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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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- Falster, Daniel, et al.
(författare)
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AusTraits, a curated plant trait database for the Australian flora
- 2021
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Ingår i: Scientific Data. - : Nature Portfolio. - 2052-4463. ; 8:1
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Tidskriftsartikel (refereegranskat)abstract
- We introduce the AusTraits database - a compilation of values of plant traits for taxa in the Australian flora (hereafter AusTraits). AusTraits synthesises data on 448 traits across 28,640 taxa from field campaigns, published literature, taxonomic monographs, and individual taxon descriptions. Traits vary in scope from physiological measures of performance (e.g. photosynthetic gas exchange, water-use efficiency) to morphological attributes (e.g. leaf area, seed mass, plant height) which link to aspects of ecological variation. AusTraits contains curated and harmonised individual- and species-level measurements coupled to, where available, contextual information on site properties and experimental conditions. This article provides information on version 3.0.2 of AusTraits which contains data for 997,808 trait-by-taxon combinations. We envision AusTraits as an ongoing collaborative initiative for easily archiving and sharing trait data, which also provides a template for other national or regional initiatives globally to fill persistent gaps in trait knowledge.
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- Kyprianou, Dimitris, et al.
(författare)
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The application of polythiol molecules for protein immobilisation on sensor surfaces
- 2010
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Ingår i: Biosensors & bioelectronics. - : Elsevier Science B.V., Amsterdam.. - 0956-5663 .- 1873-4235. ; 25:5, s. 1049-1055
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Tidskriftsartikel (refereegranskat)abstract
- The immobilisation of bio-receptors on transducer surfaces is a key step in the development of biosensors. The immobilisation needs to be fast, cheap and most importantly should not affect the biorecognition activity of the immobilised receptor. The development of a protocol for biomolecule immobilisation onto a surface plasmon resonance (SPR) sensor surface using inexpensive polythiol compounds is presented here. The method used here is based on the reaction between primary amines and thioacetal groups, formed upon reaction of o-phthaldialdehyde (OPA) and thiol compounds. The self-assembled thiol monolayers were characterised using contact angle and XPS. The possibility to immobilise proteins on monolayers was assessed by employing BSA as a model protein. For the polythiol layers exhibiting the best performance, a general protocol was optimised suitable for the immobilisation of enzymes and antibodies such as anti-prostate specific antigen (anti-PSA) and anti Salmonella typhimurium. The kinetic data was obtained for PSA binding to anti-PSA and for S. typhimurium cells with a detection limit of 5 x 10(6) cells mL(-1) with minimal non-specific binding of other biomolecules. These findings make this technique a very promising alternative for amine coupling compared to peptide bond formation. Additionally, it offers opportunity for immobilising proteins (even those with low isoelectric point) on neutral polythiol layers without any activation step. (C) 2009 Elsevier B.V. All rights reserved.
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- Orban, Jenny, et al.
(författare)
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Surface nanoengineered contact lens as a wearable point-of-care diagnostics platform
- 2014
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Ingår i: 24<sup>th </sup>Anniversary World Congress on Biosensors – Biosensors 2014. - : Elsevier.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Detection of biomarkers is essential for disease prevention, diagnosis, and prognosis of early stage treatment. Ocular fluid is an extracellular fluid excreted from the tear gland. Several important markers from ocular fluid have been identified having significant clinical diagnostic value for various diseases. The contact lens is disposable, relatively cheap and serves as a platform to obtain direct intimate contact with ocular fluid and is therefore an attractive and a promising platform for point-of-care diagnostic tests. Here, we present an innovative concept of a wearable contact lens biosensor with nanoengineered biorecognition architecture based on a Layer-by-Layer (LbL) technique on the contact lens surface. This technique enables us to deposit multiple layers of biomolecules to create biorecognition layer under a mild aqueous physiological temperature and pH conditions. The fabrication of the biorecognition layer is simply through physical adsorption and has no restrictions with respect to the substrate size and topology (i.e. contact lens). The thickness of the resulting biorecognition layer is only hundreds of nanometers, and hence has minimal influence on the overall thickness of the contact lens, thus preserving the optical properties of the contact lens for vision correction. We have demonstrated that eye inflammation biomarkers such as interleukin (IL) can be captured in vitro with the contact lens, thus facilitating colorimetric affinity bioassays to measure the amount of interleukin. An in vitro ocular model composed of hydrogel-based artificial cornea and microfluidics was developed to study the performance of the contact lens biosensing platform. The contact platform was able to detect IL-1α down to the physiological concentration, which is in the range of pg mL-1. There is a recent trend away from handheld devices towards wearable systems, illustrated by popular technology such as “Google glasses” and the “i-Watch”. We believe that the concept of wearable diagnostics holds significant promise as the next generation point-of-care diagnostic platform and that this contact lens-based approach is a convenient platform for a number of important applications.
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