blind analysis (MC = 50.8, P < 0.0001), and the MAI combined with the lymph node status, tumour stage and the cytometric features Auer type and 2cDI was the best multivariate combination (MC = 94.0). Separate analyses of the lymph node negative (LN-, n = 39) and positive (LN+, n = 53) groups showed that the 5cER and MAI were the most important in the LN- and the MAI in the LN+ patients.

Breast Neoplasms↗
PubMed1996

Cellular sociology of proliferating tumor cells in invasive ductal breast cancer.

OBJECTIVE: The topology of proliferating cells in tissue sections of breast cancers was studied as the static expression of the cellular sociology of distinct subpopulations of tumor cells. With the terms neighborhood and connectivity, an attempt was made to describe the mutual interrelations of cycling tumor cells objectively in different prognostic subgroups of invasive ductal breast carcinomas. STUDY DESIGN: For these studies the spatial distribution of Ki-67 (Mib1)-positive tumor cells was investigated by means of Voronoi tesselation, minimum spanning tree techniques, cluster analysis and second-order stereology in a series of 80 breast tumors. Thirty-nine basic features were used for describing the topology of tumor cells with different degrees of Mib-1 reactivity. RESULTS: Multivariate analyses demonstrated different proliferation patterns associated with tumor size, nodal status, histopathologic grade, DNA ploidy and flow cytometric proliferation variables. CONCLUSION: Independent of the spatial organization of the tumor cells, their tissue pattern of proliferation activity correlated with the tumor properties.

Antigens, Nuclear↗
PubMed1996

Alterations in the alveolar epithelium after injury leading to pulmonary fibrosis.

This review discusses current knowledge of the involvement of the alveolar epithelium in tissue remodelling during fibrogenesis. The purpose of the present paper is to give an overview, including the authors' own results, of knowledge of ultrastructural alterations, proliferation kinetics and phenotypic changes of pneumocytes in experimental and clinical pathology of pulmonary fibrosis. After lung injury, the alveolar epithelial cells show ultrastructural alterations, hypertrophy and hyperplasia, and a modulation of a series of structural and membrane proteins such as cytoskeletal changes, loss or de novo expression of epithelial adhesion molecules, and altered lectin binding. Furthermore, enhanced secretion of proteases, of cytokines and other soluble factors can be observed in the alveolar epithelium. These findings suggest the contribution of the epithelium in the remodelling process to be greater than expected. Estimations of the cell kinetics show that type II pneumocytes have the proliferative capacity to restore high proportions of damaged type I cells within few hours. In fibrosis this capacity also seems to be affected seriously, resulting in transitional phenotypes between type II and type I cells. Additionally, in the light of the detection of CD44 type of adhesion molecules at the foot processes of type II pneumocytes, some aspects of epithelial-fibroblast interaction are described.

Animals↗