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1.
Methods Mol Biol ; 1797: 497-505, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29896712

RESUMO

A broad range of research must be answered in order to gain a complete understanding of the histological and histochemical profile of teratological exposure in Mus musculus. Continued research is needed to track patterns of teratogen effects on the DNA expression of the embryonic brain and its variation impact. An important technique used in cell and molecular biology is Western blotting. By using a Western blot analysis and immunostaining, researchers are able to predict embryotoxicity in Mus musculus. The method uses three elements to accomplish this task: (1) Nonspecific antibody binding to a nitrocellulose membrane, (2) an incubation using a primary antibody, and (3) the antigen-antibody reaction using a secondary antibody. The proteins are further stained with a substrate/chromogen. In this chapter, the electrophoresis-based protein detection following mice embryonic exposure to a teratogen, 2-methoxyethanol, is described.


Assuntos
Western Blotting , Embrião de Mamíferos/efeitos dos fármacos , Embrião de Mamíferos/metabolismo , Imuno-Histoquímica , Testes de Toxicidade , Animais , Western Blotting/métodos , Imuno-Histoquímica/métodos , Camundongos , Testes de Toxicidade/métodos
2.
Int. j. morphol ; 37(3): 1164-1171, Sept. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1012412

RESUMO

Resveratrol is a stilbenoid, a type of natural phenol, and a phytoalexin produced by several plants in response to injury or attack by fungi. The underutilization of soybean seed coat (Glycine max (L.) Merrill.) and tempeh, cheap Indonesia fermented food thus opens up a new opportunity for developing a Resveratrol-based medicine for Plants-Derived Neuroprotective Agents purposes. In this study, it was isolated from tempeh, ordinarily well-known as Indonesian soybean fermented food, and soybean seed coat. The finding of this compound was confirmed by TLC and HPLC analysis applying fluorescence detection. From this, the Rf-value for transresveratrol is 0.64. As eluent, a mixture of chloroform, ethyl acetate, and formic acid (2.5+1+0.1, v/v) was selected. In addition, retention time for tempeh was 14.467 and for soybean seed coat was 11.977. The extraction yield of resveratrol was 65.15 % in tempeh and 55.35 % in soybean seed coat. Resveratrol isolated from Tempeh and Soybean seed coat gave prevents some reaction by modulating intracellular signaling pathways: protein kinase C (PKC), a family of 12 serine/ threonine kinases and providing a new lead molecule for neuroprotective affects in addition to has prevented cell death by apoptosis.


El resveratrol es un estilbenoide, un tipo de fenol natural, y fitoalexina producida por varias plantas en respuesta a una lesión o ataque de hongos. La subutilización de la cubierta de la semilla de soja (Glycine max (L.) Merrill.) y el tempeh, alimento fermentado barato de Indonesia, abren una nueva oportunidad para obtener un medicamento a base de resveratrol para propósitos de desarrollo de agentes neuroprotectores derivados de plantas. En este estudio, se aisló el resveratrol del tempeh, generalmente conocido como alimento fermentado de soja de Indonesia y de la cubierta de la semilla de soja. El hallazgo de este compuesto se confirmó mediante análisis de TLC y HPLC aplicando detección de fluorescencia. A partir de esto, el valor de Rf para trans-resveratrol es 0,64. Como eluyente, se seleccionó una mezcla de cloroformo, acetato de etilo y ácido fórmico (2,5 + 1 + 0,1, v / v). Además, el tiempo de retención para el tempeh fue de 14,467 y para el revestimiento de semilla de soja fue de 11,977. El rendimiento de extracción del resveratrol fue del 65,15 % en tempeh y del 55,35 % en la cubierta de la semilla de soja. El resveratrol aislado de tempeh y de la cubierta de la semilla de soja previno reacciones mediante la modulación de ciertas vías de señalización intracelular: proteína quinasa C (PKC), una familia de 12 serina/treonin quinasas, proporcionando una nueva molécula de plomo con efectos neuroprotectores, además de prevenir la muerte celular por apoptosis.


Assuntos
Animais , Camundongos , Glycine max/química , Fármacos Neuroprotetores/isolamento & purificação , Alimentos de Soja/análise , Resveratrol/isolamento & purificação , Sementes/química , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina
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