RESUMO
OBJECTIVES: To evaluate the number of AgNORs per nucleus and the expression of Ki-67 at the tumor invasion front (TIF) in relation to clinical parameters (TNM), TIF classification and the prognosis of oral squamous cell carcinomas in an Uruguayan population. MATERIAL AND METHODS: This study was conducted through a retrospective survey from 2000 to 2010 at the National Institute of Cancer Montevideo, Uruguay and included 40 patients. The samples were obtained from the resection of the tumor and the TIF was defined according with Bryne, et al.5 (1992). Expression of Ki-67 was assessed by the percentage of positive tumor cells and the AgNOR was recorded as the mean AgNOR (mAgNOR) and the percentage of AgNOR per nucleus (pAgNOR). All analyzes were performed by a blinded and calibrated observer. RESULTS: No statistically significant association was observed between immunostaining of Ki-67 and AgNOR with the different types of TIF, regional metastasis and patients prognosis, however it was observed an increase in Ki-67 expression associated with worse patient's clinical staging, although not statistically significant. CONCLUSIONS: Our results suggest that proliferation markers as AgNOR and Ki-67 are not prognostic markers at the tumor invasive front of carcinoma of oral squamous cell.
Assuntos
Antígenos Nucleares/análise , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Biomarcadores Tumorais , Proliferação de Células , Feminino , Humanos , Imuno-Histoquímica , Antígeno Ki-67/análise , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/patologia , Prognóstico , Valores de Referência , Reprodutibilidade dos Testes , Estudos Retrospectivos , Carga Tumoral , UruguaiRESUMO
Abstract Objectives To evaluate the number of AgNORs per nucleus and the expression of Ki-67 at the tumor invasion front (TIF) in relation to clinical parameters (TNM), TIF classification and the prognosis of oral squamous cell carcinomas in an Uruguayan population. Material and Methods This study was conducted through a retrospective survey from 2000 to 2010 at the National Institute of Cancer Montevideo, Uruguay and included 40 patients. The samples were obtained from the resection of the tumor and the TIF was defined according with Bryne, et al.5 (1992). Expression of Ki-67 was assessed by the percentage of positive tumor cells and the AgNOR was recorded as the mean AgNOR (mAgNOR) and the percentage of AgNOR per nucleus (pAgNOR). All analyzes were performed by a blinded and calibrated observer. Results No statistically significant association was observed between immunostaining of Ki-67 and AgNOR with the different types of TIF, regional metastasis and patients prognosis, however it was observed an increase in Ki-67 expression associated with worse patient's clinical staging, although not statistically significant. Conclusions Our results suggest that proliferation markers as AgNOR and Ki-67 are not prognostic markers at the tumor invasive front of carcinoma of oral squamous cell.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Neoplasias Bucais/patologia , Carcinoma de Células Escamosas/patologia , Antígenos Nucleares/análise , Prognóstico , Valores de Referência , Uruguai , Imuno-Histoquímica , Biomarcadores Tumorais , Reprodutibilidade dos Testes , Estudos Retrospectivos , Análise de Variância , Antígeno Ki-67/análise , Carga Tumoral , Proliferação de Células , Invasividade Neoplásica/patologiaRESUMO
OBJECTIVE: To evaluate transcallosal changes after a local ischemic injury in rats by using the monoclonal marker anti-NeuN (Mouse anti-neuronal nuclei). METHODS: Twenty-eight adult, male, Wistar rats were subjected to focal injury in the right hemisphere. The technique used was the experimental model of focal ischemic injury through intraluminal suture of the middle cerebral artery. Analyses were made for the five groups: after the lesion (control), at 24 h, 96 h, 10 days and 20 days. Exofocal neuronal damage was inferred from neuronal immunoreactivity changes to NeuN. RESULTS: In the cortex contralateral to the lesion, immunoreactivity was diminished. This finding was most notable in the supra-granular sheets 24 h post ischemia. After 96 h, there was a generalized diminishment of the inmmunoreactivity in the supra and infra-granular sheets. At 10 and 20 days, the tissue recovered some immunoreactivity to NeuN, but there were some changes in the VI layer. CONCLUSION: The immunoreactive changes to NeuN support the process of inter-hemispheric diaschisis. Changes in immunoreactivity could indicate metabolic stress secondary to the disruption in connectivity to the site of lesion.
OBJETIVO: Evaluar los cambios exofocales transcallosos después de lesión isquémica focal en ratas, mediante marcación inmunohistoquímica con el anticuerpo monoclonal anti-NeuN (Mouse Anti-Neuronal Nuclei). MÉTODOS: Se intervinieron 28 ratas machos Wistar adultas. Mediante el modelo experimental de isquemia cerebral focal del territorio de la arteria cerebral media por filamento intraluminal, se les ocasionó una lesión focal en el hemisferio derecho. Posteriormente se evaluó el hemisferio contralateral, marcando la población neuronal con el anticuerpo monoclonal anti-NeuN. Se definieron cinco grupos de evaluación: uno de control, 24 horas, 96 horas, 10 días y 20 días. Se evaluaron los cambios neuronales exofocales después de la lesión con base en la observación de los cambios en la inmunoreactividad de las neuronas al NeuN. RESULTADOS: Se redujo la inmunoreactividad en la corteza contralateral a la lesión. Este fenómeno fue más notable en las capas supragranulares después de 24 h post isquemia. Después de 96 h hubo una disminución generalizada de la inmmunoreactivity en las capas supra e infragranulares. A los 10 y 20 días, el tejido recobró alguna inmunoreactividad NeuN, estos cambios se dieron en la capa VI. CONCLUSIONES: Los cambios inmunorreactivos a NeuN apoyan el proceso de diasquisis interhemisférica. Los cambios en la inmunorreactividad podrían indicar estrés metabólico secundario a la interrupción en la conectividad con el sitio de la lesión.
Assuntos
Antígenos Nucleares/análise , Isquemia Encefálica/complicações , Corpo Caloso/patologia , Artéria Cerebral Média , Animais , Anticorpos Monoclonais , Antígenos Nucleares/imunologia , Biomarcadores , Isquemia Encefálica/patologia , Corpo Caloso/imunologia , Imuno-Histoquímica , Masculino , Necrose , Ratos , Ratos WistarRESUMO
Objective: To evaluate transcallosal changes after a local ischemic injury in rats by using the monoclonal marker anti-NeuN (Mouse anti-neuronal nuclei). Methods: Twenty eight adult, male, Wistar rats were subjected to focal injury in the right hemisphere. The technique used was the experimental model of focal ischemic injury through intraluminal suture of the middle cerebral artery. Analyses were made for the five groups: and after the lesion (control), at 24 h, 96 h, 10 days and 20 days. Exofocal neuronal damage was inferred from neuronal immunoreactivity changes to NeuN. Results: In the cortex contralateral to the lesion, immunoreactivity was diminished. This was most notable in the supragranular layers 24 h post ischemia. After 96 h, there was a generalized diminishment of the inmmunoreactivity in supra and infragranular layers. At 10 and 20 days, the tissue recovered some NeuN immunoreactivity, but there were set changes in the VI layer. Conclusion: The immunoreactive changes to NeuN support the process of interhemispheric diaschisis. Changes in immunoreactivity could indicate metabolic stress secondary to the disruption in connectivity to the site of lesion.
Objetivo: Evaluar los cambios exofocales transcallosos después de lesión isquémica focal en ratas, mediante marcación inmunohistoquímica con el anticuerpo monoclonal anti-NeuN (Mouse Anti-Neuronal Nuclei). Métodos: Se intervinieron 28 ratas machos Wistar adultas. Mediante el modelo experimental de isquemia cerebral focal del territorio de la arteria cerebral media por filamento intraluminal, se les ocasionó una lesión focal en el hemisferio derecho. Posteriormente se evaluó el hemisferio contralateral, marcando la población neuronal con el anticuerpo monoclonal anti-NeuN. Se definieron cinco grupos de evaluación: uno de control, 24 h, 96 h, 10 días y 20 días. Se evaluaron los cambios neuronales exofocales después de la lesión con base en la observación de los cambios en la inmunoreactividad de las neuronas al NeuN. Resultados: Se redujo la inmunoreactividad en la corteza contralateral a la lesión. Este fenómeno fue más notable en las capas supragranulares después de 24 h post isquemia. Después de 96 h hubo una disminución generalizada de la inmmunoreactivity en las capas supra e infragranulares. A los 10 y 20 días, el tejido recobró alguna inmunoreactividad NeuN, estos cambios se dieron en la capa VI. Conclusiones: Los cambios inmunorreactivos a NeuN apoyan el proceso de diasquisis interhemisférica. Los cambios en la inmunorreactividad podrían indicar estrés metabólico secundario a la interrupción en la conectividad con el sitio de la lesión.
Assuntos
Animais , Masculino , Ratos , Isquemia Encefálica/complicações , Corpo Caloso/patologia , Artéria Cerebral Média , Antígenos Nucleares/análise , Imuno-Histoquímica , Biomarcadores , Isquemia Encefálica/patologia , Ratos Wistar , Corpo Caloso/imunologia , Antígenos Nucleares/imunologia , Anticorpos Monoclonais , NecroseRESUMO
Oral cancer is a world health problem, and one of the highest incidence rates of oral cancer worldwide occurs in Brazil. STAG2 is part of the cohesin complex which is responsible for sister chromatid cohesion. STAG2 loss of expression was reported in a range of tumors, and STAG2 loss was found to cause chromosomal instability and aneuploidy in cancer cells. On the basis of these findings, we investigated STAG2 expression in oral cancer and potentially malignant lesions. We investigated STAG2 immunoexpression in oral cancer, lip cancer, oral leukoplakia, and actinic cheilitis, including complete clinical information. Normal oral mucosa samples were included as normal controls. STAG2 protein was highly expressed in all samples. We further tested STAG2 expression in gastric adenocarcinomas and glioblastomas, as these tumor types were previously shown to lose STAG2 expression. We found homogenous expression of STAG2 by these tumor cells. Our results suggest that STAG2 loss of expression is not a common event in oral carcinogenesis.
Assuntos
Antígenos Nucleares/análise , Queilite/genética , Neoplasias Labiais/genética , Neoplasias Bucais/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Ciclo Celular , Queilite/metabolismo , Feminino , Glioblastoma/química , Glioblastoma/genética , Humanos , Imuno-Histoquímica , Leucoplasia Oral/química , Leucoplasia Oral/genética , Neoplasias Labiais/química , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/química , Neoplasias Gástricas/química , Neoplasias Gástricas/genéticaRESUMO
AIM: To evaluate the proliferative potential and the cell proliferation rate of odontogenic epithelial cells. MATERIALS AND METHODS: Forty-two cases of pericoronal follicles of impacted third molars were submitted to silver impregnation technique for quantification of argyrophilic nucleolar organizer regions (AgNOR) and immunohistochemical staining for EGFR and Ki-67. For AgNOR quantification, the mean number of active nucleolar organizer regions per nucleus (mAgNOR) and the percentage of cells with 1, 2, 3 and 4 or more AgNORs per nucleus (pAgNOR) were quantified. Ki-67 immunolabeling was quantified, whereas for EGFR, a descriptive analysis of staining patterns (membrane, cytoplasm or membrane + cytoplasm positivity) was performed. We evaluated the reduced epithelium of the enamel organ and/or islands of odontogenic epithelium present in the entire connective tissue. RESULTS: mAgNOR were 1.43 (1.0-2.42) and were significantly different among pericoronary follicles from upper and lower teeth (p = 0.041). Immunostaining of Ki-67 was negative in all cases. EGFR immunolabeling was found mainly in the cytoplasm and was more intense in islands and cords when compared to reduced epithelium of the enamel organ. CONCLUSION: Odontogenic epithelial cells of some pericoronal follicles have proliferative potential, suggesting their association with the development of odontogenic lesions. CLINICAL SIGNIFICANCE: The authors suggest that nonerupted, especially of the lower teeth, should be monitored and if necessary removed.
Assuntos
Saco Dentário/citologia , Odontogênese/fisiologia , Adolescente , Adulto , Antígenos Nucleares/análise , Membrana Celular/ultraestrutura , Núcleo Celular/ultraestrutura , Proliferação de Células , Citoplasma/ultraestrutura , Saco Dentário/ultraestrutura , Órgão do Esmalte/citologia , Órgão do Esmalte/ultraestrutura , Células Epiteliais/fisiologia , Células Epiteliais/ultraestrutura , Receptores ErbB/análise , Feminino , Humanos , Imuno-Histoquímica , Antígeno Ki-67/análise , Masculino , Região Organizadora do Nucléolo/ultraestrutura , Adulto JovemRESUMO
This study presents the development and evaluation of an image processing software for computer-assisted cellular structure counting. The proposed software consists of a set of processing and analytical tools which allows its use in several applications of cell and cellular structure counting. A particular application in AgNOR quantitative analysis is presented. The knowledge obtained from experienced pathologists has been codified in a sequence of processing steps in order to allow automatic estimation of the mean number of AgNORs per cell in ameloblastomas. The performance of the presented software in such application was verified by comparing the data provided by visual analysis, by two observers previously calibrated and under supervision of two experienced specialists (Group 1) and by the computer program (Group 2). No statistical difference was observed (p<0.05) between the two groups. The use of the proposed method in AgNOR applications permitted attainment of accurate and precise data without the difficulties frequently found in the traditional visual analysis method (time, training and subjectivity). The developed software is an interesting tool as an aid in the study (estimation of the number of cells and cellular structures) of malignant and benign neoplasms.
Assuntos
Ameloblastoma/ultraestrutura , Antígenos Nucleares/análise , Processamento de Imagem Assistida por Computador/métodos , Neoplasias Maxilomandibulares/ultraestrutura , Região Organizadora do Nucléolo/ultraestrutura , Patologia Bucal/instrumentação , Contagem de Células/métodos , Núcleo Celular/ultraestrutura , Proliferação de Células , Intervalos de Confiança , Humanos , Imuno-Histoquímica , Patologia Bucal/métodos , Reprodutibilidade dos Testes , SoftwareRESUMO
PURPOSE: To analyze the association between nuclear medicine examination (single photon emission computed tomography [SPECT]), histology, and Argyrophilic Nuclear Organizer Region (AgNOR) count in patients with active condylar hyperplasia who have undergone condylectomy. PATIENTS AND METHODS: Eight patients with a diagnosis of active condylar hyperplasia and evidence of facial asymmetry, with progressive deformation in time and on SPECT studies, were evaluated. The relationship between the rate of technetium Tc 99 intake, cartilage layer thickness, and cellular activity measured by recounting nucleolar organizers with AgNOR was evaluated. RESULTS: The 4 pathologic layers of condylar hyperplasia (fibrous, mesenchymal, and hypertrophic chondrocyte layers and ossification layer) showed great variability and different thicknesses among the cases analyzed. As age increased, the histologic layer thickness decreased (r = -0.73, P = .04). The age of the patients was inversely related to the number of AgNOR dots (r = -0.65, P = .08). The thickness of both mesenchymal and hypertrophic chondrocyte layers was related to cartilage island depth (r = 0.81, P = .02). CONCLUSIONS: Younger patients with condylar hyperplasia had a thicker condylar layer and more cellular activity measured by AgNOR count. The histologic features of this group of patients could not be associated with their SPECT findings.
Assuntos
Antígenos Nucleares/análise , Antígenos/análise , Côndilo Mandibular/patologia , Região Organizadora do Nucléolo/patologia , Tomografia Computadorizada de Emissão de Fóton Único , Adolescente , Adulto , Fatores Etários , Cartilagem Articular/diagnóstico por imagem , Cartilagem Articular/patologia , Proliferação de Células , Condrócitos/patologia , Assimetria Facial/diagnóstico por imagem , Assimetria Facial/patologia , Assimetria Facial/cirurgia , Feminino , Fibrose , Humanos , Hiperplasia , Hipertrofia , Masculino , Côndilo Mandibular/diagnóstico por imagem , Côndilo Mandibular/cirurgia , Mesoderma/patologia , Ossificação Heterotópica/diagnóstico por imagem , Ossificação Heterotópica/patologia , Compostos Radiofarmacêuticos , Estudos Retrospectivos , Coloração pela Prata , Tecnécio , Adulto JovemRESUMO
The main purpose for studying cytological body fluids is confirmation of a benign or malignant effusion. Our cytology laboratory analyzes body fluids and results are requested urgently. The samples are stained by the Giemsa and Papanicolaou methods to give a preliminary report, then they are examined by other complementary techniques. Three hundred thirty samples of pleural and peritoneal fluids were studied to compare the sensitivity of Papanicolaou and Giemsa stains. AgNOR assay, immunocytochemistry and assessment of ploidy were used to improve the sensitivity of the cytodiagnosis. Two hundred one samples were positive, 84 negative and 45 inconclusive using the Papanicolaou stain, while 135 samples were positive, 72 negative and 123 inconclusive using Giemsa stain. The sensitivity was 79%, 53% and 83% for Papanicolaou, Giemsa, and both techniques together, respectively. Using complementary techniques, the sensitivity reached 95% for AgNOR, 87% for tumor markers (panel), and 92% for Ploidy. There were no false positive in our series; therefore specificity was 100%. The use of both Papanicolaou and Giemsa in conjunction increased the sensitivity of the cytodiagnosis in body fluids. The complementary methods, especially AgNOR assay and assessment of ploidy, diminished the number of inconclusive cases.
Assuntos
Corantes Azur/química , Líquidos Corporais/citologia , Citodiagnóstico/métodos , Imuno-Histoquímica/métodos , Coloração e Rotulagem/métodos , Idoso , Antígenos Nucleares/análise , Líquidos Corporais/química , Feminino , Humanos , Ploidias , Sensibilidade e EspecificidadeRESUMO
Traumatic brain injury (TBI) produces several cellular changes, such as gliosis, axonal and dendritic plasticity, and inhibition-excitation imbalance, as well as cell death, which can initiate epileptogenesis. It has been demonstrated that dysfunction of the inhibitory components of the cerebral cortex after injury may cause status epilepticus in experimental models; we proposed to analyze the response of cortical interneurons and astrocytes after TBI in humans. Twelve contusion samples were evaluated, identifying the expression of glial fibrillary acidic protein (GFAP) and calcium-binding proteins (CaBPs). The study was made in sectors with and without preserved cytoarchitecture evaluated with NeuN immunoreactivity (IR). In sectors with total loss of NeuN-IR the results showed a remarkable loss of CaBP-IR both in neuropil and somata. In sectors with conserved cytoarchitecture less drastic changes in CaBP-IR were detected. These changes include a decrease in the amount of parvalbumin (PV-IR) neurons in layer II, an increase of calbindin (CB-IR) neurons in layers III and V, and an increase in calretinin (CR-IR) neurons in layer II. We also observed glial fibrillary acidic protein immunoreactivity (GFAP-IR) in the white matter, in the gray-white matter transition, and around the sectors with NeuN-IR total loss. These findings may reflect dynamic activity as a consequence of the lesion that is associated with changes in the excitatory circuits of neighboring hyperactivated glutamatergic neurons, possibly due to the primary impact, or secondary events such as hypoxia-ischemia. Temporal evolution of these changes may be the substrate linking severe cortical contusion and the resulting epileptogenic activity observed in some patients.
Assuntos
Lesões Encefálicas/metabolismo , Lesões Encefálicas/patologia , Proteínas de Ligação ao Cálcio/metabolismo , Córtex Cerebral/metabolismo , Córtex Cerebral/patologia , Adolescente , Adulto , Idoso , Antígenos Nucleares/análise , Antígenos Nucleares/metabolismo , Biomarcadores/metabolismo , Lesões Encefálicas/fisiopatologia , Calbindina 2 , Calbindinas , Proteínas de Ligação ao Cálcio/análise , Córtex Cerebral/lesões , Progressão da Doença , Feminino , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/metabolismo , Gliose/patologia , Gliose/fisiopatologia , Ácido Glutâmico/metabolismo , Humanos , Hipóxia-Isquemia Encefálica/metabolismo , Hipóxia-Isquemia Encefálica/patologia , Hipóxia-Isquemia Encefálica/fisiopatologia , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Fibras Nervosas Mielinizadas/metabolismo , Fibras Nervosas Mielinizadas/patologia , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Neurônios/patologia , Parvalbuminas/análise , Parvalbuminas/metabolismo , Proteína G de Ligação ao Cálcio S100/análise , Proteína G de Ligação ao Cálcio S100/metabolismo , Adulto JovemRESUMO
This study reports the first description of the karyotype of Agonostomus monticola, a species belonging to a genus which is considered to be the most primitive among living mugilid fish. Specimens from Panama and Venezuela were cytogenetically analysed by conventional chromosome banding (Ag and base-specific-fluorochrome staining, C-banding) and by fluorescent in situ hybridization (FISH). Agonostomus monticola showed a chromosome complement of 2n = 48, composed of 23 acrocentric and one subtelocentric chromosome pairs and a pericentromeric distribution of the C-positive heterochromatin in all chromosomes. Major ribosomal genes were found to be located on the short arms of the subtelocentric chromosome pair number 24 and minor ribosomal genes in a paracentromeric position of a single medium-sized chromosome pair. All these observed cytogenetic features are similar to those previously described in four representatives of two genera, Liza and Chelon, which are considered to be among the most advanced in the family. Thus, this karyotypic form might represent the plesiomorphic condition for the mullets. This hypothesis regarding the plesiomorphic condition, if confirmed, would shed new light on the previously inferred cytotaxonomic relationships for the studied species of Mugilidae, because the karyotype with 48 acrocentric chromosomes, which has been so far regarded as primitive for the family, would have to be considered as derived.
Assuntos
RNA Ribossômico 18S/análise , RNA Ribossômico 5S/análise , Smegmamorpha/classificação , Smegmamorpha/genética , Animais , Antígenos Nucleares/análise , Bandeamento Cromossômico , Cromossomos/genética , Cromossomos/ultraestrutura , DNA Ribossômico/análise , Evolução Molecular , Heterocromatina/genética , Hibridização in Situ Fluorescente , Cariotipagem , Panamá , RNA Ribossômico 18S/genética , RNA Ribossômico 5S/genética , Especificidade da Espécie , VenezuelaRESUMO
OBJECTIVE: To determine the cell proliferation rate and possible effects of cigarette smoking on the oral mucosa lining through analysis of silver-stained nucleolar organizer regions (AgNORs) in exfoliative cytology specimens. STUDY DESIGN: Exfoliative cytology was performed on the left side of the border of the tongue and of the floor of the mouth in 25 smoking patients and 25 nonsmoking patients. The inclusion criterion for smokers was the consumption of more than 20 cigarettes per day for a minimum of 30 years. RESULTS: The slides were stained by histochemical AgNOR method. In the nonsmoking group the mean number of AgNORs per nucleus was 2.732 +/- 0.236 in the tongue border and 2.918 +/- 0.195 in the floor of the mouth. In smoking patients the mean number of AgNORs per nucleus was 3.372 +/- 0.375 in the tongue border and 3.245 +/- 0.237 in the floor of the mouth. CONCLUSION: The results suggest higher cell proliferation quantified by the histochemical AgNOR technique in exfoliative cytology specimens obtained from the oral mucosa lining of smokers presenting no clinical alterations.
Assuntos
Antígenos Nucleares/análise , Citodiagnóstico/métodos , Mucosa Bucal/química , Mucosa Bucal/citologia , Região Organizadora do Nucléolo/química , Coloração pela Prata/métodos , Fumar , Adulto , Idoso , Proliferação de Células , Humanos , Pessoa de Meia-Idade , Região Organizadora do Nucléolo/ultraestrutura , Fumar/epidemiologiaRESUMO
The argyrophylic staining of the nucleolar organizer regions (AgNOR positive response) in interphase nuclei is often related directly to the cellular demand for ribosome biogenesis and is considered of relevance in studies of tumor pathology. Transformation of human breast epithelial MCF-10A cells by the c-Ha-ras oncogene results in altered growth, invasiveness and tumorigenicity in nude mice. Since ras transformation may be associated with a more intense nucleolar activity, we examined the influence of transfection by the Ha-ras oncogene on AgNOR staining response in MCF-10A cells. Following assessment of the AgNOR response with video image analysis, the AgNOR-positive areas and the AgNOR area/nuclear area ratio, but not the number of AgNOR aggregates or dots per nucleus, were found to be much higher after ras transformation. A role of the Ha-ras transformation on the nucleolar activity of the MCF-10A is thus suggested as assessed by the AgNOR staining. Based on data in the literature, it is also hypothesized that a decreased wild-type p53 level, possibly promoted by the ras transformation, may be associated with the increased AgNOR response.
Assuntos
Antígenos Nucleares/análise , Transformação Celular Neoplásica , Células Epiteliais/metabolismo , Genes ras/genética , Processamento de Imagem Assistida por Computador/métodos , Animais , Antígenos Nucleares/metabolismo , Mama/patologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Transformada , Nucléolo Celular/metabolismo , Células Epiteliais/patologia , Humanos , Camundongos , Microespectrofotometria , Modelos Biológicos , TransfecçãoRESUMO
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.
Assuntos
Biomarcadores Tumorais , Neoplasias Maxilares/patologia , Região Organizadora do Nucléolo/patologia , Osteossarcoma/patologia , Análise de Variância , Antígenos Nucleares/análise , Carcinoma de Células Escamosas/patologia , Humanos , Invasividade Neoplásica , Proteínas Nucleares/análise , Coloração pela PrataRESUMO
OBJECTIVE: The aim of this study was to evaluate the proliferation activity by means of the quantification of the argyrophilic nucleolar organizer regions (AgNORs) and the patterns of expression of the epidermal growth factor receptor (EGFR) in ameloblastomas. METHOD: The methods of evaluation included the H/E stain for the morphologic analysis, the silver impregnation technique for quantification of the AgNORs and the immunohistochemical stain with anti-EGFR antibody in 11 cases of ameloblastoma. RESULTS: The results did not show a significant statistical difference as per quantification of the AgNORs. The expression of the EGFR on the epithelial islands of ameloblastoma was not uniform, and the location of the expression was also variable. The predominant expression was that of cytoplasm and the islands with an expression of membrane only were rare and generally smaller in size. CONCLUSION: The tumor presents an irregular growth. Smaller islands are associated with a higher proliferation activity and therefore could be responsible for tumor infiltration.
Assuntos
Ameloblastoma/química , Antígenos Nucleares/análise , Receptores ErbB/análise , Neoplasias Maxilomandibulares/química , Proteínas de Neoplasias/análise , Proteínas Nucleares/análise , Humanos , Coloração e RotulagemRESUMO
This study used Ehrlich solid tumor as an experimental model for breast cancer to investigate the effects of thyroid hormones and castration on tumor development in adult female mice. Artificial hyperthyroidism was induced in animals, and after a 30-day-treatment, they received subcutaneous injection of neoplastic cells between left plantar cushions. We measured the growth of tumor inoculated in the paws for 10 days at necropsy. Hyperthyroidism induction led to significantly increased tumor size in non-castrated animals, and alterations were less intense in association with artificial hyperthyroidism and castration (p<0.05). Histomorphologic and histomorphometric analyses and neoplastic cell characterization were carried out by measuring nuclear diameter, by evaluating AgNORs, by mitotic count, and by measuring cell proliferation using immunohistochemical marker CDC47. At the end of the experiment, we noted metabolism and a decrease in cell proliferation in groups having received l-thyroxine, which were more evident in the non-castrated group (p<0.001).
Assuntos
Neoplasias da Mama/patologia , Carcinoma de Ehrlich/patologia , Hipertireoidismo/patologia , Ovariectomia , Animais , Antígenos Nucleares/análise , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/complicações , Neoplasias da Mama/metabolismo , Carcinoma de Ehrlich/complicações , Carcinoma de Ehrlich/metabolismo , Núcleo Celular/química , Núcleo Celular/patologia , Proliferação de Células , Modelos Animais de Doenças , Feminino , Hipertireoidismo/induzido quimicamente , Hipertireoidismo/complicações , Hipertireoidismo/metabolismo , Camundongos , Índice Mitótico , Proteínas Nucleares/análise , Proteínas Nucleares/metabolismo , Tiroxina/farmacologiaRESUMO
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.
Assuntos
Humanos , Biomarcadores Tumorais , Neoplasias Maxilares/patologia , Osteossarcoma , Região Organizadora do Nucléolo/patologia , Análise de Variância , Antígenos Nucleares/análise , Carcinoma de Células Escamosas/patologia , Invasividade Neoplásica , Proteínas Nucleares/análise , Coloração pela PrataRESUMO
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.(AU)
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.(AU)
Assuntos
Humanos , Neoplasias Maxilares/patologia , Região Organizadora do Nucléolo/patologia , Osteossarcoma/patologia , Biomarcadores Tumorais , Invasividade Neoplásica , Análise de Variância , Antígenos Nucleares/análise , Carcinoma de Células Escamosas/patologia , Proteínas Nucleares/análise , Coloração pela PrataRESUMO
Nucleolar Organizer Region (AgNOR) counts have been reported as a useful, complementary diagnostic tool for several neoplasias. The morphometric evaluation of AgNOR has also been shown to contribute to the detection of incipient cellular alterations. The aim of the present study was to perform a morphometric analysis of AgNOR in keratocysts (KC), and in three rare cases of malignant transformation of keratocysts (KCm) to explore their use as a marker of the process of malignant transformation in this entity. We performed a study of the 3 cases of keratocysts that had suffered transformation into squamous cell carcinoma (SCC) that were available in our files and of 8 cases of solitary keratocysts (KCs). Paraffin sections were silver stained and employed to perform a morphometric analysis of 7 parameters related to number, size and shape of AgNORs. A comparative analysis of the epithelium of solitary keractocyts (KCs), the cystic epithelium adjacent to the area of transformation (KCm) and the epithelium of the SCC cords was performed. The epithelia of KCs and KCm exhibited statistically significant differences for four of the parameters analyzed: nuclear volume (VNUC), single AgNOR volume (VNOR), Total AgNOR per nucleus (TVNOR) and Contour Index of nuclei (CINUC). KCs and SCC exhibited statistically significant differences for all of the parameters. Given the ease with which this technique can be applied to routine histopathological material, we propose AgNOR evaluation as a diagnostic and prognostic aid in cystic entities that are suspected to have a neoplastic potential.
Assuntos
Antígenos de Neoplasias/análise , Antígenos Nucleares/análise , Biomarcadores Tumorais/análise , Carcinoma de Células Escamosas/patologia , Transformação Celular Neoplásica/patologia , Neoplasias Bucais/patologia , Proteínas Nucleares/análise , Região Organizadora do Nucléolo/ultraestrutura , Cistos Odontogênicos/patologia , Núcleo Celular/ultraestrutura , Células Epiteliais/patologia , Epitélio/patologia , Humanos , Coloração pela PrataRESUMO
OBJECTIVE: To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of thyroid cancer and invasion. STUDY DESIGN: Eighty-one consecutive cases of thyroid carcinoma were selected for DNA and AgNOR analysis. The diagnoses were: papillary carcinoma (n = 40), follicular carcinoma (n = 31), Hürthle cell adenocarcinoma (n = 4), and undifferentiated carcinoma (n = 6). Seven normal thyroids were used as controls. DNA quantitative measurement was performed with Vidas 2.0 software (Kontron Bildanalyse, Munich, Germany) connected to an MPM 210 photometer microscope (Carl Zeiss, Oberkochen, Germany). The DNA index was obtained using histograms. Counting the NORs was performed by subjectively counting the NORs in 200 malignant cells. RESULTS: DNA ploidy analysis showed all Hürthle cell adenocarcinomas, 21 (67%)follicular tumors, 23 (57%) papillary tumors and 4 (67%) undifferentiated carcinomas to be aneuploid. DNA analysis correlated with histologic type of the tumor (p = 0.032). There was no statistical significance to the AgNOR counting variables studied. Statistical analysis showed correlation between ploidy and histologic diagnosis, but not AgNOR counting, to have prognostic value. CONCLUSION: DNA ploidy is more useful than subjective counting of NORs as an adjunct method for thyroid lesion analysis.