21 research outputs found

    Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

    Get PDF

    Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

    Get PDF
    In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

    Increased anandamide induced relaxation in mesenteric arteries of cirrhotic rats. Role of cannabinoid and vanilloid receptors

    Get PDF
    Background and aims: Anandamide is an endocannabinoid that evokes hypotension by interaction with peripheral cannabinoid CB1 receptors and with the perivascular transient receptor potential vanilloid type 1 protein (TRPV1). As anandamide has been implicated in the vasodilated state in advanced cirrhosis, the study investigated whether the mesenteric bed from cirrhotic rats has an altered and selective vasodilator response to anandamide. Methods: We assessed vascular sensitivity to anandamide, mRNA and protein expression of cannabinoid CB1 receptor and TRPV1 receptor, and the topographical distribution of cannabinoid CB1 receptors in resistance mesenteric arteries of cirrhotic and control rats. Results: Mesenteric vessels of cirrhotic animals displayed greater sensitivity to anandamide than control vessels. This vasodilator response was reverted by CB1 or TRPV1 receptor blockade, but not after endothelium denudation or nitric oxide inhibition. Anandamide had no effect on distal femoral arteries. CB1 and TRPV1 receptor protein was higher in cirrhotic than in control vessels. Neither CB1 mRNA nor protein was detected in femoral arteries. Immunochemistry showed that CB1 receptors were mainly in the adventitia and in the endothelial monolayer, with higher expression observed in vessels of cirrhotic rats than in controls. Conclusions: These results indicate that anandamide is a selective splanchnic vasodilator in cirrhosis which predominantly acts via interaction with two different types of receptors, CB1 and TRPV1 receptors, which are mainly located in perivascular sensory nerve terminals of the mesenteric resistance arteries of these animals

    Evaluación tecnofuncional de la harina de tempeh de soya.

    Get PDF
    El tempeh es un alimento que se obtiene por la fermentación con hongos del género Rhizopus endistintos sustratos, siendo las semillas de soya (Glycine max L.) uno de los más utilizados. Este producto contiene gran cantidad de proteínas, no contiene colesterol y es bajo en grasas saturadas. En la actualidad, las proteínas vegetales son utilizadas para formular alimentos especializados debido a su gran valor nutricional y a sus buenas propiedades funcionales. Se determinó el contenido de proteína del tempeh el cual ha sido 31.3% en la harina desengrasada. Por otra parte, las propiedades tecno-funcionales, tales como el índice de absorción de agua y aceite en la harina sin desengrasar resultaron en promedio 81.1177±0.5887 y 90.0290%±0.9385, respectivamente, mientras que en la harina desengrasada han sido de 91.9574 ± 0.2895 y 86.5573% ± 0.0129. La capacidad emulsificante de la harina desengrasada y sin desengrasar del tempeh fue de 66.6667% ± 0.0000 para ambas muestras, mientras que los valores de la estabilidad espumante para la harina desengrasada y sin desengrasar fueron de 1.3333%±0.0000 y 1.4667%±0.0000, respectivamente

    Structural modifications of gold thin films produced by thiol-derivatized single-stranded DNA immobilization

    No full text
    Recent experiments have reported an opposite sign of the differential surface stress produced on gold-coated cantilevers by a thiol-derivatized single-stranded DNA (SH-DNA) immobilization process. The sign of the surface stress depends on the method used to evaporate the gold thin film, being compressive (negative) or tensile (positive) for e-beam or resistively deposited gold, respectively. This study investigates the origin of this effect by means of a combination of x-ray diffraction and x-ray photoelectron spectroscopy. Both e-beam and resistively grown gold thin films are characterized to find the subtle differences responsible for this intriguing stress behaviour. Somewhat remarkably, these studies show a tight relation between the surface structure of the gold overlayer and the SH-DNA immobilization efficiency. The average grain size variation seems to correlate well with the differential surface stress triggered by the SH-DNA immobilization previously reported. These results suggest that the relation of the probe molecules with the surface structure must be considered to understand surface stress changes. © 2014 IOP Publishing Ltd.JLCK and MAH acknowledge financial support from MAT2008-06330. Work at Centro de Astrobiología was supported by MICINN (grants EUI2008-00158, EUI2008-00122 and BIO2010-20696), CSIC (grant 200920I040), the European Union and Comunidad de Madrid. CR acknowledges the financial support from MAT2010-21156-C03-03.Peer Reviewe

    Estandarización del protocolo para elaborar tempeh con diversos granos (y su adaptación a la comida regional mexicana).

    Get PDF
    Se evaluó el crecimiento del hongo Rhizopus oryzae en diferentes granos, legumbres y semillas para la producción de tempeh, el cual se considera como un superalimento. Los sustratos se sometieron a diversos procesos de cocción, escurrimiento y fermentación, los cuales fueron optimizados de acuerdo a las características del sustrato seleccionado. La fermentación se llevó a cabo mediante la inoculación con la cepa R. oryzae. Sucesivamente, se mezcló el sustrato con el inóculo, y se colocó en bolsas plásticas con cierre hermético, previamente perforadas. Estas fueron incubadas a 35 ºC por un período de 24 a 48 horas. El producto obtenido en las diferentes variantes presentó las características típicas de tempeh de buena calidad

    Increased anandamide induced relaxation in mesenteric arteries of cirrhotic rats. Role of cannabinoid and vanilloid receptors

    No full text
    Background and aims: Anandamide is an endocannabinoid that evokes hypotension by interaction with peripheral cannabinoid CB1 receptors and with the perivascular transient receptor potential vanilloid type 1 protein (TRPV1). As anandamide has been implicated in the vasodilated state in advanced cirrhosis, the study investigated whether the mesenteric bed from cirrhotic rats has an altered and selective vasodilator response to anandamide. Methods: We assessed vascular sensitivity to anandamide, mRNA and protein expression of cannabinoid CB1 receptor and TRPV1 receptor, and the topographical distribution of cannabinoid CB1 receptors in resistance mesenteric arteries of cirrhotic and control rats. Results: Mesenteric vessels of cirrhotic animals displayed greater sensitivity to anandamide than control vessels. This vasodilator response was reverted by CB1 or TRPV1 receptor blockade, but not after endothelium denudation or nitric oxide inhibition. Anandamide had no effect on distal femoral arteries. CB1 and TRPV1 receptor protein was higher in cirrhotic than in control vessels. Neither CB1 mRNA nor protein was detected in femoral arteries. Immunochemistry showed that CB1 receptors were mainly in the adventitia and in the endothelial monolayer, with higher expression observed in vessels of cirrhotic rats than in controls. Conclusions: These results indicate that anandamide is a selective splanchnic vasodilator in cirrhosis which predominantly acts via interaction with two different types of receptors, CB1 and TRPV1 receptors, which are mainly located in perivascular sensory nerve terminals of the mesenteric resistance arteries of these animals
    corecore