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- Klionsky, Daniel J., et al.
(author)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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In: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Research review (peer-reviewed)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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| 4. |
- Dimkovski, Zlate, et al.
(author)
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Interference Measurements of Deposits on Cylinder Liner Surfaces
- 2011
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In: Wear. - Amsterdam : Elsevier. - 0043-1648 .- 1873-2577. ; 270:3-4, s. 247-251
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Journal article (peer-reviewed)abstract
- The accumulation of deposits in the honing grooves of the cylinder liner surfaces of internal combustion engines is undesirable as they seal the grooves (reducing their oil retention capability) and increase engine's oil consumption. As part of a long-term programme of truck engine development, after different running times and under similar conditions of load, engine speed and lubrication, a number of grey iron cylinder liners were axially sectioned, measured, inspected and a presence of deposits was discovered. These deposits were characterised in order to gain knowledge about their origin and quantities. The X-ray energy dispersive analysis revealed elements stemming from the oil/fuel (C, O and S), from the detergent (Ca and Mg), from the anti-wear additive (Zn and P), and from some contaminants (K and Si). Higher concentration of S and Ca were mostly found in the honing grooves covered with deposits suggesting a domination of the detergent additive. Deposit thickness measurements obtained by a white light interferometer revealed patchy deposit topographies concentrated at the top region reducing towards the bottom of the liner which was also confirmed by scanning electron microscope measurements. Despite the limitations of the interferometer, it has been shown that the interference measurements are sufficiently reliable for a quick and objective quantification of the overall deposit accumulation.
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| 6. |
- Rosén, Bengt Göran, 1962-, et al.
(author)
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Analysis of 3-D surfaces in forming dies and sheet metal - Today and tomorrow
- 2004
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In: Proceedings of the International Conference on Recent Advances in Manufacture and Use of Tools and Dies and Stamping of Steel Sheets. ; , s. 29-53
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Conference paper (peer-reviewed)abstract
- The effects of surface roughness on production of steel sheet are critically reviewed. New developments in the measurement and characterisation of surface roughness seem likely to improve our understanding of its effect on sheet metal production. Much useful information can still be acquired by 2D techniques but increasingly the evidence is that sheet metal surface roughness is better measured and characterised by 3D methods. The facility for direct surface visualization in 3D is shown to be particularly helpful. When measuring 3D the measurement area size and sampling space are critical and for evaluation the filtering of surface data into roughness and waviness is more complicated than for 2D profiles.
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| 7. |
- Rosén, Bengt Göran, et al.
(author)
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Topographical characterisation of artificial femoral heads : a Benchmarking study
- 2004
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In: Wear. - : Elsevier. - 0043-1648 .- 1873-2577. ; 257:12, s. 1275-1280
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Journal article (peer-reviewed)abstract
- The main functional demands on artificial joints are that they last more than 20 years. This means optimising the friction coefficient to minimise forces transmitted to the surrounding bone and the wear of the contacting surfaces. Surface roughness has an important influence on friction and wear. Traditionally, the 2D Ra-parameter has been the preferred way to characterise the joint's topography. Today ongoing development of 3D techniques are available for the research, and the aim of the present paper is to highlight the importance and possibilities using high-resolution 3D atomic force microscope ( AFM) and the optical phase-shifting interferometer for texture characterisation of eight commercially available femoral heads. The results show a significant variation of topography and surface-lay for the different femoral heads in the study. The variation and difference between heads were further underlined by the use of different characterisation techniques and measuring strategy. Results highlight the importance of the selection of measurement strategy, parameters, instrumentation, and scale of measurement in order properly to characterise the femoral heads in this study.
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| 9. |
- Rosén, Stefan, et al.
(author)
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The Stedman diagram revisited
- 2014
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In: Surface Topography : Metrology and Properties. - Bristol : Institute of Physics (IOP). - 2051-672X. ; 2:1
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Journal article (peer-reviewed)abstract
- The Stedman diagram has been used for some years to display aspects of the performance of instruments measuring surface roughness. Such diagrams are herein employed to compare the features of a range of proprietary measuring instruments, including contact and non-contact devices. An extension of the basic diagram is proposed, which would allow it to include a further aspect: the speed of data collection. Figures of merit based on the revised diagram are computed, which enable instruments to be ranked on these particular aspects of their performance. Contact instruments emerge as comparable to non-contact, as their slower rate of data acquisition can be offset by the greater area they can access in amplitude–wavelength space. © 2014 IOP Publishing Ltd.
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| 10. |
- Rosén, Stefan, et al.
(author)
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The Stedman diagram revisited
- 2013
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In: Metrology and Properties of Engineering Surfaces, 2013. ; , s. 201-206
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Conference paper (peer-reviewed)abstract
- The Stedman diagram has been used for some years to display aspects of the performance of surface roughness measuring instruments. Such diagrams are herein employed to compare the features of a range of proprietary measuring instruments, including contact and non-contact devices. An extension of the basic diagram is proposed which would allow it to include a further aspect, speed of data collection. Figures of merit based on the revised diagram are computed which enable instruments to be ranked on these particular aspects of their performance. Contact instruments emerge as comparable to non-contact, as their slower rate of data acquisition can be offset by the greater area they can access in amplitude-wavelength space.
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