Characterization of caspases from Lepidoptera and their role in apoptosis

Multicellular organisms need a tightly controlled regulation of cell proliferation and of cell specialization together with a controlled cell death to maintain their integrity. A family of evolutionarily conserved proteases, called caspases, plays a central role in regulating apoptosis in animals. The discovery of p35, a baculovirus caspase inhibitor, has led to the characterization of the first lepidopteran caspase, Sf-Caspase-1, from Spodoptera frugiperda. Studies on the Sf-Caspase-1 mode of activation suggested that apoptosis in Lepidoptera also requires a cascade of caspase activation, as demonstrated in many other species. We identified 66 sequences encoding putative caspases, distributed among 27 species. Phylogenetic analyses showed that Lepidoptera possess at least 5 caspases, for which we propose a unified nomenclature. According to their homology to their Drosophila counterparts and their primary structure, we proposed that Lep-Caspase-1, -2 and -3 are effector caspases, whereas Lep-Caspase-5 and -6 are putative initiators. The likely function of Lep-Caspase-4 remains unclear. We found that Caspase-1 and -2 have complementary expression profiles during larval development, suggesting a differential regulation. Caspase-3 and -6 are upregulated upon immune challenge, suggesting a role in the imd pathway. Caspase-5 is upregulated during pupation and upon induction with 20-hydroxyecdysone, supporting the hypothesis of Caspase-5 playing a similar role as Dronc in developmental apoptosis in Drosophila. In an attempt to obtain further insights into the molecular pathway underlying apoptosis in lepidopteran insects, we used 2D DIGE to identify 13 proteins, for which relative abundance was altered during apoptosis of H. armigera derived cells, induced by actinomycin D. Among these 13 proteins, we identified the putative effector Caspase-1, 3 chaperone proteins and several pro-apoptotic proteins, such as actin and a voltage dependant anion channel

Sialome of a Generalist Lepidopteran Herbivore: Identification of Transcripts and Proteins from Helicoverpa armigera Labial Salivary Glands

Although the importance of insect saliva in insect-host plant interactions has been acknowledged, there is very limited information on the nature and complexity of the salivary proteome in lepidopteran herbivores. We inspected the labial salivary transcriptome and proteome of Helicoverpa armigera, an important polyphagous pest species. To identify the majority of the salivary proteins we have randomly sequenced 19,389 expressed sequence tags (ESTs) from a normalized cDNA library of salivary glands. In parallel, a non-cytosolic enriched protein fraction was obtained from labial salivary glands and subjected to two-dimensional gel electrophoresis (2-DE) and de novo peptide sequencing. This procedure allowed comparison of peptides and EST sequences and enabled us to identify 65 protein spots from the secreted labial saliva 2DE proteome. The mass spectrometry analysis revealed ecdysone, glucose oxidase, fructosidase, carboxyl/cholinesterase and an uncharacterized protein previously detected in H. armigera midgut proteome. Consistently, their corresponding transcripts are among the most abundant in our cDNA library. We did find redundancy of sequence identification of saliva-secreted proteins suggesting multiple isoforms. As expected, we found several enzymes responsible for digestion and plant offense. In addition, we identified non-digestive proteins such as an arginine kinase and abundant proteins of unknown function. This identification of secreted salivary gland proteins allows a more comprehensive understanding of insect feeding and poses new challenges for the elucidation of protein function

A comprehensive characterization of the caspase gene family in insects from the order Lepidoptera

<p>Abstract</p> <p>Background</p> <p>The cell suicide pathway of apoptosis is a necessary event in the life of multicellular organisms. It is involved in many biological processes ranging from development to the immune response. Evolutionarily conserved proteases, called caspases, play a central role in regulating apoptosis. Reception of death stimuli triggers the activation of initiator caspases, which in turn activate the effector caspases. In Lepidoptera, apoptosis is crucial in processes such as metamorphosis or defending against baculovirus infection. The discovery of p35, a baculovirus protein inhibiting caspase activity, has led to the characterization of the first lepidopteran caspase, Sf-Caspase-1. Studies on Sf-Caspase-1 mode of activation suggested that apoptosis in Lepidoptera requires a cascade of caspase activation, as demonstrated in many other species.</p> <p>Results</p> <p>In order to get insights into this gene family in Lepidoptera, we performed an extensive survey of lepidopteran-derived EST datasets. We identified 66 sequences distributed among 27 species encoding putative caspases. Phylogenetic analyses showed that Lepidoptera possess at least 5 caspases, for which we propose a unified nomenclature. According to homology to their <it>Drosophila </it>counterparts and their primary structure, we determined that Lep-Caspase-1, -2 and -3 are putative effector caspases, whereas Lep-Caspase-5 and -6 are putative initiators. The likely function of Lep-Caspase-4 remains unclear. Lep-Caspase-2 is absent from the silkworm genome and appears to be noctuid-specific, and to have arisen from a tandem duplication of the Caspase-1 gene. In the tobacco hawkmoth, 3 distinct transcripts encoding putative Caspase-4 were identified, suggesting at least 2 duplication events in this species.</p> <p>Conclusions</p> <p>The basic repertoire of five major types of caspases shared among Lepidoptera seems to be smaller than for most other groups studied to date, but gene duplication still plays a role in lineage-specific increases in diversity, just as in Diptera and mammals.</p

Extracorporeal Membrane Oxygenation for Severe Acute Respiratory Distress Syndrome associated with COVID-19: An Emulated Target Trial Analysis.

RATIONALE: Whether COVID patients may benefit from extracorporeal membrane oxygenation (ECMO) compared with conventional invasive mechanical ventilation (IMV) remains unknown. OBJECTIVES: To estimate the effect of ECMO on 90-Day mortality vs IMV only Methods: Among 4,244 critically ill adult patients with COVID-19 included in a multicenter cohort study, we emulated a target trial comparing the treatment strategies of initiating ECMO vs. no ECMO within 7 days of IMV in patients with severe acute respiratory distress syndrome (PaO2/FiO2 <80 or PaCO2 ≥60 mmHg). We controlled for confounding using a multivariable Cox model based on predefined variables. MAIN RESULTS: 1,235 patients met the full eligibility criteria for the emulated trial, among whom 164 patients initiated ECMO. The ECMO strategy had a higher survival probability at Day-7 from the onset of eligibility criteria (87% vs 83%, risk difference: 4%, 95% CI 0;9%) which decreased during follow-up (survival at Day-90: 63% vs 65%, risk difference: -2%, 95% CI -10;5%). However, ECMO was associated with higher survival when performed in high-volume ECMO centers or in regions where a specific ECMO network organization was set up to handle high demand, and when initiated within the first 4 days of MV and in profoundly hypoxemic patients. CONCLUSIONS: In an emulated trial based on a nationwide COVID-19 cohort, we found differential survival over time of an ECMO compared with a no-ECMO strategy. However, ECMO was consistently associated with better outcomes when performed in high-volume centers and in regions with ECMO capacities specifically organized to handle high demand. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Restauration de Sapho ou Le Chant de Raoul Verlet

L'étude de Sapho ou Le Chant, modèle de fonderie en plâtre élaboré par Raoul Verlet et conservé au Musée d'Angoulême depuis 1926, a permis d'aborder un sujet courant, celui des plâtres fracturés par l'expansion de la corrosion d'armatures internes causée par l'humidité. La principale intervention s'est axée sur le remontage des fragments nécessitant la conception d'une structure de remontage en acier inoxydable. Les interventions  structurelles ont été complétées par des collages simples ou renforcés des fragments de grandes dimensions.The study of Sapho ou Le Chant, a plaster cast foundry created by Raoul Verlet and preserved in the Angoulême Museum since 1926, has allowed to approach a common problematic, that one of split plaster cast by the expansion of steel reinforcements corrosion, caused by high humidity conservation conditions. The main intervention concerned the fragments reconstruction requiring the design and the execution of a stainless steel pedestal. The structural interventions was completed with simple and reinforced stickings of the larger fragments

Characterization of Caspases from Lepidoptera and their role in Apoptosis

Multicellular organisms need a tightly controlled regulation of cell proliferation and of cell specialization together with a controlled cell death to maintain their integrity. A family of evolutionarily conserved proteases, called caspases, plays a central role in regulating apoptosis in animals. The discovery of p35, a baculovirus caspase inhibitor, has led to the characterization of the first lepidopteran caspase, Sf-Caspase-1, from Spodoptera frugiperda. Studies on the Sf-Caspase-1 mode of activation suggested that apoptosis in Lepidoptera also requires a cascade of caspase activation, as demonstrated in many other species. We identified 66 sequences encoding putative caspases, distributed among 27 species. Phylogenetic analyses showed that Lepidoptera possess at least 5 caspases, for which we propose a unified nomenclature. According to their homology to their Drosophila counterparts and their primary structure, we proposed that Lep-Caspase-1, -2 and -3 are effector caspases, whereas Lep-Caspase-5 and -6 are putative initiators. The likely function of Lep-Caspase-4 remains unclear. We found that Caspase-1 and -2 have complementary expression profiles during larval development, suggesting a differential regulation. Caspase-3 and -6 are upregulated upon immune challenge, suggesting a role in the imd pathway. Caspase-5 is upregulated during pupation and upon induction with 20-hydroxyecdysone, supporting the hypothesis of Caspase-5 playing a similar role as Dronc in developmental apoptosis in Drosophila. In an attempt to obtain further insights into the molecular pathway underlying apoptosis in lepidopteran insects, we used 2D DIGE to identify 13 proteins, for which relative abundance was altered during apoptosis of H. armigera derived cells, induced by actinomycin D. Among these 13 proteins, we identified the putative effector Caspase-1, 3 chaperone proteins and several pro-apoptotic proteins, such as actin and a voltage dependant anion channel

Inventaire et collecte des contes en Guyane : état des lieux et prospective

Pierre Courtiade, chargé d\u27étude à la Compagnie Zoukouyanyan, dresse un état des lieux et les prospectives envisagés dans le cadre de l\u27inventaire et de la collecte des contes en Guyane

Analyse expérimentale des écoulements haute vitesse et instationnaires dans un compresseur multi-étages à forte charge aérodynamique

The present work is a result of collaboration between the LMFA (Laboratoire de Mécanique des Fluides et d’Acoustique, Ecole Centrale de Lyon – France), Snecma and the Cerfacs. It aims at studying the flow in the 3.5-stages high-speed axial compressor CREATE (Compresseur de Recherche pour l’Etude des effets Aérodynamique et TEchnologique - rotation speed: 11543 RPM, Rotor 1 tip speed: 313 m/s), designed and built by Snecma and investigated at LMFA on a 2-MW test rig. Steady measurements, as well as laser velocimetry, fast-response wall static and total pressure measurements have been used to experimentally investigate the flow. The analysis focuses on two main aspects: the study of the flow at stable operating points, with a special interest on the rotor-stator interactions, and the study of the instabilities arising in the machine at low mass flow rates.The description of the unsteady flow field at stable operating points is done through measurements of wall-static pressure, total pressure and velocity, but also total temperature, entropy and angle of the fluid. It is shown that the complexity and unsteadiness of the flow in a multistage compressor strongly increases in the rear part of the machine, because of the interactions between steady and rotating rows. Therefore, a modal analysis method developed at LMFA and based on the decomposition of Tyler and Sofrin is presented to analyze these interactions. It is first applied to the pressure measurements, in order to extract the contributions of each row. It shows that all the complex pressure interactions in CREATE can be reduced to three main types of interactions. The decomposition method is then applied to the entropy field extracted from URANS CFD calculations performed by the Cerfacs, in order to evaluate the impact of the interactions on the performance of the machine in term of production of losses.The last part of this work is devoted to the analysis of the instabilities arising in CREATE at low mass flows. It shows that rotating pressure waves appear at stable operating points, and increase in amplitude when going towards the surge line, until reaching a critical size provoking the onset a full span stall cell bringing the machine to surge within a few rotor revolutions. The study of these pressure waves, and the understanding of their true nature is achieved through the experimental results and the use of some analytical models. A precise description of the surge transient through wall-static pressure measurements above the rotors is also provided, as well as a description of a complete surge cycle. An anti-surge control system based on the detection of the amplitude of the pressure waves is finally proposed.Ce travail est le produit d’une collaboration entre le LMFA (Laboratoire de Mécanique des Fluides et d’Acoustique, École Centrale de Lyon – France), Snecma et le Cerfacs. Il vise à étudier l’écoulement se développant dans le compresseur haute-vitesse axial de 3.5 étages CREATE (Compresseur de Recherche pour l’Etude des effets Aérodynamique et TEchnologique – vitesse de rotation : 11543 tr/min, vitesse en tête de rotor 1 : 313 m/s), conçu et construit par Snecma et étudié au LMFA sur un banc d’essai de 2 MW. Pour étudier l’écoulement, des mesures stationnaires de pression et température, de la vélocimétrie laser et des mesures rapides de pression statique et totale ont été utilisées. L’analyse se concentre sur deux aspects principaux : l’étude de l’écoulement aux points de fonctionnement stables, avec un intérêt tout particulier pour les interactions rotor-stator, et l’étude des instabilités apparaissant dans la machine à faibles débits.La description de l’écoulement instationnaire aux points stables est faite par le biais de mesures de pression statique en parois, de pression totale et de vitesse, mais également de température totale, entropie et angle d’incidence du fluide. Il est montré que la complexité et l’instationnarité de l’écoulement dans un compresseur multiétagé augmente fortement à l’arrière de la machine à cause des interactions entre les roues fixes et mobiles. Ainsi, une méthode d’analyse modale basée sur la décomposition de Tyler et Sofrin a été développée pour analyser ces interactions. Elle est d’abord appliquée aux mesures de pression afin d’extraire les contributions de chaque roue. Il est ainsi montré que les interactions complexes de pression dans CREATE peuvent être réduites à trois principaux types d’interactions. La méthode de décomposition est enfin appliquée au champ d’entropie dans toute la machine extrait de calculs CFD URANS réalisés par le Cerfacs, afin d’évaluer l’impact des interactions sur les performances de CREATE en terme de génération de pertes.La dernière partie de ce travail est dédié à l’analyse des instabilités apparaissant dans CREATE à faible débit. Il est montré que des ondes de pression tournantes apparaissent aux points stables et augmentent en amplitude à mesure qu’on se rapproche de la ligne de pompage, jusqu’à atteindre une taille critique induisant l’apparition d’une cellule de décollement tournant sur toute la hauteur de veine. Cette cellule entraîne la machine en pompage en seulement quelques tours. L’étude de ces ondes de pression, et la compréhension de leur véritable nature sont réalisées grâce à l’application d’un modèle analytique aux mesures expérimentales. Une description précise du déclenchement et du cycle du pompage est également faite grâce aux mesures de pression statique au dessus des rotors. Un système de contrôle anti-pompage développé au laboratoire et basé sur la détection de l’amplitude des ondes de pression est finalement décrit