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At least 1,063 records · Page 59Linked to original sources
PubMed1997

DNA ploidy in malignant melanoma, skin cancer and pigmented nevi.

The determination of DNA content in human cancers is the subject of increasing interest, particularly in view of its potential clinical applications. There are relatively few studies which describe DNA content of skin neoplasms and pigmented nevi. These studies have shown conflicting results. In the present investigation the authors measured DNA ploidy using flow and video-imaging cytometry in 51 malignant melanomas, 20 skin cancers and 48 pigmented nevi. For DNA measurement paraffin embedded tissues and fresh cell smears were used. Clinical and histological data of malignant melanomas were recorded and correlated with DNA ploidy. DNA histograms were examined for DNA aneuploidy by DNA Index. DNA ploidy in primary lesions of melanomas and their metastases were compared. The aneuploidy rate, found in our observation, was significantly higher in whole malignant melanoma group, in clinical Stage II and III, in tumors with thickness greater then 1.5 mm, tumors with Clark level III, IV and V. Another clinical and histological factors did not show significant correlation with ploidy. Aneuploidy was found in 8 of 20 (45.0%) skin cancers. In the whole population of pigmented nevi aneuploid DNA content was identified in 10 nevi (20.1%). The results of this study suggest that aneuploidy seems to be connected with advanced stage of malignant melanoma but it does not replace other prognostic factors. Both cytometric methods can be used for routine DNA ploidy analysis. Ploidy studies are not useful for predicting metastatic potential of primary melanoma. Results obtained from fresh cell smears and paraffin embedded tissues were identical.

Aneuploidy↗
PubMed1998

[Barrett esophagus with severe dysplasia in argon beam therapy].

A curative therapy of Barrett's esophagus is not established, yet. However, in prospective clinical research trials, a combined therapy of potent pharmacological acid suppression with superficial coagulation of metaplastic epithelium is examined. By local coagulation using photodynamic therapy, laser or argon plasma coagulation, a complete reconstitution of squamous epithelium should be achieved. We report on a case of a patient with Barrett's esophagus and low-grade dysplasia, who developed high-grade dysplastic cell formations under argon plasma coagulation therapy. As part of a clinical study, the patient was treated over a period of one year monthly. We monitored the result of the therapy over the period of one year with endoscopy, histolgical investigations and DNA image cytometry. Ultimately a decision in favour of a further surgical management was made. The diagnostic instruments to identify patients with a high risk for the development of adenocarcinoma are discussed.

Adenocarcinoma↗
PubMed1998

Quantification of eumelanin and pheomelanin: stereologic image analysis method.

OBJECTIVE: To develop an ultrastructural stereologic image analysis method allowing quantification of intracellular melanization. STUDY DESIGN: First, in the field of image analysis, a newly elaborated segmentation method, SEM2, was compared with a previously described method based on gray level histograms. Only SEM2 allows the segmentation of melanin in micrographs of poorly melanized melanocytes. Second, in the field of stereology, estimation of cell volume remains problematic in the case of mixed cell populations. This problem is approached by the comparison of stereologic alternatives and cytochemistry (L-3, 4-dihydroxyphenylalanine reaction) in epidermal melanocytes and melanoma cells from several in vitro experiments. The cytochemical approach was found to be the best choice. RESULTS: Concerning the quantification of eumelanin and pheomelanin, the alkali elution method, permitting the specific dissolution of pheomelanin on ultrathin sections, was validated in normal human follicular melanocytes. CONCLUSION: These results allow us to envisage the stereologic quantification of eumelanin and pheomelanin at the ultrastructural level. At present our method is undergoing evaluation by comparison with the present method of reference based on high-performance liquid chromatography.

Adolescent↗
PubMed1998

Automated feature extraction and identification of colon carcinoma.

OBJECTIVE: To assess an automated algorithm, developed for the classification of normal and cancerous colonic mucosa, using geometric analysis of features and texture analysis. STUDY DESIGN: Twenty-one images were analyzed, 10 from normal and 11 from cancerous mucosa. The classification was based on a regularity index dependent on shape, object orientation for establishing parallelism and five texture features derived using the co-occurrence image analysis method. RESULTS: Geometric analysis yielded an overall classification accuracy of 80%. The corresponding sensitivity and specificity were 94% and 64%, respectively. Using texture analysis, the overall classification accuracy was 90%, with a sensitivity and specificity of 82% and 100%, respectively. CONCLUSION: This initial study demonstrated that geometric and texture analysis techniques show promise for automated analysis of colon cancer.

Algorithms↗
PubMed1998

Machine vision in the detection of prostate lesions in histologic sections.

OBJECTIVE: To explore the utility of N-gram encoding for the automated detection and delineation of regions of histologic abnormality in tissue sections of prostate. STUDY DESIGN: Digitized imagery of tissue sections from normal prostate glandular tissue, stroma and regions of well- and poorly differentiated lesions was recorded and successively subdivided into square subregions of 256 x 256 to 16 x 16 pixels. N-grams of N = 2 to N = 6 were computed, with each element assuming a value representing an optical density interval 0.30 units wide, covering the range from optical density = 0.0 to 1.80. Then, from a large database, prototype frequency histograms of the different N-grams were established. For each subregion the Euclidean distances to the different prototype histograms were computed and defined as "distance to prototype" features. Standard discriminant analyses and a nonparametric classifier were used to assign subregions to the different tissue categories. RESULTS: Classification of subregions was achieved for most discrimination tasks at a correct recognition rate ranging from 85% to 100% on both training set and test set data, with a few exceptions. N-grams of N > 4 had considerable discriminatory power. CONCLUSION: N-gram encoding has the potential to provide highly discriminating, texture-based characterization of subregions of digitized imagery of prostate lesions and may be very useful in the development of decision procedures for the automated detection of prostate lesions by a machine vision system.

Algorithms↗
PubMed1998

Statistical histometry of the basal cell/secretory cell bilayer in prostatic intraepithelial neoplasia.

OBJECTIVE: To delineate the sampling requirements for a histometric assessment of progression in low grade and high grade prostatic intraepithelial neoplasia (PIN) lesions. STUDY DESIGN: Images of whole glands from normal prostates, low grade PIN lesions and high grade PIN lesions were digitized. The images were processed by a machine vision system and automatically segmented, and a number of histometric characteristics descriptive of the disruption of the basal cell layer were extracted. Next, high-resolution images of secretory cell nuclei still facing or no longer facing intact segments of the basal cell layer were recorded and karyometrically analyzed. RESULTS: For the characterization of an individual lesion a minimum of 20-30 glands should be analyzed to provide an estimate of a progression index. Then, a change in progression, or due to regression, of approximately 16% can be documented. The disruption of the basal cell layer is accompanied by statistically highly significant changes in the chromatin texture and spatial distribution in secretory cell nuclei no longer facing an intact segment of that layer. CONCLUSION: Automated histometry by machine vision can provide valuable quantitative data for diagnostic assessment and for monitoring the efficacy of chemopreventive treatment.

Analysis of Variance↗
PubMed1997

[Studies on biological effects of kappa-selenocarrageenan on human breast cancer cell line BCaP-37].

Proliferation, cell cycle, total amount of DNA, area of cell nucleus, as well as epidermal growth factor receptors (EGFR) and expression of oncogene C-erbB2 mRNA of Chinese breast cancer cell line (BCaP-37) after being treated with kappa-selenocarrageenan were determined by cell culture technique, image cytometry (ICM) and northern blot to explore its anti-tumor mechanism. Results revealed 3.0-120 mg/L selenocarrageenan could inhibit proliferation of BCaP-37, with a response of time and dose dependence. The areas of nuclei were significantly lower with ICM in cells treated with 15 or 60 mg/L selenocarrageenan for four days than those in controls (P < 0.01). Levels of EGFR and expression of C-erbB2 mRNA were significantly inhibited in cells treated with 60 mg/L selenocarrageenan. It suggests that selenocarrageenan can inhibit proliferation of breast cancer cells through regulation of the levels of EGFR and expression of C-erbB2 mRNA.

Antineoplastic Agents↗
PubMed1998

Morphometric analysis of human gingival elastic fibres degradation by human leukocyte elastase protective effect of avocado and soybean unsaponifiables (ASU).

Degradation of preelastic fibres (oxytalan and elaunin) and mature elastic fibres by human leukocyte elastase (HLE) was investigated using automated image analysis. Specimens from two young healthy adults were used. Although HLE hydrolyzed both fibre types, mature elastic fibres exhibited greater susceptibility to this effect than preelastic fibres. Avocado and soybean unsaponifiables are widely prescribed in rheumatology and parodontology and have also been the focus of ex vivo experiments aimed at determining whether they protect elastic fibres against degradation by HLE. Findings from the present study indicate that avocado and soybean unsaponifiables protect all types of gingival elastic fibres from degradation by HLE. Avocado and soybean unsaponifiables may be beneficial in patients with gingival inflammation and parodontitis, since HLE plays a major role in these disease states.

Adult↗
PubMed1998

Quantification of tissue eosinophils and lymphocytes in histologic sections.

During a study of eosinophil-predominant gallbladder disease, an image analysis (IA) technique was developed for quantification of eosinophils and lymphocytes in routine formalin-fixed tissue sections. Alternating sections were stained with hematoxylin and eosin for eosinophils and with monoclonal CD45 antibody visualized with diaminobenzidine by an avidin-biotin procedure for lymphocytes. A protocol was then developed using a commercially available image analyzer and two well-defined macro routines. The system was validated with cell block sections prepared from peripheral blood samples with known eosinophil and lymphocyte counts. The eosinophil counts obtained by this IA technique showed excellent correlation with the absolute counts from a peripheral blood analyzer (r2 = 0.987). The lymphocyte counts obtained by IA showed good correlation with the absolute counts (r2 = 0.820). This IA-based technique provides a sensitive, reproducible, and substantiated means of quantifying inflammatory cells in tissue sections. This rapid and easily learned technique adds quantification as a complementary dimension to the subjective assessment of tissue morphology.

Eosinophils↗
PubMed1992

Predicting outcome for patients with node negative breast cancer: a comparative study of the value of flow cytometry and cell image analysis for determination of DNA ploidy.

This study was aimed at determining whether tumour DNA content measured by cell image analysis could provide additional prognostic information when compared to that provided by flow cytometry. Sections cut from paraffin blocks of tumours from 101 patients with node negative breast cancer were analysed by both methods and the results related to other prognostic variables and to patient relapse and overall survival. DNA ploidy measured by flow cytometry classified 46 tumours as diploid and 55 as aneuploid, whereas by cell image analysis 30 were diploid and 71 aneuploid (P less than 0.002). There were 20 tumours with discrepancies between the two methods; 18 of these were tumours with only one peak in flow analysis, but determined to be aneuploid with image analysis. DNA content as measured by both methods was significant for predicting relapse and survival by log-rank test, as were tumour histological grade, c-erbB-2 expression and tumour size. Multivariate analysis showed DNA ploidy measured by flow cytometry to be the only variable of independent significance (P less than 0.02) for both relapse and overall survival. Compared with cell image analysis, flow cytometry demonstrated a significantly higher proportion of diploid tumours, which may be related to differences in the internal standards applied to each method. We suggest that cell image analysis techniques can provide more sensitive information on the DNA content of tumour cells by direct measurement of nuclear DNA density of both normal lymphocytes and tumour cells in the same section. However, although image analysis appears to be more sensitive than flow cytometry in detecting DNA aneuploidy, the image technique appears to lack the specificity of flow cytometry in correlation with clinical outcome.

Adult↗
PubMed1992

Methodologic sources of errors in image and flow cytometric DNA assessments of the malignancy potential of prostatic carcinoma.

Considerable controversy exists about the value of using cytometric assessments of nuclear DNA ploidy patterns from neoplastic parenchymal cells of prostatic carcinomas as a supplement to conventional clinicopathologic data in an assessment of malignancy potential in neoplastic disease. To a great extent, the controversy is of methodologic origin. By revealing common pitfalls in this kind of DNA cytometry and by suggesting means to avoid or at least to reduce them, a realistic assessment can ultimately be made of the prognostic values of this technique: (1) the results of DNA assessments using flow cytometry and image cytometry should be combined (advantages and disadvantages with the two techniques outbalance each other); (2) the cytometric DNA ploidy pattern must always be correlated with results of histopathologic assessments and clinical data; (3) a standardization of DNA histograms obtained by means of both flow DNA cytometry and image DNA cytometry must be made with precise definitions of "diploidy," "tetraploidy," and "aneuploidy"; (4) the subjective component in classifying DNA histograms can be reduced by means of a computerized evaluation technique; (5) the degree of intratumoral variation in DNA ploidy patterns of neoplastic cell nuclei must be established by means of assessments of multiple biopsy specimens from widely different parts of a carcinoma; and (6) foci of a previously poor recognized neuroendocrine phenotype of neoplastic parenchymal cells are ubiquitous in prostatic carcinomas, particularly in anaplastic adenocarcinomas. Today, the DNA ploidy pattern of this phenotype can be assessed by means of a new image DNA cytometry technique.

DNA, Neoplasm↗
PubMed1989

Comparison between image and flow DNA cytometry in non-Hodgkin's lymphomas.

To assess the reliability of DNA estimation in cytological material, Feulgen lymph node imprints from 22 cases of malignant non-Hodgkin's lymphomas were examined by image cytometry (ICM) for both ploidy and cell kinetics, and the results obtained were compared with flow cytometry (FCM). The DNA distribution pattern was less accurate with ICM than with FCM; however, a high correlation was found between proliferative indices (r = 0.91) and between aneuploidy rates (agreement in about 86% of the cases) determined by FCM and ICM. Moreover, DNA tetraploid (or near-tetraploid) stem lines were more easily detected by ICM, due to the morphological selection of lymphomatous cells. The proliferation rate and the aneuploidy frequency according to morphological classification were in agreement with larger, previously reported studies with FCM. Therefore, ICM appears to supply additional complementary information to that obtained with FCM, particularly for the study of heterogeneous cell populations, which may be usefully applied to refine the large cell lymphoma subclassification.

DNA, Neoplasm↗
PubMed1989

Comparison of image analysis of imprints with flow cytometry for DNA analysis of solid tumors.

Quantitative analysis of cellular DNA content may be clinically useful for several solid tumors. The technology for this analysis by flow cytometry or image analysis has existed for several years but has not been widely used, except in a handful of specialized research institutions. Recently, however, relatively inexpensive image analyzers intended for use by hospital pathologists have been introduced that can analyze DNA content from cytology or imprint specimens which are readily obtainable from solid tumors. We report here an assessment of this technology for analysis of tumor imprint specimens, using flow cytometry of tissue blocks as the standard for comparison. We used image analysis equipment on Feulgan-stained imprint preparations from 31 tumors and compared the histograms with those obtained by flow cytometric analysis of archival tissue blocks from the same tumors. The ploidy descriptors (diploid, tetraploid, and aneuploid) were concordant for the two methods in 27 specimens, with three specimens yielding discordant results and one specimen considered unevaluable by image analysis. The image analysis method using imprints appeared to have several advantages over flow cytometry, including lower instrument cost, no need to dissociate paraffin blocks or fresh tissue, and ability to analyze very small samples. Somewhat lower resolution of the histograms, extremely localized tissue sampling, and possibly greater risk of occasionally obtaining unevaluable preparations were disadvantages. Microcomputer-based image analysis performed on imprints appeared to be a viable alternative to flow cytometric analysis of tissue blocks for quantitative DNA analysis of tumor specimens.

Aneuploidy↗
PubMed1997

DNA ploidy analysis in breast carcinoma. Comparison of unfixed and fixed tissue analyzed by image and flow cytometry.

OBJECTIVE: To form a methodologic basis for DNA analysis of ductal carcinoma in situ (DCIS) and invasive carcinoma (IC) of the breast, including very small lesions, by comparison of flow cytometric (FCM) and image cytometric (ICM) methods for DNA quantitation. STUDY DESIGN: The material consisted of 41 DCIS lesions and 26 ICs. FCM DNA analysis of unfixed, frozen samples were compared to (1) FCM of formalin-fixed, paraffin-embedded tissue; (2) ICM of imprints; and (3) ICM of paraffin-embedded tissue sections. RESULTS: FCM of unfixed tissue showed higher DNA measurement precision and a higher number of DNA nondiploid clones as compared to the other three methods. For the classification of DNA diploid/nondiploid cases, high concordance rates were found between the methods. Discordant cases were predominantly DNA neardiploid by FCM of unfixed tissue but DNA diploid by the other methods. The reproducibility of the DNA index (DI) was best in the interval 1.2 < DI < or = 2.2; it was 74% for FCM of fixed tissue and 79% for ICM of imprints. Clones with DI > 3 were found almost exclusively by ICM of imprints. For ICM of tissue sections, DI could not be reliably estimated. By ICM, contrary to FCM, a combined DNA diploid and nondiploid pattern was found frequently. CONCLUSION: Each of the methods has its own advantages and limitations. If possible, FCM should be combined with ICM. FCM of unfixed tissue is superior to the other methods with respect to precise DI estimation. Alternatively, FCM of fixed tissue and ICM of imprints may both give a reliable estimate of DI. ICM of tissue sections can discriminate DNA diploid from nondiploid clones, except for neardiploid subpopulations, and permits the analysis of very small lesions.

Breast Neoplasms↗
PubMed2003

A feature set for cytometry on digitized microscopic images.

Feature extraction is a crucial step in most cytometry studies. In this paper a systematic approach to feature extraction is presented. The feature sets that have been developed and used for quantitative cytology at the Laboratory for Biomedical Image Analysis of the GSF as well as at the Center for Image Analysis in Uppsala over the last 25 years are described and illustrated. The feature sets described are divided into morphometric, densitometric, textural and structural features. The latter group is used to describe the eu- and hetero-chromatin in a way complementing the textural methods. The main goal of the paper is to bring attention to the need of a common and well defined description of features used in cyto- and histometrical studies. The application of the sets of features is shown in an overview of projects from different fields. Finally some rules of thumb for the design of studies in this field are proposed. Colour figures can be viewed on http://www.esacp.org/acp/2003/25-1/rodenacker.htm.

Algorithms↗
PubMed1989

Image and flow DNA cytometry of small cell carcinoma of the lung.

Both image and flow DNA cytometry were performed in isolated nuclei from paraffin-embedded tumor tissue of patients with small cell carcinoma of the lung (SCCL). In 14 patients tissue was obtained by surgery from the primary tumor. From 14 patients tissue was taken by autopsy. From two patients tissue obtained by both surgery and later autopsy were available. From the autopsy patients tissue was taken only from the primary tumor (n = 6), from a metastasis (n = 1) and from the primary tumor and distant metastases (n = 7). Twelve of the tumors obtained by surgery were diploid, and two multiploid (two stem lines present). This was found both with image and flow cytometry. The group of patients could clearly be subdivided in short survivors (less than 9 months, n = 6) and long survivors (greater than 16 months, n = 8); since in both groups one multiploid and the remainder diploid cases were present, ploidy did not seem to be a good prognosticator for survival. In most (n = 26) of the tissues measured from the autopsy patients, again, a good correlation between image and flow DNA cytometry was obtained, the histograms being either (near) diploid or multiploid. In six cases, however, flow cytometry showed multiploidy whereas image showed aneuploidy (one single peak clearly deviating from diploidy). This discrepancy is caused because normal diploid (nonneoplastic) cells in the preparations could not be discarded from the flow cytometry measurements. Using the image cytometry data of the primary tumors, five diploid, three aneuploid, and four multiploid tumors were found. In five of the seven patients of whom tissue was obtained from the primary tumor and multiple metastases, differences between the histograms were found, mostly showing two malignant cell populations in one tissue and only one of them in another. Of one of the two patients of whom tissue was obtained by surgery and later autopsy, a change in histogram pattern was observed. It is concluded that although there is a high similarity between image and flow DNA cytometry, for an optimal interpretation of the histogram pattern, image measurements are more reliable. Ploidy determination does not seem to be of use in prediction of survival, and care should be taken in interpreting DNA histograms of metastases in SCCL patients because of the variability in histogram pattern.

Adult↗
PubMed1991

Quantitative DNA analysis in breast carcinomas: a comparison between image analysis and flow cytometry.

We compared the results of quantitative DNA analysis of fresh tumor tissue from 50 invasive breast carcinomas by image analysis and flow cytometry. For image analysis, Feulgen-stained slides of tumor imprints and of disaggregated tumor cytospin preparations were evaluated with the CAS-200 image analyzer. For flow cytometry, propidium iodide-stained disaggregated tumor cells were analyzed with the Coulter EPICS-C flow cytometer. The two methods yielded comparable results. The DNA indices obtained by the two methods showed close correlation by linear regression analysis (r = 0.86, P less than 0.001). There were 26 diploid (52%) and 24 nondiploid (48%) carcinomas. The ploidy pattern between the two methods showed agreement in 41 carcinomas (82%) and discordance in two (4%). Three tumors (6%) were equivocal by flow cytometry and four (8%) by image analysis. The equivocal cases presented potential sources of error in the evaluation of histograms in the near-tetraploid region by flow cytometry and in the near-diploid region by image analysis. Image analysis required smaller tissue samples and permitted direct visualization and selection of tumor cells. It also detected more tetraploid carcinomas. In contrast, flow cytometry analyzed larger cell samples and provided histograms with better resolution. It more readily detected the presence of multiple aneuploid peaks and also the presence of aneuploid peaks in the near-diploid range. The presence of aneuploidy was significantly related to the loss of hormone receptor expression, high mitotic rate, and high histologic and nuclear grades. Our study indicates that image analysis and flow cytometry provide comparable results in a majority of breast carcinomas.(ABSTRACT TRUNCATED AT 250 WORDS)

Breast Neoplasms↗