Streptococcus pyogenes Cas9 ribonucleoprotein delivery for efficient, rapid and marker-free gene editing in Trypanosoma and Leishmania

Kinetoplastids are unicellular eukaryotic flagellated parasites found in a wide range of hosts within the animal and plant kingdoms. They are known to be responsible in humans for African sleeping sickness ( Trypanosoma brucei ), Chagas disease ( Trypanosoma cruzi ), and various forms of leishmaniasis ( Leishmania spp.), as well as several animal diseases with important economic impact (African trypanosomes, including T. congolense ). Understanding the biology of these parasites necessarily implies the ability to manipulate their genomes. In this study, we demonstrate that transfection of a ribonucleoprotein complex, composed of recombinant Streptococcus pyogenes Cas9 ( Sp Cas9) and an in vitro -synthesized guide RNA, results in rapid and efficient genetic modifications of trypanosomatids, in marker-free conditions. This approach was successfully developed to inactivate, delete and mutate candidate genes in various stages of the life cycle of T. brucei and T. congolense , and Leishmania promastigotes. The functionality of Sp Cas9 in these parasites now provides, to the research community working on these parasites, a rapid and efficient method of genome editing, without requiring plasmid construction and selection by antibiotics. Importantly, this approach is adaptable to any wild-type parasite, including field isolates

The James Webb Space Telescope Mission

Twenty-six years ago a small committee report, building on earlier studies, expounded a compelling and poetic vision for the future of astronomy, calling for an infrared-optimized space telescope with an aperture of at least $4m$. With the support of their governments in the US, Europe, and Canada, 20,000 people realized that vision as the $6.5m$ James Webb Space Telescope. A generation of astronomers will celebrate their accomplishments for the life of the mission, potentially as long as 20 years, and beyond. This report and the scientific discoveries that follow are extended thank-you notes to the 20,000 team members. The telescope is working perfectly, with much better image quality than expected. In this and accompanying papers, we give a brief history, describe the observatory, outline its objectives and current observing program, and discuss the inventions and people who made it possible. We cite detailed reports on the design and the measured performance on orbit.Comment: Accepted by PASP for the special issue on The James Webb Space Telescope Overview, 29 pages, 4 figure

Comprendre les leviers de développement territorial par des projets de transition énergétique

The purpose of this report is to present the 6 months of internship and to analyze a territorial issue. The "centrales villageoises", through the deployment of renewable energies, are tools for the energy transition for the territories. They are levers for territorial development. They promote inter-territorial relations, the main issues studied by Transitions.Ce rapport vise à présenter le stage effectué pendant 6 mois ainsi qu’à analyser une question territoriale. Les centrales villageoises, par le déploiement d’énergies renouvelables, sont des outils à la transition énergétique pour les territoires. Elles sont des leviers de développement territorial. Elles favorisent, entre autres, les relations inter-territoriales, principales problématiques étudiés par Transitions

Le miracle du Muqaṭṭam à travers les siècles

Le miracle du déplacement du Muqaṭṭam circule au sein de la communauté copte depuis un millier d’années. Aujourd’hui encore, il est souvent évoqué par les fidèles de l’Église copte comme un exemple de victoire de la foi chrétienne contre l’islam. Ce récit, même s’il possède des traits structurels fixes, a pourtant connu d’importantes variantes au fil des siècles, au départ de sa première attestation dans le corpus copto-arabe de l’Histoire des Patriarches d’Alexandrie. De manière générale, il a surtout vu se développer ses aspects apologétiques, finissant par inclure la conversion au christianisme du calife al‑Muʿizz, qui fonda la ville du Caire à la fin du xe s. Au xxe s., cette affirmation fit l’objet de débats virulents dans la presse. À partir des années 1970, un jeune prêcheur redonna une nouvelle jeunesse à ce récit en entreprenant de fonder un énorme lieu de culte au Muqaṭṭam, commémorant le miracle. Fruit d’une collaboration entre un historien de l’époque contemporaine et de deux philologues, cet article entend proposer une approche décloisonnée, où l’histoire des textes dialogue avec l’analyse des pratiques contemporaines. Cette recherche montre que, si de nombreuses traditions ont d’anciennes racines, elles n’en sont pas moins toujours réinventées en fonction des contextes successifs au sein desquels elles s’inscrivent.The miracle tale of the Moving Muqaṭṭam mountain has been circulating among the Coptic community for a thousand years. Nowadays, members of the Coptic Church still refers to it as an example of victory of the Christian faith over Islam. This narrative, even if it bears some fixed structural features, went through numerous variations throughout times. On the basis of its first attestation in the Coptic‑Arabic text of the History of the Patriachs of Alexandria, it was later further developped, mainly under an apologetic angle. Finally, it included the episode of the Caliph al‑Muʿizz—founder of Cairo in the 10th century—converting to christianity. In the 20th century, this was subject to considerable debate in the press. From the 1970’s onwards, a young preacher revigorated the famous narrative by founding a huge place of worship at the Muqaṭṭam, commemorating the miracle. This paper, which is the result of a collective work by a historian and two philologists, intends to offer a collaborative approach, where the history of texts is combined with the analysis of contemporary practices. This research shows that, even if numerous traditions have very old roots, they are always reinveted, depending on successive contexts in which they take place

Le miracle du Muqaṭṭam à travers les siècles

International audienc

A geospatial model of nature-based recreation for urban planning: Case study of Paris, France

International audienceIncorporating nature-based recreation into urban planning analyses requires understanding the accessibility, quality, and demand for urban greenspace (UGS) across a city. Here, we present a novel tool that lowers the barriers to such information by (i) providing a spatially-explicit assessment of recreational UGS supply and demand; (ii) differentiating results by population group or UGS type; and (iii) using an accessible open-source software platform that facilitates scenario comparison and communication. In a case study in Paris, France, we demonstrate how the tool helps address important urban planning questions. We show that between 42% and 55% of the population is currently below the UGS target of 10 m2 per person, depending on the accessibility criteria used. Using revealed preference data, we demonstrate that older adults are disproportionately affected by the UGS deficit. Our assessment of future scenarios reveals that UGS targets set by public policies are largely insufficient (500–2800 ha are planned by 2030, while more than 4000 ha are needed to meet the policy target). By combining the strengths of established geospatial methods, the tool helps researchers and practitioners produce a more nuanced analysis of the recreation benefits of UGS implementation

Interaction of human 3-phosphoglycerate kinase with Its two substrates : Is substrate antagonism a kinetic advantage ?

International audienceSubstrate antagonism has been described for a variety of enzymes with more than one substrate and is characterized by a lowering of the affinity of one substrate in the presence of the other(s). 3-Phosphoglycerate kinase (PGK) catalyzes phosphotransfer from 1,3-bisphosphoglycerate (bPG) to ADP to give 3-phosphoglycerate (PG) and ATP, and is subject to substrate antagonism. Because of the instability of bPG, antagonism has only been described between PG and ATP or ADP. Here, we show that antagonism also occurs between bPG and ADP. Using the stopped-flow method, we show that the dissociation constant for one substrate increases in the presence of the other, and that this decrease in affinity is mainly due to an increase in the dissociation rate constant. As a consequence, there is an increase in the overall interaction kinetics. Interestingly, in the presence of the mirror image of natural D-ADP, L-ADP (a good substrate for PGK), antagonism is absent. Using rapid-quench-flow, we studied the kinetics of ATP formation. The time courses present the following: (1) a lag with L-ADP, but not with D-ADP, the kinetics of which were similar to the interaction kinetics measured by stopped-flow; (2) a burst that is directed by the phosphotransfer; and (3) a steady-state that is rate limited by the release of product kinetics. Structural explanations for these results are proposed by analyzing the crystallographic structure of the fully closed conformation of PGK in complex with L-ADP, PG, and the transition-state analogue AlF4 − compared to previously determined structures

Prognostic factors of toxicity of immune checkpoint inhibitors in nonsmall cell lung cancer and small cell lung cancer patients: The ToxImmune study

Abstract Background Immunotherapy alone or in combination has clearly improved the survival of patients with lung cancer. However, it may also be responsible for adverse events impacting these patients' quality of life. The ToxImmune study aims to identify prognostic factors that can help to predict immune‐related adverse events. Methods We included all patients aged 18 years and older who had received at least one dose of immune checkpoint inhibitors, with or without other therapy, between June 2015 and December 2020 and were diagnosed with nonsmall cell lung cancer or small‐cell lung cancer. Patients' baseline demographic characteristics, biological blood markers, and imaging by PET‐scanner were collected from electronic medical records. All adverse events (AEs) and immune‐related AEs (irAEs) were recorded (Common Terminology Criteria For Adverse Events V.5.0). Results Sixty‐four patients were included, of whom 60 (94%) presented at least one irAE. The incidence of Common Terminology Criteria for Adverse Events (CTCAE) grade 2 and grade 3–4 was 34% and 8% respectively. Female sex, Primitive Tumor Standardized Uptake Value Max (SUVmax) <5, number of metastases ≥3 and immunotherapy received after the first line were found to be significant risk factors for immune‐related adverse events. Based on the number of risk factors, the ToxImmune score predicts the risk of having a grade ≥2 adverse event (primitive tumor SUV ≥ 5 = 0 vs. primitive tumor SUV <5 = 1, number of metastases <3 = 0 vs. number of metastases ≥3 = 1 and L1 = 0 vs. L1 ≥ 1). The incidence of grade ≥2 adverse events was 20%, 55% and 90% with ToxImmune scores 0, 1 and = 2 respectively (p = .003). Median progression‐free survival (PFS) times were 19.2 months, 6.64 months and 2.63 months for ToxImmune scores 0, 1 and = 2 respectively, p = .13. Median overall survival times were 22.6 months, 16.4 months and 9.8 months for ToxImmune scores 0, 1 and ≥2 respectively, p = .24. The disease control rate (DRR) was 78% in ToxIummune score 0 group, and 50% in ToxImmune score 1 and ≥2 groups (p = .363). Conclusion The ToxImmune score, which is grounded on objective clinical parameters, indicates that cases with a high score had an advanced threat of severe adverse events. The ToxImmune score could therefore be used in clinical practice to identify patients treated for lung cancer with immunotherapy and at risk of severe AE