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- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)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.
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- Ramaswamy, Babu Rajendran, et al.
(författare)
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GC-MS analysis and ecotoxicological risk assessment of triclosan, carbamazepine and parabens in Indian rivers.
- 2011
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Ingår i: Journal of hazardous materials. - : Elsevier BV. - 1873-3336 .- 0304-3894. ; 186:2-3, s. 1586-93
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Tidskriftsartikel (refereegranskat)abstract
- Pharmaceutical and personal care products are used extensively worldwide and their residues are frequently reported in aquatic environments. In this study, antiepileptic, antimicrobial and preservative compounds were analyzed in surface water and sediment from the Kaveri, Vellar and Tamiraparani rivers, and in the Pichavaram mangrove in India by gas chromatography-mass spectrometry (GC-MS). The mean concentration of carbamazepine recorded in the Kaveri River water (28.3 ng/L) was higher than in the other rivers and the mangrove. Because carbamazepine is used only in human drugs, this may reflect the relative contributions of human excretions/sewage in these rivers. The mean triclosan level in the Tamiraparani River (944 ng/L) was an order of magnitude greater than in the other water systems, and the concentrations at two of the sites reported here (3800-5160 ng/L) are, to our best knowledge, among the highest detected in surface waters. Sediment levels were, however, comparable with other sites. We conclude that industrial releases are likely major contributors of triclosan into this river system. Among parabens, ethyl paraben was predominantly observed. Hazard Quotients suggest greater environmental risks for triclosan than for carbamazepine and parabens. This is the first study on antiepileptic, antimicrobial and preservatives in rivers and mangroves from India.
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