| 1. |
- 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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| 2. |
- Ananías, Rubén A., et al.
(author)
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Collapse of Eucalyptus nitens Wood after Drying Depending on the Radial Location Within the Stem
- 2014
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In: Drying Technology. - : Informa UK Limited. - 0737-3937 .- 1532-2300. ; 32:14, s. 1699-1705
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Journal article (peer-reviewed)abstract
- Collapse is almost certain to occur in the industrial drying of Eucalyptus nitens, and as such this prevents the lumber manufacturing industry in Chile from producing commercial solid wood products from this species. This problem is still unsolved, and different studies to reduce collapse are currently underway. In this exploratory study, shrinkage and collapse after drying of Eucalyptus nitens was measured for boards cut from different radial locations within the stem (core, transition and outer wood from pith to bark) and having different annual ring orientation (flat-sawn and quarter-sawn). Even though exploratory, the results appear to confirm that pieces that were cut from the center of the trees were less susceptible to collapse than the pieces cut from the transition zone between the center and the periphery. On average, collapse in transition wood was approximately 50% higher than the collapse observed in wood cut from the central zone of the trees.
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| 3. |
- Ananías, Rubén A., et al.
(author)
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Drying Schedules for Canelo Wood
- 2013
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In: Drying Technology. - : Informa UK Limited. - 0737-3937 .- 1532-2300. ; 31:3, s. 282-285
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Journal article (peer-reviewed)abstract
- Canelo wood is a highly valued native species in Chile that shows delicate marbling patterns with a pinkish soft silver luster. Due to its decorative qualities, canelo wood is dried for the manufacture of furniture and musical instruments. However, canelo wood lacks vessels cells that typically transport the water in hardwoods. Per its drying behavior, canelo wood is considered a transition species between hardwoods and softwoods. Therefore, this article reports drying schedules that were developed for drying 25-mm and 50-mm canelo lumber. In addition, this article reports experimental overall mass transfer coefficients, so that drying times for each of the drying stages can be easily estimated
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| 4. |
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| 5. |
- Ananías, Rubén A., et al.
(author)
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Energy Consumption in Industrial Drying of Radiata Pine
- 2012
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In: Drying Technology. - : Informa UK Limited. - 0737-3937 .- 1532-2300. ; 30:7, s. 774-779
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Journal article (peer-reviewed)abstract
- This article reports the results of an exploratory study designed to reduce heat and electricity consumption in industrial drying of radiata pine. The experiments were performed with slight modifications of the standard drying schedules used by the sawmill to dry radiata pine in 100-m 3 industrial kilns. The heat and electricity consumption were determined with data collected during the drying runs and calculations based on mathematical models. The results showed that depending on the case, heat and power consumption were respectively reduced by up to 14 and 35%.
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| 6. |
- Ananías, Rubén A., et al.
(author)
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Testing New In-Kiln Meter for Monitoring Lumber Moisture Content during Drying
- 2013
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In: Drying Technology. - : Informa UK Limited. - 0737-3937 .- 1532-2300. ; 31:3, s. 277-281
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Journal article (peer-reviewed)abstract
- The objective of this study was to test a new in-kiln sensor for monitoring lumber moisture content during industrial drying. The theoretical foundation of the technology was already known, because it is based on electrical conductivity, but the mechanism of implementation was new and required validation. For this reason, the technology was compared with two other widely used methods for assessing lumber moisture content, namely, the oven-drying and electrical capacitance methods. The tests were performed in a 120-m3 industrial kiln operated by a sawmill in the eighth region of Chile, and the results showed that the average moisture content at the end of drying was satisfactorily determined by the new in-line sensor. As predicted by theory, the sensor was not able to accurately measure moisture content above 25%, but it was still able to provide the equivalent of a drying curve for monitoring of the drying process
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| 7. |
- Rangel-Salazar, Ruben, et al.
(author)
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Human native lipoprotein-induced de novo DNA methylation is associated with repression of inflammatory genes in THP-1 macrophages
- 2011
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In: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 12
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Journal article (peer-reviewed)abstract
- Background: We previously showed that a VLDL-and LDL-rich mix of human native lipoproteins induces a set of repressive epigenetic marks, i. e. de novo DNA methylation, histone 4 hypoacetylation and histone 4 lysine 20 (H4K20) hypermethylation in THP-1 macrophages. Here, we: 1) ask what gene expression changes accompany these epigenetic responses; 2) test the involvement of candidate factors mediating the latter. We exploited genome expression arrays to identify target genes for lipoprotein-induced silencing, in addition to RNAi and expression studies to test the involvement of candidate mediating factors. The study was conducted in human THP-1 macrophages. Results: Native lipoprotein-induced de novo DNA methylation was associated with a general repression of various critical genes for macrophage function, including pro-inflammatory genes. Lipoproteins showed differential effects on epigenetic marks, as de novo DNA methylation was induced by VLDL and to a lesser extent by LDL, but not by HDL, and VLDL induced H4K20 hypermethylation, while HDL caused H4 deacetylation. The analysis of candidate factors mediating VLDL-induced DNA hypermethylation revealed that this response was: 1) surprisingly, mediated exclusively by the canonical maintenance DNA methyltransferase DNMT1, and 2) independent of the Dicer/microRNA pathway. Conclusions: Our work provides novel insights into epigenetic gene regulation by native lipoproteins. Furthermore, we provide an example of DNMT1 acting as a de novo DNA methyltransferase independently of canonical de novo enzymes, and show proof of principle that de novo DNA methylation can occur independently of a functional Dicer/micro-RNA pathway in mammals.
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