Role of Ki-67 and Argyrophilic Nucleolar Organiser Region (AgNoR) Counts in Squamous Cell Carcinoma

Background: Squamous cell carcinoma of the cervix is the most common subtype of cervical cancer and it usually accounts for 80-90% of the cases. These carcinomas mostly grow at the squamocolumnar junction (SCJ). The tumour tends to grow outward showing an exophytic growth especially at the squamocolumnar junction outside the external uterine wall whereas cancer tends to grow along the cervical canal showing an endophytic growth pattern where the SCJ is located within the cervical canal. This study has experimentally shown the role of AgNOR and Ki-67 scores in tumour proliferation. Methods: Cervical specimens were obtained and prepared in a specified fashion as described. The samples were de-waxed and staining was done with AgNOR. Then, the AgNOR was counted and scoring was done. Again, the samples were also stained with Ki-67. Results: This study has included 235 cases from the Department of Pathology of Ganesh Shankar Vidyarthi Memorial Medical College, Kanpur, India. The mean value of AgNOR count in the Control group came to be 10.62�45 while the mean AgNOR count was found to be 10.62�45, 15.10�79, 18.39�67, 19.75�74 and 19.59�59 for in-Situ SCC, well-differentiated SCC, moderately differentiated SCC, poorly differentiated SCC, SCC with basaloid differentiation, respectively. Conclusion: The study confirmed that AgNOR count increases with a higher grade of malignancy. It was concluded that AgNOR and Ki-67 scores can be used as an efficient predictor of tumour proliferation.

ROLE OF TUMOR MARKERS IN CERVICAL CARCINOGENESIS

Various types of tumor markers are currently being investigated to ascertain their capability in discriminating pre-cancerous lesions of cervix who have tendency for progression. The adequate treatment of such cases will check any chances of occurrence of carcinoma cervix in the population. The micro- RNAs are sensitive tumor markers but their high cost and sophisticated technique make them not feasible to be introduced in any cervical cancer screening program under Indian setup. Other tumor markers like claudins, p16, Ki67 etc are also very expensive. AgNOR pleomorphic counts and micronuclei counts are cheaper, the farmer being more reliable can be introduced in cytological screening program to identify high risk cases and can easily replace costly Human papilloma virus (HPV)- DNA testing.

Chromosome analysis in Saccodon wagneri (Characiformes) and insights into the karyotype evolution of Parodontidae

Parodontidae is a relatively small group of Neotropical characiform fishes consisting of three genera (Apareiodon, Parodon, and Saccodon) with 32 valid species. A vast cytogenetic literature is available on Apareiodon and Parodon, but to date, there is no cytogenetic data about Saccodon, a genus that contains only three species with a trans-Andean distribution. In the present study the karyotype of S. wagneri was described, based on both conventional (Giemsa staining, Ag-NOR, C-bands) and molecular (repetitive DNA mapping by fluorescent in situ hybridization) methods. A diploid chromosome number of 2n = 54 was observed in both sexes, and the presence of heteromorphic sex chromosomes of the ZZ/ZW type was detected. The W chromosome has a terminal heterochromatin band that occupies approximately half of the long arm, being this band approximately half the size of the Z chromosome. The FISH assay showed a synteny of the 18S-rDNA and 5S-rDNA genes in the chromosome pair 14, and the absence of interstitial telomeric sites. Our data reinforce the hypothesis of a conservative karyotype structure in Parodontidae and suggest an ancient origin of the sex chromosomes in the fishes of this family.(AU)

Parodontidae é um grupo relativamente pequeno de peixes caraciformes neotropicais que consiste em três gêneros (Apareiodon, Parodon e Saccodon) com 32 espécies válidas. Uma vasta literatura citogenética está disponível sobre Apareiodon e Parodon, mas até o momento não há dados citogenéticos sobre Saccodon, um gênero que contém apenas três espécies com distribuição transandina. No presente estudo foi descrito o cariótipo de S. wagneri, baseado em métodos convencionais (coloração de Giemsa, Ag-NOR, bandas C) e moleculares (mapeamento de DNA repetitivo por hibridização fluorescente in situ). Um número cromossômico diplóide de 2n = 54 foi observado, e a presença de cromossomos sexuais heteromórficos do tipo ZZ/ZW foi revelada. O cromossomo W possui uma banda terminal heterocromática que ocupa aproximadamente metade do braço longo, sendo esta banda aproximadamente a metade do tamanho do cromossomo Z. O ensaio FISH mostrou uma sintenia dos genes 18S-rDNA e 5S-rDNA no par de cromossomos 14, e a ausência de sítios teloméricos intersticiais. Nossos dados reforçam a hipótese de uma estrutura cariotípica conservadora em Parodontidae e sugerem uma origem ancestral dos cromossomos sexuais nos peixes desta família.(AU)

Cervical cancer screening in rural India: Status & current concepts

Cervical carcinoma is one of the most common and dreaded diseases of women, and in India, it accounts for 16 per cent of total cervical cancer cases occurring globally. The situation is more alarming in the rural areas where the majority of women are illiterate and ignorant about the hazards of cervical cancer. Different screening strategies such as rural cancer registries and camp approach for cancer detection have been found useful in minimizing the problem of cervical cancer in the villages. Various screening techniques such as visual inspection with acetic acid, visual inspection with Lugol's iodine, visual inspection with magnification devices-magnavisualizer, Pap smear and HPV-DNA testing have been suggested and tried under low-resource settings of our country, and cervical cytology screening has been found effective in reducing incidence of the disease. In the present review, feasibility of different screening methods has been assessed to find out the most suitable mode applicable at the rural level. Single lifetime screening particularly of high-risk women along with analysis of cost-effective tumour markers such as Argyrophilic nucleolar organizer regions (AgNOR) counts to discriminate high-risk dysplasia cases appears to be an appropriate approach in fighting against cervical cancer.

Tratamento de feridas excisionais de coelhos com extrato de barbatimão associado a células mononucleares autólogas da medula óssea / Treatment of excisional wound in rabbits with barbatiman extracts associated with autologous bone marrow mononuclear cells

Objetivou-se com este estudo avaliar o processo de cicatrização de feridas de coelhos tratadas com extrato de barbatimão (S. adstringens) associado a células mononucleares autólogas da medula óssea (CMMO). Utilizaram-se 20 coelhos, distribuídos em quatro grupos: B, extrato de barbatimão; CB, CMMO com extrato de barbatimão; CS, CMMO com solução fisiológica; S, solução fisiológica. Foi avaliada a presença de crosta, hiperemia, secreção, hemorragia, reepitelização, área da ferida e tempo de cicatrização. No terceiro, sétimo, 14º e 21º dias pós-operatório, realizou-se a biópsia das feridas e avaliaram-se os indicadores dos processos de inflamação e de reparo, com destaque para o colágeno, na coloração picrosírius, bem como de proliferação celular, na coloração AgNOR. Houve maior deposição de fibras colágenas nos grupos B e CB (P=0,00003) e formação de crostas mais espessas no sétimo dia, com fibras colágenas mais organizadas no 21º dia. Conclui-se que o barbatimão estimula a produção de fibras colágenas e promove a formação de crostas mais espessas sobre a ferida na fase inicial da cicatrização e, na fase de remodelação, favorece a orientação das fibras colágenas. Além disso, a associação desse fitoterápico com CMMO não estimula a cicatrização de feridas.(AU)

This study aimed to evaluate the healing process of wounds of rabbits in response to treatment with barbatiman extract (S. adstringens) associated with autologous mononuclear bone marrow cells (BM-MNC). We used 20 rabbits, divided into four groups: B, 10% barbatiman extract with 9.48% of total tannins; CB, BM-MNC with barbatiman extract; CS, BM-MNC with NaCl 0.9% solution; S, NaCl 0.9% solution. Clinical evaluation was performed by observing the presence of crust, redness, discharge, bleeding, re-epithelialization, the wound area and healing time in days. In the third, seventh, 14th and 21st postoperative days wounds were biopsied for microscopic evaluation of inflammation and repair process indicators, especially collagen, in picrosirius staining, and cell proliferation, in AgNOR staining. There was a greater deposition of collagen fibers in groups B and CB (p=0.00003) on the seventh day and formation of thicker crusts, and more organized collagen fibers on the 21st day in these groups. In conclusion, in the initial phase of healing, barbatiman extract stimulates the production of collagen fibers and promotes the formation of more exuberant crusts on the wounds and remodeling phase favors the orientation of collagen fibers, but when combined with BMMC does not stimulate wound healing in rabbits.(AU)

Evaluation of cell proliferation and endometrial thickness of bitches in different periods of diestrus

ABSTRACT This study aimed to determine the histological features of the endometrium of bitches, as well as the cell proliferation at specific moments of diestrus, 10, 20, 30, 40, 50 and 60 days post ovulation, correlating the endometrial thickness with the uterine cell proliferation and the metabolic state (weight, blood glucose and plasma cholesterol) of the animals. Therefore, the right and left uterine horns of 26 clinically healthy bitches submitted to ovariohysterectomy were histologically analyzed 10, 20, 30, 40, 50 and 60 days post ovulation. The hematoxylin-eosin and AgNOR staining techniques were performed. All parameters were evaluated by ANOVA and post-hoc Tukey test (p<0.05). The correlation between endometrial thickness and uterine cell proliferation, weight, blood glucose and plasma cholesterol of animals was observed using the Pearson method (p<0.05). In the present study, it is concluded that endometrial thickness does not differ at any of the moments analyzed in diestrus. The endometrial thickness is not influenced by hormones, weight, blood glucose or serum cholesterol of bitches in this phase of the estrous cycle. However, there is greater cell proliferation in the endometrium at day 40 compared to day 60 post ovulation under the influence of the endocrine profile.

Relación entre conducta y activación de áreas cerebrales: Empleo de la técnica de AgNOR en Psicología Comparada / Relationship between behavior and activation of brain areas: Using the AgNOR technique in comparative psychology

Se presenta una revisión de distintos estudios que emplearon la técnica de AgNOR en el campo de las neurociencias del comportamiento. AgNOR es una técnica histológica que permite estudiar la activación cerebral de una región específica. Esta técnica ha sido utilizada en distintos grupos zoológicos que comprenden desde peces y anfibios hasta mamíferos, incluyendo actividad neural tanto en procesos normales como patológicos. En conjunto, la técnica de AgNOR tiene tres notables ventajas con respecto a otras técnicas histológicas como las inmunohistoquímicas. En primer lugar, su bajo costo. En segundo lugar, su robustez. Finalmente, la posibilidad de ser aplicada con facilidad a cualquier especie animal, siendo esto último particularmente relevante en el campo de la Psicología Comparada. Paradójicamente a pesar de estas ventajas, es una técnica poco conocida y empleada. El propósito de este artículo es contribuir a su difusión dentro de la comunidad de científicos del comportamiento.

In this paper we review the use of AgNOR technique in the field of behavioral neuroscience. AgNOR is a histological technique used to study the activation of a specific brain region. NORs (nucleolar organizer regions) are the cell nucleus compartments where part of the ribosomal synthesis occurs. When a neuron enhances its protein synthesis demand, for example during a learning process, NORs increase their size. Staining with silver nitrate, colors the cell nucleus in brown and the cell nucleolus in dark brown, nearly black. Thus, the size of the nucleus and the nucleolus can be easily measured in histological preparations treated with silver nitrate through any appropriated image processing software. In this manner, AgNOR technique consist in staining the tissue of subjects that have received different treatments and comparing the average size of the nucleolus of a particular brain region, which is an indirect marker of cellular activity. The use of AgNOR technique in the study of neural basis of learning, typically involves: (1) training animals in a particular task, (2) after acquisition, animals in the trained group along with animals from a control condition are sacrificed in order to obtain a brain sample, (3) brains of animals from both conditions are processed and compared. This procedure was successfully used to study telencephalic activation during spatial learning in fish (Carassius auratus). The task consisted in using visual cues to identify the correct exit from a trap. In the case of fish, the histological analysis showed an increase of the nucleolus size in the lateral pallium, proposed as homologous to the mammalian hippocampus, observing that this area is involved in spatial memory. In our laboratory this technique was also used to analyze the neural activity after spatial learning in the terrestrial toad Rhinella arenarum. We conducted several studies where dehydrated animals had to use visual cues to locate the correct position of a water source. In accordance with fish’s data, our results indicated a higher activity of the hippocampal homologous region (the amphibian medial pallium), showing a high degree of functional equivalence. On the other hand, we also studied in toads the telencephalic activity during an aversive learning task. Here, it was observed an increase in the average of NORs size in the ventrolateral telencephalic region, the striatum (proposed as homologous to the mammalian amygdala), also showing a highly preserved functional equivalence. In mammals, AgNOR technique was used with different purposes, such as the study of changes in hippocampal activity during development in rats, the comparison of the basal activity of two regions of the rats hippocampus, the CA1 and the CA3, both strongly related with learning, or the the hippocampal activity in two different situations in which the liver activity was altered (by an artificial cholestasis, or by alcohol consumption). Finally, AgNOR technique was also used in the study of human brain activity of subjects that had committed suicide. On the whole, the AgNOR technique has three distinct advantages over other histological techniques such as the immunohistochemical. First, their low cost. Second, their robustness. Finally, the fact that it can be easily applied to any animal species, which makes it particularly attractive to the field of the Comparative Psychology. But paradoxically, despite these advantages it is a poorly known and used technique. The aim of this paper is to contribute to the diffusion of the AgNOR technique in the community of behavioral scientists, showing that its use has provided a valuable contribution to research in different fields and animal species.

First cytogenetic record for a species of Otothyropsis Ribeiro, Carvalho & Melo, 2005 (Loricariidae, Hypoptopomatinae)

Hypoptopomatinae is a monophyletic subfamily that includes 147 species, distributed in 20 genera. Otothyropsis is a genus of Hypoptopomatinae, recently described. Here, we provided the first cytogenetic information of Otothyropsis . The specimens were collected from córrego Dourado, a small tributary of rio Iguatemi, which flows into rio Paraná. The specimens of Otothyropsis cf. polyodon were analyzed with respect to diploid number, C-Band and Ag-NOR patterns. The diploid number was 54 chromosomes, distributed in 18 metacentric, 28 submetacentric, and 8 subtelocentric chromosomes, with single Ag-NOR and conspicuous heterochromatic blocks on the short and long arms of the 24th pair of chromosomes. Our study highlights the conservation trend of the diploid number (2n=54) and fundamental number (FN = 108) among the species of Hypoptopomatinae. However, the karyotype formula (18m+28sm+8st) seems to be specific to O. cf. polyodon , considering the other Hypoptopomatinae species already analyzed.

Hypoptopomatinae é uma subfamília monofilética que inclui 147 espécies distribuídas em 20 gêneros, sendo Otothyropsis um gênero recentemente descrito. Aqui, fornecemos a primeira informação citogenética do gênero Otothyropsis . Espécimes foram coletados no córrego Dourado, um pequeno tributário do rio Iguatemi, o qual deságua no rio Paraná. Espécimes de Otothyropsis cf. polyodon foram analisados em relação ao número diploide e padrões de Banda-C e Ag-NOR. O número diploide foi de 54 cromossomos, distribuídos em 18 metacêntricos, 28 submetacêntricos e 8 subtelocêntricos, com Ag-NOR simples e blocos heterocromáticos evidentes no braços curto e longo do par de cromossomos 24. Nosso estudo destaca a tendência de conservação do número diploide (2n=54) e número fundamental (NF=108) entre as espécies de Hypoptopomatinae. Entretanto, a fórmula cariotípica (18m+28sm+8st) parece ser específica para O. cf. polyodon, considerando as outras espécies de Hypoptopomatinae já analisadas.

Avaliação da atividade proliferativa celular das lesões bucais causadas pelo uso de próteses totais através da impregnação tecidual pela prata / Proliferative cellular activity evaluation of lesions caused by dentures through tissue silver impregnation

Introdução: As próteses totais visam conservar a função do sistema estomatognático do paciente totalmente edêntulo. Porém, na mucosa bucal podem aparecer manifestações cuja principal causa são as próteses totais mal adaptadas. Objetivo: o presente estudo objetiva investigar a proliferação tecidual das lesões causadas por próteses totais removíveis através do método de impregnação pela prata (AgNOR), com isso facilitando o tratamento e a determinação do prognóstico das lesões a serem estudadas. Metodologia: foram selecionados todos os casos das lesões bucais mais comumente associadas à utilização de próteses totais registradas no Serviço de Diagnóstico Histopatológico do ICB­UPF nos anos de 2012 e 2013, tendo sido encontrados 5 casos de granuloma piogênico, 5 casos de hiperplasia de fundo de sulco, 5 casos de fibroma de irritação e 2 casos de fibroma ossificante periférico. Os cortes histopatológicos das lesões foram impregnados pela prata (método AgNOR), tendo sido obtido, com auxílio do programa Image Tool®, o número de NORs de 100 células de cada caso, resultando numa média de NORs em cada grupo de lesões. Resultados: os resultados obtidos foram tabulados em planilha eletrônica e a comparação do número médio de NORs de cada grupo foi realizado por meio do teste estatístico ANOVA, 5% de significância. Resultados: o grupo das hiperplasias de fundo de sulco mostrou média de 2,41 NORs por núcleo, o grupo dos granulo mas piogênicos mostrou 2,44, o fibroma de irritação 2,22, e o fibroma ossificante periférico mostrou média de 1,89 NORs por núcleo celular, diferindo estatisticamente esta lesão das anteriormente mencionadas (p = 0,002). Conclusão: o fibroma ossificante periférico mostrou ser a lesão causada por prótese total removível com a menor atividade proliferativa celular. Tal estudo precisa ser complementado por futuros estudos clínicos.

Introduction: the total dentures are aimed at preserving the function of the stomatognathic system of the fully edentulous patient. However, in the oral mucosa may appear manifestations whose main cause are the totally unsuitable dentures. Objective: this study aims to investigate the proliferation of tissue lesions caused by removable dentures by impregnation method for silver (AgNOR), thereby facilitating the treatment and determining the prognosis of the lesions to be studied. Methodology: we selected all cases of oral lesions most commonly associated with the use of dentures recorded in Histopathological Diagnostic Service ICB-UPF in the years 2012 and 2013, having been found 5 cases of pyogenic granuloma, 5 cases of hyperplasia, 5 cases of irritation fibroma and 2 cases of peripheral ossifying fibroma. Histopathological lesions cuts were impregnated by silver (AgNOR method), having been obtained with the aid of the program Image Tool®, the number of NOR cells 100 in each case, resulting in an average NORs in every group of lesions. Results: the results were tabulated in a spreadsheet and comparing the average number of NORs of each group was conducted through ANOVA, 5% significance level. Results: The group of hyperplasias showed average of 2.41 NORs per nucleus, the group of pyogenic granulomas showed 2.44, the irritation fibroma 2.22, and peripheral ossifying fibroma showed average of 1.89 NORs for cell nucleus, differing significantly from that of the aforementioned lesions (p = 0.002). Conclusion: the peripheral ossifying fibroma proved the injury caused by removable dentures with lower cell proliferative activity. This study needs to be complemented by future clinical studies.

Activation of Epidermal Growth Factor Receptor (pEGFR) and Expression of Argentophilic Nucleolus Organizer Regions (AgNOR) in Seminomas.

Activation of epidermal growth factor receptor (p-EGFR) is one of the triggers in the development of many malignant tumors, and the measurement of Argentophilic Nucleolus Organizer Regions (AgNOR) area is used as a marker of tumor proliferation. Alpha actin smooth muscle (αASM) of peritubular cells (PT) and Vimentin expression are modified during the expansion of the CIS /intratubular germ cell neoplasia (ITGCN) to solid seminoma. We evaluated the expression pEGFR, the AgNOR areas, the expression of αASM in PT and the Vimentin in neoplastic cells from CIS/ ITGCN to solid seminoma. 28 formalin-fixed paraffin-embedded archival tissue blocks of Seminomas from the Departments of Pathology of Clinical Hospital and Ramos Mejía Hospital (CABA, Argentina) were used in this study. pEGFR was expressed in CIS/ ITGCN with membranous pattern (8/9), switching to a cytoplasmic pattern in the solid seminoma (15/19). AgNOR areas of atypical gonocytes were increasing from CIS (3.5±0.3 µ2) to intratubular seminoma (3.8±0.4µ2) (p<.5), until solid seminoma (5.3±0.7 µ2) (p<.01). Increase of AgNOR areas is proportional to the expression of pEGFR. Conclusions: pEGFR was expressed in CIS/ ITGCN with membranous pattern, switching to a cytoplasmic pattern in the solid seminoma. The activation of this receptor could be the first step in the mitogenic signal transduction. AgNOR areas complement diagnosis of testicular CIS, but not differentiate between CIS and intratubular seminoma. The value of AgNOR areas is maximum in the invasive seminoma, and statistically different from intratubular seminoma. The intensity of αASM in PT decreased as the tumor progressed and Vimentin was negative in neoplastic cells.

Avaliação da atividade proliferativacelular da leucoplasia bucal por meio do método AgNOR / Assessment of cell proliferative activity of oral leukoplakia through the AgNOR method

A leucoplasia é considerada uma importante lesão bucal cancerizável. Objetivo: realizar um estudo epidemiológico dos casos de leucoplasia bucal registrados no Serviço de Diagnóstico Histopatológico do Instituto de Ciências Biológicas da Universidade de Passo Fundo (SDH/ICB-UPF) entre 1989 e 2012, aplicando a tais casos o método AgNOR para avaliar a atividade proliferativa celular desses. Materiais e método: o trabalho consistiu num estudo epidemiológico-histoquímico transversal de 44 casos de leucoplasia. Após a confirmação diagnóstica e a tabulação dos dados clínicos e histopatológicos dos casos de leucoplasia, novas secções histológicas (3 µm) das lesões foram impregnadas pela prata por meio do método AgNOR. As lâminas obtidas foram fotografadas em imersão, tendo sido contabilizadas as NORs de 100 células de cada caso. Tal procedimento foi realizado com o auxílio do software Image Tool®. Os resultados foram comparados levando-se em conta as diferentes características clínicas e histopatológicas dos casos de leucoplasia. Resultados: a faixa etária mais frequente foi de 61 a 76 anos (19 casos - 43,2%), 22 pacientes pertenciam ao sexo masculino (50%) e a etnia predominante foi a leucoderma (36 casos - 81,8%). No que diz respeito aos hábitos nocivos, 23 pacientes (52,3%) informaram ser tabagistas. Ao se comparar o número médio de NORs dos casos de leucoplasia a diferentes características clínicas das lesões, não foi observada uma relação estatisticamente significativa. Conclusão: conclui-se que a leucoplasia acomete principalmente homens leucodermas, fumantes e com mais de 60 anos de idade. Além disso, na amostra ora estudada, a relação da atividade proliferativa celular da leucoplasia bucal com as características clínicas/ histopatológicas das lesões não foi estatisticamente significativa.

Avaliação do metabolismo epitelial em cistos radiculares pela técnica de AgNORS / Epithelial metabolism evaluation in radicular cysts by the AGNOR technique

Introdução: A formação do Cisto Radicular (CR) está associada à proliferação dos restos epiteliais de Malassez por estímulos inflamatórios, provenientes da proliferação bacteriana do canal radicular de um dente não vital. Quando o dente é removido, esse cisto passa a ser denominado Cisto Residual (CRe). O tratamento de escolha para o CR é endodôntico, com o objetivo de eliminar a inflamação presente no periápice. No entanto, em alguns casos, o cisto pode continuar a crescer, necessitando de tratamento cirúrgico, o que ocorre na maioria dos casos de CRe. Objetivo: Avaliar o metabolismo do epitélio de revestimento de CR e CRe, utilizando a quantificação das AgNORs, e verificar a influência da presença de inflamação sobre o crescimento desses cistos. Material e método: Vinte casos de CR e dez de CRe foram submetidos à técnica de AgNOR. A análise quantitativa das NORs foi realizada utilizando-se o software 'Contando células'. O teste estatístico pós-hock de Newman-keuls foi realizado para a comparação do número médio de AgNORs entre CR e CRe, e entre áreas inflamadas e não inflamadas. Resultado: Diferença estatisticamente significante (p=0,0094) foi observada entre áreas inflamadas (1,86±0,26) e não inflamadas (1,65±0,20). Na comparação entre CR (1,81±0,28) e CRe (1,73±0,16), não houve diferença estatisticamente significante (p=0,37). Conclusão: A inflamação interfere no metabolismo epitelial de CR e CRe, o que reflete a ação de fatores de crescimento na proliferação do epitélio, contribuindo para o crescimento do cisto, independentemente da presença do fator etiológico associado com a origem da lesão. .

Introduction: Radicular Cyst (RC) development is associated with the activation and proliferation of epithelial rests of Malassez. This occurs due to inflammatory stimuli resulting from bacteria proliferation in the root canal of a non-vital tooth. When the tooth is removed, the cyst becomes known as Residual Cyst (ReC). The RC common treatment is the endodontic therapy, with the aim of eliminating the inflammation in the periapical region. Nonetheless, this treatment is not always effective and the cyst may continue to grow, what happens in most cases of ReC. Objective: To evaluate the metabolism of the RC and ReC epithelial lining through the technique of AgNOR quantification and of the influence of the presence of inflammation in the growth of these cysts. Material and method: 20 cases of RC and 10 of ReC, were processed by the AgNOR technique. The AgNOR quantitative analysis was performed using the "Counting cells" software. The statistical post-hock Newman-Keuls test was applied to compare the RC and ReC mean number of AgNORs in inflamed and non-inflamed areas. Result: Statistically significant difference (p=0.0094) was observed between inflamed (1.86±0.26) and non-inflamed (1.65±0.20) areas. No statistically significant difference (p=0.37) was observed in the comparison between RC (1.81±0.28) and ReC (1.73±0.16). Conclusion: Inflammation interferes in the epithelial metabolism of RC and ReC, reflectings the action of growth factors in epithelial proliferation and contributing to the growth of the cyst, regardless the presence of the etiologic factor associated with the injury origin. .

Significance of AgNOR in Salivary Gland Tumours.

Diagnosis of Salivary gland tumours is challenging, because of wide variation in differentiation and overlapping morphological features. Sometimes, the difficulty encountered in distinguishing between pleomorphic adenoma, adenoid cystic carcinoma and adenocarcinoma. The objective is to study the application of AgNOR pattern in differentiating benign and malignant tumours of the salivary glands on Fine needle aspirates and their correlation with histopathology. Material and method: Cytological material was obtained by FNAC from forty three patients of salivary gland tumours. MGG and Pap stained smears were prepared for cytological interpretation. Histopathological study was done on routine formalin fixed and Haematoxylin & Eosin stained sections. Smears and sections were stained with Silver colloid stain for study of AgNOR counting. Results: AgNOR in benign tumours were small, round, uniform and less in number (1.02-1.97) while in malignant tumours they were very large, irregular, haphazardly distributed with high counts (1.23-16). Conclusion: Present study shows that count as well as morphology of AgNOR dots was helpful in differentiating between benign and malignant tumours and their grading of malignancy .

Diversity and chromosomal evolution in the genus Ancistrus Kner, 1854 (Loricariidae: Ancistrini) from three hydrographic basins of Mato Grosso State, Brazil

Cytogenetic analyses were carried out in 117 specimens of seven species of the genus Ancistrus from three hydrographic in Mato Grosso State: Paraguay, Araguaia-Tocantins and Amazon basins. Conventional cytogenetic techniques were used to obtain mitotic chromosomes. C-banding was performed to detect heterochromatic regions and silver nitrate staining was used to identify nucleolar organizer regions (Ag-NORs). The counted and paired chromosomes revealed diploid numbers ranging from 2n = 40 to 2n = 54 with karyotype formulae varying from FN = 80 to FN = 86. Single marks in distinct chromosomes identified the nucleolar organizer regions. The constitutive heterochromatin was scarce in the diploid number from 2n = 50 to 2n = 54 and conspicuous blocks were observed in a single species with 2n = 40 chromosomes. These data corroborate the hypotheses of reduction of diploid number in species with derived features such as presence of sex chromosomes and polymorphisms, besides allowing inferences about the evolutionary mechanisms and the ancestor karyotype that favored the diversification of this important genus in the tribe Ancistrini.

Foram realizadas análises citogenéticas de 117 espécimes do gênero Ancistrus de três bacias hidrográficas do estado de Mato Grosso: Paraguai, Araguaia-Tocantins e Amazônica, utilizando as técnicas de citogenética convencional para obtenção de cromossomos mitóticos, visualização de regiões heterocromáticas e regiões organizadoras de nucléolos. Os cromossomos pareados revelaram uma variação no número diploide de 2n = 40 a 2n = 54 e número fundamental de NF = 80 a NF = 86. As regiões organizadoras de nucléolos foram evidenciadas em um único par de cromossomos para todas as espécies e a heterocromatina é escassa nas espécies com números diploides elevados (2n = 50 a 2n = 54). Os blocos heterocromáticos mais evidentes foram observados nos pares portadores das AgRONs e em cromossomos da espécie com 2n = 40. Estes dados contribuem para a hipótese de redução do número diploide nas espécies que apresentam polimorfismos cromossômicos e cromossomos sexuais, além de contribuir para inferências sobre os mecanismos de evolução cariotípica que favoreceu a diversificação do gênero Ancistrus, o mais representativo na tribo Ancistrini.

Description of the karyotype of Geophagus cf. proximus (Perciformes, Cichlidae, Geophaginae) / Descrição do cariótipo de Geophagus cf. proximus (Perciformes, Cichlidae, Geophaginae)

This study described the karyotype of Geophagus cf. proximus. Specimens were collected in Água Preta Lake, Parque Ambiental de Belém, Pará State, Brazil. The karyotype were 2n = 48 chromosomes (FN = 60: 12M/SM+36ST/A) and no sexual chromosome differentiation. C-banding showed centromeric staining in all chromosomes. The first chromosome pair, besides centromeric coloration, presented a totally heterochromatic long arm. The nucleolus organizer regions (NORs) were studied by means of AgNO3. NORs were located at the short arm of the second chromosome pair.

Este estudo descreve o cariótipo de Geophagus cf. proximus. Espécimes foram coletados no lago Água Preta, Parque Ambiental de Belém, Estado do Pará, Brasil. O cariótipo obtido apresentou 2n = 48 cromossomos (NF = 60: 12M/SM+36ST/A), sem diferenciação de cromossomos sexuais. O bandeamento C mostrou marcações centroméricas em todos os cromossomos. O primeiro par cromossômico, além da coloração centromérica, apresentou o braço longo totalmente heterocromático. As regiões organizadoras do nucléolo (RONs) foram estudadas por meio da coloração de AgNO3. As RONs foram encontradas no braço curto do segundo par de cromossomos.

Study of AgNOR count in FNAC of breast lesions.

Background:ArgyrophilicNucleolar Organizer Region technique has a potential value in the diagnosis of malignancy and can be used in cases with equivocal and inconclusive cytological picture. The purpose of this study was to evaluate mean ArgyrophilicNucleolar Organizer Region count and ArgyrophilicNucleolar Organizer Region Pattern Assessment score in fine needle aspirates of breast lumps.Materials and Methods: The present study consists of 80 cases of AgNOR count done in fine needle aspiration cytology of various breast lesions in patients on O.P.D. basis or those admitted in Guru Govindsingh Hospital attached to Shri M.P. Shah Medical College, Jamnagar during the period between August '98 to May 2000. Fine Needle Aspiration smears were studied by conventional methods and silver staining for ArgyrophilicNucleolar Organizer Regions. Histopathologic diagnosis was taken as the gold standard. Results:ArgyrophilicNucleolar Organizer Region count and ArgyrophilicNucleolar Organizer Region Pattern Assessment score were helpful in differentiating benign from malignant lesions. Mean ArgyrophilicNucleolar Organizer Region count and ArgyrophilicNucleolar Organizer Region Pattern Assessment score were 2.63 ± 1.36 and6.26 ± 1.19 respectively in benign lesions while they were 8.42 ± 2.53 and 10.05 ± 2.22 respectively in malignant lesions.With few exceptions, cases with high counts had high scores. Conclusion: Mean ArgyrophilicNucleolar Organizer Region AgNOR count and Subjective ArgyrophilicNucleolar Organizer Region Pattern Assessment score provide useful information regarding cellular proliferation. Both count and score have comparable diagnostic potential but the latter is a more convenient and rapid method for ArgyrophilicNucleolar Organizer Region evaluation.

Effects of dietary high fat on prostate intraepithelial neoplasia in TRAMP mice / 한국실험동물학회지

Increased fat intake is known to be a major cause of prostate cancer. In this study, we investigated the effect of dietary high fat on prostate intraepithelial neoplasia using transgenic adenocarcinoma mouse prostate (TRAMP) mice. Six-week-old male TRAMP mice were fed AIN93G (control group, 4.0 kcal/kg, n=6) and AIN93G-HFD (experimental group, 4.8 kcal/kg, n=7) for 10 weeks. Prostate histopathology, urogenital tract (UGT) weight, epididymal white adipose tissue weight, argyrophilic nucleolar organizer regions (AgNORs) counts, and serum leptin levels were examined. AIN93G-HFD fed group showed progressed neoplastic lesions in the prostate (P<0.05) compared to AIN93G fed group. AIN93G-HFD intake resulted in a increase in the weight of UGT (P<0.05) and epididymal white adipose tissue. The number of Ag-NOR positive dots significantly increased in each prostate lobe and final serum leptin levels in AIN93G-HFD fed group were about twice those of AIN93G fed group (P<0.05). Dietary high fat was related to the prostate cancer progression in the early stage of TRAMP mice and increased serum leptin levels, suggesting that the regulation of dietary components could delay the progression of prostate cancer.

Quantification of AgNOR expression in exfoliated oral mucosal cells of tobacco chewers with and without lesion.

Background: Nucleolar organizer regions (NOR) are associated with proliferative activity and represent a diagnostic and prognostic marker. Materials and Methods: Smears were taken from smokers, tobacco chewers, oral squamous cell carcinoma patients, and normal subjects and evaluated by 2 silver-staining nucleolar organizer region (AgNOR) counting methods: (1) mean number of AgNORs per nucleus (mAgNOR); and (2) percentage of nuclei with >3 and >5 AgNORs (pAgNOR). Results: A statistically significant difference was observed between normal subjects, smokers, tobacco chewers, and oral cancer patients and between tobacco chewers with and without lesion. No significant difference was observed between tobacco chewers and smokers except in the percentage of >5 criteria. Conclusions: AgNOR enumeration using noninvasive methods, such as the cytobrush appears to be useful technique in distinguishing between normal mucosa, mucosa with and without lesions exposed to carcinogens, such as tobacco and frank oral carcinoma.

Evaluation of proliferative activity in oral squamous cell carcinoma by the AgNOR staining method / Avaliação da atividade proliferativa do carcinoma espinocelular pelo método AgNOR

The objective of this study was to evaluate the cellular proliferative activity of argyrophilic nucleolar organizer regions (AgNORs) in a series of oral squamous cell carcinoma (OSCC) cases and to correlate this activity with the histopathologic grade of the tumor defined by the World Health Organization (WHO).Methodology: twenty cases of OSCC obtained from the Diagnostic Histopathology Service of UPF database were studied. Age, gender and ethnic origin of the patients and oral site affected by the tumor were obtained directly from the histopathologic reports. The AgNOR technique was used for histochemical analysis of the tumors.Results: there was a predominance of OSCC in male patients in the sixth decade of life. The lip was the most affected oral site and WHO grade I was the predominant histologic grade. The mean number of NORs per nucleus in the sample was 2.00 (standard deviation: 0.58). Pearson’s correlation test showed that the number of NORs in OSCC was directly correlated with histologic tumor grade (p = 0.021).Conclusion: the AgNOR method was useful as a diagnostic tool and prognostic indicator of OSCC when combined with conventional histopathologic analysis...

Avaliar a atividade proliferativa celular das regiões organizadoras nucleolares (AgNORs) em uma série de casos de carcinoma espinocelular da cavidade bucal (CECB) e correlacionar esta atividade com o grau histológico do tumor, definida pela Organização Mundial da Saúde (OMS). Metodologia: vinte casos de CECB estudados foram obtidos a partir de diagnóstico do Serviço de Diagnóstico Histopatológico da UPF. Dados relativos à idade, sexo e origem étnica dos pacientes, além da localização da lesão na cavidade bucal, foram obtidos a partir dos relatórios histopatológicos. A técnica de AgNOR foi utilizada para análise histoquímica dos tumores.Resultados: houve predomínio de CECB em pacientes do sexo masculino na sexta década de vida. O lábio foi o local mais afetado e o grau histológico I foi o predominante. O número médio de NORs por núcleo na amostra foi de 2,00 (desvio padrão: 0,58). O teste de correlação de Pearson mostrou que o número de NORs no CECB foi diretamente correlacionado com o grau histológico do tumor (p = 0,021). Conclusão: o método AgNOR foi útil como uma ferramenta de diagnóstico e indicador de prognóstico do CECB, quando combinado com a análise histopatológica convencional...

Cell proliferation markers in the transplanted canine transmissible venereal tumor / Marcadores de proliferação celular no tumor venéreo transmissível canino transplantado

Adult male mongrel dogs were subcutaneously transplanted with the canine transmissible venereal tumor (TVT) on the hypogastric region. Twelve specimens of tumors were collected, half during the proliferative phase and the other half during the regressive phase. Fragments of the tumor were fixed in 10 percent buffered formalin and routinely processed for light microscopy. Sections of 4µm were stained by Schorr or AgNOR or either immunostained for MIB1 (Ki67). Schorr stain, AgNOR and MIB1 showed an increased proliferative activity through mitotic index, nuclear argyrophilic protein stain and cycling tumoral cells in the growing tumors, respectively. All of the three cell proliferation markers were able to distinguish the TVT in both evolution phases. MIB1 monoclonal antibody was the best in the morphologic evaluation of growth and regression of TVT. This resulted in higher values than AgNORs counting and mitotic index. MIB1 immunostaining was the most effective parameter of the proliferative activity of TVT. However, a significant correlation has been detected only between mitosis counting and AgNORs.

Cães machos, adultos, mestiços, foram transplantados com células do tumor venéreo transmissível canino (TVTC), na região hipogástrica. Foram coletados doze espécimes do TVTC, sendo metade durante a fase proliferativa e metade durante a fase regressiva. Fragmentos do tumor foram fixados em formol a 10 por cento, tamponado e processado rotineiramente para microscopia de luz. Secções de 4µm foram coradas pelo Shorr, ou pela AgNOR, ou ainda, imunocorado para MIB 1 (Ki67). As colorações pelo Shorr, AgNOR, ou MIB 1 mostraram um aumento do índice mitótico, coloração da proteína argirofílica nuclear e células tumorais ciclando em tumores em crescimento, respectivamente. Todos os três marcadores de proliferação celular foram capazes de distinguir o TVTC em ambas as fases de evolução. O anticorpo monoclonal MIB 1 foi o melhor na avaliação morfológica, de crescimento e regressão do TVTC. Isto resultou em um valor maior que a contagem de AgNOR e do índice mitótico. A imunomarcação com MIB1 foi o parâmetro mais efetivo da atividade proliferativa. No entanto, só foi observada uma correlação positiva entre a contagem de mitose e a AgNOR.