AgNOR Dots Research Articles

Association between Ag-NOR dots and immunohistochemical markers in different grades of breast ductal carcinoma

Association between Ag-NOR dots and immunohistochemical markers in different grades of breast ductal carcinoma

Evaluation of Oxidative Stress and Histiocytic Behaviour of Canine TVT using Alpha Antitrypsin

Background: There is scanty literature on evaluation of oxidative stress and alpha antitrypsin marker expression in canine TVT to know its histiocytic behaviour. Since venereal transmissible tumor is one of most important tumor affecting large population of canine, thus, study was conducted to investigate the above based on clinicopathology. Methods: The study was conducted on dogs suffering from transmissible venereal tumor (TVT) for evaluation of clinicopathological alterations, oxidative stress and expression pattern of alpha antitrypsin. Ten bitches and five male dogs with tumour masses seen on the external genitalia were used for the study. Detailed alterations due to TVT with respect to haematobiochemistry, oxidative stress, pathomorphology, proliferation and expression pattern of alpha antitrypsin were assessed. Result: Grossly tumors on the genital area were either pedunculated or sessile and of various shape such as cauliflower, filiform and irregular. Hematological alterations such as normocytic normochromic type of anaemia, thrombocytopenia, neutropenia and lymphocytopenia in TVT cases indicated immunosuppression. Significant alterations in liver enzymes, hypercalcemia and hypoglycaemia were also observed in TVT affected dogs. Increased level of MDA and decreased in concentration of antioxidant enzymes indicated association of oxidative stress in proliferation of tumor and reduction in number of T lymphocytes. Cytological examination revealed characteristic punctuate basophilia and vacuolation in tumour cells. Histopathological examination showed characteristic round cells with a little fibrous septa in a sheet like manner. AgNOR staining indicated moderate proliferative capacity as evidenced by scattered AgNOR dots. TVT cells showed moderate type of immunoreactivity with alpha anti-trypsin confirming its histiocytic origin upon immunohistochemistry.

Study of Association of AgNOR vs Ki-67 in Thyroid Lesions

The thyroid is an important endocrine gland, which plays a major role in the functions of almost all normal metabolic processes. In present study, by focusing the sections at very high magnification (100x, oil immersion level) AgNOR dots are visible as black dots inside the nucleus. Our studies which have reported that thyroid lesions with less AgNOR counts are more likely to regress as compared to those with high AgNOR counts and that showed its prognostic significance. Thus, the AgNOR technique can be useful as a supportive tool in both the prognosis and therapeutic value in thyroid lesions most commonly in malignant lesions that are stained by routinely performed haematoxylin and eosin. The Present Study is aim to find out the prevalence of thyroid lesions in the patients and also to Study and find out the different types of neoplastic and non- neoplastic thyroid lesions and their relation with AgNOR and Ki-67 proliferation makers. The Present Study focuses on to evaluate the association of AgNOR vs Ki-67 in the prognostic value of thyroid lesions.

Diagnostic and Prognostic Significance of AgNOR Counts in Before and After Radiotherapy-Treated Cervical Carcinogenesis is Compared with Cervical Cytology and Histology

Nucleolar organizer region-associated proteins are argyrophilic and visualized by silver stains. There are two types of dots are present. One is the presence of single dots, and another is pleomorphic dots. Pleomorphic dots are few, increase with severity of cervical lesions and hence indicate a more severe prognosis. The present study aimed at investigating the diagnostic value of both pleomorphic and single AgNOR dots in cervical carcinogenesis in different cytological stages of cervix before and after radiotherapy. The results were correlated with cervical cytology and histopathology. The study was conducted on the samples of 37 untreated cases of patients coming to the outpatient door of AHRCC. In two slides, samples were taken for PAP smear cytology and AgNOR staining. From histopathological detection, various grades of squamous cell carcinoma of the cervix were determined. Samples were divided into cytological/histological normal/controls (N = 10), inflammation (N = 7), LSIL (N = 8), HSIL (N = 7) and invasive carcinoma (N = 5). AgNOR staining was performed slides containing cervical smears. Samples from same patients [LSIL (N = 8), HSIL (N = 7) and invasive carcinoma (N = 5)] that were subjected to radiotherapy were assessed again for AgNOR scoring. The rate of increase in cell counts of pleomorphic dots was higher than the rate of increase in cell counts of single dots, and there is a positive and significant correlation of cell counts of both pleomorphic and single dots with disease severity. The results were correlated with PAP stained cervical cytology and histopathology to find out the diagnostic and prognostic significance. In squamous cells of the normal/control group, the AgNORs were fine and centrally placed in the nucleoli. AgNORs in the case of LSIL, HSIL and invasive carcinoma of the cervix before radiotherapy were large and variable in size and shape. The cases studied for AgNORs count after 6 month of radiotherapy showed less number of AgNORs dots which were single and fewer in numbers as compared to the pre-radiation group. AgNOR pleomorphic dot counts increase with severity of cervical lesions and may be used as an effective tool along with cervical cytology and histopathology for detection of cervical carcinogenesis.

Significance of Silver Stained Nucleolar Organiser Region (AgNOR) in Fine Needle Aspiration Smears of Breast Lesions

Background: Breast cancer is the leading cause of cancer - related deaths in Asia. The number of intra-nuclear silver stained structures, termed AgNORs, is significantly higher in malignant cells than in normal, reactive or benign cells. The purpose of this study was to evaluate the AgNOR scores in FNA smears of breast lesions and their correlation with histopathology. Aims & Objective: To establish AgNOR staining as a diagnostic and prognostic tool in management of various breast lesions. Methods: This was a prospective study conducted over a period of 18 months may 2015 to November 2016. A total of 100 cases were included in the study. AgNOR stain was done in both cytology and scoring was done and analysed. Results: In FNAC aspirates, Mean AgNOR count ranged from 5 to 9.9 per high power field with a mean value of 2.888±2.553. The AgNOR dots morphology was homogenous, symmetric with regular contours in FNAC slides of benign breast lesion. In malignant breast lesions, the dots were asymmetric with irregular contours and were aggregated, smaller and more scattered. Conclusions: The present study showed that fine needle aspiration cytology is a useful modality for diagnosis of breast lesions. It has a high concordance with the histopathology . AgNOR count assessment provides a useful objective measure for segregation of different grades of tumor with 100% accuracy for detection of higher grade of lesions, as observed in present study. For differentiation of benign from malignant lesions too it has a high sensitivity as well as specificity. The usefulness of FNAC to evaluate nodal involvement also showed a 90% sensitivity and 95.6% specificity. On the basis of present study, it could be concluded that AgNOR count estimation using fine needle aspiration is a useful method to differentiate and diagnose breast lesions.

How Useful is the AgNOR Staining Method for the Diagnosis of Salivary Gland Tumors?

Objectives: The purpose of this study was to determine the numbers and mean areas of the AgNOR dots in the 3 most common salivary gland tumors. Materials and Methods: One hundred and seventy paraffin blocks histologically diagnosed as pleomorphic adenoma, adenoid cystic carcinoma, and mucoepidermoid carcinoma were selected from the archives of the pathology department of Loghman hospital, Tehran, Iran and then cut and stained using the standard AgNOR method. The number and area of AgNOR dots were evaluated. Chi-square and Pearson correlation (2-tailed) tests were used for statistical analysis. The significance level was P < 0.05. Results: The chi-square test showed significant differences between tumor types and the numbers of AgNORs (P = 0.000) and between tumor types and areas of AgNORs (P = 0.000). Conclusions: The AgNOR technique can be used to diagnose salivary gland tumors.

Prognostic Significance of Two Dimensional AgNOR Evaluation in Local Advanced Rectal Cancer Treated with Chemoradiotherapy.

The prognostic significance of AgNOR proteins in stage II-III rectal cancers treated with chemoradiotherapy was evaluated. Silver staining was applied to the 3μm sections of parafin blocked tissues from 30 rectal cancer patients who received 5-FU based chemoradiotherapy from May 2003 to June 2006. The microscopic displays of the cells were transferred into the computer via a video camera. AgNOR area (nucleolus organizer region area) and nucleus area values were determined as a nucleolus organizer regions area/total nucleus area (NORa/ TNa). The mean NORa/TNa value was found to be 9.02±3.68. The overall survival and disease free survival in the high NORa/TNa (>9.02) patients were 52.2 months and 39.4 months respectively, as compared to 100.7 months and 98.4 months in the low NORa/TNa (<9.02) cases. (p<0.001 and p<0.001 respectively). In addition, the prognosis in the high NORa/TNa patients was worse than low NORa/TNa patients (p<0.05). In terms of overall survival and disease-free survival, a statistically significant negative correlation was found with the value of NORa/TNa in the correlations tests. Cox regression analyses demostrated that overall survival and disease-free survival were associated with lymph node status (negative or positive) and the NORa/TNa value. We suggest that two-dimensional AgNOR evaluation may be a safe and usable parameter for prognosis and an indicator of cell proliferation instead of AgNOR dots.

Quantitative analysis of agnor counts of buccal mucosal cells of chewers and non chewers of gutkha: A comparative cytologic study.

The present study was taken up to evaluate the AgNOR counts in the buccal mucosa cells of gutkha chewers and compare that with the sex-matched controls. In all, 100 gutkha chewers and 50 sex-matched non-chewers (controls) were chosen. None of the patients in both groups had any clinical oral lesions or systemic diseases. After rinsing with 0.9% sodium chloride, cytologic smears were prepared and stained using the AgNOR method and observed in immersion oil at 1000 × magnification. Finally, 50 cells were selected at random; AgNOR dots were counted and their mean was recorded. The student t-test was used for analysis of data. Comparison between mean AgNOR counts of gutkha chewers (2.68 ± 0.23) and non-chewers (2.01 ± 0.14) was found to be statistically significant. Cytology associated with AgNOR staining can effectively detect the early molecular changes within buccal mucosa cells of oral mucosa.

Evaluation of AgNoR scores in aspiration cytology smears of breast lesions and their correlation with histopathology

Background: Breast cancer is the leading cause of cancer-related deaths in Asia. The number of intra-nuclear silver stained structures, termed AgNORs, is significantly higher in malignant cells than in normal, reactive or benign cells. The main purpose of this study was to evaluate the AgNOR scores in aspiration cytology smears of breast lesions and their correlation with histopathology.Materials and Methods: This was a prospective study conducted over a period of 24 months from October, 2009 to 2011. A total of 40 cases were included in the study, including fine needle aspiration and biopsy. AgNOR stain was done in both cytology and histopathology slides and scoring was done and analyzed.Results: The mean AgNOR counts were 1.734 for benign cases and 4.508 for malignant cases. Statistically significant ‘P’ value &lt; 0.05 for both benign and malignant cases was observed. The AgNOR dots morphology was homogenous, symmetric with regular contours in both FNAC and histopathology slides of benign breast lesion. In malignant breast lesions, the dots were asymmetric with irregular contours and were aggregated, smaller and more scatteredConclusion: The mAgNOR counts were significantly higher in smears from malignant breast lesions than in those from benign. These results were similar to those obtained for tissue sections and were comparable to established data. Both AgNOR counts and SAPA score gave similar results done in this study, indicating that SAPA is also as convenient, reproducible and rapid method of AgNOR evaluation.DOI: http://dx.doi.org/10.3126/jpn.v4i8.11592 Journal of Pathology of Nepal; Vol.4,No. 8 (2014) 649-653

Reduction of the argyrophilic nucleolar organizing region associated protein synthesis with age in buccal epithelial cells of healthy individuals.

Nucleolus organizer regions (NORs) consist of the rRNA coding gene family (rDNA) in the cell nucleus. The argyrophilic proteins are selectively stained with silver nitrate and bind these regions. It was reported that NOR (rDNA) activity decreases in human lymphocytes, fibroblasts and bone marrow with age. However, to our knowledge there have not been any studies related to the NORs in oral epithelial cells of healthy individuals. Our aim is to detect any correlation between age and Total AgNOR area/Total nucleus area (TAA/TNA) values in buccal epithelial cells of healthy individuals. Oral epithelial cells from 50 healthy individuals (age range of 2-80 years old) were spread onto a clean glass slide, air dried and fixed. Then the AgNOR staining protocol was performed on these cells. TAA/TNA ratio and AgNOR dots were calculated using software. From each person 50 oral epithelial cells were evaluated. Statistically significant correlations were found between mean TAA/TNA values and age (Rsq = 0.534, p < 0.001 for linear and Rsq = 0.728, p < 0.0001 for polynominal regression), and between AgNOR number and age (Rsq = 0.621, p < 0.001 for linear and Rsq = 0.693, p < 0.0001 for polynominal regression). There is a significant correlation between age and AgNOR amount (ribosome biosynthesis rate) in buccal epithelial cells of healthy individuals. AgNORs in buccal epithelial cells may be used for detection of age.

Silver binding nucleolar organizer regions dots in oral leukoplakia with epithelial dysplasia and oral squamous cell carcinoma: an in vivo study.

Silver binding nucleolar organizer regions (AgNOR) in normal oral mucosa (NOM), oral leukoplakia with epithelial dysplasia (ED), and oral squamous cell carcinoma (OSCC) were studied. The mean AgNOR count per nucleus increased from NOM to ED to OSCC. Tissue showing ED in oral leukoplakia and OSCC cases showed higher counts, wider scatter, and smaller size of AgNOR dots in the nuclei. The study seems to suggest that time method has some potential in distinguishing between NOM and oral leukoplakia with ED and OSCC. Studies of larger numbers are needed to arrive at more substantial conclusions.

Significance of AgNORs and Ki-67 Proliferative Markers in Differential Diagnosis of Thyroid Lesions

We aimed to assess the utility of quantitative analysis of AgNORs and Ki67 labeling index (LI) in the differential diagnosis of different thyroid lesions. This study included: 25 papillary carcinomas, 7 follicular carcinomas, 21 follicular adenomas and 27 nodular goiters. Using a semiautomatic image analysis system, Ag NORs parameters were measured and calculated including: total area of AgNORs, mean Ag NOR number in nuclei, nuclear area, mean area of AgNOR dots per each nucleus, number of central and marginal AgNOR dots, and the relative ratio of total area of AgNOR dots/total area of nucleus. Ki67 immunostaining was performed and the LI was determined. There was a significant difference between groups of thyroid lesions regarding total area of AgNORs, Ag NOR number and number of marginal Ag NOR dots. According to receiver operating characteristic curve, Ag NORs number =2.91 and marginal Ag NORs = 2.67 were useful cut off values above which follicular carcinoma can be diagnosed with 100% sensitivity, 79% specificity, 76% PPV, 100% NPV and 85% diagnostic accuracy for both parameters. Mean Ki67 LI in our study was 14.12 ± 2.29, 61.42 ± 3.77, 34.90 ± 3.49 and 18.60 ± 1.96 for papillary carcinoma, follicular carcinoma, follicular adenoma and nodular goiter respectively. Ki67 LI showed statistically significant difference between follicular carcinoma and follicular adenoma (p = 0.026) and between papillary carcinoma and follicular adenoma (p = 0.007). Quantification of Ag NORs and Ki67 LI could be used as helpful ancillary methods in the differentiation between different thyroid lesions.

Histopathological study comparing upstream binding factor expression and AgNOR staining

AgNOR staining has been in past years, the subject of numerous publications, which have failed to reach agreement regarding its usefulness as a proliferation marker. This silver staining method does not react with NORs (actual chromosome regions containing rRNA (ribosomal RNA) genes), but with proteins associated with them, whose quantity increases in parallel with ribosome biogenesis. The transcription factor UBF (upstream binding factor) is associated with NORs and has an important regulatory role in rRNA synthesis as cofactor of RNA polymerase I. Recent research has revealed an additional cytoarchitectural function of UBF in decondensing r-chromatin (ribosomal-chromatin). Immune detection of UBF expression and AgNOR counts are closely correlated as both techniques identify substrates in or closely adjacent to NORs. However, contrary to AgNOR dots, the UBF signal disappears in cells which undergo apoptosis or terminal differentiation. These features imply that UBF evaluation would reflect tumour cell proliferation (growth fraction) more accurately than AgNOR counts. Here we also show that immunohistochemical staining of UBF may reveal distinct active NORs with open, decondensed chromatin and we hypothesize that the large stretches of decondensed r-chromatin revealed by UBF staining may correspond to clusters seen after silver staining and, conversely, shorter areas of decondensed r-chromatin should match the small AgNOR grains typically found in some tumour types. The length of decondensed r-chromatin may be a reflection of the ratio of active to silent r-RNA genes.

Comparative morphometric study of AgNORs in variants of ameloblastoma

Aims:To compare the area and number of AgNORs (silver stained nucleolar organizer regions) by morphometry between follicular and plexiform variants of ameloblastoma in order to analyze their cell proliferation rates.Materials and Methods:This retrospective study was carried out on 30 cases each of follicular and plexiform ameloblastoma. The sections were obtained and stained with silver staining technique to identify the nucleolar organizer regions. AgNORs were quantified using two parameters; manual tag for the number of AgNORs and area measurement using the image analyzer software, Image-Pro-Express.Results:Morphometric area measurements of AgNOR were significantly higher for Plexiform ameloblastoma (0.831μm2) than follicular ameloblastoma (0.528μm2). Enumeration of the number of AgNORs showed a significantly higher number of AgNOR for follicular ameloblastoma (1.71) than plexiform ameloblastoma (1.43). Among the groups studied, follicular ameloblastoma was more aggressive than plexiform ameloblastoma, as it showed smaller AgNOR area and higher AgNOR number.Conclusion:The combination of counting the number and measuring the area of AgNOR dots showed a significant overall difference between AgNOR profiles of follicular and plexiform variants of ameloblastoma.

Comparison of Ki67 and modified AgNOR in dysplastic epithelial lesion, primary squamous cell carcinoma and metastatic squamous cell carcinoma of head and neck

ABSTRACTThe variations in nucleolar organizer regions (AgNOR) and the increase in Ki67 expression and correlation with histological findings in 64 specimen squamous epithelium from normal epithelium to overt carcinoma were evaluated. Mean AgNOR counts (mAgNOR), proliferative index (pAgNOR), variation in size and dispersion of AgNOR dots in cells were graded and compared in normal, dysplastic, primary and metastatic squamous cell carcinoma (SCC). The Ki67 percentage is significantly increased from normal squamous to SCC group, and the reactivity of staining were related to histological differentiation. The mAgNOR counts were high in all 44 cases of primary and metastatic SCC and low in normal squamous tissue and increased in dysplastic lesions. The mAgNOR count in SCC increased from well differentiated to poorly differentiated. PAgNOR counts were gradually increased in different grade of SCC, 1.4% in well differentiated, 6% in moderately differentiated, 19% in poorly differentiated. There was only significant difference between well differentiated versus poorly differentiated cases. Only significant correlation between Ki67 percentage and mAgNOR and between Ki67 percentage and pAgNOR in primary SCC group was noticed. Therefore, Ki67 is gold standard method for evaluation of proliferation activity. AgNOR quantity is proportional to the proliferative activity of the cell but does not necessarily indicate malignancy.

Oral epithelial atypical changes in apparently healthy oral mucosa exposed to smoking, alcohol, peppers and hot meals, using the AgNOR and Papanicolaou staining techniques

To evaluate cytological atypical changes in apparently healthy oral mucosa exposed to smoking, alcohol, hot meals, and peppers using the AgNOR and Papanicolaou methods. A total of 180 individuals were evaluated, of which 60 were smokers, 34 were alcohol users, 52 were habitual peppers and hot meal (exposed) consumers, 24 were non-exposed, and 10 were patients with Oral Squamous Cell Carcinoma (OSCC), as an internal control. Cytological materials were obtained by brushing of buccal mucosa, on the border of the tongue and on the floor of the mouth, and participants underwent the Papanicolaou test for cytological changes and AgNOR staining for evaluation of the mean number of AgNOR dots per nucleus. SPSS program was used to perform the Pearson chi-square test. The 95% confidence level, Odds Ratio (OR), and the 95% Confidence Intervals (CI) were used. The features of cytological atypia were verified among 10 individuals, including 5 smokers, 2 alcohol users, 2 hot meals and peppers consumers, and one non-exposed. For atypia among tobacco smokers, the adjusted Odds Ratio (OR) and the 95% CI were found to be 2 (0.246-16.24). Increased keratinization was detected among 27 (45%) of the smokers (P < 0.0001), 17 (32.7%) of the pepper and hot meals consumers (P < 0.005), 4 (11.8%) of the alcohol consumers, and among 2 (3.7%) of the non-exposed group. Statistical analyses revealed a greater mean number of AgNORs per nucleus in smokers (3.68) followed by (2.82) alcohol consumers, compared to the habitual peppers and hot meal consumers (2.28) and the non-exposed group (2.00). What's more, 80% of the smears with cytological atypia were identified with 6 +/- 2 AgNOR mean count. The increase of the variables suggests that the evaluation of epithelial atypical changes in individuals exposed to smoking and alcohol carcinogens may be a useful screening tool. While hot meals and peppers did not seem to be a risk for oral mucosal proliferation, they increased the potency of keratinization and infection.

PERBEDAAN INDEKS PROLIFERASI SEL KISTA DENTIGEROUS, KISTA DENTIGEROUS YANG TIMBUL BERSAMA DENGAN AMELOBLASTOMA PLEKSIFORM DAN AMELOBLASTOMA PLEKSIFORM DENGAN MENGGUNAKAN PEWARNAAN AGNOR

The purpose of the study to seek the differences of cell proliferation index among dentigerous cyst, dentigerous with ameloblastoma and ameloblastoma plexiform. This study utilized retrospective method, which was achieved by collecting data from medical record in oral surgery clinic, Cipto Mangunkusumo Hospital and Pathology Anatomy Laboratory at Faculty of Medicine University of Indonesia from Januari 1998 to April 2002. Histological examination were prepared from 34 samples; consist of 15 dentigerous cysts, 11 ameloblastoma and 8 dentigerous cyst which arise with ameloblastoma. In each case, intra-nuclear AgNOR dots were counted in 100 consecutive basal nuclei. Statistic analysis using ANOVA show the difference among study objects (p<0.05). AgNOR an ameloblastoma is significantly higher than in dentigerous cyst. The differences may indicate the variation of metabolic, proliferation or transcriptional activities of the cell. The study found the difference of AgNOR count in cell proliferation index among dentigerous cyst, dentigerous cyst which arise with ameloblastoma and ameloblastoma. The finding can be used to determine the diagnoses in doubtfull cases where dentigerous and ameloblastoma cannot be distinguished clinically and pathologically.

9p21 Deletion in the diagnosis of malignant mesothelioma in serous effusions additional to immunocytochemistry, DNA-ICM, and AgNOR analysis

The diagnosis of malignant mesothelioma (MM) in serous effusions is difficult but may be achieved by the application of adjuvant methods. The authors cytologically diagnosed 33 effusions as suspicious or positive for MM cells by using DNA-image cytometry (DNA-ICM), immunocytochemistry and AgNOR analysis. The authors further detected 9p21 deletions by chromosomal fluorescence in situ hybridization (FISH). In addition, 31 cases of metastatic carcinomas and 39 of tumor cell-negative effusions were investigated. All diagnoses were confirmed by histologic and/or clinical follow-up. DNA aneuploidy was found in 71% of MMs, 100% of metastatic carcinomas, and in none of the negative effusions. Calretinin was positive in 100% of MMs, in none of the metastatic carcinomas, and in 94.9% of negative effusions. BerEP4 showed positivity in 15.6% of MMs, 87.1% of metastatic carcinomas, and in none of the negative effusions. With AgNOR analysis, 89.3% of MMs and 96.7% of metastatic carcinomas showed >or=2.5 AgNOR dots as satellites and >or=4.5 as total AgNOR counts. 9p21 deletions were demonstrated in 90.9% of MM cases, 45.2% of metastatic carcinomas, and in none of the negative effusions. By cytology alone, 81.8% of MMs were identified unequivocally. Addition of DNA-ICM improved the prevalence of tumor cell detection to 87.9% and of AgNOR analysis to 97%. The introduction of 9p21 deletions by FISH improved this prevalence to 100%. Because of these results, the authors propose the sequential application of immunocytochemistry, DNA-ICM, and AgNOR analysis to establish a cytological diagnosis of malignant mesothelioma in serous effusions. In persistent doubtful diagnoses, the authors recommend fluorescence in situ hybridization to analyze the 9p21 deletion.

Image analysis of the AgNOR response in ras-transformed human breast epithelial cells

The argyrophylic staining of the nucleolar organizer regions (AgNOR positive response) in interphase nuclei is often related directly to the cellular demand for ribosome biogenesis and is considered of relevance in studies of tumor pathology. Transformation of human breast epithelial MCF-10A cells by the c-Ha-ras oncogene results in altered growth, invasiveness and tumorigenicity in nude mice. Since ras transformation may be associated with a more intense nucleolar activity, we examined the influence of transfection by the Ha-ras oncogene on AgNOR staining response in MCF-10A cells. Following assessment of the AgNOR response with video image analysis, the AgNOR-positive areas and the AgNOR area/nuclear area ratio, but not the number of AgNOR aggregates or dots per nucleus, were found to be much higher after ras transformation. A role of the Ha-ras transformation on the nucleolar activity of the MCF-10A is thus suggested as assessed by the AgNOR staining. Based on data in the literature, it is also hypothesized that a decreased wild-type p53 level, possibly promoted by the ras transformation, may be associated with the increased AgNOR response.

Usefulness of ploidy, AgNOR and immunocytochemistry for differentiating benign and malignant cells in serous effusions

The objective of this study was to establish the value of different markers in differentiating reactive mesothelial cells from metastatic adenocarcinomatous cells in serous effusions (SE). Forty-five SE were processed for morphological examination (Papanicolaou stain), assessment of ploidy, AgNOR counting and immunocytochemical assay of carcinoembryonic antigen (CEA), epithelial membrane antigens (EMA), Ber-EP4 and Leu-M1. Ploidy was established in an image-analyser in smears stained by the Feulgen stain method. AgNOR dots were counted in the smears stained with the silver nitrate assay for non-histone proteins present in the nucleolar organizer region. CEA, EMA, Ber-EP4 and Leu-M1 were evaluated by immunocytochemistry using the streptavidin-biotin complex method. All the smears with positive cytology were aneuploid. Three false negatives by morphological studies were aneuploid, with AgNOR values in two of them corresponding to the neoplastic group. CEA and Leu-M1 showed a low specificity; EMA and Ber-EP4 showed moderate sensitivity. The assessment of ploidy and the study of AgNOR were better methods than immunocytochemistry for distinguishing between reactive mesothelial cells and adenocarcinomatous cells in serous fluid.