RESUMO
INTRODUCTION: Oral cytopathology is able to detect incipient cellular alterations, but it is not routinely applied to this purpose. We aimed to establish a model to screen individuals with no oral lesion exposed to smoking/alcohol, by means of the nuclear area, cell proliferation rate, and analysis of genetic damage. METHODS: In this cross-sectional pilot study, 90 patients were allocated into 3 groups: oral cancer group (patients with oral squamous cell carcinoma), tobacco/alcohol group (patients without oral lesions and exposed to these risk factors), and control group (individuals with no lesion and not exposed to tobacco and alcohol). The cytological smears performed in these individuals were stained with Papanicolaou, a silver-staining and a Feulgen reaction. The nuclei of cells were measured, and AgNORs/nucleus and micronuclei (MN) were quantified. The cutoff values were stipulated evaluating the healthy mucosa (control group) and the cancerization field mucosa (oral cancer group). RESULTS: Cutoff values for the screening of individuals exposed to carcinogens were ≥8% of nuclei larger than 100 µm2, ≥3.38 AgNOR/nucleus, and ≥3 MN per 1,000 cells. CONCLUSIONS: Nuclear area measurement and AgNORs/nucleus and MN quantification identified the incipient phase of oral carcinogenesis. A screening model for individuals without oral lesion exposed to smoking/alcohol was proposed.
Assuntos
Carcinoma de Células Escamosas , Neoplasias Bucais , Carcinógenos/farmacologia , Carcinoma de Células Escamosas/patologia , Estudos Transversais , Humanos , Mucosa Bucal/patologia , Neoplasias Bucais/induzido quimicamente , Neoplasias Bucais/diagnóstico , Neoplasias Bucais/patologia , Região Organizadora do Nucléolo/patologia , Projetos Piloto , Coloração pela PrataRESUMO
SUMMARY: Nucleolus Organizer Regions (NORs) are defined as nucleolar components containing argyrophilic proteins selectively stained by silver methods (AgNORs). Several investigations have shown the AgNOR quantity and area represent a valuable parameter of cell kinetics, since they reflect the level of activity and cellular proliferation. This article addresses an evaluation of the functional activity and relation between days of pregnancy and proliferative capacity of trophoblastic mononucleate and binucleate cells from bovine placentomes. Both the number and size of AgNORs were determined in different phases of gestation by silver nitrate staining in conventional histological slides. The results showed a significant increase (from 1 to 12 AgNORs) in the number of AgNORS per trophoblastic mononucleate cell in the 3rd trimester, with predominance of 4-6 AgNORs/cell. In the 1st and 2nd trimesters, the number ranged between 1 and 9 AgNORs/cell, with predominance of 1-3 AgNORs. No significant differences were observed between the 2nd and 3rd trimesters, but in the first, in binucleate cells (19-27 and 10-18 AgNORs/cell, respectively) - this number was higher than the one registered in trophoblastic mononucleate cells in the same period. Thus, AgNORs can be used as markers of the proliferative placental cell cycle and established a relation between number of AgNORs and days of gestation. This relation can be used for diagnoses and prognoses of several placental pathologies, including pregnancy losses from manipulated embryos.
RESUMEN: Las Regiones Organizadoras de Nucléolos (NOR) se definen como componentes nucleolares que contienen proteínas argirofílicas teñidas selectivamente por métodos de plata (AgNOR). Varias investigaciones han demostrado que la cantidad y el área de AgNOR representan un parámetro importante de la cinética celular, ya que reflejan el nivel de actividad y proliferación celular. Este trabajo analiza la actividad funcional y la relación entre los días de preñez y la capacidad proliferativa de las células trofoblásticas mononucleadas y binucleadas de placentomas bovinos. Tanto el número como el tamaño de los AgNOR se determinaron en diferentes fases de la gestación mediante tinción con nitrato de plata en portaobjetos histológicos convencionales. Los resultados mostraron un aumento significativo (de 1 a 12 AgNOR) en el número de AgNORS por célula mononucleada trofoblástica en el tercer trimestre, con predominio de 4-6 AgNOR / célula. En el primer y segundo trimestre, el número osciló entre 1 y 9 AgNOR / célula, con predominio de 1-3 AgNOR. No se observaron diferencias significativas entre el 2do y 3er trimester; en el primer trimestre, en células binucleadas (19-27 y 10-18 AgNORs / célula, respectivamente) - este número fue superior a la cantidad registrada en células mononucleadas trofoblásticas en el mismo período. Por tanto, los AgNOR se pueden utilizar como marcadores del ciclo celular placentario proliferativo y se establece una relación entre el número de AgNOR y los días de gestación. Esta relación puede ser útil en el diagnóstico y pronóstico de varias patologías placentarias, incluidas las pérdidas de preñeces de embriones manipulados.
Assuntos
Animais , Feminino , Gravidez , Bovinos , Placenta/metabolismo , Proliferação de Células , Região Organizadora do Nucléolo/metabolismoRESUMO
OBJECTIVES: The purpose of the study is to investigate whether there is any relationship between mean argyrophilic nucleolar organizing regions (AgNOR) number and total AgNOR area/total nuclear area (TAA/TNA) ratio and the levels of brain hypoxia after exposure to different acute doses of carbon monoxide (CO) gas. METHODS: Each experimental group was exposed to CO gas (concentrations of 1,000, 3,000 and 5,000 ppm). Then, the rats were anesthetized, and blood samples were taken from the right jugular vein for carboxyhemoglobin levels detection. The rats were sacrificed on seventh day. AgNOR staining was applied to brain tissues. TAA/TNA and mean AgNOR number were detected for each nucleus. RESULTS: Significant differences were detected among all groups for TAA/TNA ratio, mean AgNOR number and carboxyhemoglobin level. According to a double comparison of groups, the differences between control and 1,000 ppm, control and 3,000 ppm, control and 5,000 ppm, and between 1,000 and 5,000 ppm were significant for TAA/TNA ratio. When mean AgNOR number was considered, significant differences were detected between control and 1,000 ppm, control and 3,000 ppm, control and 5,000 ppm, and between 1,000 and 3,000 ppm. CONCLUSION: AgNOR proteins may be used for early detection of the duration, intensity, and damage of brain injury caused by CO poisoning. Thus, effective treatment strategies can be developed for the prevention of hypoxic conditions.
OBJETIVOS: El objetivo del estudio es investigar si existe alguna relación entre el número medio de regiones organizadoras nucleolares argirófilas (AgNOR) y la proporción de área total de AgNOR/área nuclear total (TAA/TNA) y los niveles de hipoxia cerebral en la exposición a diferentes dosis agudas de gas monóxido de carbono (CO). MÉTODOS: Cada grupo experimental fue expuesto a gas CO (concentraciones de 1,000, 3,000 y 5,000 ppm). Luego las ratas fueron anestesiadas, se tomaron muestras de sangre de la vena yugular derecha para la detección de los niveles de carboxihemoglobina. Las ratas se sacrificaron el séptimo día. Se aplicó tinción con AgNOR en los tejidos cerebrales. Se detectaron el TAA/TNA y el número medio de AgNOR para cada núcleo. RESULTADOS: Se detectaron diferencias significativas entre todos los grupos para la relación TAA/TNA, el número medio de AgNOR y el nivel de carboxihemoglobina. Según la doble comparación de grupos, las diferencias entre control y 1,000 ppm, control y 3,000 ppm, control y 5,000 ppm y 1,000 y 5,000 ppm fueron significativas para la relación TAA/TNA. Cuando se consideró el número de AgNOR medio, se detectaron diferencias significativas entre control y 1,000ppm, control y 3,000ppm, control y 5,000 ppm y 1,000 y 3,000 ppm. CONCLUSIÓN: Las proteínas AgNOR pueden usarse para la detección temprana de la duración, intensidad y daño de la lesión cerebral causada por la intoxicación por CO. Por lo tanto, se pueden desarrollar estrategias de tratamiento efectivas para la prevención de condiciones hipóxicas.
Assuntos
Intoxicação por Monóxido de Carbono , Hipóxia Encefálica , Animais , Antígenos Nucleares , Biomarcadores , Intoxicação por Monóxido de Carbono/diagnóstico , Hipóxia Encefálica/diagnóstico , Região Organizadora do Nucléolo , RatosRESUMO
AIM: This study relates the average number of nucleolar organizing regions (NORs) obtained in a series of cases of oral leukoplakia, with three methods of histopathological classification of such lesion. MATERIALS AND METHODS: This is a histopathological-histochemical laboratory cross-sectional study. The 18 cases of leukoplakia analyzed were filed at Pathology Service of the Biological Sciences Institute of the University of Passo Fundo, Rio Grande do Sul, Brazil (SDH/ICB/UPF) (2017 and 2018), from which epidemiological data were extracted. New histological sections were performed for impregnation by the argyrophilic nucleolar organizer regions (AgNOR) technique. The histopathological slides were analyzed by photon microscopy (1,000×), and the nuclei of 100 epithelial cells were photographed to count the number of NORs. Three methods were used for the lesions' histopathological classification [World Health Organization (WHO), Brothwell, and binary system]. The means of NORs were compared with the three histopathological classifications by means of the t or analysis of variance (ANOVA) statistical tests, at a significance level of 5%. RESULTS: According to the WHO classification method, most cases (11-61.1%) had a moderate classification. Evaluations by the Brothwell method showed moderate and mild classification in 50 and 38.9% of cases, respectively. According to the binary system, most cases (10-55.6%) had low risk. The average NORs found in 100 nuclei of each of the 18 lesions ranged from 2 to 4. When crossing the average NORs with the histopathological classification methods of the lesions by means of the t test or ANOVA, no significant relationship was noted. CONCLUSION: The average of NORs is not associated with the histological classifications of leukoplakias. Thus, the AgNOR method should be used with caution when differentiating different histological grades of leukoplakias. CLINICAL SIGNIFICANCE: The AgNOR method should be used with caution to determine the clinical treatment of oral leukoplakias, since no agreement was observed between this method and the histopathological classifications available for such lesion.
Assuntos
Leucoplasia Oral , Região Organizadora do Nucléolo , Análise de Variância , Brasil , Estudos Transversais , Humanos , Leucoplasia Oral/genéticaRESUMO
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.
Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/patologia , Região Organizadora do Nucléolo/patologia , Prognóstico , Coloração pela Prata , Estatísticas não ParamétricasRESUMO
OBJECTIVE: Evaluate the relationships between AgNORs polymorphisms and squamous intraepithelial lesions (SIL) and squamous cell carcinoma (SCC) with HPV infection. MATERIALS AND METHODS: A study was carried out on sixty women from the state of Guerrero, Mexico. HPV detection was performed by PCR. AgNORs were identified by argentic impregnation. One hundred cells per slide were counted and classified according to the polymorphism of AgNORs dots; typical (spherical) and atypical (large, kidney-shaped and clustered). RESULTS: A total of 100 percent of the cases were positive for HPV infection. Nine different high-risk HPV genotypes were found, type16 was the most common (48.6 percent). The AgNORs showed a significant decrease in spherical shape according to neoplastic development. The three atypical shapes showed a significant increase in SIL and SCC (p-trend<0.001). CONCLUSIONS: AgNORs polymorphism rises progressively according to the grade of histological lesions that can be useful as a prognosis for progression of SCC.
OBJETIVO: Evaluar la relación entre los polimorfismos de AgNORs con las lesiones intraepiteliales escamosas (LIE) y carcinoma de células escamosas (CCE). MATERIAL Y MÉTODOS: Se estudiaron sesenta mujeres del estado de Guerrero, México. La detección del VPH fue por PCR y los AgNORs por impregnación argéntica; se contaron 100 células y se clasificaron por tipo de polimorfismo de AgNORs: típico (esférico) y atípicos (largo, forma de riñón o de racimo). RESULTADOS: El 100 por ciento de los casos presentaron infección por VPH, se encontraron nueve genotipos diferentes de VPH de alto riesgo, el 16 fue el más común (48.6 por ciento). La forma esférica de los polimorfismos de AgNORs mostró una disminución con el desarrollo neoplásico y las atípicas incrementaron progresivamente con SIL y SCC (p-tendencia<0.001). CONCLUSIONES: Los polimorfismos AgNORs se incrementan progresivamente con el grado de lesión histológica, y pueden ser útiles en el pronóstico de progresión del carcinoma cervical.
Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Carcinoma de Células Escamosas/ultraestrutura , Região Organizadora do Nucléolo/ultraestrutura , Infecções por Papillomavirus/patologia , Neoplasias do Colo do Útero/ultraestrutura , Cervicite Uterina/patologia , Alphapapillomavirus/classificação , Alphapapillomavirus/genética , Alphapapillomavirus/isolamento & purificação , Carcinoma de Células Escamosas/virologia , Displasia do Colo do Útero/ultraestrutura , Displasia do Colo do Útero/virologia , Sondas de DNA de HPV , Progressão da Doença , Genótipo , Coloração pela Prata , Neoplasias do Colo do Útero/virologia , Cervicite Uterina/virologia , Adulto JovemRESUMO
Growth is one of the most important aspects in the genetic improvement of cultured fish species. Consequently, genetic parameters related to this feature and their response to selection have been the focus of most research in this area. Such research indicates that, in general, there is enough additive genetic variance related to growth, justifying the use of selection. Based on the usefulness of cytogenetic and molecular markers in the fish culture, the aim of the present work was to analyze the possible relationships among cytogenetic characteristics, specifically the NOR phenotypes, and the increase in length and weight in specimens of the rainbow trout (Oncorhynchus mykiss), resultant from directed mating between homozygous females and heterozygous males according to their NOR phenotypic patterns. The equations of the relationship between length and weight of the analyzed specimens followed the model Wt = a Lt b, showing b values higher than 3, determinant of a positive allometric growth. The results showed that the different NOR phenotypes were not related with the growth values for length and weight in any statistical test.
O crescimento é um dos mais importantes aspectos considerados no melhoramento genético de espécies de peixes cultivadas. Conseqüentemente, a ênfase das pesquisas na área tem sido avaliar os parâmetros genéticos relacionados com esta característica e sua resposta à seleção. Essas pesquisas indicam, em geral, haver variância genética aditiva suficiente para justificar o uso da seleção. Considerando que a utilização de marcadores citogenéticos ou moleculares pode ser de grande valia para a piscicultura, o presente trabalho teve como objetivo analisar a possível relação entre as características citogenéticas, de modo específico os padrões fenotípicos das NORs e o crescimento em comprimento e em peso dos exemplares de trutas arco-íris (Oncorhynchus mykiss) resultantes de acasalamentos dirigidos entre fêmeas homozigotas e machos heterozigotos, conforme caracterização do padrão fenotípico da NOR. As equações da relação comprimento e peso dos indivíduos analisados seguiram o modelo onde Wt = a Lt b, mostrando valores de b maiores que 3, determinante de um crescimento do tipo alométrico positivo. Os resultados mostraram que os padrões fenotípicos da NOR dos indivíduos homozigotos e dos heterozigotos não apresentaram diferenças estatisticamente significativas entre seus valores de crescimento em comprimento ou peso.
Assuntos
Animais , Masculino , Feminino , Aquicultura , Região Organizadora do Nucléolo/genética , Oncorhynchus mykiss/genética , Fenótipo , Seleção Genética , Aumento de Peso/genética , Análise Citogenética , Heterozigoto , Homozigoto , Modelos Genéticos , Oncorhynchus mykiss/crescimento & desenvolvimentoRESUMO
Growth is one of the most important aspects in the genetic improvement of cultured fish species. Consequently, genetic parameters related to this feature and their response to selection have been the focus of most research in this area. Such research indicates that, in general, there is enough additive genetic variance related to growth, justifying the use of selection. Based on the usefulness of cytogenetic and molecular markers in the fish culture, the aim of the present work was to analyze the possible relationships among cytogenetic characteristics, specifically the NOR phenotypes, and the increase in length and weight in specimens of the rainbow trout (Oncorhynchus mykiss), resultant from directed mating between homozygous females and heterozygous males according to their NOR phenotypic patterns. The equations of the relationship between length and weight of the analyzed specimens followed the model Wt = a Lt b, showing b values higher than 3, determinant of a positive allometric growth. The results showed that the different NOR phenotypes were not related with the growth values for length and weight in any statistical test.
O crescimento é um dos mais importantes aspectos considerados no melhoramento genético de espécies de peixes cultivadas. Conseqüentemente, a ênfase das pesquisas na área tem sido avaliar os parâmetros genéticos relacionados com esta característica e sua resposta à seleção. Essas pesquisas indicam, em geral, haver variância genética aditiva suficiente para justificar o uso da seleção. Considerando que a utilização de marcadores citogenéticos ou moleculares pode ser de grande valia para a piscicultura, o presente trabalho teve como objetivo analisar a possível relação entre as características citogenéticas, de modo específico os padrões fenotípicos das NORs e o crescimento em comprimento e em peso dos exemplares de trutas arco-íris (Oncorhynchus mykiss) resultantes de acasalamentos dirigidos entre fêmeas homozigotas e machos heterozigotos, conforme caracterização do padrão fenotípico da NOR. As equações da relação comprimento e peso dos indivíduos analisados seguiram o modelo onde Wt = a Lt b, mostrando valores de b maiores que 3, determinante de um crescimento do tipo alométrico positivo. Os resultados mostraram que os padrões fenotípicos da NOR dos indivíduos homozigotos e dos heterozigotos não apresentaram diferenças estatisticamente significativas entre seus valores de crescimento em comprimento ou peso.