RESUMEN
The Buccal Micronucleus Cytome Assay (BMCyt) has become an important biomonitoring tool for assessing cytogenetic damage in many studied populations. Each laboratory applies protocols that vary according to the method of collecting and preparing samples. Besides, Brazil is a country of great territorial extensions that received immigrants from various parts of the world with different genetic backgrounds. Therefore, the present study aimed to evaluate the inter-laboratory variation in scoring the same set of slides using the more comprehensive scoring criteria, to standardize the BMCyt protocol, to observe the basal alterations in populations of different Brazilian regions and to compare it with other places around the world. Our results showed that a valuable number of laboratories participated, ten laboratories from different regions of the country, for the validation of the BMCyt in human biomonitoring studies, resulting in the 804 healthy individuals. This was possible because we observed: a range of measures needs to be considered, such as the baseline frequency of DNA damage and cell death in non-exposed individuals; age when grouped showed an influence on DNA damage, although when evaluated by group we did not see an influence; association between smoking habit and all endpoints of the BMCyt (except karyolytic cells) was evident; the basal MN frequency, in the majority of groups, follows those around the world; and the BMCyt was confirmed as a good health status biomarker. We emphasize the need for constant discussions on the parameters of cell death due to greater difficulty among the analyzers.
Asunto(s)
Bioensayo/normas , Núcleo Celular/genética , Células Epiteliales/ultraestructura , Laboratorios/normas , Micronúcleos con Defecto Cromosómico , Pruebas de Micronúcleos/normas , Mucosa Bucal/citología , Adolescente , Adulto , Bioensayo/métodos , Brasil , Muerte Celular/genética , Núcleo Celular/ultraestructura , Daño del ADN , Femenino , Humanos , Masculino , Micronúcleos con Defecto Cromosómico/estadística & datos numéricos , Pruebas de Micronúcleos/métodos , Persona de Mediana Edad , Mucosa Bucal/ultraestructura , Adulto JovenRESUMEN
Soybean farmers are exposed to various types of pesticides that contain in their formulations a combination of chemicals with genotoxic and mutagenic potential. Therefore, the objective of this paper was to evaluate the genetic damages caused by this pesticide exposure to soybean producers in the state of Mato Grosso (Brazil), regarding biochemical, genetic polymorphic and in silico analyses. A total of 148 individuals were evaluated, 76 of which were occupationally exposed and 72 were not exposed at all. The buccal micronucleus cytome assay (BMCyt) detected in the exposed group an increase on DNA damage and cell death. No inhibition of butyrylcholinesterase (BchE) was observed within the exposed group. The detection of inorganic elements was made through the particle-induced X-ray emission technique (PIXE), which revealed higher concentrations of Bromine (Br), Rubidium (Rb) and Lead (Pb) in rural workers. A molecular model using in silico analysis suggests how metal ions can cause both DNA damage and apoptosis in the exposed cells. Analysis of the compared effect of X-ray Repair Cross-complement Protein 1 (XRCC1) and Paraoxonase 1 (PON1) genotypes in the groups demonstrated an increase of binucleated cells (exposed group) and nuclear bud (non-exposed group) in individuals with the XRCC1 Trip/- and PON1 Arg/- genes. There was no significant difference in the telomere (TL) mean value in the exposed group in contrast to the non-exposed group. Our results showed that soybean producers showed genotoxic effect and cell death, which may have been induced by exposure to complex mixtures of agrochemicals and fertilizers. In addition, XRCC1 Arg/Arg could, in some respects, provide protection to individuals.
Asunto(s)
Mezclas Complejas/toxicidad , Daño del ADN , Fertilizantes/toxicidad , Micronúcleos con Defecto Cromosómico/inducido químicamente , Exposición Profesional/efectos adversos , Plaguicidas/toxicidad , Polimorfismo Genético , Adulto , Apoptosis/efectos de los fármacos , Arildialquilfosfatasa/efectos de los fármacos , Brasil , Simulación por Computador , Células Epiteliales/efectos de los fármacos , Agricultores , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Mucosa Bucal/citología , Exposición Profesional/análisis , Proteína 1 de Reparación por Escisión del Grupo de Complementación Cruzada de las Lesiones por Rayos X/genéticaRESUMEN
BACKGROUND: Gap junctions between ß-cells participate in the precise regulation of insulin secretion. Adherens junctions and their associated proteins are required for the formation, function and structural maintenance of gap junctions. Increases in the number of the gap junctions between ß-cells and enhanced glucose-stimulated insulin secretion are observed during pregnancy. In contrast, protein restriction produces structural and functional alterations that result in poor insulin secretion in response to glucose. We investigated whether protein restriction during pregnancy affects the expression of mRNA and proteins involved in gap and adherens junctions in pancreatic islets. An isoenergetic low-protein diet (6% protein) was fed to non-pregnant or pregnant rats from day 1-15 of pregnancy, and rats fed an isocaloric normal-protein diet (17% protein) were used as controls. RESULTS: The low-protein diet reduced the levels of connexin 36 and ß-catenin protein in pancreatic islets. In rats fed the control diet, pregnancy increased the levels of phospho-[Ser(279/282)]-connexin 43, and it decreased the levels of connexin 36, ß-catenin and beta-actin mRNA as well as the levels of connexin 36 and ß-catenin protein in islets. The low-protein diet during pregnancy did not alter these mRNA and protein levels, but avoided the increase of levels of phospho-[Ser(279/282)]-connexin 43 in islets. Insulin secretion in response to 8.3 mmol/L glucose was higher in pregnant rats than in non-pregnant rats, independently of the nutritional status. CONCLUSION: Short-term protein restriction during pregnancy prevented the Cx43 phosphorylation, but this event did not interfer in the insulin secretion.
Asunto(s)
Comunicación Celular/fisiología , Diabetes Gestacional/dietoterapia , Dieta con Restricción de Proteínas , Uniones Intercelulares/metabolismo , Islotes Pancreáticos/metabolismo , ARN Mensajero/metabolismo , Actinas/metabolismo , Uniones Adherentes/metabolismo , Análisis de Varianza , Animales , Glucemia/análisis , Conexina 43/metabolismo , Conexinas/metabolismo , Diabetes Gestacional/prevención & control , Femenino , Uniones Comunicantes/metabolismo , Glucosa/administración & dosificación , Insulina/metabolismo , Secreción de Insulina , Embarazo , Ratas Wistar , Reacción en Cadena en Tiempo Real de la Polimerasa , beta Catenina/metabolismo , Proteína delta-6 de Union ComunicanteRESUMEN
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Asunto(s)
Animales , Femenino , Embarazo , Comunicación Celular/fisiología , Dieta con Restricción de Proteínas , Diabetes Gestacional/dietoterapia , Uniones Intercelulares/metabolismo , Islotes Pancreáticos/metabolismo , ARN Mensajero/metabolismo , Análisis de Varianza , Actinas/metabolismo , Uniones Adherentes/metabolismo , Glucemia/análisis , /metabolismo , Conexinas/metabolismo , Diabetes Gestacional/prevención & control , Uniones Comunicantes/metabolismo , Glucosa/administración & dosificación , Insulina/metabolismo , Insulina , Ratas Wistar , Reacción en Cadena en Tiempo Real de la Polimerasa , beta Catenina/metabolismoRESUMEN
In this study, we analysed the frequency of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) and evaluated mutagen-induced sensitivity in the lymphocytes of patients chronically infected with hepatitis B virus (HBV) or hepatitis C virus (HCV). In total, 49 patients with chronic viral hepatitis (28 HBV-infected and 21 HCV-infected patients) and 33 healthy, non-infected blood donor controls were investigated. The frequencies () of MN, NPBs and NBUDs in the controls were 4.41 ± 2.15, 1.15 ± 0.97 and 2.98 ± 1.31, respectively. The frequencies of MN and NPBs were significantly increased (p < 0.0001) in the patient group (7.01 ± 3.23 and 2.76 ± 2.08, respectively) compared with the control group. When considered separately, the HBV-infected patients (7.18 ± 3.57) and HCV-infected patients (3.27 ± 2.40) each had greater numbers of MN than did the controls (p < 0.0001). The HCV-infected patients displayed high numbers of NPBs (2.09 ± 1.33) and NBUDs (4.38 ± 3.28), but only the HBV-infected patients exhibited a significant difference (NPBs = 3.27 ± 2.40, p < 0.0001 and NBUDs = 4.71 ± 2.79, p = 0.03) in comparison with the controls. Similar results were obtained for males, but not for females, when all patients or the HBV-infected group was compared with the controls. The lymphocytes of the infected patients did not exhibit sensitivity to mutagen in comparison with the lymphocytes of the controls (p = 0.06). These results showed that the lymphocytes of patients who were chronically infected with HBV or HCV presented greater chromosomal instability.
Asunto(s)
Núcleo Celular/virología , Hepatitis B Crónica/virología , Hepatitis C Crónica/virología , Linfocitos/virología , Micronúcleos con Defecto Cromosómico/estadística & datos numéricos , Adulto , Factores de Edad , Análisis de Varianza , Núcleo Celular/ultraestructura , Distribución de Chi-Cuadrado , Inestabilidad Cromosómica , Daño del ADN , Femenino , Humanos , Linfocitos/ultraestructura , Masculino , Pruebas de Micronúcleos , Persona de Mediana Edad , Factores Sexuales , Adulto JovenRESUMEN
In this study, we analysed the frequency of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) and evaluated mutagen-induced sensitivity in the lymphocytes of patients chronically infected with hepatitis B virus (HBV) or hepatitis C virus (HCV). In total, 49 patients with chronic viral hepatitis (28 HBV-infected and 21 HCV-infected patients) and 33 healthy, non-infected blood donor controls were investigated. The frequencies (‰) of MN, NPBs and NBUDs in the controls were 4.41 ± 2.15, 1.15 ± 0.97 and 2.98 ± 1.31, respectively. The frequencies of MN and NPBs were significantly increased (p < 0.0001) in the patient group (7.01 ± 3.23 and 2.76 ± 2.08, respectively) compared with the control group. When considered separately, the HBV-infected patients (7.18 ± 3.57) and HCV-infected patients (3.27 ± 2.40) each had greater numbers of MN than did the controls (p < 0.0001). The HCV-infected patients displayed high numbers of NPBs (2.09 ± 1.33) and NBUDs (4.38 ± 3.28), but only the HBV-infected patients exhibited a significant difference (NPBs = 3.27 ± 2.40, p < 0.0001 and NBUDs = 4.71 ± 2.79, p = 0.03) in comparison with the controls. Similar results were obtained for males, but not for females, when all patients or the HBV-infected group was compared with the controls. The lymphocytes of the infected patients did not exhibit sensitivity to mutagen in comparison with the lymphocytes of the controls (p = 0.06). These results showed that the lymphocytes of patients who were chronically infected with HBV or HCV presented greater chromosomal instability.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Núcleo Celular/virología , Hepatitis B Crónica/virología , Hepatitis C Crónica/virología , Linfocitos/virología , Micronúcleos con Defecto Cromosómico/estadística & datos numéricos , Factores de Edad , Análisis de Varianza , Distribución de Chi-Cuadrado , Inestabilidad Cromosómica , Núcleo Celular/ultraestructura , Daño del ADN , Linfocitos/ultraestructura , Pruebas de Micronúcleos , Factores SexualesRESUMEN
INTRODUCTION: The progression of hepatic disease in chronic viral hepatitis is accompanied by an increased production of reactive oxygen species (ROS), as well as an accumulation of oxidative DNA damage, which is primarily repaired through base excision repair. XRCC1 (X-ray repair cross complementing protein 1) is one of the most important proteins involved in this repair pathway. The present study was carried out to verify the possible association of the XRCC1 rs25487 polymorphism with cirrhosis in patients from Central-West Brazil. MATERIAL AND METHODS: A total of 227 individuals with viral hepatitis, 53 cirrhotic and 174 non-cirrhotic, were genotyped for the XRCC1 rs25487 polymorphism using PCR-RFLP. RESULTS: There were significantly higher frequencies of both the Arg/Gln genotype and of individuals with at least one Gln allele (Arg/ Gln+Gln/Gln) among cirrhotic patients (56.6% and 69.8%) compared with non-cirrhotic patients (25.8% and 37.9%). Both conditions were significantly associated with cirrhosis, independent of age, sex, alcohol intake or tobacco use (adjusted OR = 3.5, CI = 1.7-7.4, p = 0.001 and adjusted OR = 3.1, CI = 1.5-6.3, p = 0.002, respectively). Similar results were obtained for a group of HCV-infected patients but not for HBV-infected patients. CONCLUSIONS: The XRCC1 rs25487 polymorphism may influence the development of cirrhosis in viral hepatitis patients, and additional investigation will be necessary.