Henan University of Technology

About: Henan University of Technology is a(n) education organization based out in Zhengzhou, China. It is known for research contribution in the topic(s): Catalysis & Starch. The organization has 7648 authors who have published 6503 publication(s) receiving 73067 citation(s). The organization is also known as: Hénán Gōngyè Dàxué.

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Abstract: In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure flux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation, it is imperative to target by gene knockout or RNA interference more than one autophagy-related protein. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways implying that not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular assays, we hope to encourage technical innovation in the field.

Guidelines for the use and interpretation of assays for monitoring autophagy

Abstract: In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.

Transesterification of soybean oil catalyzed by potassium loaded on alumina as a solid-base catalyst

Abstract: Biodiesel fuel, consisting of methyl esters of long chain fatty acids produced by transesterification of vegetable oils or animal fats with methanol, is a promising alternative diesel fuel regarding the limited resources of fossil fuels and the environmental concerns. In this work, an environmentally benign process for the transesterification of soybean oil to methyl esters using alumina loaded with potassium as a solid base catalyst in a heterogeneous manner was developed. The catalyst loaded KNO 3 of 35 wt.% on Al 2 O 3 , after being calcined at 773 K for 5 h, it was found to be the optimum catalyst, which can give the highest basicity and the best catalytic activity for this reaction. The effects of various reaction variables such as the catalyst loading, oil to methanol ratio, reaction time and temperature on the conversion of soybean oil were investigated. The catalysts were characterized by means of XRD, IR and Hammett titration method. The results indicated that K 2 O derived from KNO 3 at high temperature and that the Al–O–K groups were, probably, the main reasons for the catalytic activity towards the reaction. The catalyst activity was correlated closely with its basicity as determined by the Hammett method.

Calcined Mg–Al hydrotalcites as solid base catalysts for methanolysis of soybean oil

Abstract: Methyl ester of fatty acids, derived from vegetable oils or animal fats and known as biodiesel, is a promising alternative diesel fuel regarding the limited resources of fossil fuel and the environmental concerns. In this work, an environmentally benign process for the methanolysis of soybean oil to methyl esters using calcined Mg–Al hydrotalcites as solid base catalysts in a heterogeneous manner was developed. When the reaction was carried out at reflux of methanol, with a molar ratio of soybean oil to methanol of 15:1, a reaction time 9 h and a catalyst amount 7.5%, the oil conversion was 67%. The calcined hydrotalcite with an Mg/Al ratio of 3.0 derived from calcination at 773 K was found to be the optimum catalyst that can give the highest basicity and the best catalytic activity for this reaction. The catalysts were characterized with SEM, XRD, IR, DTA-TG and Hammett titration method. The activity of the catalysts for the methanolysis reaction was correlated closely with their basicity as determined by the Hammett method.

Alumina-supported potassium iodide as a heterogeneous catalyst for biodiesel production from soybean oil

Abstract: Biodiesel fuel, a promising alternative diesel fuel produced by a catalytic transesterification of vegetable oils, has become more attractive recently because of its environmental concerns and the fact that it is made from renewable resources. In this work, the transesterification of soybean oil with methanol has been studied in a heterogeneous system, using alumina loaded with potassium iodide as a solid base catalyst. After loading KI of 35 wt.% on alumina followed by calcination at 773 K for 3 h, the catalyst gave the highest basicity and the best catalytic activity for this reaction. The catalysts were characterized by means of XRD, IR, SEM and the Hammett indicator method. Moreover, the dependence of the conversion of soybean oil on the reaction variables such as the catalyst loading, the molar ratio of methanol to oil and the reaction time was studied. The conversion of 96% was achieved under the optimum reaction conditions. Besides, a correlation of the catalyst activity for the transesterification reaction with its basicity was proposed.