Éditeur(s) : HAL CCSD Landes Bioscience
Résumé : International audience
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.
Autophagy

hal-00735751
https://hal.archives-ouvertes.fr/hal-00735751
PUBMED : 22966490

Éditeur(s) : HAL CCSD Elsevier Masson
Résumé : Fire-free forest conversion with organic inputs as an alternative to slash-and-burn could improve agro-ecosystem sustainability. We assessed soil carbon mass changes in a sandy-clayey and well-drained soil in French Guiana after forest clearing by the chop-and-mulch method and crop establishment. At the experimental site of Combi, native forest was cut down in October 2008; woody biomass was chopped and incorporated into the top 20 cm of soil. After about one year of legume and grass cover, three forms of land management were compared: grassland (Urochloa ruziziensis), maize/soybean crop rotation with disk tillage and in direct seeding without tillage. There were four replicates. We measured 14.16 kg m(-2) of carbon in 2 mm-sieved soil down to 2 m depth for the initial forest. Forest clearing did not induce significant soil compaction; neither did any specific agricultural practice. In converted soils, C stocks were measured in the 0-30 cm layer after each crop for three years. Carbon mass changes for soil fractions <2 mm (soil C stock) and >2 mm (soil C pool) in the 0-5, 5-10, 10-20 and 20-30 cm soil layers were assessed on an equivalent soil mass basis. One year and 1.5 years after deforestation, higher C stocks (+0.64 to 1.16 kg C m(-2) yr(-1)) and C pools (+0.52 to 0.90 kg C m(-2) yr(-1)) were measured in converted soils, compared to those of the forest into the top 30 cm of soil. However, the masses of carbon in these converted soils declined later. The highest rates of carbon decrease were measured between 1.5 and 2 years after forest conversion in the <2 mm soil fraction, from 0.46 kg Cm-2 yr(-1) (in grassland soils) to 0.71 kg C m(-2) yr(-1) (in cropland under no tillage). The carbon pool declined during the third year at rates of 0.41 kg C m(-2) yr(-1) (cropland under disk tillage) to 0.76 kg C m(-2) yr(-1) (grassland soils). Three years after forest conversion, C masses in the top 30 cm of soils for grassland showed similar values than for forest. In comparison, the carbon stock in cropped soils managed under no tillage in direct seeding (without mulch) was significantly 17% and 16% lower than in forest and grassland soils, respectively. None of the studied agricultural practices succeeded in accumulating carbon from the chopped forest biomass.
ISSN: 0167-8809

hal-01032438
https://hal.archives-ouvertes.fr/hal-01032438
DOI : 10.1016/j.agee.2013.11.009

Éditeur(s) : HAL CCSD Le Cherche midi CNRS
Résumé : 192 pages
National audience
https://hal.archives-ouvertes.fr/hal-01118878

ISBN : 978-2-7491-4075-9
hal-01118878
https://hal.archives-ouvertes.fr/hal-01118878

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
The ‘International Intercomparison Exercise of fCO₂ Systems’ was carried out in 1996 during the R/V Meteor Cruise 36/1 from Bermuda/UK to Gran Canaria/Spain. Nine groups from six countries (Australia, Denmark, France, Germany, Japan, USA) participated in this exercise, bringing together 15 participants with seven underway fugacity of carbon dioxide (fCO₂) systems, one discrete fCO₂ system, and two underway pH systems, as well as systems for discrete measurement of total alkalinity and total dissolved inorganic carbon. Here, we compare surface seawater fCO₂ measured synchronously by all participating instruments. A common infrastructure (seawater and calibration gas supply), different quality checks (performance of calibration procedures for CO2, temperature measurements) and a common procedure for calculation of final fCO₂ were provided to reduce the largest possible amount of controllable sources of error. The results show that under such conditions underway measurements of the fCO₂ in surface seawater and overlying air can be made to a high degree of agreement (±1 μatm) with a variety of possible equilibrator and system designs. Also, discrete fCO₂ measurements can be made in good agreement (±3 μatm) with underway fCO₂ data sets. However, even well-designed systems, which are operated without any obvious sign of malfunction, can show significant differences of the order of 10 μatm. Based on our results, no “best choice” for the type of the equilibrator nor specifics on its dimensions and flow rates of seawater and air can be made in regard to the achievable accuracy of the fCO₂ system. Measurements of equilibrator temperature do not seem to be made with the required accuracy resulting in significant errors in fCO₂ results. Calculation of fCO₂ from high-quality total dissolved inorganic carbon (CT) and total alkalinity (AT) measurements does not yield results comparable in accuracy and precision to fCO₂ measurements.
ISSN: 0304-4203

hal-01245605
https://hal.archives-ouvertes.fr/hal-01245605
DOI : 10.1016/S0304-4203(00)00080-3