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. .

Histologic grading and nucleolar organizer regions in oral squamous cell carcinomas

OBJECTIVE: The purposes of this study were to histologically assess different types of oral squamous cell carcinoma and the silver-binding nucleolar organizer region (AgNOR) morphology in neoplastic cells, as well as to quantify the number of AgNORs in each type of carcinoma in order to relate AgNOR count and histologic grading. MATERIAL AND METHODS: Twenty-eight cases of oral squamous cell carcinoma were divided into 4 groups, namely well-differentiated, moderately differentiated, poorly differentiated, and undifferentiated. For NOR study, 3-µm-thick sections were stained with 50 percent aqueous silver nitrate solution. The predominant microscopic pattern of NORs was determined. Quantitative analyses of NORs were obtained of all cells present on each histological field using a 0.025 mm² eyepiece graticule. Different histological fields were analyzed until the total number of NORs was 120 cells for each tumor. Kruskall-Wallis test was applied to compare the groups of sample data at a significance level of p=0.05. RESULTS: The mean number of AgNORs per nucleus was 3.20 for the well-differentiated group, 5.33 for the moderately differentiated one, 8.27 for the poorly differentiated one, and 10.08 for the undifferentiated one. AgNOR count was significantly different (p<0.05) among all of the studied groups. CONCLUSION: AgNOR staining technique seems to be a useful diagnostic tool since differences in AgNOR numeric values can be identified in the different types of oral squamous cell carcinoma. This technique is easy to handle and inexpensive, thus justifying its large use in histopathology.

Effect of finasteride on serum prostate-specific antigen (PSA) and on prostate of hamster Mesocricetus auratus (hMa) / Efeito da finasterida no antígeno prostático específico (PSA) sérico e na próstata do hamster Mesocricetus auratus (hMa)

PURPOSE: Evaluate the effects of finasteride on the serum PSA and on the prostate of hamster-Mesocricetus auratus(hMa). METHODS: Twenty hMa male adults were split in groups control and experimental (n=10). Animals of the experimental group received 7.14ng/mL of finasteride, subcutaneously (SC) on the back three times per week, during 90 days. The finasteride dose was equivalent to 5.0mg administered to a 70kg man. At the end of the experiment the mean age for the animals in the control group was 15.2±1.13months and for the experimental group was 17.7±0.67 months. There was a statistically significant difference between mean ages of both groups (t value=5.98; p=0.001). The animals of the control group weighted 129.0±18.8g and the experimental group weighted 145.0±15.5g, t=1.88 e p=0.0514. The serum PSA was assessed through ELISA method. Prostates of those animals were collected and processed to histology and morphometry: the diameter of the acinous glands and the acinous epithelium, apoptosis, AgNORs and cellularity were assessed in both groups. RESULTS: Serum PSA decreased in the experimental group, 0.003ng/mL versus 0.763ng/mL, H= 7.982 e p= 0.0047. Decrease in the acinous area occurred in animals that received finasteride, 238.000±24.600 μm² versus 398.600±55.320 μm²; t= 2.653; p= 0.0122. A remarkable decrease in the area of the acinous epithelium occurred in the animals that received finasteride, 111.900±12.820 μm² versus 160.400±18.430 μm² t= 2.162; p= 0.0361. AgNORs were less expressed in finasteride treated animals, 2.846±0.877 versus 3.68 ±1.07 argyrophilic clusters for μm², p= < 0.0001. Apoptosis was more intense in the experimental group, 53.62±1.389 than in controls, 14.76 ± 2.137, p= 0.0408. However, there was no statistical difference in the cellularity between both groups, 74.75±5.5 cells, in controls versus 65.07±13.24, in treated animals, p=0.5105. CONCLUSIONS: Use of finasteride decreased serum ...

OBJETIVO: Avaliar o efeito da finasterida no PSA sérico e na próstata do hamster-Mesocricetus auratus (hMa). MÉTODOS: 20 hMa adultos machos foram divididos em grupos de 10 animais. No experimento foram administrados 7,14 ng/mL de finasterida, subcutâneo (SC), no dorso, três vezes por semana, por 90 dias, dose equivalente a 5,0 mg usada em homem de 70Kg. Ao final da pesquisa, grupo experimento apresentou idade média de 17,7 ± 0,67 meses. O grupo controle apresentou idade média de 15,2 ± 1,13 meses. O valor de t na comparação das médias das idades entre os dois grupos foi de 5,98 e p=0.0001. Os animais-controle pesaram em média 129,0 ± 18,8g e o experimento 145,0 ± 15,5g; t=1,88 e p=0,0514. Na microscopia óptica de luz e estudo morfométrico: avaliaram-se o diâmetro dos ácinos e epitélio acinar prostáticos, a apoptose, a expressão AgNORs e a celularidade. RESULTADOS: O grupo-experimento apresentou média de PSA de 0,003 ng/mL e o grupo-controle de 0,763 ng/mL, H=7,982 e p=0,0047. A área dos ácinos do grupo-experimento foi de 238,000±24,600 μm² versus 398,600±55,320 μm²; t= 2,653; p= 0,0122. A área do epitélio acinar no grupo-experimento foi de 111,900±12,820 μm² versus 160,400±18,430 μm² t= 2,162; p= 0,0361. A expressão de AgNORs foi menor no grupo-experimento 2,846±0,877 versus 3,68 ±1,07 grumos argilófilos por μm², p= < 0,0001. A apoptose foi mais freqüente no grupo-experimento, 53,62±1,389 versus controle, 14,76 ± 2,137, p= 0,0408. Não houve diferença na celularidade entre os grupos de animais, 74,75±5,5 células no grupo-controle versus 65,07±13,24, no grupo-experimento, p= 0,5105. CONCLUSÕES: A finasterida diminuiu o PSA sérico, a área do lúmen, o epitélio acinar, a expressão de AgNORs e promoveu a apoptose nos ácinos da próstata dos hamsteres experimento e não houve diferença na celularidade acinar entre os animais estudados.

Argyrophilic nucleolar organizer regions in inflammatory, premalignant, and malignant oral lesions: a quantitative and qualitative assessment.

BACKGROUND AND OBJECTIVE: Argyrophilic nucleolar organizer regions (AgNORs) have found widespread application in the past, especially in tumor histopathology. This study was undertaken to evaluate the significance of various AgNOR parameters and to assess their role in differentiating hyperplastic, premalignant, and malignant lesions. MATERIALS AND METHODS: The study sample consisted of archival biopsy specimens of ten squamous cell carcinomas, ten premalignant lesions, and five inflammatory lesions. Two biopsies from normal mucosa acted as control. AgNORs were assessed both quantitatively and qualitatively. The data were analyzed using Student's independent t-test, one-way analysis of variance (ANOVA), and multiple range test (Tukey-HSD). RESULTS: Quantitatively significant difference existed in the number of AgNORs between the normal mucosa, inflammatory lesions, and carcinomas, but the premalignant lesions failed to differ significantly from the normal mucosa. The number of AgNORs was found to be related to epithelial proliferation. Qualitatively, in terms of size, shape, and pattern of distribution, the normal mucosa and inflammatory lesion were alike, but the premalignant and malignant lesions differed significantly from the normal, with a marked degree of AgNOR pleomorphism being observed in carcinomas. CONCLUSIONS: AgNOR quantity is strictly proportional to the proliferative activity of the cell and does not necessarily indicate malignancy. It is the qualitative characteristics of AgNOR that help to differentiate hyperplastic, premalignant, and malignant lesions.

The diagnostic and prognostic implications of silver-binding nucleolar organizer regions in periodontal lesions.

BACKGROUND: The periodontal lesions with cellular proliferation can be assessed by various methods. One of the most recent methods to determine the proliferative activity is silver-staining nucleolar organizer region (AgNOR) staining. The purpose of the present study was to evaluate, if AgNOR count can act as a proliferative marker and can aid in the diagnosis and prognosis of periodontal lesions. MATERIALS AND METHODS: For this study, subjects with healthy gingival status, non-neoplastic lesions, neoplastic lesions, and plaque-induced gingivitis were included. Following the provisional diagnosis of clinical entity, biopsies were taken from the respective selected sites for histopathological diagnosis. In plaque-induced gingivitis cases, a second biopsy was taken from the selected sites 3 weeks following scaling. After histological confirmation, one more section was prepared, which was subjected to AgNOR staining, and AgNOR numbers were counted by individual and cluster counts and statistically analyzed. RESULTS: Results showed the highest AgNOR count in neoplastic lesions. Non-neoplastic lesions showed a higher AgNOR count as compared to clinically healthy gingiva. Plaque-induced gingivitis showed a considerable reduction in AgNOR count after treatment. CONCLUSION: Results of this study confirmed that AgNOR count reflects the cellular proliferation and has a limited diagnostic value. However, the prognostic value of AgNOR for periodontal lesions is dependable.

Morphometric Analysis of Nucleus and Nucleolar Organizer Regions (NORs) in Tongue Squamous Cell Carcinoma (SCC) / Análisis Morfométrico de Núcleos y Regiones del Organizador Nucleolar (NORs) en Carcinomas de Células Escamosas de la Lengua (SCC)

The tongue is one of the most likely places for the primary squamous cell carcinoma (SCC) to arise in the western world.This work intends to do a morphometric evaluation of the cellular nucleus, as well as a morphometric analysis of the nucleolar organizer regions (NORs). The biopsy specimen was removed from patients with tongue SCC, staged T2N0M0, before they underwent treatment. They were, then, subject to hemiglossectomy and supraomohyoid neck dissection (SOHND) and presented different clinical evolution. According to that, they were subdivided in three groups: Group A, control group, formed by patients without tongue neoplasm (the histopathologic specimen was obtained from the Department of Pathology at Hospital de Base and Faculdade de Medicina de Sao José do Rio Preto - FAMERP-HB); Group B, T2N0M0+ hemiglossectomy + SOHND, survival of only 36 months; Group C, T2N0M0 + hemiglossectomy + SOHND, survival of 5 years. The morphometric analysis included the study of larger diameter, smaller diameter, medium diameter, the relation between larger/smaller diameters, perimeter, area, volume, the relation between volume/area, idiosyncrasy, shape coefficient and the nuclear contour index of the tumor cells from patients with tongue SCC, as well as the same measurements of the nucleolar organizer regions. The analysis was aleatory and double-blind. All the morphometric data were compiled and statistically evaluated by the Kruskall-Wallis test and by the ANOVA test, and a 5% alpha error rate was adopted.

La lengua es probablemente, el lugar en donde más frecuentemente surge el carcinoma primario de células escamosas (SCC), en el mundo occidental. Este estudio propone realizar la evaluación morfométrica del núcleo celular, así como el análisis morfométrico de las regiones de los organizadores nucleolares (NORs). La muestra de biopsia fue removida de pacientes con SCC de lengua, etapificado en T2N0M0, antes de recibir tratamiento. Los pacientes fueron sometidos a una hemiglosectomía y una disección supraomohiodea del cuello (SOHND) y presentaron diferente evolución clínica. De acuerdo a esto, fueron subdividos en tres grupos: Grupo A, grupo control formado por pacientes sin neoplasia de lengua (la muestra fue obtenida del Departamento de Patología del Hospital Base de la Facultad de Medicina de Sao José do Rio Preto - FAMERP-HB, Brasil); Grupo B, T2N0M0 + hemiglosectomía + SOHND, sobrevida de 36 meses; Grupo C, T2N0M0 + hemiglosectomía + SOHND, sobrevida 5 años. El estudio morfométrico incluyó los análisis del diámetro mayor, diámetro menor, diámetro medio, la relación entre el diámetro mayor/menor, perímetro, área, volumen, relación volumen/área, idiosincrasia, coeficiente de forma y el índice de contorno nuclear de células tumorales en pacientes con SCC de lengua, así también las mismas mediciones de las regiones nucleolares organizadas. El análisis fue aleatorio y doble ciego. Todos los datos morfométricos fueron recopilados y evaluados estadísticamente con el test de Kruskall-Wallis y el Test ANO VA, con un valor alfa de un 5%.

Evaluation of nucleolar organizer regions in maxillary osteosarcoma

Maxillary osteosarcomas are a relatively frequent malignant tumor of the oral cavity. Similarly to other skeletal osteosarcomas, they exhibit different cellular differentiation patterns, i.e. chondroblastic, osteoblastic, or fibroblastic. Although their histological features resemble those of osteosarcomas of the long bones, their pattern of evolution usually differs. Morphometric variations in silver stained Nucleolar Organizer Regions (AgNOR) have proved of value to study the biology of several tumors. However, information on the analysis of AgNOR in maxillary tumors is scarce. The aim of the present study was to analyze the variations of different morphological parameters related to AgNOR in a series of 32 cases of maxillary osteosarcoma. In each case we analyzed 100 nuclei corresponding to the prevalent cellular differentiation type, selecting the most aggressive area. We employed software previously developed at our laboratory that yields information on different AgNOR-related parameters. The results were compared with those previously reported in a study on 12 cases of osteosarcoma of long bones. Six cases of oral mucosa squamous cell carcinoma were also included for comparative purposes. Single AgNOR volume proved to be the most discriminatory and informative parameter. The value of single AgNOR volume was considerably lower in mandible osteosarcomas than in osteosarcomas of the upper maxilla (p=0.02). The values were significantly lower in maxillary osteosarcomas than in long bone osteosarcomas and in oral carcinomas. This finding would suggest a slower rate of cell activity in maxillary osteosarcomas, associated in turn to its known lower degree of aggressiveness. The present results suggest that the analysis of AgNOR is a valuable and easily applicable marker to determine the degree of malignancy and biology of maxillary osteosarcomas.

Los osteoscaromas de maxilares son entidades relativamente frecuentes entre los tumores malignos de la cavidad bucal. Al igual que los osteosarcomas de otras localizaciones del esqueleto, pueden presentar diferentes patrones de diferenciación celular (condroblástico, osteoblástico o fibroblástico). Si bien sus características histológicas son similares, tienen generalmente un comportamiento evolutivo diferente al de los huesos largos. Las variaciones morfométricas de las regiones organizadoras del nucleoloidentificadas por impregnación argéntica (AgNOR) han demostrado ser marcadores útiles para el estudio de la biología de diversas entidades tumorales, pero hay muy escasa información de su análisis en tumores de los huesos maxilares. El objetivo de este trabajo fue analizar las variaciones de diferentes parámetros morfológicos de las AgNOR en una serie de 32 casos de osteosarcomas de maxilar. En cada caso se analizaron 100 núcleos en el patrón de diferenciación celular predominante, seleccionando la zona de mayor agresividad. Se utilizó un programa que aporta información sobre diferentes parámetros de AgNOR, desarrollado previamente en nuestro laboratorio. El parámetro más indicativo resultó ser el volumen individual de las AgNOR. Este parámetro en los osteosarcomas con localización mandibular fue considerablemente menor que aquellos localizados en maxilar superior (p=0.02). En los osteosarcomas de maxilar los valores fueron significativamente menores que en los de huesos largos y en los carcinomas bucales. Ellos podría ser indicativo de una menor actividad celular, a su vez asociada a su reconocida menor agresividad. Estos resultados sugieren que el análisis de AgNOR podría ser considerado como un marcador de utilidad y de fácil aplicación para determinar el grado de malignidad en osteosarcomas de maxilar y estimar su comportamiento biológico.

Usefulness of argyrophilic nucleolar organizer regions in detection of lung cells alterations after benzo[a]pyrene instillation

PURPOSE: To verify the relationship between AgNOR expression and lung tissues changes of Wistar rats after pulmonary instillation of benzo[a]pyrene (B[a]P). METHODS: Male Rattus norvegicus albinus,Wistar lineage were given a single intrapulmonary instillation of B[a]P at doses of 10 and 20 mg/kg in a volume of approximately 0,3 ml. After 7 and 21 days the rats were killed and the lung slices submitted to a histological technique of AgNOR. AgNOR dots were quantified and the result analyzed by statistical tests; p <= 0,05 was considered significant. RESULTS: The mean values of AgNOR dots for the experimental groups 10/7 (1,51±0,86) and 10/21 (1,84±0,13) were statistically different (p = 0,009). Among the groups 20/7 (1,63±0,11) and 20/21 (2,48±0,28) was observed statistically significant difference (p = 0,003). CONCLUSION: The AgNOR technique can be useful in identification of cells changes induced by B[a]P.

OBJETIVO: Verificar a relação entre a expressão de AgNOR e alterações teciduais pulmonares em ratos Wistar após instilação pulmonar de benzo[a]pireno (B[a]P). MÉTODOS: Rattus norvegicus albinus, linhagem Wistar machos foram submetidos à instilação pulmonar única de B[a]P em doses de 10 e 20mg/kg, em um volume aproximado de 0,3 ml. Os animais foram sacrificados após 7 e 21 dias e o tecido pulmonar submetido a técnica histológica de AgNOR. Os pontos AgNOR foram quantificados e os resultados analisados estatisticamente; foram considerados significantes os valores de p <= 0,05. RESULTADOS: Os valores médios de pontos AgNOR no grupo experimental 10/7 (1,51±0,86) e 10/21 (1,84±0,13) foram estatisticamente significantes (p = 0,009). Entre os grupos 20/7 (1,63±0,11) e 20/21 (2,48±0,28) a diferença observada foi também considerada significante (p = 0,003). CONCLUSÃO: A técnica de AgNOR pode ser útil na identificação de alterações celulares induzidas pelo B[a]P.

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

The number of argyrophilic nucleolar organizer regions [AgNOR] correlates with cellular proliferative activity. Comparing nonrecurrent, recurrent, atypical and malignant meningiomas we studied the value of the routine applicability of the AgNOR count in the prognostication of this tumor. Two 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. Silverstained slides were prepared and mean median and standard deviation of AgNOR dots determined. There 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. This 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

Regiões organizadoras nucleolares argirofílicas no sarcoma sinovial / Argyrophilic nucleolar organizer regions in synovial sarcoma

Regiões organizadoras nucleolares (NORs) correspondem a alças do DNA contendo genes responsáveis pela transcrição do RNA ribossômico, de 18S e 28S, situados no nucléolo da célula. Por possuírem reatividade com a prata são chamadas de AgNORs. O presente estudo investigou a correlação entre o número de AgNORs por núcleo e o prognóstico em dez sarcomas sinoviais diagnosticados no Hospital A. C. Camargo, entre 1972 e 1986, com informações confiáveis de seguimento clínico. A análise dos resultados não revelou correlação entre as variáveis estudadas, mostrando que o número de casos investigados e sua homogeneidade morfológica não permitem conclusões acerca do valor prognóstico dos AgNORs nos sarcomas sinoviais. Outrossim, o método se revelou barato, rápido e de aplicação simples para todo e qualquer serviço de patologia cirúrgica, podendo ser útil para avaliação da proliferação celular na rotina de laboratórios de pequeno porte.

Congenital glaucoma associated with 22p+ variant in a dysmorphic child.

A case of congenital glaucoma with developmental delay and several dysmorphic features showing 22p+ chromosomal variant is reported.

Proliferation in squamous cell carcinoma of cervix: a comparative assessment by two markers.

The study was aimed at assessing the cell proliferation in various grades of squamous cell carcinoma of cervix using two proliferation markers: AgNORs and Ki-67. 18 biopsy sections of various grades of SCC of cervix were chosen. AgNORs staining was done by single step staining technique using gelatin. Ki-67 immunostaining was done by Avidin--Biotin technique using DAB as a chromogen. Highest mean number of AgNORs was found in poorly differentiated squamous cell carcinoma and statistically significant difference was observed between well and poorly differentiated SCC and between moderately and poorly differentiated SCC. Highest mean Ki-67 LI was seen in poorly differentiated SCC. We observed correlation between histological grade and Ki-67 and AgNORs separately but no statistically significant correlation was found between Ki-67 and AgNOR counts. AgNORs and Ki-67 are both simple and easily performed techniques. They may prove to be a useful adjunct in estimating tumor proliferation and hence in determining the management strategy of the patients.

Proliferative activity in squamous intraepithelial and invasive lesions of cervix: analysis by AgNOR staining.

Proliferation potential of squamous intraepithelial lesions of cervix is the main factor affecting the biological aggressiveness of the lesion. The present study was aimed at assessing the proliferation in preinvasive and invasive squamous epithelial lesions of cervix using a simple and inexpensive proliferation marker: AgNOR (Argyrophilic nucleolar organizer regions). Fifty biopsy sections of various grades of preinvasive and invasive squamous epithelial lesions of cervix were taken for AgNOR staining. Single step staining technique using gelatin in formic acid solution was employed. Counting was done with the help of 'CAMERA LUCIDA' drawings. AgNOR counts/nucleus in parabasal and basal layer showed progressive rise in their mean value with increasing grade of lesion. Amongst invasive malignancies, highest mean of AgNORs/nucleus was observed in poorly differentiated squamous cell carcinoma of cervix. In conclusion, AgNORs can prove to be a simple inexpensive and reliable proliferation marker in lesions of cervix.

Oral exfoliative cytology of smokers at discrete clinical stages using AgNOR staining.

OBJECTIVE: This study was designed to evaluate oral exfoliative cytology of smokers without any clinically evident lesion and smokers with leukoplakia or oral cancer using AgNOR staining. STUDY DESIGN: Cytological smears of 30 smokers without lesion, 30 smokers with leukoplakia, 30 smokers with oral cancer and 30 non-smokers (control group) were studied using one step silver staining method. The AgNOR count was established on 100 cells. Mean AgNOR count and mean % of cells with 5 or more AgNORs was evaluated. The count values of groups were compared and analysed using Student's unpaired t-test. RESULTS: The mean AgNOR count for control group was 2.94 +/- 0.325, smokers without lesion 3.79 +/- 0.480 smokers with leukoplakia 3.89 +/- 0.433 and oral cancer 4.96 +/- 0.467. Mean % of cells with 5 or more AgNORs was 11.7, 26.5, 30.2 and 55.8 for control group, smokers without lesion, smokers with leukoplakia and oral cancer respectively. CONCLUSION: Analysis of AgNORs suggest that smoking influences proliferative activity in cells of smokers without any clinical lesion and that oral cancer shows highest proliferative activity.

Study of nucleolar organizer regions (NORs) in lesions of gallbladder.

Tissue sections from 100 specimens of gallbladder including 40 cases of chronic cholecystitis, 30 cases of epithelial hyperplasia and 30 cases of well, moderately and poorly differentiated adenocarcinoma of gallbladder were studied for argyrophillic nucleolar organizer regions (AgNORs). The arithmetic mean +/- S.D. were calculated for each category. This showed NOR counts of chronic cholecystitis--(1.89+/-0.96), epithelial hyperplasia--(3.99+/-1.03), well differentiated adenocarcinoma (WDCA)--(7.04+/-1.34), moderately differentiated adenocarcinoma (MDCA)--(7.52+/-0.97) and poorly differentiated adenocarcinoma (PDCA)--(8.37+/-1.27). The study revealed increasing number of AgNORS from cases of chronic cholecystitis to epithelial hyperplasia to carcinoma gallbladder, but a considerable overlap in AgNOR counts of individual cases was observed suggesting that though AgNOR count cannot act as a specific diagnostic parameter for diagnosis of early carcinoma & dysplasia in isolated cases, they may prove to be a good adjunct to presently available imaging techniques and cytology.

AgNOR counts in cervical lesions.

Present study was undertaken with a view to assess the utility of AgNOR counts in differentiating cervical lesions. One hundred cases of cervical lesions including normal, chronic cervicitis, CIN (I, II, III) and carcinoma were taken up for study during the period December 1995 to November 1996. Three micron thick sections of paraffin were subjected to AgNOR staining as described by Crocker and Smith with a little modification of 0.01% safranin counterstain. Statistical counts of AgNORs were done. The mean AgNOR counts in cervical epithelium showed a progressive and statistically significant increase from normal to chronic cervicitis to CIN, I, II and III (P<0.001). Scores in carcinoma also exceeded that of CIN (P<0.05). This can prove to be a useful adjunct to routine histopathology to evaluate cervical lesions.

AgNOR counts are not altered in alcoholic cirrhosis.

One hundred and two liver biopsy specimens were stained for Argyrophilic nucleolar organizer regions and associated proteins to assess its utility in differentiating normal, cirrhotic and neoplastic liver tissue. A statistically significant (p < 0.001) difference was observed between mean AgNOR counts of normal (1.53 +/- 0.4), post-hepatitic cirrhosis (3.65 +/- 0.53) and hepatocellular carcinoma (7.94 +/- 1.18). In contrast the mean AgNOR count of biopsies with alcoholic cirrhosis (1.57 +/- 0.06) was significantly less (p < 0.001) than post-hepatitic cirrhosis and was similar to that of normal liver tissue. It is concluded that AgNORs can act as a good adjuvant to histology in diagnosing liver diseases especially in differentiating post-hepatitic and alcoholic cirrhosis.

Técnica de detección de la región del organizador nucleolar en células exfoliadas de cuello uterino / Technique for the assay of nucleolus organizer region in exfoliated cells from uterine cervix

La técnica de detección de la región del organizador nucleolar (AgNOR) fue aplicada a 22 extendidos citológicos de exo y endocervix: 5 con células endocervicales, 2 hipotróficos (controles), 2 atróficos, 4 inflamatorios, 2 con células metaplásicas, 4 con células metaplásicas reactivas y 3 de lesiones intraepiteliales de bajo grado. Los promedios de partículas de AgNOR y el área coloreada aumentaron desde los controles hasta las células metaplásicas reactivas. Esta técnica permite identificar células exfoliadas del cuello uterino con mayor capacidad proliferativa, lo que resulta de utilidad en el citodiagnóstico de lesiones sospechosas en las coloraciones de rutina con el método de Papanicolaou

Helicobacter pylori status and cell proliferation activity in chronic antral gastritis.

BACKGROUND: Helicobacter pylori is known to cause antral gastritis and multifocal atrophic gastritis. In addition to its inflammatory effect, H. pylori has a direct effect on gastric mucosa. Increased epithelial proliferation, which may be an early biologic change in the development of gastric carcinoma, can be measured using silver stain for nuclear organizer regions (AgNOR). AIM: To detect the relation between H. pylori colonization and AgNOR index. METHODS: One hundred and twenty consecutive antral endoscopic biopsy specimens from patients with dyspepsia were examined for H. pylori colonization, polymorphonuclear infiltrate, mononuclear infiltrate, germinal center formation, mucus depletion and AgNOR index. RESULTS: AgNOR indices were not significantly related to grades of H. pylori colonization and chronic and active inflammation. The index increased significantly (p=0.03; ANOVA) with increasing mucin depletion. CONCLUSION: H. pylori colonization and presence of gastric antral inflammation are not related to cell proliferation activity; the latter is associated with mucin depletion.

Prognostic utility of silver stained nucleolar organiser region counting in non Hodgkin's lymphoma.

Silver stained Nucleolar Organiser Region (AgNOR) counting is a well established method of estimating the proliferative activity of a tumour. Their utility in prognostication in Non Hodgkin's Lymphoma (NHL) has not been fully investigated. We analysed hundred cases of NHL, all of which had a minimum of two and a half years of follow up, using histology by International Working Formulation (IWF) and their AgNOR scores. We found the IWF grading to have a good correlation with the outcome. The two and a half year survival for low grade NHL was 90 percent, for intermediate grade NHL was 47 percent and for high grade NHL was 30 percent. AgNOR scores correlated well with IWF grading and classification, with average AgNOR score of 1.82 (SD 0.34) in low grade NHL; 4.47 (SD 2.25) in intermediate grade NHL and 7.07 (SD 4.40) in high grade NHL. Over histology, AgNOR counting yielded additional prognostic information only in the diffuse small cleaved cell NHL (DSC), where finely dispersed NORs were seen. Of the 22 case of DSC included, 11 cases with AgNOR scores less than three had a much better outcome with 75 percent survival at two and a half years. Cases of DSC with AgNOR scores greater than three had a significantly worse outcome with only 25 percent survival at two and a half years (p=0.0103). Such sub-stratification could not be done in any other catergory of NHL using AgNOR scores. Thus, counting of AgNORs in lymphomas with finely dispersed NORs as in DSC is of prognostic value.