Argyrophilic Nucleolar Organizer Region Dots Research Articles

Evaluation of proliferation activity of oral lichen planus and oral lichenoid reactions using quantitative and qualitative analysis of AgNORs (Argyrophilic Nucleolar Organizer Regions)

Oral lichenoid reactions (OLR) and oral lichen planus (OLP) are clinically and histopathologically similar but their therapeutic approach are different. Due to the presence of dysplasia in oral lichen planus which effects prognosis, the aim of study was the evaluation of proliferation activity of OLPs and OLRs and their subtypes. AgNOR technique that is inexpensive and easy to perform was used to differentiate OLPs and OLRs and their subtypes. Materials & Methods: in this cross sectional , retrospective study, samples consisted of 45 OLPs, 30 OLRs 15 normal oral mucosa (N) from healthy persons (as control group). The samples were stained by AgNOR technique. AgNOR dots were counted on 100 cells from basal and para basal layer . The mean AgNOR dost per nucleus were calculated. Proliferation index (number of cells with AgNOR dots≥ 5) was also determined. Variation in size of AgNOR dots was categorized into 2 groups. Result: There were significant differences between OLP(and its subtypes) and N and also between OLR(and erosive type) and N . Such a difference was also between reticular OLP and reticular OLR.with regard to AgNOR count. Conclusion: Since reticular OLP had significantly higher AgNOR count than reticular OLR in our study, we can suggest the use of AgNOR technique for differentiation of reticular type of OLP from OLR on challenging histopathologic cases

Pleomorphism of argyrophilic nucleolar organizer regions in oral submucous fibrosis and oral squamous cell carcinoma.

Background:Argyrophilic nucleolar organizer regions (AgNORs) is demonstrated to be useful in diagnostic pathology, mainly to distinguish benign lesions from their malignant counterparts. We aimed to correlate AgNORs pleomorphism with the severity of lesion in oral submucous fibrosis (OSMF) and oral squamous cell carcinoma (OSCC) using a retrospective study on 45 archival tissues.Materials and Methods:Silver nitrate staining was performed on archival tissues consisting of 20 OSMF and 20 OSCC. Five biopsies from normal oral mucosa acted as a control. One hundred cells per slide were observed for AgNORs dots, which were classified as typical (spherical) and atypical (large, kidney-shaped and clustered).Results:A positive and significant correlation was found between increased atypical shapes and increasing grades of OSMF and OSCC.Conclusions:AgNORs pleomorphism can be a reliable criterion to assess disease severity and progression in OSMF and OSCC.

Evaluation of Proliferation Activity in Dysplastic and Nondysplastic Oral Lichen Planus Through the Analysis of Argyrophilic Nucleolar Organizer Regions

Lichen planus is a mucocutaneous disease, and its potential for malignant transformation is a subject of controversy. We found dysplastic changes in slide review of about 11% of oral lichen planus (OLP) in our previous research. Dysplastic changes can be an initial phase for carcinogenesis. The question is whether it is possible to detect early malignant changes in OLP through the evaluation of proliferation activity. The aim of this study was to compare the cell proliferation activity in dysplastic and nondysplastic oral lichen planus using the quantitative and qualitative analysis of argyrophilic nucleolar organizer region (AgNOR) and to compare candidal infection in these 2 groups. In this cross-sectional, observational, and analytical study, all 14 cases of confirmed dysplastic OLPs and 17 nondysplastic OLPs were stained with an AgNOR technique. Analysis of argyrophilic nucleolar organizer region dots were counted in 100 cells of basal and parabasal layers, and proliferation index (cells with ≥ 5 dots in nucleus) and variations in size of AgNOR dots were also evaluated. Periodic acid-Schiff staining was performed for detection of candidal infection. Mean AgNOR count in dysplastic OLP (2.39 ± 0.2) was more than nondysplastic OLP (2.27 ± 0.59). Candidiasis was found in 14.3% of dysplastic OLP and in 12.5% of nondysplastic OLP. There was no statistically significant difference between the 2 groups in mean AgNOR count (P = 0.62), variation in size of AgNOR dots (P = 1), proliferation index (P = 0.53), and candidiasis (P = 1). The current study showed slight difference in proliferation rate and candidal infection between dysplastic and nondysplastic OLP.

Argyrophilic nucleolar organizer regions of thyroid lesions on fi ne needle aspiration smears

Background: Argyrophilic nucleolar organizer region is higher in malignant nuclei as compared to reactive and benign nuclei. The main objective of this study was to evaluate the role of Argyrophilic nucleolar organizer region and its extent in differentiating benign and malignant lesions, particularly with respect to follicular neoplasms of the thyroid. Materials and Methods: This was a prospective, cross sectional study which included 110 patients presenting with thyroid swelling. The fi ne needle aspiration smears were studied by conventional methods and silver staining for Argyrophilic nucleolar organizer region. In 50 cases, diagnosis was confi rmed by histopathology. The Argyrophilic nucleolar organizer region count was done and the mean Argyrophilic nucleolar organizer region count per nucleus was calculated along with pattern of distribution. Results: Mean Argyrophilic nucleolar organizer region count was higher in neoplastic as compared to non-neoplastic and infl ammatory lesions. Papillary carcinoma had the highest mean Argyrophilic nucleolar organizer region count (5.40 ± 0.86) and the lowest count was seen in Hashimoto’s thyroiditis (2.10 ± 0.36). Higher mean Argyrophilic nucleolar organizer region count was recorded in follicular carcinoma (4.84 ± 1.42) as compared to follicular adenoma (3.5 ± 0.27) which was found to be statistically signifi cant (P<0.05). Argyrophilic nucleolar organizer region dots were arranged in clusters in colloid goiter whereas scattered discrete black dots were seen in neoplastic lesions. Conclusion: Argyrophilic nucleolar organizer region study of thyroid can be used as an additional diagnostic adjunct with cytomorphological features to differentiate benign and malignant follicular neoplasms. Journal of Pathology of Nepal (2013) Vol. 3, No.1, Issue 5, 361-366 DOI: http://dx.doi.org/10.3126/jpn.v3i5.7859

The Role of Argyrophilic Nucleolar Organizer Region (AgNOR) Study in Cytological Evaluation of Fluids, Especially for Detection of Malignancy

Serous effusion smears reported as (suspicious for malignancy) pose problems in clinical management. Silver staining for argyrophilic nucleolar organizer regions (AgNOR) has proved useful in making a cytopathologic differential diagnosis between benign and malignant cells. Nucleolar organizer regions(NORs) are loops of DNA located in acrocentric chromosomes. These NORs are visualized by silver staining technique that recognizes these argyrophilia associated proteins which are increased in malignancy. This study aimed to distinguish reactive mesothelial cells from malignant cells in serous effusions using these NORs. A total of 174 serous effusions received at the Department of Pathology, TUTH, during a period of one year were included in the study. Smears were studied by conventional Papanicolaou and Giemsa stains. AgNOR counts, variation in size and dispersion of AgNOR dots in smears were graded and compared in malignant and non-malignant effusions. Mean AgNOR counts of 10.43+\-0.73 and 10.21+\-0.51 in malignant peritoneal and pleural effusions, respectively, were significantly (p < 0.0001) greater as compared with counts of 2.12 +\- 0.54 and 2.11 +\- 0.54 in non-malignant effusions. The AgNORs were irregular in shape in malignant effusions whereas they were comparatively larger, single dots in benign effusions. AgNOR size and dispersion were of higher grade in significantly greater proportion of malignant as compared with non malignant effusions (p < 0.0001). Of the cytologically suspicious samples, nine were in the malignant range and one was in the benign range. AgNOR study appears to be clinically useful as an additional diagnostic tool for use in serous effusion when the cytologic diagnosis is difficult.

Quantitative analysis of AgNOR proteins in buccal epithelial cells of Indian street boys addicted to gasp ‘golden glue’

The effect of glue snuffle on the expression of argyrophilic nucleolar organizer regions (AgNORs), an indicator of ribosome biosynthesis, in epithelial cells of oral mucosa has been investigated. AgNOR was evaluated by cytochemical staining in 148 Indian street boys (median age 12 year) who had different bad addictions like tobacco smoking, chewing and most importantly inhaling glue and 20 age- and body mass index-matched school boys who had no such type of bad habit. Compared with school boys, glue addicted street boys showed remarkably increased number of AgNOR dots per nucleus (9.38 ± 1.84 vs. 3.12 ± 0.87, p < 0.001), AgNOR size (1.34 ± 0.52 vs. 0.43 ± 0.02 μm 2, p < 0.001) and percentage of AgNOR occupied nuclear area (9.38 ± 2.12 vs. 0.99 ± 0.03%, p < 0.001). Increase in number and size of the dots is also higher in tobacco smokers and chewers when compared with school boys but a remarkable difference was recorded in glue addicted boys. The changes in AgNOR expression were positively associated with years of addiction after controlling potential confounders. Thus, glue snuffle appeared to be a risk factor for abnormal cell growth via up-regulation of ribosome biogenesis.

Effect of Indoor Air Pollution from Biomass Fuel Use on Argyrophilic Nuclear Organizer Regions in Buccal Epithelial Cells

This study investigated the effect of indoor air pollution from biomass-fuel use on the expression of argyrophilic nucleolar organizer regions (AgNORs), an indicator of ribosome biosynthesis, in epithelial cells of oral mucosa. AgNORs were evaluated using cytochemical staining in 62 nonsmoking indian women (median age, 34 years), who cooked exclusively with biomass, and 55 age-matched women, who were from a similar neighborhood and cooked with relatively clean liquefied petroleum gas (LPG). Concentrations of particulate pollutants in indoor air were measured using a real-time aerosol monitor. Compared to the LPG-using controls, biomass-fuel users showed a remarkably increased number of AgNOR dots per nucleus (6.08 +/-2.26 vs 3.16 +/-0.86, p < 0.001), AgNOR size (0.85 +/-0.19 vs 0.53 +/-0.15 mum2, p < 0.001), and percentage of AgNOR-occupied nuclear area (4.88 +/-1.49 vs 1.75 +/-0.13%, p < 0.001). Biomass-using households had 2 to 4 times more particulate pollutants than that of LPG-using households. The changes in AgNOR expression were positively associated with PM10 and PM2.5 levels in indoor air after controlling for potential confounders such as age, kitchen location, and family income. Thus, biomass smoke appears to be a risk factor for abnormal cell growth via upregulation of ribosome biogenesis.

Evaluation of argyrophilic nucleolar organizer region staining in predicting the behavior of meningiomas.

BACKGROUNDThe number of argyrophilic nucleolar organizer regions (AgNOR) correlates with cellular proliferative activity. Comparing non-recurrent, recurrent, atypical and malignant meningiomas we studied the value of the routine applicability of the AgNOR count in the prognostication of this tumor.PATIENTS AND METHODSTwo hundred and thirty-eight meningiomas were reviewed blindly and graded using WHO grading schema. Eighty-one cases were selected and arranged in six groups according to clinical data and grading: 14 benign non-recurrent meningiomas; 14 primary benign recurrent meningiomas and their subsequent benign recurrences; 14 atypical; 11 malignant and 14 spinal meningiomas. Silver-stained slides were prepared and mean, median and standard deviation of AgNOR dots determined.RESULTSThere was a proportionate increase of AgNOR dots with increasing tumor grade. There was a significant difference between benign non-recurrent tumors versus benign recurrent (P<0.0001) and atypical or malignant (P<0.0001) tumors. A difference was also noted between the recurring tumors versus malignant ones (P= 0.002) but no significant difference was seen between recurrent and atypical; atypical and malignant; intracranial and intraspinal; and primary of recurring meningiomas with their subsequent recurrences. A mean AgNOR count of <2.3 could separate benign tumors from atypical or malignant meningiomas with 93% specificity; and 93% of tumors with benign histology had no recurrence potential if their mean AgNOR count was less than 1.8.CONCLUSIONThis study indicates that in meningioma, the AgNOR count has a good correlation with tumor grading and recurrence, which may aid pathologists and clinicians in predicting tumor behavior.

Application of argyrophilic nucleolar organizer region (AgNOR) staining for cytology of biliary tract carcinomas.

Increased numbers of argyrophilic nucleolar organizer region (AgNOR) dots reflect higher proliferating activity of malignant cells. To determine whether AgNOR staining is suitable for cytology of bile in biliary diseases as an ancillary diagnostic method, we examined the mean number of AgNOR dots (MNA) in cells of both bile smear and tissue sections in 14 benign biliary diseases and 25 malignancies of the biliary tract. The malignant diseases consisted of 11 gallbladder cancers, 10 bile duct cancers, and 4 ampulla cancers in patients who underwent surgical resection. Results of AgNOR staining were available in 15 min, and the MNA in the nucleus was counted in 50 cells/specimen. The MNAs in malignant cells in bile smear (9.6 +/- 3.8) and tissue sections (9.2 +/- 3.5) were significantly higher than those in the corresponding cells in benign biliary diseases (4.1 +/- 1.0 and 2.9 +/- 0.8, respectively; P< 0.01). The MNA of bile smear for both benign and malignant cells correlated significantly with that of tissue sections ( r= 0.915; P< 0.0001). When the cutoff value of MNA for bile smear was set at 7.0, benign diseases could be discriminated from malignancy, and the MNA of 20 (80%) malignant specimens was higher than this value. Our results suggest that the AgNOR dot count of cells in bile smears would be a useful diagnostic tool, in combination with conventional cytological diagnosis, before and during surgery.

Modulation of carcinogenic response and antioxidant enzymes of rats administered with 1,2-dimethylhydrazine by Picroliv

The effect of Picroliv treatment on the carcinogenic response and, hepatic and renal antioxidant enzymes of rats administered with 1,2-dimethylhydrazine hydrochloride (DMH) was studied in male Sprague–Dawley rats. DMH-induced hepatic carcinogenic response and necrosis were inhibited by oral administration of Picroliv (40 and 200 mg/kg). Liver γ-glutamyl transpeptidase, which was elevated to 0.41±0.06 nmol/mg protein by DMH administration was found to be reduced to 0.22±0.04 and 0.18±0.03 nmol/mg protein by Picroliv treatment 40 and 200 mg/kg, respectively. Elevated number of Argyrophilic Nucleolar Organizer Region dots and clusters, an index of proliferation, of DMH treated rat liver was reduced by Picroliv treatment. DMH-induced depletion of hepatic and renal antioxidant enzymes such as catalase and superoxide dismutase levels were restored to normal by Picroliv treatment. Picroliv treatment reduced the DMH-induced elevation of lipidperoxidation in liver, kidney and serum. Elevated levels of serum total bilirubin by DMH administration was reduced by Picroliv treatment. Depleted renal glutathione S-transferase and hepatic glutathione levels after DMH administered rats were found to be significantly increased by Picroliv treatment. Histological analysis of the DMH administered rat liver showed hepatic cell necrosis, coalescent nodular areas and cystic hyperplasia of the bile ducts with inflammation. Picroliv treated liver resembled normal liver except the presence of a few degenerating cells. Renal anatomy was not altered by DMH administration.

Increased Cell Proliferation of Urothelium in Rat Bladder Following Unilateral Nephrectomy

Purpose The aim of this study is to investigate the effect of unilateral nephrectomy on cell proliferation of normal urinary bladder epithelium in adult rats. Materials and Methods Following unilateral nephrectomy or sham operation 5 rats for each time point were sacrificed and their bladders were removed at 6 and 12 hours and also on 1, 2, 4, 7, 14, 28, and 56 days. Another 5 rats were not operated and served as the control. The percent number of proliferating cell nuclear antigen (PCNA) positive cells and the number of argyrophilic nucleolar organizer regions (AgNORs) were determined for each bladder. Results In the nephrectomized groups, a significant increase of PCNA expression in urothelial cells of the bladder was found at each sacrifice time with the exception of 28 and 56 days when compared with that of the control and corresponding sham-operated groups. Percent PCNA labeling index (PCNA-LI) reached a peak value of approximately 9.1% on day 2 post operation. From day 4 post nephrectomy, however, PCNA-LI decreased gradually reaching control levels by day 28. No significant difference was found between the control and sham-operated groups. The mean number of AgNOR dots in nephrectomized rats sacrificed on day 1, 2, 4, 7 and 14 post operation was significantly larger than that of the control and corresponding sham-operated groups. Conclusion The results suggest that unilateral nephrectomy temporarily enhances normal urothelial cell proliferation in adult rat bladder.

Argyrophilic nucleolar organizer regions in colorectal malignancy: application of typing by their density.

Colorectal neoplasms obtained at colonoscopy were examined by argyrophilic nucleolar organizer region (AgNOR) staining to evaluate the usefulness of AgNOR as a discriminant for malignancy. AgNOR dots were divided into two kinds: 'structures' (larger and less-densely stained) corresponding to the nucleolus, and 'units' (smaller and densely stained) presumed to be true AgNOR within the structure. The number of structures per nucleus did not differ between the adenoma and carcinoma groups, whereas the number of units per nucleus showed a significant difference. However there were several cases showing an overlap between the adenoma and carcinoma groups, leading to a difficulty in deciding whether any given case was benign or malignant. Three types of AgNOR patterns were categorized based on the ratio of units to structure. Type I was defined as the unit being indistinguishable from the structure, Type II as each structure having one to five units, and Type III as at least one structure having six or more units, irrespective of total number of units per nucleus. The colorectal lesions in which more than half of the neoplastic cells showed Type III coincided well with carcinomas histologically diagnosed, with the exception of adenomas with severe atypia. Labeling index of proliferating cell nuclear antigen (PCNA LI) differed between the adenoma and carcinoma groups with a considerable extent of overlap, and correlated to some extent with the AgNOR values. These results showed that the AgNOR staining was useful for determining malignancy and its usefulness appeared superior to PCNA LI.

Comparison of Ureteropelvic Transitional Cell Carcinoma with Bladder Transitional Cell Carcinoma Using an Image Analyzer

To elucidate whether a significant difference in malignant potential between ureteropelvic transitional cell carcinoma (TCC) and bladder TCC is present, the argyrophilic nucleolar organizer region (AgNOR), nuclear DNA content and mean nuclear area (MNA) were evaluated in 32 ureteropelvic TCCs and 60 bladder TCCs, and were also compared with histological grade and stage. The parameters used were the mean AgNOR count (C-AgNOR), the percentage of cells exhibiting more than 3 AgNOR dots within nuclei (P-AgNOR), 2c deviation index (2cDI), 5c exceeding rate (5cER) and MNA. No significant difference in these 5 parameters was found between ureteropelvic and bladder TCCs. In addition, within each histological grade of tumor, no significant difference in each parameter between ureteropelvic and bladder TCCs was noted. Similarly, within pTa or pT1 TCCs, there was no significant difference in the parameters between ureteropelvic and bladder TCCs. On the other hand, invasive TCCs (pT2 and higher), the C-AgNOR and P-AgNOR were significantly higher in ureteropelvic than in bladder TCC (p < 0.05), while no significant difference in 2cDI, 5cFR and MNA was noted. These findings suggest that: (i) as a whole, the difference in malignant potential between ureteropelvic and bladder TCCs appears to be less significant, and (ii) factors other than the malignant potential of a tumor might contribute to the unfavorable clinical outcome in patients with ureteropelvic TCC.

Argyrophilic nucleolar organizer region (AgNOR) area per nucleus as a prognostic factor in breast cancer.

Whether argyrophilic nucleolar organizer region (AgNOR) is a predictor of prognosis in breast cancer is controversial. The semiquantitative procedure used to calculate interphasic argyrophilic dots optically is neither reliable nor reproducible. We measured the mean area of AgNOR dots, the mean AgNOR area per nucleus (MA), and the mean AgNOR number per nucleus (MN) by automated image analysis in specimens from 131 patients with breast cancer. Higher MA (> 7.41 microns2), detected in 22 (16.8%) of 131 patients, did not correlate with clinicobiologic variables, except for DNA ploidy status. Overall survival was significantly better in patients with lower MA than in those with higher MA. On multivariate analysis, MA was a significant independent factor, followed by nodal status. These findings indicate that expression of AgNOR should be evaluated in terms of area occupied and that the mean area of AgNOR per nucleus (MA) is an important prognostic factor in the overall survival of breast cancer patients.

Proliferative activity in normal salivary gland and pleomorphic adenoma. A study by argyrophilic nucleolar organizer region (AgNOR) staining.

Proliferative activity of the structural components of normal salivary glands (27 cases) and characteristic histological areas in salivary pleomorphic adenoma (51 cases) was examined by visualization of argyrophilic nucleolar organizer regions (AgNORs). In the normal salivary gland, there were clusters of cells, each with two or three AgNOR dots, within the intercalated ducts. The mean number of AgNORs in this portion was 1.80, which was significantly higher than in the other epithelial parts. This indicates vigorous proliferative activity of the intercalated ducts, and thus the ducts may play an important role as reserve cells in neoplastic transformation. In pleomorphic adenoma, solid nests had a high AgNOR count (1.61), showing a significant difference from the tumor cells scattered in the "stroma". Thus the decrease in proliferative ability might occur concurrently with formation of the extracellular matrix.