Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Métabolisme de l'ARN chez les archées : dialogue entre l'hélicase à ARN ASH-Ski2, l'exoribonucléase 5'- 3' aRNase J et l'exosome 3'- 5' à ARN, à proximité du ribosome

A deep understanding of RNA metabolism, which includes any event in the life cycle of RNA molecules, such as their synthesis, folding/unfolding, modification, processing and degradation, is critical. The continuous breakdown and synthesis of mRNAs allow cells to rapidly adapt to changing environment. RNA biology became an important and fast-evolving field at the center of post-transcriptional regulation of gene expression. Although numerous regulation pathways have been established in Eukarya and Bacteria, these processes have been hardly examined in Archaea. Despite fragmented information gathered on the proteins that maturate and degrade RNAs within archaeal cells, many open questions remain and RNA pathways in Archaea are, for the most part, still a mystery. Archaea are considered to form a separate domain of life, distinct from Eukarya and Bacteria. In-depth phylogenomic studies give strong evidences that Eukarya and Archaea are sister groups with closely related genetic information systems (translation, transcription, replication, DNA recombination and reparation). Archaea are now viewed as models with universal importance to study the molecular mechanisms that are conserved between Eukarya and Archaea. The "RNA biology of Archaea" group has developed an innovative area of research to identify RNA processing machineries in Archaea. In this context, the thesis work consisted (i) in listing the RNase families (structure and function) known in the Archaea for the writing of a review (Clouet-d'Orval et al 2018) and (ii) in identifying and characterizing the protein partners of exoRNase 5'-3' of the family ß-CASP, aRNase J, in Thermococcales. The results obtained, using proteomic, biochemical and enzymatic approaches, show that aRNase J is specifically engaged with the ASH-Ski2 helicase and the RNA exosome in the vinicity of the ribosome (published results: Phung et al NAR 2020) and that the ASH-Ski2 partner is an RNA helicase, with a 3'-5' polarity, whose predominant activity is strand assembly (annealing) (writing in progress: Batista et al, in preparation). All the data obtained, which are supported by phylogenomic approaches, offer new prospects for the identification of new molecular machinery involved in RNA metabolism in Archaea.La compréhension du métabolisme de l'ARN, c'est-à-dire tout événement du cycle de la vie des molécules d'ARN, de leur synthèse à leur dégradation, est fondamentale. Ces mécanismes permettent aux cellules de s'adapter rapidement à un environnement variable. Bien que de nombreuses voies de régulation aient été établies chez les eucaryotes et les bactéries, ces processus ont été peu examinés chez les archées. Malgré les informations fragmentaires disponibles sur les protéines qui assurent la maturation et la dégradation des ARN, de nombreuses questions demeurent en suspens et les voies métaboliques de l'ARN dans les cellules d'archées restent, pour la plupart, à découvrir. Les archées sont considérées comme formant un domaine du vivant singulier, distinct des eucaryotes et des bactéries, qui est désormais considéré comme constituant des modèles d'une importance universelle pour l'étude des mécanismes moléculaires conservés. L'équipe d'accueil "RNA biology of Archaea" développe un domaine de recherche innovant pour identifier les machines moléculaires et les enzymes impliquées dans le métabolisme de l'ARN chez les Archées. Ainsi les ribonucléases (RNases) de type ß-CASP ont été identifiées comme des enzymes ubiquitaires chez les Archées. Dans ce contexte le travail de thèse a consisté (i) à répertorier les familles de RNases (structures et fonction) connues chez les Archées pour la rédaction d'une revue (Clouet-d'Orval et al 2018) et (ii) à identifier et caractériser les partenaires protéiques de l'exoRNase 5'-3' de la famille ß-CASP, aRNase J, chez les Thermococcales. Les résultats obtenus, par des approches de protéomiques, biochimiques et enzymatiques, montrent que la aRNase J est engagée spécifiquement avec l'hélicase ASH-Ski2 et l'exosome à ARN à proximité du ribosome (résultats publiés : Phung et al NAR 2020) et que le partenaire ASH-Ski2 est une hélicase à ARN, avec une polarité 3'- 5' dont l'activité prépondérante est l'assemblage de brins (annealing) (rédaction en cours : Batista et al, en préparation). L'ensemble des données obtenues qui sont confortées par des approches phylogénomiques, offrent de nouvelles perspectives quant à l'identification de nouvelles machineries moléculaires impliquées dans le métabolisme de l'ARN chez les Archées

RNA metabolism in Archaea Dialogue between the ASH-Ski2 RNA helicase, the 5’-3’ exoribonuclease aRNase J and the RNA exosome at the vinicity of the ribosome

La compréhension du métabolisme de l'ARN, c'est-à-dire tout événement du cycle de la vie des molécules d'ARN, de leur synthèse à leur dégradation, est fondamentale. Ces mécanismes permettent aux cellules de s'adapter rapidement à un environnement variable. Bien que de nombreuses voies de régulation aient été établies chez les eucaryotes et les bactéries, ces processus ont été peu examinés chez les archées. Malgré les informations fragmentaires disponibles sur les protéines qui assurent la maturation et la dégradation des ARN, de nombreuses questions demeurent en suspens et les voies métaboliques de l'ARN dans les cellules d'archées restent, pour la plupart, à découvrir. Les archées sont considérées comme formant un domaine du vivant singulier, distinct des eucaryotes et des bactéries, qui est désormais considéré comme constituant des modèles d'une importance universelle pour l'étude des mécanismes moléculaires conservés. L'équipe d'accueil "RNA biology of Archaea" développe un domaine de recherche innovant pour identifier les machines moléculaires et les enzymes impliquées dans le métabolisme de l'ARN chez les Archées. Ainsi les ribonucléases (RNases) de type ß-CASP ont été identifiées comme des enzymes ubiquitaires chez les Archées. Dans ce contexte le travail de thèse a consisté (i) à répertorier les familles de RNases (structures et fonction) connues chez les Archées pour la rédaction d'une revue (Clouet-d'Orval et al 2018) et (ii) à identifier et caractériser les partenaires protéiques de l'exoRNase 5'-3' de la famille ß-CASP, aRNase J, chez les Thermococcales. Les résultats obtenus, par des approches de protéomiques, biochimiques et enzymatiques, montrent que la aRNase J est engagée spécifiquement avec l'hélicase ASH-Ski2 et l'exosome à ARN à proximité du ribosome (résultats publiés : Phung et al NAR 2020) et que le partenaire ASH-Ski2 est une hélicase à ARN, avec une polarité 3'- 5' dont l'activité prépondérante est l'assemblage de brins (annealing) (rédaction en cours : Batista et al, en préparation). L'ensemble des données obtenues qui sont confortées par des approches phylogénomiques, offrent de nouvelles perspectives quant à l'identification de nouvelles machineries moléculaires impliquées dans le métabolisme de l'ARN chez les Archées.A deep understanding of RNA metabolism, which includes any event in the life cycle of RNA molecules, such as their synthesis, folding/unfolding, modification, processing and degradation, is critical. The continuous breakdown and synthesis of mRNAs allow cells to rapidly adapt to changing environment. RNA biology became an important and fast-evolving field at the center of post-transcriptional regulation of gene expression. Although numerous regulation pathways have been established in Eukarya and Bacteria, these processes have been hardly examined in Archaea. Despite fragmented information gathered on the proteins that maturate and degrade RNAs within archaeal cells, many open questions remain and RNA pathways in Archaea are, for the most part, still a mystery. Archaea are considered to form a separate domain of life, distinct from Eukarya and Bacteria. In-depth phylogenomic studies give strong evidences that Eukarya and Archaea are sister groups with closely related genetic information systems (translation, transcription, replication, DNA recombination and reparation). Archaea are now viewed as models with universal importance to study the molecular mechanisms that are conserved between Eukarya and Archaea. The "RNA biology of Archaea" group has developed an innovative area of research to identify RNA processing machineries in Archaea. In this context, the thesis work consisted (i) in listing the RNase families (structure and function) known in the Archaea for the writing of a review (Clouet-d'Orval et al 2018) and (ii) in identifying and characterizing the protein partners of exoRNase 5'-3' of the family ß-CASP, aRNase J, in Thermococcales. The results obtained, using proteomic, biochemical and enzymatic approaches, show that aRNase J is specifically engaged with the ASH-Ski2 helicase and the RNA exosome in the vinicity of the ribosome (published results: Phung et al NAR 2020) and that the ASH-Ski2 partner is an RNA helicase, with a 3'-5' polarity, whose predominant activity is strand assembly (annealing) (writing in progress: Batista et al, in preparation). All the data obtained, which are supported by phylogenomic approaches, offer new prospects for the identification of new molecular machinery involved in RNA metabolism in Archaea

Insights into RNA-processing pathways and associated RNA-degrading enzymes in Archaea

International audienc

The conserved yeast protein Knr4 involved in cell wall integrity is a multi-domain intrinsically disordered protein

International audienceKnr4/Smi1; Saccharomyces cerevisiae; cell wall integrity; integrative structural biology; intrinsically disordered protein

Cold-conserved hybrid immature embryos for efficient wheat transformation

Transgenesis through biolistic of immature embryos is the most convenient way to introduce artificially new genes in bread wheat (Triticum aestivum L.). However, only a few genotypes can be efficiently transformed. To improve the transformation of wheat varieties, we stored immature seeds at room temperature or 4 degrees C during 4 or 7 days and extracted immature embryos prior to transformation. Shelling stops the embryo's growth and almost all the embryos formed a callus on selective media when stored at 4 degrees C for 4 or 7 days (respectively 87% and 99%). We also used hybrid immature embryos derived from a cross between a transformable line (Courtot) and a non-transformable line (Chinese Spring) for biolistic transformation. Hybrid embryos showed the same response to biolistic than the responsive parent. All together, these results improve significantly the biolistic protocol for wheat transformation by reducing the number of mother plants in the greenhouse, and improve the transformation of additional genotypes through hybrid transformation

Phylogenetic Diversity of Lhr Proteins and Biochemical Activities of the Thermococcales aLhr2 DNA/RNA Helicase

International audienceHelicase proteins are known to use the energy of ATP to unwind nucleic acids and to remodel protein-nucleic acid complexes. They are involved in almost every aspect of DNA and RNA metabolisms and participate in numerous repair mechanisms that maintain cellular integrity. The archaeal Lhr-type proteins are SF2 helicases that are mostly uncharacterized. They have been proposed to be DNA helicases that act in DNA recombination and repair processes in Sulfolobales and Methanothermobacter. In Thermococcales, a protein annotated as an Lhr2 protein was found in the network of proteins involved in RNA metabolism. To investigate this, we performed in-depth phylogenomic analyses to report the classification and taxonomic distribution of Lhr-type proteins in Archaea, and to better understand their relationship with bacterial Lhr. Furthermore, with the goal of envisioning the role(s) of aLhr2 in Thermococcales cells, we deciphered the enzymatic activities of aLhr2 from Thermococcus barophilus (Tbar). We showed that Tbar-aLhr2 is a DNA/RNA helicase with a significant annealing activity that is involved in processes dependent on DNA and RNA transactions

Evolutionary and functional insights into the Ski2-like helicase family in Archaea: a comparison of Thermococcales ASH-Ski2 and Hel308 activities

RNA helicases perform essential housekeeping and regulatory functions in all domains of life by binding and unwinding RNA molecules. The Ski2-like proteins are primordial helicases that play an active role in eukaryotic RNA homeostasis pathways, with multiple homologs having specialized functions. The significance of the expansion and diversity of Ski2-like proteins in Archaea, the third domain of life, has not yet been established. Here, by studying the phylogenetic diversity of Ski2-like helicases among archaeal genomes and the enzymatic activities of those in Thermococcales, we provide further evidence of the function of this protein family in archaeal metabolism of nucleic acids. We show that, in the course of evolution, ASH-Ski2 and Hel308-Ski2, the two main groups of Ski2-like proteins, have diverged in their biological functions. Whereas Hel308 has been shown to mainly act on DNA, we show that ASH-Ski2, previously described to be associated with the 5′-3′ aRNase J exonuclease, acts on RNA by supporting an efficient annealing activity, but also an RNA unwinding with a 3′-5′ polarity. To gain insights into the function of Ski2, we also analyse the transcriptome of Thermococcus barophilus ΔASH-Ski2 mutant strain and provide evidence of the importance of ASH-Ski2 in cellular metabolism pathways related to translation

RNA processing machineries in Archaea: the 5′-3′ exoribonuclease aRNase J of the β-CASP family is engaged specifically with the helicase ASH-Ski2 and the 3′-5′ exoribonucleolytic RNA exosome machinery

A network of RNA helicases, endoribonucleases and exoribonucleases regulates the quantity and quality of cellular RNAs. To date, mechanistic studies focussed on bacterial and eukaryal systems due to the challenge of identifying the main drivers of RNA decay and processing in Archaea. Here, our data support that aRNase J, a 5′-3′ exoribonuclease of the β-CASP family conserved in Euryarchaeota, engages specifically with a Ski2-like helicase and the RNA exosome to potentially exert control over RNA surveillance, at the vicinity of the ribosome. Proteomic landscapes and direct protein–protein interaction analyses, strengthened by comprehensive phylogenomic studies demonstrated that aRNase J interplay with ASH-Ski2 and a cap exosome subunit. Finally, Thermococcus barophilus whole-cell extract fractionation experiments provide evidences that an aRNase J/ASH-Ski2 complex might exist in vivo and hint at an association of aRNase J with the ribosome that is emphasised in absence of ASH-Ski2. Whilst aRNase J homologues are found among bacteria, the RNA exosome and the Ski2-like RNA helicase have eukaryotic homologues, underlining the mosaic aspect of archaeal RNA machines. Altogether, these results suggest a fundamental role of β-CASP RNase/helicase complex in archaeal RNA metabolism