Adapted technique for demonstrating argyrophilic nucleolar organizer regions in the cytologic samples of canine mast cell tumors.

INTRODUCTION: Argyrophilic nucleolar organizer regions (AgNORs) are nonhistone argyrophilic nucleolar proteins associated with ribosomal genes found in the nucleolar organizer region that reflect cell proliferation and have an affinity for silver. AgNOR staining may be useful to evaluate prognosis in several neoplasms because higher AgNOR counts are related to higher grade tumors, metastases, and shorter survival times. OBJECTIVE: We aimed to report on a quick and practical technique to identify AgNORs adapted for use in routine cytology. MATERIALS AND METHODS: The cytopathologic diagnosis of mast cell tumor (MCT) in samples collected by fine-needle aspiration (FNA) was determined. Next, slides were impregnated with a solution containing silver nitrate; the main modification of our technique included incubation of these slides at a controlled temperature of 25 °C. Some slides were previously stained with Diff-Quik and others were only fixed with methanol. The slides were analyzed under a microscope, and the number of blackened intranuclear points (AgNORs) was counted. RESULTS: Slides prestained with Diff-Quik were easily counted compared with slides only fixed in methanol. Technical issues encountered with the methanol-fixed slides included insufficient cellularity, background precipitation, and an absence of silver impregnation. CONCLUSIONS: The technique reported in this study showed satisfactory results for AgNOR counting in cytologic smears from MCT, such as good impregnation and the elimination of background interferents. Further evaluation of this method comparing AgNOR counts with histologic examinations, tumor grades, other prognostic markers, and survival times are needed to fully evaluate the benefit of this technique.

Cytological Screening Model of Normal Oral Mucosa Exposed to Carcinogens: A Pilot Study.

INTRODUCTION: Oral cytopathology is able to detect incipient cellular alterations, but it is not routinely applied to this purpose. We aimed to establish a model to screen individuals with no oral lesion exposed to smoking/alcohol, by means of the nuclear area, cell proliferation rate, and analysis of genetic damage. METHODS: In this cross-sectional pilot study, 90 patients were allocated into 3 groups: oral cancer group (patients with oral squamous cell carcinoma), tobacco/alcohol group (patients without oral lesions and exposed to these risk factors), and control group (individuals with no lesion and not exposed to tobacco and alcohol). The cytological smears performed in these individuals were stained with Papanicolaou, a silver-staining and a Feulgen reaction. The nuclei of cells were measured, and AgNORs/nucleus and micronuclei (MN) were quantified. The cutoff values were stipulated evaluating the healthy mucosa (control group) and the cancerization field mucosa (oral cancer group). RESULTS: Cutoff values for the screening of individuals exposed to carcinogens were ≥8% of nuclei larger than 100 µm2, ≥3.38 AgNOR/nucleus, and ≥3 MN per 1,000 cells. CONCLUSIONS: Nuclear area measurement and AgNORs/nucleus and MN quantification identified the incipient phase of oral carcinogenesis. A screening model for individuals without oral lesion exposed to smoking/alcohol was proposed.

Quantitative analysis of AgNOR counts and pap stain in exfoliative cytology specimens of oral mucosa in bidi smokers and nonsmokers.

Background: Bidi smoking is a serious health hazard which is common throughout South Asia and parts of the Middle East. It has been strongly implicated to various benign and malignant lesions of oral cavity and oropharynx. These tobacco-filled leaves deliver more than three times the amount of nicotine, carbon monoxide, and tar as cigarettes which exert injurious effects on cells reflected in terms of accelerated proliferative activity in normal oral mucosal cells. Aim: This study aimed to compare the exfoliated cells from the oral mucosa of bidi smokers and nonsmokers, with emphasis on proliferative activity. Materials and Methods: Exfoliative smears were obtained from the oral mucosa of forty participants (twenty nonsmokers and twenty smokers) with age group ranging from 30-80 years, in and around Barwala (Haryana). The cytologic smears were evaluated using Papanicolaou (PAP) stain and AgNOR in order to evaluate the presence of cytological alterations suggestive of inflammation, dysplasia, keratinization, and proliferative activity of epithelial cells. Only PAP Class I and Class II smears were observed. Results: Comparison of the mean number of AgNORs showed a significant difference between nonsmokers and smokers. Inflammatory alterations were found in 70% of smokers and 55% of nonsmokers. A significant difference in proliferative activity was observed between smokers and nonsmokers classified as PAP Class II. Conclusion: A significant difference of AgNORs/nucleus was observed between bidi smokers and nonsmokers.

RésuméContexte: Bidi fumeurs est un grave danger pour la santé qui est commune dans toute l'Asie du Sud et certaines parties du Moyen-Orient. Il a été fortement impliqué dans diverses lésions bénignes et malignes de la cavité buccale et l'oropharynx. Ces feuilles de tabac offrent plus de trois fois la quantité de nicotine, monoxyde de carbone et de goudron que les cigarettes qui exercent des effets préjudiciables sur les cellules reflétés sous la forme d' une accélération de l'activité proliférative des cellules de la muqueuse buccale normale. Objectif: Cette étude visait à comparer les cellules exfoliées de la muqueuse orale des bidis fumeurs et non fumeurs, avec l'accent sur l'activité proliférative. Matériels et méthodes: frottis Exfoliative ont été obtenus à partir de la muqueuse orale de 40 participants (20 non-fumeurs et fumeurs) avec 20 Groupe d'âge allant de 30-80 ans, dans et autour de Barwala (Haryana). Le frottis cytologique ont été évalués à l'aide de la coloration de Papanicolaou (PAP) et d'AgNOR afin d'évaluer la présence d' altérations cytologiques évocateurs d'infl ammation, dysplasie, la kératinisation, et l'activité proliférative des cellules épithéliales. PAP uniquement les catégories I et II de Papanicolaou n'a été observé. Résultats: comparaison du nombre moyen d'AgNORs ont montré une différence entre les non-fumeurs et les fumeurs. Des modifications ont été trouvés infl ammatory dans 70% des fumeurs et 55% des non-fumeurs. Une différence dans l'activité proliférative a été observée entre les fumeurs et les non-fumeurs PAP, le niveau d'emploi de la classe II. Conclusion: une différence de AgNORs/noyau a été observée entre fumeurs et non-fumeurs bidi. Mots-clés: AgNOR, frottis exfoliative, les fumeurs.

The use of cytopathology to identify disturbances in oral squamous cell carcinoma at early stage: A case report.

Early detection of oral squamous cell carcinoma (OSCC) is a challenge for oral surgeons, as clinical features are not always classical. Cytopathological assays can help identify alterations at the cellular level. This article reports a case of OSCC in a young male adult without exposure to classical risk factors. The histopathological examination showed a micro invasive carcinoma invading the connective tissue. Cytopathological results showed a higher percentage of cells in deeper epithelial layers; the cytomorphometric examination revealed a nuclear/cytoplasmic ratio of 0.14; the mean number of Nucleolar Organiser Regions which have a high affinity for silver (AgNOR) per nucleus was 2.86, and the mean percentage of nuclei with >2 AgNOR was 58%. The micronucleus test found 3 micronucleated cells and several metanuclear aberrations. These findings support the hypothesis that cytological examination is an important tool to identify early changes in oral smears and thus help in the early clinical detection of suspicious malignant oral lesions that should be more rigorously followed.

The role of argyrophilic nucleolar organizing region-associated proteins in clinical exacerbation of chronic obstructive pulmonary disease.

OBJECTIVE: To investigate whether argyrophilic nucleolar organizing region-associated protein (AgNOR) parameters can be used as a biomarker that could potentially help with the management and clinical prognosis of chronic obstructive pulmonary disease (COPD) exacerbation. METHODS: This case-control study enrolled patients with COPD who were admitted to the Emergency Department and healthy sex- and age-matched control subjects. Peripheral blood samples were collected at hospital admission and the peripheral lymphocytes were silver-stained to investigate the quantity and distribution of AgNOR proteins. Fifty nuclei per patient were viewed and the total AgNOR area/total nuclear area (TAA/TNA) ratio and the mean AgNOR number for each patient were calculated. RESULTS: A total of 20 patients with COPD exacerbation and 17 healthy control subjects were recruited to the study. The TAA/TNA ratio and the mean AgNOR number were significantly higher in the patients with COPD exacerbation compared with the healthy control subjects. The mean AgNOR number showed a positive correlation with the pCO2 levels on admission. CONCLUSION: AgNOR protein levels were elevated during a COPD exacerbation compared with healthy control subjects and there was a positive correlation between pCO2 levels and mean AgNOR number.

[Role of the Nucleolus in Rearrangements of the IGH Locus].

The review summarizes the results from a series of studies focusing on the role that the nucleolus plays in maturation of the IGH locus and the choice of its partner genes in leukemia-associated translocations. The role of nuclear compartmentalization and nuclear localization of translocated oncogenes in ectopic activation of their transcription is discussed.

Analysis of silver binding nucleolar organizer regions in exfoliative cytology smears of potentially malignant and malignant oral lesions.

Nucleolar organizer regions are nucleolar components that contain proteins that are stained selectively by silver methods; they can be identified as black dots throughout the nucleolus and are known as silver binding nucleolar organizer regions (AgNOR). The number of AgNOR is related to the cell cycle and the proliferative activity of the cells. We investigated AgNOR using exfoliative cytology smears of potentially malignant oral lesions. Eighty individuals were divided into four equal groups: healthy controls, oral leukoplakia, oral submucous fibrosis and oral squamous cell carcinoma. The mean number of AgNOR in each study group gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. The proliferative index was increased in the oral premalignant and malignant patients compared to normal subjects. The mean AgNOR size gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. Spherical shaped AgNOR were most common in controls, whereas large, clustered and kidney shapes were most common in oral squamous cell carcinoma. Multiparameter analysis of AgNOR in oral exfoliative smears is a simple, sensitive and cost-effective method for differentiating premalignant from malignant lesions and can be used in conjunction with routine cytomorphological evaluation.

Correlation between ploidy status using flow cytometry and nucleolar organizer regions in benign and malignant epithelial odontogenic tumors.

OBJECTIVE: Differentiation between the aggressive benign odontogenic tumors and their malignant counterparts is controversial and difficult. While flow cytometry (FCM) allowed DNA analysis in neoplasia, argyrophilic organizer regions (AgNORs) number and/or size in a nucleus are correlated with the ribosomal gene activity and therefore with cellular proliferation. The aim of this research was to study the diagnostic accuracy of FCM and AgNORs staining in differentiating between benign and malignant epithelial odontogenic tumors and to correlate between these two interventions. DESIGN: Sixteen benign cases [8 cases of ameloblastoma (AB) and 8 cases of keratocystic odontogenic tumor (KCOT)] and 13 malignant epithelial odontogenic tumors [8 cases of ameloblastic carcinoma (ABC) and 5 cases of clear cell odontogenic carcinoma(CCOC)] were included in the current study. For FCM analysis, a single cell suspension from Formalin fixed paraffin-embedded (FFPE) tumors was prepared according to a modified method described by Hedley (1989) and AgNORs staining were performed in accordance to the Ploton protocol (1986). Analysis of AgNORs was performed using both quantitative and qualitative methods. RESULTS: The work revealed that all the examined tumors were diploid, except for 40% of CCOC cases. The S-phase fraction (SPF) value, AgNORs count and AgNORs area/cell showed statistically significant difference on comparing benign and malignant groups. A weak positive correlation was observed between SPF and AgNORs count. CONCLUSION: The SPF value was considered to be more sensitive and specific in differentiation between aggressive benign and malignant epithelial odontogenic tumors in comparison to AgNORs counting.

Decreased argyrophilic nucleolar organiser region (AgNOR) expression in Purkinje cells: first signal of neuronal damage in sudden fetal and infant death.

AIMS: The nucleolus is an important cellular component involved in the biogenesis of the ribosome. This study was performed in order to validate the introduction of the argyrophilic nucleolar organiser region (AgNOR) stain technique, specific for the nucleoli detection, in neuropathological studies on sudden fetal and infant death. METHODS: In a wide set of fetuses and infants, aged from 27 gestational weeks to eight postnatal months and dead from both known and unknown causes, an in-depth neuropathological study usually applied at the Lino Rossi Research Center of the Milan University was implemented by the AgNOR method. RESULTS: Peculiar abnormalities of the nucleoli, as partial or total disruption above all in Purkinje cells (PCs), were exclusively found in victims of sudden fetal and infant death, and not in controls. The observed nucleolar alterations were frequently related to nicotine absorption in pregnancy. CONCLUSIONS: We conclude that these findings represent early hallmarks of PC degeneration, contributing to the pathophysiology of sudden perinatal death.

Are there any effects of chronic carbon monoxide exposure on argyrophilic nucleolar-organizing region-associated protein synthesis in rat myocardium?

The aims of the study are to detect whether there are any possible effects of chronic carbon monoxide (CO) exposure on the argyrophilic nucleolar-organizing region (AgNOR)-associated protein synthesis and evaluate any possible relationship between the amount of AgNOR protein and the level of myocardial injury also and between AgNOR and histopathological evaluation methods. Adult male albino Wistar rats (n = 18) were randomly divided into three groups (groups A, B, and C). Group A served as control, while groups B and C were rats exposed to CO gas chronically (1000 and 3000 ppm CO concentration with a flow rate of 4 L/min for 30 min/day for 7 days, respectively). Total AgNOR area/nuclear area (TAA/NA) and the mean AgNOR numbers for each myocyte nucleus were determined. There were significant differences among all groups for TAA/NA ratio. These differences were not significant for mean AgNOR numbers. According to the histopathological evaluation scores, there were significant differences between the groups. The differences were significant among the groups for loss of sarcomere pattern. A strong positive correlation between histopathological injury scores and TAA/NA ratio was found (Rsq = 0.48; p = 0.002), however, the correlation was not significant for mean AgNOR numbers (Rsq = 0.08; p = 0.25). In conclusion, TAA/NA ratio can be used as an indicator for obtaining information about the level of myocardial damage instead of histopathological evaluation scores.

Apoptosis and expression of argyrophilic nucleolus organizer regions in epithelial neoplasms of the larynx / Apoptose e expressão de regiões argirófilas organizadoras de nucléolos em neoplasias epiteliais da laringe

INTRODUCTION: Occurrence of apoptosis and expression of proliferative markers are powerful tools to establish a prognosis in the follow-up of cancer. OBJECTIVE: To evaluate the growth fraction in papillomas and laryngeal squamous cell carcinomas with three degrees of differentiation through apoptosis and the expression of nucleolus organizer regions. METHODS: Retrospective study from which paraffin material was submitted to microtomy and hematoxylin-eosin and silver staining. Stained slides were used to quantify the apoptotic index and the number of nucleolus organizer regions by morphometry. RESULTS: Apoptosis was significantly more frequent in well differentiated carcinomas and in papillomas, and a higher growth fraction of expressed nucleolus organizer regions and cells that expressed a greater than average number of nucleolus organizer regions were more frequently noted in undifferentiated carcinomas. CONCLUSIONS: Thus, it was possible to verify that a high apoptotic index was associated with a lower chance of tumor differentiation in carcinomas, while a greater number of total nucleolus organizer regions, cells expressing nucleolus organizer regions above average and a higher growth fraction were associated with greater likelihood of abnormal cell proliferation and increased tumor differentiation. .

INTRODUÇÃO: A ocorrência de apoptose e a expressão de marcadores proliferativos são ferramentas poderosas no estabelecimento do prognóstico do câncer. OBJETIVO: Avaliar a fração de crescimento de papilomas e carcinomas laríngeos de células escamosas, com três graus de diferenciação, através da apoptose e expressão de regiões organizadoras de nucléolo. MÉTODO: Estudo retrospectivo, cujo material em blocado em parafina foi submetido à microtomia e coloração em hematoxilina-eosina, e pela prata. As lâminas coradas foram utilizadas para quantificar o índice apoptótico e o número de regiões organizadoras de nucléolo (NORs) através da morfometria. RESULTADOS: A apoptose foi significativamente mais frequente em carcinomas bem diferenciados e em papilomas; enquanto que uma maior fração de crescimento, de NORs expressos e de células que expressaram maior número de NORs, foram mais frequentes nos carcinomas indiferenciados. CONCLUSÕES: Foi possível verificar que o índice apoptótico elevado indica menores chances de diferenciação tumoral nos carcinomas, enquanto que um maior número de NORs totais e células expressando NORs acima da média, e uma maior fração de crescimento, determinam maiores chances de proliferação celular anormal e maior diferenciação tumoral. .

Apoptosis and expression of argyrophilic nucleolus organizer regions in epithelial neoplasms of the larynx.

INTRODUCTION: Occurrence of apoptosis and expression of proliferative markers are powerful tools to establish a prognosis in the follow-up of cancer. OBJECTIVE: To evaluate the growth fraction in papillomas and laryngeal squamous cell carcinomas with three degrees of differentiation through apoptosis and the expression of nucleolus organizer regions. METHODS: Retrospective study from which paraffin material was submitted to microtomy and hematoxylin-eosin and silver staining. Stained slides were used to quantify the apoptotic index and the number of nucleolus organizer regions by morphometry. RESULTS: Apoptosis was significantly more frequent in well differentiated carcinomas and in papillomas, and a higher growth fraction of expressed nucleolus organizer regions and cells that expressed a greater than average number of nucleolus organizer regions were more frequently noted in undifferentiated carcinomas. CONCLUSIONS: Thus, it was possible to verify that a high apoptotic index was associated with a lower chance of tumor differentiation in carcinomas, while a greater number of total nucleolus organizer regions, cells expressing nucleolus organizer regions above average and a higher growth fraction were associated with greater likelihood of abnormal cell proliferation and increased tumor differentiation.

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 estimation of AgNORs in normal, dysplastic and malignant oral mucosa.

AIMS: Silver stainable nucleolar organizer regions (AgNORs) have received a great deal of attention recently as their frequency within the nuclei is significantly higher in malignant cells than in normal, reactive or benign neoplastic cells. The objective of this study was to carry out a quantitative assessment of large and small AgNORs in oral normal mucosa, precancerous lesions and infiltrating squamous cell carcinomas. METHODS: The study comprised 110 formalin-fixed, paraffin-embedded oral mucosal biopsies consisting of 30 oral dysplasia, 60 oral squamous cell carcinomas and 20 normal oral mucosa. AgNORs were counted in each nucleus, categorized as small, large and total number of AgNORs in each cell and their means were calculated. RESULTS: The mean value of small AgNORs, large AgNORs and total AgNORs increased gradually from normal mucosa to dysplastic lesions to squamous cell carcinomas. The study clearly indicates that in oral squamous cell carcinomas, AgNORs diminish in size as they increase in number. Further, AgNOR counts increase as the degree of malignant potential of the cell increases. CONCLUSIONS: By combining both the enumeration of AgNORs and their size, good distinction can be made between normal, dysplastic and infiltrating squamous cell carcinomas. This could help in the early diagnosis and prognosis of dysplastic mucosal lesions and their malignant transformation.

Value of argyrophilic nucleolar organizing region protein determinations in nondiagnostic fine needle aspiration samples (due to insufficient cell groups) of thyroid nodules.

OBJECTIVE: To evaluate the diagnostic value of argyrophilic nucleolar organizing regions (AgNORs) in the nondiagnostic fine needle aspiration biopsy (NFNAB) specimens (due to insufficient cell groups) of cases with thyroid nodules. STUDY DESIGN: Thirty-five patients with NFNAB were included in the study. FNAB stained with Giemsa was faded using the McKee technique, then stained for AgNOR detection according to a specific protocol. One hundred nuclei per individual were evaluated to detect the AgNOR count/nucleus and total AgNOR area/ nuclear area (TNA/NA) of individual cells by using a computer program. RESULTS: The AgNOR values of patients with NFNAB were between 1.2-2.4% for AgNOR count/ nucleus and between 3.1-9.1%for TNA/NA. When the cutoff values were taken as > 3 for AgNOR count/nucleus and > 8 for TNA/NA, the sensitivity ratios were 100% and 97% for AgNOR count/nucleus and TNA/ NA for discriminating benign/malignant lesions. CONCLUSION: By using some cutoff values obtained from a modified AgNOR staining method, benign/malignant potential of thyroid aspirations with NFNAB may be estimated. (Anal Quant Cytopathol Histopathol

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.

Long-term analysis to objectify the tumour grading by means of automated microscopic image analysis of the nucleolar organizer regions (AgNORs) in the case of breast carcinoma.

BACKGROUND: Apart from a number of cases of inaccurate prognosis in regard to individual patients, the inter- and intra-observer variability of the classical, histological prognosis parameters have been under repeated discussion. For this reason, a long-term analysis was carried out in regard to overall survival by means of automated microscopic image analysis of the nucleolar organizer regions (AgNORs) to objectify tumour grading in the case of breast carcinoma.This consists of a selective representation of argyrophilic proteins that are associated with the nucleolus organising regions. METHODS: The evaluation included 244 female patients with an average age of 59.3 years. The characterisation of the histological sections was carried out on the basis of the AMBA/R system. With this software the histometric characterisation level was evaluated in terms of the nucleolus organizer regions. The post-observation data were obtained from the clinical register and were complemented by mortality data from the cancer registers and by data supplied by the residents' registration office of Berlin. RESULTS: The average post-observation period was 106.6 months. With the Cox-Regression the influence of the co-variables (conventional prognosis parameters and AgNOR parameters) were examined. In the model, only the parameters pN, G and various AgNOR parameters remain present. CONCLUSION: There is a strong correlation between survival and selected AgNOR parameters. These could replace the conventional grading as the standard measure for the mitosis rate together with the pleomorphism level. Instead of the-time consuming AMBA/R system originally used, a new implementation of AgNOR quantification with modern VM systems could be applied. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1449591192859058.

Exfoliative cytological assessment of apparently normal buccal mucosa among quid chewers using argyrophilic nucleolar organizer region counts and Papanicolaou staining.

OBJECTIVE: Quid chewing is associated with an increased risk of oral cancer. This study aims to analyze argyrophilic nucleolar organizer region (AgNOR) counts along with Papanicolaou (PAP) staining in exfoliative smears of quid chewers and non-chewers to correlate quid chewing habits with possible early cytological changes in apparently normal buccal mucosa. STUDY DESIGN: Exfoliative smears were obtained from normal buccal mucosa of 30 male quid chewers and non-chewers. The smears were stained using the AgNOR silver staining technique to evaluate the proliferative activity and PAP for cytological appearance. RESULTS: Statistically higher AgNOR counts were observed in chewers as compared to non-chewers. The difference in the mean percentage of nuclei having ≥5 AgNORs in both groups was statistically significant (p = 0.001). In chewers, PAP showed 77% with class I and the remaining 23% were class II, while the non-chewers showed only class I cytology. AgNOR counts between chewers and non-chewers having class I cytological appearance demonstrated a greater mean AgNOR count in chewers (p = 0.0001). CONCLUSION: Quid chewing seems to have a definite role in promoting proliferative activity of apparently normal buccal mucosal cells. Exfoliative cytological assessment of a combination of AgNOR counts and PAP has the potential for prediction of early quid-associated cellular changes before the appearance of clinical premalignant and malignant lesions.

Antiproliferative efficacy of hesperetin (citrus flavanoid) in 1,2-dimethylhydrazine-induced colon cancer.

Cancer is the second leading cause of death worldwide and is increasing at an alarming rate. The present study was to evaluate the antiproliferative effects of hesperetin, a flavonoid commonly found in many herbal medicines and foods, on aberrant crypt foci (ACF), argyrophylic nucleolar organizer regions (AgNORs) and proliferating cell nuclear antigen (PCNA) in 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats. Rats were given subcutaneous injections of DMH (20 mg/kg body weight) weekly for 15 weeks to induce carcinogenesis, and hesperetin was administered orally at the dose of 20 mg/kg body weight. DMH exposure alone produced a high incidence of ACF and showed positive staining for PCNA and AgNORs in colonic tissues. Supplementation with hesperetin lowered the PCNA labeling index and suppressed the formation of ACF in the rats with colon cancer. These results clearly reveal that dietary hesperetin possesses antiproliferative ability against chemically induced colon tumourigenesis.

From conventional fluid cytology to unusual histological diagnosis: report of four cases.

Malignant small round cell tumors represent a diagnostic challenge for cytologists and pathologists. This case series describes four cases of unusual metastasis of small round cell tumors subtypes into body cavities generating effusions in which fluid cytological examination suggested the neuroendocrine origin of the tumors. Tumor diagnosis (Ewing sarcoma/primitive neuroectodermal tumor and desmoplastic small round cell tumors) were unknown at the cytological evaluation. We can highlight the importance of the accurate analyses of body fluids, both for early diagnosis of metastatic disease, and for the diagnosis of primary tumor when serous effusion is the first manifestation of the neoplasia.