Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
The use of recombinant antigens has been shown to improve both the sensitivity and the standardization of the serological diagnosis of Farmer's lung disease (FLD). The aim of this study was to complete the panel of recombinant antigens available for FLD serodiagnosis with antigens of Lichtheimia corymbifera, known to be involved in FLD. L. corymbifera proteins were thus separated by 2D electrophoresis and subjected to western blotting with sera from 7 patients with FLD and 9 healthy exposed controls (HEC). FLD-associated immunoreactive proteins were identified by mass spectrometry based on a protein database specifically created for this study and subsequently produced as recombinant antigens. The ability of recombinant antigens to discriminate patients with FLD from controls was assessed by ELISA performed with sera from FLD patients (n = 41) and controls (n = 43) recruited from five university hospital pneumology departments of France and Switzerland. Forty-one FLD-associated immunoreactive proteins from L. corymbifera were identified. Six of them were produced as recombinant antigens. With a sensitivity and specificity of 81.4 and 77.3% respectively, dihydrolipoyl dehydrogenase was the most effective antigen for discriminating FLD patients from HEC. ELISA performed with the putative proteasome subunit alpha type as an antigen was especially specific (88.6%) and could thus be used for FLD confirmation. The production of recombinant antigens from L. corymbifera represents an additional step towards the development of a standardized ELISA kit for FLD diagnosis. © 2016 Rognon et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
ISSN: 1932-6203

hal-01367148
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01367148
DOI : 10.1371/journal.pone.0160888

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
Résumé : International audience
The barrier reef tract of New Caledonia is the second largest in the world enclosing lagoons of variable width. The Late Quaternary evolution of these barrier reefs and lagoons is poorly known. A recent high resolution seismic and multibeam bathymetry survey was carried out in the south-western lagoon from shoreline to open ocean across the barrier reef pass systems to perform sequence stratigraphy and to better understand the Quaternary evolution of the lagoon. Two depositional sequences bounded by type-1 erosional surfaces can be identified. The bounding surfaces delineate the bottom of two generations of incised valleys. System tracts comprising a complete succession of regressive, lowstand, transgressive and highstand tracts were observed only in the upper sequence. Chronostratigraphic and sedimentological interpretations of the seismic data are based on correlations with the chronology defined in the nearby cores drilled through the reefs and in the gravity cores previously recovered in the inner lagoon. Interpretation of seismic data is also carried out on the basis of correlation with multibeam bathymetric data. Our results indicate that the history of the New Caledonian lagoon probably does not exceed 200ka marked by two or three brief episodes each of immersion of about ten thousand years. The bathymetric data reveal that during successive low sea level stands, the lagoon was dependent upon erosional fluvial networks. The deep fluvial valleys were connected to the continental slope through the passes. Seismic records reveal that upstream palaeo-channels are not connected to the modern coastal rivers. In contrast, many aggrading deposits, including numerous stacking channels, and extending across the proximal part of the outer lagoon, can be identified. Examination of drainage network morphology suggests a progressive tilt of the outer lagoon. Such evolution reveals that the Nouméa lagoon which is one of the widest in the world is an exceptional modern example of a rimmed shelf providing data to analyze the past counterparts.
ISSN: 0031-0182

insu-00338953
https://hal-insu.archives-ouvertes.fr/insu-00338953
DOI : 10.1016/j.palaeo.2008.08.012