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PubMed1997

DNA ploidy and pS2 protein expression in breast cancer.

The DNA content of ductal breast carcinomas of varying histological grade was measured using static image cytometry and correlated with pS2 expression in the tumour cells. Our study was performed on imprint of surgical biopsies of 60 women with ductal breast cancer. A statistically significant difference was observed between pS2+ expression and grade of malignancy (P < 0.001). The percentage of euploid tumours significantly decreased from grade I to grade II to grade III (P = 0.01). The percentage of aneuploid tumours increased from pS2+ to pS2- breast tumours (P < 0.001). These findings may be indicative of pS2 and DNA ploidy alterations and tumour aggressiveness.

Aneuploidy↗
PubMedDate not supplied

DNA ploidy and proliferative index of cardiac myxoma.

Cardiac myxoma is the commonest tumour of heart. The histogenesis of this lesion is controversial as its origin has often been debated between the thrombogenic and neoplastic theories. We analysed DNA ploidy and proliferation indices of 30 cardiac myxomas which include 25 sporadic and five familial cases by image cytometry and proliferating cell nuclear antigen immunostaining. Of the 25 cases, 18 were aneuploid, three diploid and four tetraploid. Four of the five familial cases including the recurrent lesions were aneuploid. Poor tissue preservation precluded ploidy analysis in one familial case. The proliferation index of the sporadic cases ranged from 0.4 to 36.1 percent. The familial cases showed proliferation index between 10.2 and 22 percent. In addition to cardiac myxoma, proliferation index was assessed in 10 cardiac thrombi where it ranged from three to 58 percent. This study suggests that cardiac myxoma can be best interpreted as a neoplasm with a slow growth potential.

Adolescent↗
PubMed1987

Cytometry of breast carcinoma: significance of ploïdy balance and proliferation index.

Image cytometry of DNA distribution in fine needle biopsies of breast carcinomas at first diagnosis was performed to see if there were significant differences in DNA histograms between patients having very different outcome but same tumor histological typing and similar therapy. Two groups of patients were considered retrospectively: the first (20 patients) with survival time shorter than 5 years and the second (20 patients) with survival time longer than 10 years. Seven benign tumors were used as controls. Ten ploïdy classes were defined. The frequencies of cells in those classes were used as independent features in a supervised multivariate analysis. The advantages of this approach was pointed out with respect to the four-type classification of Auer. The scattering of DNA histograms within the feature space showed that a subgroup of patients with poor prognosis was clearly separated from a subgroup of patients with good prognosis but both long survival patients and short survival patients were scattered in between. In order to replace the multivariate classification of histograms by a simpler approach, two parameters were computed which explained most of the scattering in the feature space: the ploïdy balance (difference between the percentages of euploid and aneuploïd cells) and the proliferation index (percentage of cells between peaks). The scattergram of patients according to these parameters showed again that some DNA distributions were specific for either good or bad prognosis. But the separation was uncertain for seven short-survival patients and six long-survival patients. For six patients, the DNA distributions were very similar between long and short survival times. Those patients thus could not be separated even by means of discriminant analysis. The main conclusion of this study was that, for a significant number of patients, the objective multivariate classification of tumors DNA profiles is of little assistance to the pathologist who has to give a prognosis for the one patient under consideration.

Aneuploidy↗
PubMed2000

AgNORs in breast tumours.

There is evidence that the quantitative distribution of AgNOR proteins is a proliferation-related parameter that can be used as a prognostic index in tumour pathology. In breast cancer, some authors found a significant prognostic correlation of AgNOR protein quantity, whereas other did not. However, in all the reports dealing with AgNOR area (as opposed to count) this parameter was always turned out to be an independent prognostic indicator. The present study tests the significance of AgNOR proteins in a large series of primary breast carcinomas, exploring the associations between the AgNOR protein amount, as evaluated by image cytometry, and the other well-established prognostic markers commonly considered for breast cancer, along with patients' survival. Our results demonstrated a highly significant association between AgNOR protein quantity and tumour prognosis. Moreover, when the AgNOR area values were entered in multivariate analysis together with the other predictive parameters commonly considered in breast carcinomas, they showed an independent prognostic value together with Ki67-labelling index (LI), N-status and tumour size. Considering node-negative and -positive cases separately, the AgNOR protein area and Ki67-LI both come out as a independent predictors only in the latter group: the short follow-up time of our series (36 months median) could be responsible for this discrepancy.

Adenocarcinoma↗
PubMed1996

Cytometric measurement of cell proliferation in echo-guided biopsies from focal lesions of the liver.

Increased proliferative activity determined in surgical specimens of hepatocellular carcinoma (HCC) has been associated with tumor grade and patient survival. The measurement of cell proliferation in echo-guided biopsies of small focal liver lesions might provide useful information for the early recognition of malignancy and for predicting the aggressiveness of small HCCs. We assessed the diagnostic and prognostic value of cell proliferation in 91 echo-guided needle biopsies of focal liver lesions using the monoclonal antibody Ki-67, which detects a human nuclear antigen that is present in proliferating cells. Measurements were performed by image cytometry as the percentage of Ki-67 positive hepatocytes nuclei over total hepatocyte nuclei in the biopsy. At the histological examination, 27 lesions were diagnosed as chronic hepatitis, 10 as cirrhosis, 11 as macroregenerative nodule, and 43 as HCC in cirrhotic liver. Although the highest Ki-67 values (> 20%) were found in less-differentiated HCCs, most well-differentiated HCCs and nine borderline nodules were completely devoid of Ki-67-positive hepatocytes. A sustained Ki-67 labeling (up to 16%) was found in hepatitis and cirrhosis, similar to that found in several malignant tumors. In the HCC subset, Ki-67 labeling was strongly correlated to the Edmondson-Steiner histological grade. However, survival analysis did not indicate a better outcome for those patients with low-proliferating tumors.

Biopsy, Needle↗
PubMedDate not supplied

p53 expression, DNA ploidy and mitotic index as prognostic factors in patients with epithelial ovarian carcinoma.

AIMS AND BACKGROUND: Biological variables linked to genomic instability were examined and related to survival in 52 patients affected by ovarian carcinoma and nine patients with low malignant potential tumors (LMP). METHODS: DNA ploidy was measured by image cytometry in isolated neoplastic cells; the mitotic index was measured in Feulgen-toluidine blue-stained sections and p53 was investigated by immunohistochemistry. RESULTS: Twenty-five tumors (4 LMP) were peridiploid (ploidy < 2.25c), 22 tumors (4 LMP) were hyperdiploid (2.25c > ploidy < 2.9c) and 14 (1 LMP) had high ploidy (> or = 2.9c). MI ranged from 0.3 to 24.2 with a mean of 1.8 for LMP and 6.8 for carcinomas (P<0.001). Widespread p53 overexpression was detected in 49% of carcinomas and in none of the LMP tumors. CONCLUSIONS: Survival analysis performed in patients with carcinomas indicated that, of the examined biological variables, only MI was moderately associated with survival in a subgroup of early-stage patients.

Adult↗
PubMed1999

Prognostic significance of tumor ploidy and histomorphological parameters in adenocarcinoma of Barrett's esophagus.

BACKGROUND: Despite recent advances in surgical and multidisciplinary treatment, the prognosis for patients with adenocarcinoma of Barrett's esophagus remains poor. The low prognostic accuracy of even surgical pathologic TNM staging suggests that additional parameters are necessary in determining the prognosis. METHOD: In a retrospective analysis of 50 patients who underwent transhiatal or transthoracic esophageal resection due to adenocarcinoma of Barrett's esophagus, a quantitative DNA analysis using image cytometry was performed in addition to the TNM classification and usual morphological criteria. At the time of DNA analysis the histomorphological parameters and survival time were not known. RESULTS: The main prognostic parameter was the curativity (R classification) of the operation. Considering only patients after R0 resection, a multivariate analysis identified the DNA ploidy, the depth of tumor infiltration and distant metastases of prognostically independent factors. Furthermore, within pT2 and pT3 tumors, which account for 80% of all the tumors, DNA ploidy allows an additional prognostic differentiation which is not possible with pT stage alone. CONCLUSION: Patients with a diploid or tetraploid tumor without distant metastasis and a tumor stage pT1-pT3 should have curative (R0) resection. In case of an aneuploid DNA content or a pT4 tumor resection alone shows no advantage as compared to palliative nonoperative procedures.

Adenocarcinoma↗
PubMed1999

Wilms' tumor. Role of fine needle aspiration and DNA ploidy by image analysis in prognostication.

OBJECTIVE: To evaluate the utility of cytomorphologic features and DNA ploidy estimation in fine needle aspirates (FNAs), from Wilms' tumors for prognostication. STUDY DESIGN: Twenty-three cases of Wilms' tumor having FNA and follow-up data were selected. Cytomorphology was analyzed by two observers. DNA ploidy was determined in 19 cases by image cytometry by destaining Papanicolaou-stained slides and restaining with Feulgen stain. Various parameters and patient outcomes were compared, and statistical evaluation was done. RESULTS: Poor outcome (12/23 cases) was associated with age < 2 years (P = .01), severe pleomorphism of blastemal cells (4/23 cases, P < .05), very large nucleoli (5/23 cases, P = .075), atypical mitosis (6/23 cases, P = .032) and aneuploidy/tetraploidy of tumor cells (6/29 cases, P = .005). Term unfavorable cytology is proposed when a combination of severe pleomorphism, very large nucleoli and atypical mitosis is seen in FNA smears. Four Wilms' tumor FNAs were characterized as showing unfavorable cytology, and all had a poor outcome (P = .0351). Three of the six cases with aneuploid/tetraploid features also showed unfavorable cytology. CONCLUSION: Unfavorable cytology and aneuploidy/tetraploidy in FNA smears of Wilms' tumor are associated with a poor prognosis.

Age Factors↗
PubMed2006

Correlation of protein expression, Gleason score and DNA ploidy in prostate cancer.

The prognosis of prostate cancer correlates with tumor differentiation. Gleason score and DNA ploidy are two prognostic factors that correlate with prognosis. We analyzed differences in protein expression in prostate cancer of high and low aggressiveness according to these measures. From 35 prostatectomy specimens, 29 cancer samples and 10 benign samples were harvested by scraping cells from cut surfaces. DNA ploidy was assessed by image cytometry. Protein preparations from cell suspensions were examined by 2-DE. Protein spots that differed quantitatively between sample groups were identified by MS fingerprinting of tryptic fragments and MS/MS sequence analysis. We found 39 protein spots with expression levels that were raised or lowered in correlation with Gleason score and/or DNA ploidy pattern (31 overexpressed in high-malignant cancer, 8 underexpressed). Of these, 30 were identified by MS. Among overexpressed proteins were heat-shock, structural and membrane proteins and enzymes involved in gene silencing, protein synthesis/degradation, mitochondrial protein import (metaxin 2), detoxification (GST-pi) and energy metabolism. Stroma-associated proteins were generally underexpressed. The protein expression of prostate cancer correlates with tumor differentiation. Potential prognostic markers may be found among proteins that are differentially expressed and the clinical value of these should be validated.

Aged↗
PubMed1997

AgNOR protein quantity of cervical smears correlates with that of histological sections in cervical intraepithelial neoplasia.

The quantitative distribution of AgNOR proteins has been determined by image cytometry in 52 cervical smears obtained from normal cervix (n = 20), grade I CIN (cervical intraepithelial neoplasia) (n = 3), grade II CIN (n = 5) and grade III CIN (n = 24). No significant difference was demonstrated in the mean AgNOR protein area values between normal cervix, CIN I and CIN II, while AgNOR protein scores of CIN III were significantly greater than those of normal cervix (p < 0.05). AgNOR protein quantity was also determined in 17 colposcopic directed biopsies of patients with CIN lesions (3 CIN II and 14 CIN III) already studied by cytological analysis. When AgNOR protein values of histological sections and corresponding cytological smears were compared by linear regression analysis, a significant correlation was found (r = 0.74, p < 0.05). Our results demonstrate that AgNOR protein quantity of cervical smears actually reflects that of the cervical epithelium in situ and may therefore be useful for the cytological diagnosis of cervical lesions.

Biopsy↗
PubMed1996

DNA-aneuploidy as marker for neoplasia in G1-urothelial carcinomas.

Biopsies from 60 grade-1 urothelial carcinomas and 50 normal bladder mucosae have been investigated by DNA-image-cytometry after enzymatic cell separation to determine the diagnostic sensitivity of DNA-aneuploidy for the identification of low grade urothelial neoplasia. Two different modes to detect DNA-aneuploidy were compared: the conventional application of a threshold at 2.2c for the modal DNA value and the use of the Kolmogoroff-Smirnow (KS) test to compare GO/1 phase fractions of the cells under analysis and the internal reference cells. Whereas the specificity for both types of analysis was 100%, the sentitivity of the latter method was 73.3% compared to 48.3% of the conventional procedure. Therefore, the application of the KS-test for the identification of DNA- stemlines as aneuploid results in an increased detection rate was compared with the conventional threshold procedure. The high prevalence of DNA- stemline aneuploidy even in low grade urothelial cancers may qualify this marker as a possible aid in the cytologic evaluation of urine and bladder washings.

Aneuploidy↗
PubMed2000

p120 expression provides a reliable indication of the rapidity of cell duplication in cancer cells independently of tumour origin.

p120 is a nucleolar protein that has been immunocytochemically detected in rapidly proliferating cells of a variety of human malignancies. In the present study, the relationship between p120 expression and the rapidity of cell proliferation was evaluated in 48 human tumours of different origins. The cell proliferation rate of cancer cells was determined by quantitative analysis of AgNOR proteins. p120 immunostaining and AgNOR protein quantity were measured by image cytometry and a highly significant correlation was found between the two variables, as evaluated by linear regression analysis (r=0.98, p<0.0001). The relationship between p120 expression and the rapidity of cell duplication was also studied in vitro, in six human cancer cell lines derived from different tumour types, characterized by various doubling times (ranging from 20 to 77 h). p120 expression was determined on western blots using specific anti-p120 monoclonal antibodies. Densitometric analysis revealed a highly significant inverse correlation between the integrated optical density values of the chemoluminescence bands at 120 kD and the cell line doubling times (r=-0.93; p=0.007). The same result was obtained in situ by correlating p120 immunostaining of the cytological preparations obtained from the six cancer cell lines with the corresponding doubling time (r=-0.98, p<0.0001). These results indicate that in cancer cells, the quantitative expression of p120 is directly related to the rapidity of cell duplication, independently of the tumour origin.

Antibodies, Monoclonal↗
PubMed1999

Time-course analysis of type II cell hyperplasia and alveolar bronchiolization in rats treated with different particulates.

In this study, we compared the morphological reaction patterns in rat lungs following a single intratracheal instillation of 20 mg quartz, 20 mg coal mine dust (15.3% quartz), or 25 mg talc. Control animals received a single dose of 0.5 ml saline solution intratracheally. Investigations by light microscopy, morphometry, and DNA image cytometry were carried out 3, 6, 12, and 18 mo after dust administration. During the investigation period, we observed a temporary increase in the number, area, and proliferative activity of the type II cells, which differed in intensity among the three dusts. After 18 mo, however, type II cells in treated animals did not differ from control animals. On the other hand, the expansion of a multifocal alveolar bronchiolization as putative preneoplastic lesion had progressed enormously by the end of the test (1-3% of the investigated lung area). Consistent with this, the proliferative activity of the epithelial cells in terminal bronchi of the coal mine dust- and quartz-treated animals was enhanced by the end of the 18-mo investigation period, while the reaction to talc was minimal (0.2% of totally investigated lung area). Our data suggest that in bronchiolo-alveolar regions, especially in the epithelium of terminal bronchi, there is an overshoot regeneration after cell damage that leads to an alveolar bronchiolization.

Air Pollutants↗
PubMed2003

Lung cancer screening: a different paradigm.

Thoracic computed tomography (CT) is a sensitive method for detecting early lung cancer but has a high false-positive rate and is not sensitive for detecting central preinvasive and microinvasive cancer. Our hypothesis was that automated quantitative image cytometry (AQC) of sputum cells as the first screening method may improve detection rate by identifying individuals at highest risk for lung cancer. A total of 561 volunteer current or former smokers 50 years of age or older, with a smoking history of more than or equal to 30 pack/years, were studied. Among these, 423 were found to have sputum atypia defined as five cells or more with abnormal DNA content using AQC. Noncalcified pulmonary nodules were found in 46% (259/561). Of the 14 detected cancers, 13 were detected in subjects with sputum atypia-nine by CT and four carcinoma in situ/microinvasive cancers by autofluorescence bronchoscopy. One cancer was detected by CT alone. AQC of sputum cells improved the detection rate of lung cancer from 1.8 to 3.1%. CT scan alone would have missed 29% of the cancers. This screening paradigm shift has the additional potential of reducing the number of initial CT scans by at least 25% with further savings in follow-up investigations and treatment.

Aged↗
PubMed1999

[The accessory diagnosis of rectal carcinoma by ICM and oncogene expressions].

OBJECTIVE: To enhance the diagnostic accurate rate of preoperative biopsy on rectal carcinoma. METHODS: The paraffin embedded biopsy tissues of 17 cases of preoperatively unconfirmed rectal carcinoma and 15 cases of normal rectal mucosa (control group) were examined by routine pathology, image cytometry (ICM), and the expressions of protein products of oncogene p53 and ras. RESULTS: In the experimental group, 53% specimens were classified as non-diploid by ICM, and the positive rates of p53 and ras expressions were 71% and 47% respectively. The positive rates were higher than those of routine pathology and the differences were significant. In the control group, the results of ICM and oncogene expressions were negative. In the experimental group, there were 11 cases of atypical hyperplasia and 5 cases of chronic inflammation only. Non-diploid, and positive expressions of p53 and ras were found in 8, 11 and 7 cases of atypical hyperplasia respectively; however, they were all negative in chronic inflammation. The differences between atypical hyperplasia and chronic inflammation were significant. ICM was correlated well with expressions of p53 and ras oncogene. CONCLUSIONS: The techniques of ICM and detecting the expressions of protein products of oncogene p53 and ras can be a supplementary tool for routine pathology to improve the diagnostic rate of rectal carcinoma.

Biomarkers, Tumor↗
PubMed2002

Usefulness of DNA ploidy measurement on liquid-based smears showing conflicting results between cytology and high-risk human papillomavirus typing.

To improve the positive predictive value (PPV) for high-risk human papillomavirus (HR-HPV) in primary screening, DNA ploidy was measured on the same liquid-based sample by image cytometry in 984 cases showing discrepancies between cytology and HR-HPV testing. Of the conflicting results, 14.5% corresponded to a cytologic lesion (from atypical squamous cells of undetermined significance to high-grade squamous intraepithelial lesion [HSIL]) without HPV detected, and 85.5% of smears were within normal limits but revealed an HR-HPV infection. A suspect DNA profile was associated significantly with a lesion. In 497 patients who underwent repeated HPV testing, a normal DNA profile at the first smear predicted the clearance of HPV infection (sensitivity, 81.5%; specificity, 45.4%; PPV, 69%; negative predictive value, 62.4%). In persistent HR-HPV infection, a suspect DNA profile at the first smear increased the PPVfrom 10.8% to 22.7% for the detection of a histologically proven HSIL with a sensitivity of 95.2%. DNA ploidy can be used to select smears with high risk of HSIL, especially in cases of persistent HR-HPV infection.

Adolescent↗
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↗
PubMed2005

Cytologically malignant lymphoid pericardial effusion with benign clinical outcome.

BACKGROUND: Isolated malignant pericardial effusion is a manifestation of primary cardiac lymphoma (PCL) and primary effusion lymphoma (PEL), rare types of non-Hodgkin's lymphoma (NHL). The diagnosis is based on different cytological methods and analyses including DNA-image cytometry (ICM-DNA). DNA-aneuploidy has been reported to be highly specific for malignancy. CASE DESCRIPTIONS AND RESULTS: A 75-year-old man and a 66-year-old woman underwent urgent pericardiocentesis for cardiac tamponade due to large pericardial effusion. In both patients pericardial fluid analysis showed highly atypical blastic lymphoid cells expressing CD45 (both patients) and CD20 (assessed only in one patient), and ICM-DNA revealed significant DNA-aneuploidy (2c deviation index 9.22 and 10.73 respectively, 75% and 60% respectively of the target nuclei in aneuploid areas). Extensive staging examinations did not identify any other tumour manifestation. Although in neither of the two patients systemic chemotherapy was administered, both were free of cancer after a follow-up of ten and nine years respectively. CONCLUSIONS: Despite the highly atypical cytomorphology including unequivocal DNA aneuploidy, long-term survival in both patients strongly suggests that pronounced reactive lymphocytic changes are probably due to viral pericarditis rather than PCL or PEL as underlying conditions. It seems that DNA-aneuploidy may be not absolutely specific for the detection of malignant lymphoid cells in pericardial fluid.

Aged↗