and used in both pink mutation assay and comet assay using cellular nuclei from Tradescantia staminal hairs. The observation of staminal hair was realized along eight days (6-14) after treatment), flowers produced day 14 after treatment were utilized done according to Underbrink. In previous reports on plants, were comet assay was used, breaking cellular wall and separating by centrifugation gradient are necessary. Here, nuclei from staminal hairs were obtained by squashing the cells (is not necessary to utilize to break special procedure cellular wall), collected using a nylon mesh of 80Mm and next the comet assay was applied. Student's T test was the statistical test used for analyzing the comet assay data. Results. Both assays showed a great sensitivity to the studied mutagens. A relationship between the dose-pink event and the dose-tail length was evident. Even
ResumenObjetivo. Evaluar la genotoxicidad de N-nitroso dietilamina (NDEA), hidrazida málica (MH) y etil metano sulfonato (EMS), en núcleos de Tradescantia (clona 4430) por medio de la prueba del cometa y de la prueba de mutación rosa, en los pelos estaminales de la misma planta. Material y métodos. Las plantas de Tradescantia (clon 4430) fueron obtenidas del Laboratorio de Citogenética y Mutagénesis del Centro de ciencias de la Atmósfera de la Universidad Nacional Autónoma de México, tratadas con NDEA a 1, 5, 10 mM, MH a 1, 5, 10 mM y EMS a 15, 30 y 45 mM, y utilizadas en la prueba de mutación rosa y en la del cometa, en núcleos celulares de los pelos estaminales. En la primera, la lectura de los pelos estaminales se realizó de acuerdo con el método de Underbrink. En otros estudios, que han aplicado la prueba del cometa en plantas, existe la necesidad de romper la pared celular y separar los núcleos por gradiente de centrifugación; en este caso, los núcleos de las células de los pelos estaminales fueron extraídos por aplastamiento sin aplicar un procedimiento especial para romper la pared, colectados por filtración en una malla de nylon y sometidos a la prueba del cometa. La prueba t de Student se usó para analizar los datos obtenidos. Resultados. Ambas pruebas presentaron una gran sensibilidad a los mutá-genos estudiados y hubo una relación evidente dosis-eventos rosa / longitud de la cauda. Aunque la prueba de mutación rosa en Tradescantia fue muy sensible a MH y EMS, no se
Hibiscus sabdariffa (Roselle) is in high demand worldwide due to its beneficial health properties owing to the polyphenols content, mainly in the flower calyx. The objective of this study was to find the best conditions (time and liquid: solid ratio) to extract polyphenols from Roselle using Ultrasound-Assisted Extraction (UAE) (40 kHz, 180 W), with ethanol how solvent; as well as determine the yield of phenols, anthocyanin, flavonoids, tannins, antioxidant activity (DPPH) and antigenotoxic effect (comet assay). A traditional solid-liquid extraction was applied as a reference. Extraction times of 40 and 60 min resulted in the highest polyphenols (13.019 mg GAE/g dry weight (dw)), flavonoids (4.981 CE/g dw), anthocyanins (1.855 mg Cya3GE/g dw), and tannins (0.745 CE/g dw) recoveries and an antioxidant activity (DPPH) of 74.58%. Extracts from white calyces contained similar amounts of phenols and flavonoids, but very little condensed tannins (0.049 CE/g dw) and practically no anthocyanins. Extracts from red and white calyces, showed antigenotoxic activity and repaired capacity of damage caused by mutagens in human lymphocytes.
There is considerable controversy with regard to the genotoxicity of glyphosate, with some reports stating that this compound is non-toxic for fish, birds and mammals. In this work, we used the comet assay to examine the genotoxicity of glyphosate isopropylamine (0.7, 7, 70 and 700 μM) in human lymphocytes, erythrocytes of Oreochromis niloticus and staminal nuclei of Tradescantia (4430) in vitro and in vivo. Cells, nuclei and fish that had and had not been exposed to 5 mM N-nitrosodiethylamine (NDEA) were used as positive and negative controls, respectively. Significant (p < 0.01) genetic damage was observed in vivo and in vitro in all cell types and organisms tested. Human lymphocytes and Tradescantia hairs showed lower genetic damage in vivo compared to in vitro, possibly because of efficient metabolization of the herbicide. In O. niloticus erythrocytes, significant (p < 0.001) genotoxicity was observed at ≥ 7 μM, whereas in vitro, glyphosphate was genotoxic in human lymphocytes and Tradescantia hairs at ≥ 0.7 μM. These results indicate that glyphosate is genotoxic in the cells and organisms studied at concentrations of 0.7–7 μM.
In some species, in which the human is included, the influence of age in the variation in the number of micronucleated erythrocytes (MNE) is known. In the present work we show how the process of aging influences the number of spontaneous MNE in the gray squirrel (Sciurus aureogaster). Because of the difficulty of knowing the age of each animal, 69 animals were weighed at their arrival to the laboratory and at the start of sample taking, with the supposition that the heaviest animals were the oldest and those with the lightest weight were the youngest. The major number of MNE was found in the younger animals, whereas the adults displayed less MNE (P < 0.0001). A group of 11 animals were sampled every 15 days over a period of 6 months, and the number of MNE were found to decrease with an increment in the weight in conformity with the time elapsed. These results showed that in the gray squirrel, the number of spontaneous MNE in peripheral blood depend on age. An additional interesting datum about the increment of MNE after the administration of colchicine is shown.
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