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ImportanceNursing home (NH) residents are particularly vulnerable to SARS-CoV-2 infections and coronavirus disease 2019 (COVID-19) lethality. However, excess deaths in this population have rarely been documented.ObjectivesThe primary objective was to assess the number of excess deaths among NH residents during the first wave of the COVID-19 pandemic in France. The secondary objectives were to determine the number of excess deaths as a proportion of the total excess deaths in the general population and determine whether a harvesting effect was present.DesignWe studied a cohort of 494,753 adults (as of March 1st, 2020) aged 60 and over in 6,515 NHs in mainland France. This cohort was exposed to the first wave of the COVID-19 pandemic (from March 1st to May 31st, 2020) and was compared with the corresponding, reference cohorts from 2014 to 2019 (using data from the French National Health Data System).Main outcome and measuresThe main outcome was all-cause death. Weekly excess deaths and standardized mortality ratios (SMRs) were estimated.ResultThere were 13,505 excess deaths among NH residents. Mortality increased by 43% (SMR: 1.43). The mortality excess was higher among males than among females (SMR: 1.51 and 1.38, respectively) and decreased with age (SMRs in females: 1.61 in the 60-74 age group, 1.58 for 75-84, 1.41 for 85-94, and 1.31 for 95 or over; Males: SMRs: 1.59 for 60-74, 1.69 for 75-84, 1.47 for 85-94, and 1.41 for 95 or over). We did not observe a harvesting effect (up until August 30th, 2020). By extrapolating to all NH residents nationally (N=570,003), the latter accounted for 51% of the total excess deaths in the general population (N=15,114 out of 29,563).ConclusionNH residents accounted for about half of the total excess deaths in France during the first wave of the COVID-19 pandemic. The excess death rate was higher among males than females and among younger residents than among older residents. We did not observe a harvesting effect. A real-time mortality surveillance system and the identification of individual and environmental risk factors might help to design the future model of care for older dependent adults.Key pointsDuring the first wave of the COVID-19 pandemic in France, the mortality among nursing home residents increased by 43%.Nursing home residents accounted for 51% of the total excess deaths in France.The excess mortality was higher among younger residents than among older residents.The excess mortality was higher among males than among females.We did not observe a harvesting effect during the study period (ending on August 30th, 2020, i.e., three months after the end of the first wave).

BackgroundTumor infiltrating lymphocyte (TIL) products made from tumor digests showed a high overall response rate (ORR; 67%) and complete response (CR) rate (19%) and a safety profile consistent with lymphodepletion and high-dose interleukin (IL)-2 in a retrospective analysis of a single center experience of TILs for compassionate use treatment of advanced cutaneous melanoma (n=21; Hawkins, et al. AACR 2021. ePoster LB150). This subanalysis assesses outcomes for patients who received TILs after prior checkpoint inhibition, a subset with limited treatment options.MethodsPatients with advanced cutaneous melanoma and no standard of care treatment options received lymphodepleting chemotherapy (cyclophosphamide ×2 days; fludarabine ×5 days) followed by TIL infusion and post TIL high-dose IL-2. Safety was assessed by clinically significant adverse events (AEs). Efficacy assessments included ORR, CR rate, and overall survival (OS).ResultsOf 21 patients who underwent treatment between October 2011 and August 2019, median age was 45 years, median number of disease sites was 4, 100% of patients had M1c or M1d disease (33% with M1d), average number of prior therapies was 3 (any checkpoint inhibitor, 91%; BRAF inhibitor [BRAFi], 52%; and MEKi, 24%), and 52% were BRAF-mutated. Twelve patients received prior PD-1i therapy and are reported herein. Baseline characteristics were similar between the overall and prior PD-1i subgroup populations. All patients in the prior PD-1i subgroup received prior CTLA-4i, and all BRAF-mutated patients received prior BRAFi alone ± MEKi. The most commonly reported AEs post-TIL infusion were consistent between the overall and prior PD-1i subgroup populations and included thrombocytopenia (62% and 75%, respectively), pyrexia (57% and 50%), and rigors (43% and 50%). No treatment-related deaths occurred. With a median follow-up of 45.5 months, the ORR and CR rate for the prior PD-1i subgroup were 58% and 8%, respectively. At data cutoff, 2 of 12 patients (17%) had durable ongoing responses (>30 months post-TIL infusion). Median OS in the prior PD-1i subgroup and overall population was 21.3 months.ConclusionsIn this subanalysis of patients with relapsed advanced melanoma after both PD-1i and CTLA-4i, and for some, BRAFi, outcomes of unselected autologous TILs were similar to those observed in all treated patients, with high response rates and a safety profile consistent with that of TIL therapy. Unselected TILs may address the unmet medical need for the poor-risk subset of patients with advanced melanoma who experience disease progression following checkpoint inhibition and, if applicable, targeted therapy.AcknowledgementsThe authors would like to thank all the staff within The Christie NHS Foundation Trust and The Christie Clinic who worked tirelessly to provide high–quality care to all the patients in this report. Medical writing support was provided by Christopher Waldapfel, PharmD, of Instil Bio, Inc. and Jennifer Yang, PhD, of Nexus GG Science, with funding from Instil Bio, Inc.Ethics ApprovalAs a compassionate use study, the treatment was approved by institutional review board and National Health Service commissioning.

Un des principaux obstacles à l’évaluation de la créativité est que ses critères tendent à changer en fonction de ce qui est évalué et de celui ou celle qui fait l’évaluation. Dans cet article, nous avançons qu’il est possible de résoudre ce problème en adoptant une ontologie relationnelle ; c’est-à-dire une ontologie selon laquelle les êtres du monde acquièrent leurs propriétés en entrant en relation avec d’autres êtres. Premièrement, nous montrons les conséquences de cette approche ontologique pour l’évaluation de la créativité : (1) on peut rendre compte de la créativité d’un objet donné (ex. une personne, un produit, ou un processus) en retraçant les êtres (y compris les critères) qui le relient à sa créativité supposée. (2) On peut évaluer la créativité de cet objet en regardant le nombre d’êtres qui participent à cette relation ; c’est-à-dire en regardant ce que nous proposons d’appeler « le degré de solidité » de la relation. (3) On peut ainsi à la fois rendre compte de la spécificité des diverses formes de créativité (notamment leurs critères spécifiques) et les comparer en termes de solidité. En nous appuyant sur ces postulats ontologiques, nous présentons ensuite une nouvelle technique d’évaluation, la Technique de détection des objections (TDO), avant de la mettre à l’épreuve avec un extrait tiré d’une séance de brainstorming. Arguably, one of the main obstacles to creativity assessment is that creativity criteria are likely to change depending on what is assessed and who is making the assessment. In this paper, we argue that we can solve this problem by adopting a relational ontology, i.e., an ontology according to which beings of the world acquire their properties by relating to other beings. First, we present the main consequences of this ontological approach for creativity assessment: (1) Accounting for the creativity of a given object (e.g., a person, a product or a process) involves retracing the beings (including criteria) that relate it to its alleged creativity. (2) One can assess the creativity of this object by looking at the number of beings that substantiate this relation, i.e., by looking at what we propose to call the “degree of solidity” of the relation. (3) One can thus account for the specificity of various forms of creativity (including their specific criteria) and, at the same time, compare them in terms of solidity. Building on these ontological assumptions, we then present a new assessment technique, the Objection Counting Technique (OCT), before putting it to the test using an excerpt taken from a naturally-occurring brainstorming session.

Introduction: A minority of patients (pts) with advanced melanoma achieve long-term survival with immunotherapy and those who relapse following immune checkpoint inhibition (ICI) or BRAFi have limited treatment options. The intrinsic anti-tumor activity and broad neoantigen-specific TCR repertoire of unselected autologous TIL may provide advantages over other treatments in solid tumors, including ICI-refractory melanoma. Methods: This is a retrospective analysis of a single-center experience of TIL for treatment of advanced cutaneous melanoma. Unselected autologous young TIL derived from digested tumor tissue were manufactured under a MHRA Manufacturing Specials license. Pts with advanced cutaneous melanoma and no other treatment options received non-myeloablative lymphodepleting chemotherapy (cyclophosphamide 60 mg/kg/d x 2d, fludarabine 25 mg/m2/d x 5 d [Cy/Flu]), followed by TIL infusion and post-TIL high-dose (HD) IL-2 (600,000-720,000 IU/Kg) on a compassionate use basis. Efficacy for 15 imaging-evaluable pts was reported by investigator assessment of CT/MRI per RECIST 1.1; 6 additional pts were followed using non-RECIST 1.1 imaging (PET) and clinical monitoring. Clinically significant adverse events (AEs) with onset post-TIL infusion were reported for all treated pts. Data cutoff date: 31-DEC-2019. Results: Between OCT-2011 and AUG-2019, 21 pts with advanced cutaneous melanoma were treated with Cy/Flu, TIL (median 31.9 x109 cells infused), and HD IL-2 (median 8 doses). All had high-risk metastatic disease (median 4 sites, stage IV M1d [38%]), an average of 3 prior therapies (any ICI [91%], PD-1 [57%], BRAFi [52%], and MEKi [24%]). With a median follow up of 52.2 mos, the response rate in imaging-evaluable pts (n=15) was 53% with a CR rate of 13%; DCR was 73% including 3 (20%) pts with stable disease. Additional durable responses were observed in the 6 pts followed by PET and clinical monitoring. Responses were generally consistent across subgroups including age, number of disease sites, tumor burden, brain metastases, number of prior lines of therapies, prior PD-1 blockade, prior BRAFi, and prior MEKi. For all treated pts, the median survival was 21.3 mos. Toxicity was generally self-limited and consistent with Cy/Flu and HD IL-2. Common AEs (≥20%, any grade) were thrombocytopenia (62%), pyrexia (57%), rigors (43%), neutropenia (29%), tachycardia (29%), pulmonary edema (24%), and vascular leak (24%); no treatment-related deaths were observed. Conclusion: The high response rate observed in this series exceeds the 41% ORR estimated for TIL in advanced cutaneous melanoma (meta-analysis by Dafni et al. Ann Oncol., 2019) and highlights the successful bench-to-bedside application of unselected autologous TIL to address unmet medical need in advanced melanoma. Use of tumor digests as starting material for manufacturing of TIL demonstrates feasibility of this approach. A multicenter Phase 2 trial of this therapy in advanced melanoma is planned for 2021. Citation Format: Robert E. Hawkins, Yizhou Jiang, Paul C. Lorigan, Fiona C. Thistlethwaite, Manon Pillai, Martine Thomas, Natalia Kirilova, John S. Bridgeman, Gray Kueberuwa, Ryan S. Guest, Zachary J. Roberts. Clinical Feasibility and Treatment Outcomes with Unselected Autologous Tumor Infiltrating Lymphocyte Therapy in Patients with Advanced Cutaneous Melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB150.

Summary Arabidopsis thaliana SNF1-Related-Kinase1 (SnRK1)-Activating Kinase 1 and 2 (AtSnAK1 and 2) have been shown to phosphorylate in vitro and activate the energy signalling integrator, SnRK1. To clarify this signalling cascade in planta, a genetic- and molecular-based approach was developed. Homozygous single AtSnAK1 and AtSnAK2 T-DNA insertional mutants did not display an apparent phenotype. Crossing of the single mutants did not allow the isolation of double mutant plants while self-pollinating the S1-/- S2+/- sesquimutant specifically gave approximatively 22% individuals in their offspring that, when rescued on sugar-supplemented media in vitro, were shown to be AtSnAK1-AtSnAK2 double mutants. Interestingly, this was not obtained in the case of the other sesquimutant, S1+/- S2-/-. Although reduced in size, the double mutant had the capacity to produce flowers, but not seeds. Immunological characterization established the T-loop of the SnRK1 catalytic subunit to be non-phosphorylated in the absence of both SnAKs. When the double mutant was complemented with a DNA construct containing an AtSnAK2 open reading frame driven by its own promoter, a normal phenotype was restored. Therefore, wild-type plant growth and development is dependent on the presence of SnAK in vivo and this is correlated to SnRK1 phosphorylation. These data show that both SnAKs are kinases phosphorylating SnRK1 and thereby they contribute to energy signalling in planta. This article is protected by copyright. All rights reserved.

The CDPK-SnRK superfamily consists of seven types of serine-threonine protein kinases: calcium-dependent protein kinase (CDPKs), CDPK-related kinases (CRKs), phosphoenolpyruvate carboxylase kinases (PPCKs), PEP carboxylase kinase-related kinases (PEPRKs), calmodulin-dependent protein kinases (CaMKs), calcium and calmodulin-dependent protein kinases (CCaMKs), and SnRKs. Within this superfamily, individual isoforms and subfamilies contain distinct regulatory domains, subcellular targeting information, and substrate specificities. Our analysis of the Arabidopsis genome identified 34 CDPKs, eight CRKs, two PPCKs, two PEPRKs, and 38 SnRKs. No definitive examples were found for a CCaMK similar to those previously identified in lily (Lilium longiflorum) and tobacco (Nicotiana tabacum) or for a CaMK similar to those in animals or yeast. CDPKs are present in plants and a specific subgroup of protists, but CRKs, PPCKs, PEPRKs, and two of the SnRK subgroups have been found only in plants. CDPKs and at least one SnRK have been implicated in decoding calcium signals in Arabidopsis. Analysis of intron placements supports the hypothesis that CDPKs, CRKs, PPCKs and PEPRKs have a common evolutionary origin; however there are no conserved intron positions between these kinases and the SnRK subgroup. CDPKs and SnRKs are found on all five Arabidopsis chromosomes. The presence of closely related kinases in regions of the genome known to have arisen by genome duplication indicates that these kinases probably arose by divergence from common ancestors. The PlantsP database provides a resource of continuously updated information on protein kinases from Arabidopsis and other plants.

On cherche ici à poser les bases conceptuelles et méthodologiques de l'application de la notion d'écosystème d'affaire à l'analyse des Média Participatifs. On considère les Média Participatifs comme des organisations d'édition dont la viabilité dépend directement des relations de concurrence et de coopération qu'elles entretiennent avec d'autres organisations. Il s'agit alors de dégager les différents facteurs de viabilité s'appliquant aux Média Participatifs (MP). On distingue (pour le moment) deux grands ensembles de facteurs. (1) On isole d'abord des facteurs s'exerçant sur toute organisation d'édition. Il s'agit de facteurs liés à la nature discursive de n'importe quelle publication. On avance ainsi que la qualité des contenus édités par une organisation d'édition X détermine les relations de concurrence et de coopération « naturelles » entre cette organisation X et les autres organisations d'édition ainsi que la capacité de l'organisation X à satisfaire des utilisateurs. (2) On isole ensuite des facteurs s'exerçant spécifiquement sur les MP. Il s'agit de facteurs liés aux technologies d'édition/diffusion sur lesquelles reposent les MP. On considère ainsi que la viabilité d'un MP dépend de : 1) Sa capacité à surmonter le double problème de lisibilité du contenu et d'organisation du travail d'édition inhérent aux technologies d'Edition Continue Interactive. 2) Sa capacité à tirer profit du World Wide Web en visant un public de niche et en coopérant via des liens hypertextuels avec d'autres publications Web. 3) Sa capacité à maximiser les effets de synergie qui caractérisent son processus d'édition. L'identification de ces différents facteurs de viabilité permet alors de dessiner les contours d'une méthode permettant d'évaluer la viabilité d'un MP donné et ce faisant d'explorer le fonctionnement de l'écosystème dans lequel il s'inscrit.

The recent gain in knowledge concerning enzymes involved in signal transduction pathways is a direct consequence of the considerable advances made in molecular biology. Protein kinases and protein phosphatases, the two major enzymes implicated in post-translational modifications, have been studied in particular. The number of characterized plant genes and/or cDNAs encoding these enzymes is increasing everyday. Since 1991, 26 genes and cDNAs coding for plant protein phosphatases have been isolated and characterized. The huge number of protein kinases (estimated at several thousands) makes it impossible to give an exhaustive list of the genes already identified, but a classification of these enzymes, based on phylogenetic criteria, allows us to appreciate the range of functions this protein family may play in plants.

International audience; SNF1-related protein kinase-1 (SnRK1), the plant kinase homolog of mammalian AMP-activated protein kinase (AMPK), is a sensor that maintains cellular energy homeostasis via control of anabolism/catabolism balance. AMPK-dependent phosphorylation of p27(KIP1) affects cell-cycle progression, autophagy and apoptosis. Here, we show that SnRK1 phosphorylates the Arabidopsis thaliana cyclin-dependent kinase inhibitor p27(KIP1) homologs AtKRP6 and AtKRP7, thus extending the role of this kinase to regulation of cell-cycle progression. AtKRP6 and 7 were phosphorylated in vitro by a recombinant activated catalytic subunit of SnRK1 (AtSnRK11). Tandem mass spectrometry and site-specific mutagenesis identified Thr152 and Thr151 as the phosphorylated residues on AtKRP6- and AtKRP7, respectively. AtSnRK1 physically interacts with AtKRP6 in the nucleus of transformed BY-2 tobacco protoplasts, but, in contrast to mammals, the AtKRP6 Thr152 phosphorylation state alone did not modify its nuclear localization. Using a heterologous yeast system, consisting of a cdc28 yeast mutant complemented by A.thaliana CDKA;1, cell proliferation was shown to be abolished by AtKRP6(WT) and by the non-phosphorylatable form AtKRP6(T152A), but not by the phosphorylation-mimetic form AtKRP6(T152D). Moreover, A.thaliana SnRK11/KRP6 double over-expressor plants showed an attenuated AtKRP6-associated phenotype (strongly serrated leaves and inability to undergo callogenesis). Furthermore, this severe phenotype was not observed in AtKRP6(T152D) over-expressor plants. Overall, these results establish that the energy sensor AtSnRK1 plays a cardinal role in the control of cell proliferation in A.thaliana plants through inhibition of AtKRP6 biological function by phosphorylation.

International audience; This article is a call to describe Participatory Design (PD) projects in the making, i.e. to show how the heterogeneous elements in the field are gradually organised in a participatory manner as the projects progress. It is based on two arguments. The first is a negative argument. Very often, PD projects are not described in the making. As a result, the landmarks to be used to evaluate them remain unclear or invisible. The second argument is of a more positive nature. The articles that do describe projects in the making enable landmarks to be defined that can be effectively used to evaluate PD projects. The notion of emerging groups is one of these landmarks.

The perception of nutrient and energy levels inside and outside the cell is crucial to adjust growth and metabolism to available resources. The signaling pathways centered on the conserved TOR and SnRK1/Snf1/AMPK kinases have crucial and numerous roles in nutrient and energy sensing and in translating this information into metabolic and developmental adaptations. In plants evidence is mounting that, like in other eukaryotes, these signaling pathways have pivotal and antagonistic roles in connecting external or intracellular cues to many biological processes, including ribosome biogenesis, regulation of translation, cell division, accumulation of reserves and autophagy. Data on the plant TOR pathway have been hitherto rather scarce but recent findings have shed new light on its roles in plants. Moreover, the distinctive energy metabolism of photosynthetic organisms may reveal new features of these ancestral eukaryotic signaling elements.

Knowledge management relies – willingly or unwillingly – on three components: organizational structures, communities of practice, and material settings. In an organization, those three components are interdependent, that is, each of them enters into the composition of the others. They are also resistant to each other, which means that each of them marks the limits of the others‟ action. This relationship allows us to understand why a knowledge management system, if conceived strictly from the point of view of organizational structures, does not benefit from the support of communities of practice. It also suggests that emphasis should be put on the settings that reinforce both organizational control and the communities‟ autonomy. Finally, this allows us to understand that the appropriation of such a setting by communities of practice implies a slight modification of organizational structures.

Arabidopsis thaliana sucrose nonfermenting 1-related protein kinase 1 complexes belong to the SNF1/AMPK/SnRK1 protein kinase family that shares an ancestral function as central regulators of metabolism. In A. thaliana, the products of AtSnAK1 and AtSnAK2, orthologous to yeast genes, have been shown to autophosphorylate and to phosphorylate/activate the AtSnRK1.1 catalytic subunit on Thr(175). The phosphorylation of these kinases has been investigated by site-directed mutagenesis and tandem mass spectrometry. The autophosphorylation site of AtSnAK2 was identified as Thr(154), and it was shown to be required for AtSnAK catalytic activity. Interestingly, activated AtSnRK1 exerted a negative feedback phosphorylation on AtSnAK2 at Ser(261) (Ser(260) of AtSnAK1) that was dependent on AtSnAK autophosphorylation. The dynamics of these reciprocal phosphorylation events on the different kinases was established, and structural modeling allowed clarification of the topography of the AtSnAK phosphorylation sites. A mechanism is proposed to explain the observed changes in the enzymatic properties of each kinase triggered by these phosphorylation events.

The AKin10 gene from Arabidopsis thaliana encoding a putative Ser/Thr protein kinase (PK) has been isolated and characterized. The AKin10-encoding gene is located on a genomic 5.4-kb BamHI fragment and contains ten introns, one being located in the 5' untranslated region. The deduced amino acid sequence of AKin10 is 65% identical over the catalytic domain to the yeast PK (SNF1). SNF1 is essential for the derepression of many glucose-repressible genes, including Suc2 which encodes invertase. Southern blot hybridization experiments suggested the presence of one copy of the gene per haploid genome of A. thaliana. Northern hybridization experiments indicated that this gene is expressed in roots, shoots and leaves. AKin10 may play an important role in a signal transduction cascade regulating gene expression and carbohydrate metabolism in higher plants.

International audience; Every knowledge management relies – willingly or unwillingly – on three components: organizational structures, communities of practice, and material settings. Those three components are both interdependent and resistant to each other, that is to say none of them can substitute another. In the perspective of improving an organization's knowledge management, this relationship suggests that emphasis should be put on the settings that reinforce both organizational control and the communities' autonomy. This is what Participative Document Spaces enable by facilitating access to the in vivo working of the different communities of practice. This relationship also allows understanding that putting in place such a setting implies to slightly modify the organizational structures as well as the practices of the communities.; Toute gestion des connaissances repose volontairement ou involontairement sur trois composantes : les structures organisationnelles, les communautés de pratique, et les dispositifs matériels. Ces trois composantes sont à la fois interdépendantes et résistantes les unes aux autres, c'est-à-dire qu'aucune d'elles ne peut se substituer à une autre. Dans la perspective d'améliorer la gestion des connaissances d'une organisation, ces relations invitent ainsi à s'intéresser aux dispositifs permettant de renforcer à la fois le contrôle organisationnel et l'autonomie des communautés. C'est ce que permettent de faire les Espaces documentaires Participatifs en facilitant l'accès au travail in vivo des différentes communautés de pratique. Ces relations permettent également de comprendre que la mise en place d'un tel dispositif implique de modifier légèrement les structures organisationnelles autant que les pratiques des communautés.

International audience; Cet article propose de mettre en évidence les défis de la gestion des connaissances en explorant les relations d'interdépendance entre les structures organisationnelles, les communautés de pratique et les dispositifs matériels. L'analyse de ces relations fait d'abord apparaître la formation d'agrégats de communautés de pratique entre lesquels des pratiques – et des connaissances – communes se développent difficilement. Le premier défi de la gestion des connaissances est alors de favoriser le développement de telles pratiques. L'analyse de ces relations fait également apparaître que le problème des agrégats de communautés est un problème chronique. Apparaît alors un second défi, enchâssé dans le premier, qui est de développer une gestion des connaissances suffisamment souple pour s'adapter à l'émergence continuelle de nouveaux agrégats de communautés.

International audience; Cet article vise à faire apparaître les conditions d'une gestion efficace des connaissances. Ces conditions peuvent être déduites des relations entre trois grandes composantes : les structures organisationnelles, les communautés de pratiques et les dispositifs matériels. Les structures organisationnelles restreignent l'autonomie des communautés de pratique, notamment en renforçant les interactions entre certaines communautés au détriment d'autres communautés. Les structures organisationnelles compensent ainsi les limites inhérentes aux dispositifs matériels permettant les interactions entre communautés. Ce faisant, elles tendent aussi à pérenniser les limites de ces dispositifs matériels. Dans ces conditions, nous pensons qu'une gestion plus efficace des connaissances doit modifier les dispositifs matériels existants de façon à renforcer à la fois l'autonomie des communautés et le contrôle organisationnel.

The proteins kinases SNF1/AMPK/SnRK1 are a subfamily of serine/threonine kinases that act as metabolite sensors to constantly adapt metabolism to the supply of, and demand for, energy. In the yeast Saccharomyces cerevisiae, the SNF1 complex is a central component of the regulatory response to glucose starvation. AMP activated protein kinase (AMPK) the mammalian homologue of SNF1, plays a central role in the regulation of energy homeostasis at the cellular as well as the whole-body levels. In Arabidopsis thaliana, SnRK1.1 and SnRK1.2 have recently been described as central integrators of a transcription network for stress and energy signalling. In this study, biochemical analysis established SnRK1.1 as the major SnRK1 isoform both in isolated cells and leaves. In order to elucidate the function of SnRK1.1 in Arabidopsis thaliana, transgenic plants over-expressing SnRK1.1 were produced. Genetic, biochemical, physiological and molecular analyses of these plants revealed that SnRK1.1 is implicated in sugar and ABA signalling pathways. Modifications of the starch and soluble sugar content were observed in the 35S:SnRK1.1 transgenic lines. Our studies also revealed modifications of the activity of essential enzymes such as nitrate reductase or ADP-glucose pyrophosphorylase, and of the expression of several sugar-regulated genes, confirming the central role of the protein kinase SnRK1 in the regulation of metabolism.

The architectural excavations and studies of the chapel showed the existence of an earlier edifice. In fact, the present chapel is the choir of a much more complex structure, part of which was rediscovered in excavating, and another part found in the wall of an adjacent building. These different elements permitted an hypothetical reconstitution; it appears to be an edifice of a square plan, built around a central hall in the shape of a Greek cross. Excavations have not permitted the exact dating of this monument. However, the presence of sepulchers and the relative chronology allow for the taking of the construction of this building to the Early Middle Ages, and perhaps to the Caroligian Era. The excavations showed the architectural evolution of this monument until the French Revolution., Les fouilles et l'étude architecturale de la chapelle Saint-Jean-de-Taravon à Volonne (Alpes-de-Haute-Provence) ont montré l'existence d'un édifice antérieur. En effet, la chapelle actuelle se trouve être l'ancien chœur d'un édifice beaucoup plus complexe, dont une partie a été retrouvée en fouille et une autre reconnue en élévation dans un bâtiment mitoyen. Ces différents éléments ont permis de proposer une hypothèse de restitution: il s'agirait d'un édifice de plan carré, organisé autour d'une salle centrale en croix grecque. Les fouilles n'ont pu apporter une datation précise de ce monument. Néanmoins, la présence de sépultures et la chronologie relative permettent de situer la construction de cet édifice au haut Moyen Age, et peut-être à l'époque carolingienne. Les fouilles ont également montré l'évolution architecturale de ce monument, devenu prieuré, jusqu'à la Révolution française., Thomas Martine. Un site du Haut Moyen Age provençal : Saint-Jean-de-Taravon (Volonne, Alpes-de-Haute-Provence). In: Revue archéologique de Narbonnaise, tome 24, 1991. pp. 235-254.

International audience; The SNF1/AMPK/SnRK1 kinases are evolutionary conserved kinases involved in yeast, mammals, and plants in the control of energy balance. These heterotrimeric enzymes are composed of one alpha-type catalytic subunit and two gamma- and beta-type regulatory subunits. In yeast it has been proposed that the beta-type subunits regulate both the localization of the kinase complexes within the cell and the interaction of the kinases with their targets. In this work, we demonstrate that the three beta-type subunits of Arabidopsis (Arabidopsis thaliana; AKINbeta1, AKINbeta2, and AKINbeta3) restore the growth phenotype of the yeast sip1Deltasip2Deltagal83Delta triple mutant, thus suggesting the conservation of an ancestral function. Expression analyses, using AKINbeta promoterbeta-glucuronidase transgenic lines, reveal different and specific patterns of expression for each subunit according to organs, developmental stages, and environmental conditions. Finally, our results show that the beta-type subunits are involved in the specificity of interaction of the kinase with the cytosolic nitrate reductase. Together with previous cell-free phosphorylation data, they strongly support the proposal that nitrate reductase is a real target of SnRK1 in the physiological context. Altogether our data suggest the conservation of ancestral basic function(s) together with specialized functions for each beta-type subunit in plants.

International audience; La gestion des connaissances participative est un processus de gestion ouvert sur la diversité des acteurs impliqués et sur la multiplicité de leurs savoirs quand il est difficile d’anticiper les besoins qui seront ceux des destinataires futurs des informations prises en considération. Cette situation d’incertitude est caractéristique du projet dans lequel nous sommes impliqués, qui vise à redocumenter et à redocumentariser un certain nombre de résultats d’études techniques dans la perspective de leur exploitation ultérieure (études consacrées à la gestion de déchets radioactifs à longue durée de vie).

The mechanism underlying the accumulation of specific RNA encoding the G form (photosynthetic) of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) has been studied during the greening process of Sorghum and maize leaves. From a Γgt11 cDNA library, a cDNA clone for Sorghum leaf PEPC was identified, sequenced and found to correspond to the photosynthetic type of the protein by means of monoclonal antibodies. ‘Run off transcription experiments showed that Sorghum or maize leaf nuclei extracted from light-grown plants produced considerably more PEPC mRNAs than those obtained from dark-grown plants. Ribosomal RNA and LHCP mRNAs increased concomitantly. ‘Run off’ analyses also indicated that transcription of these mRNAs was phytochrome-mediated. In plants grown in normal photoperiodic conditions, the PEPC-RNA pool followed a rhythmical behaviour, being low at night and recovering its maximum level the following day. However, after 24 h of dark treatment, both the RNA pool and the transcription capacity were found to be strongly decreased. In contrast, the PEPC protein remained constant during the same developmental period.

The SNF1-related kinases are considered to be crucial elements of transcriptional, metabolic and developmental regulation in response to stress. In yeast, SNF1 is one of the main regulators in the shift from fermentation to aerobic metabolism; AMPK, its mammalian counterpart, is a master metabolic regulator involved in a variety of metabolic disorders such as diabetes and obesity. The aim of this review is to examine the literature concerning SnRK1 proteins, the SNF1 homologues in plants. The remarkable structural similarities between the plant complexes and those of yeast and mammalian suggest the existence of a common ancestral function in the regulation of energy and carbon metabolism. We will also highlight some distinctive features acquired by the plant proteins during evolution.

International audience; The sucrose nonfermenting-1 protein kinase (SNF1)/AMP-activated protein kinase subfamily plays a central role in metabolic responses to nutritional and environmental stresses. In yeast (Saccharomyces cerevisiae) and mammals, the beta- and gamma-noncatalytic subunits are implicated in substrate specificity and subcellular localization, respectively, and regulation of the kinase activity. The atypical betagamma-subunit has been previously described in maize (Zea mays), presenting at its N-terminal end a sequence related to the KIS (kinase interacting sequence) domain specific to the beta-subunits (Lumbreras et al., 2001). The existence of two components, SNF1-related protein kinase (SnRK1) complexes containing the betagamma-subunit and one SnRK1 kinase, had been proposed. In this work, we show that, despite its unusual features, the Arabidopsis (Arabidopsis thaliana) homolog AKINbetagamma clearly interacts with AKINbeta-subunits in vitro and in vivo, suggesting its involvement in heterotrimeric complexes located in both cytoplasm and nucleus. Unexpectedly, a transcriptional analysis of AKINbetagamma gene expression highlighted the implication of alternative splicing mechanisms in the regulation of AKINbetagamma expression. A two-hybrid screen performed with AKINbetagamma as bait, together with in planta bimolecular fluorescence complementation experiments, suggests the existence of interactions in the cytosol between AKINbetagamma and two leucine-rich repeats related to pathogen resistance proteins. Interestingly, this interaction occurs through the truncated KIS domain that corresponds exactly to a GBD (glycogen-binding domain) recently described in mammals and yeast. A phylogenetic study suggests that AKINbetagamma-related proteins are restricted to the plant kingdom. Altogether, these data suggest the existence of plant-specific SnRK1 trimeric complexes putatively involved in a plant-specific function such as plant-pathogen interactions.

The SNF1/AMPK/SnRK1 heterotrimeric kinase complex is involved in the adaptation of cellular metabolism in response to diverse stresses in yeast, mammals and plants. Following a model proposed in yeast, the kinase targets are likely to bind the complex via the non-catalytic beta-subunits. These proteins currently identified in yeast, mammals and plants present a common structure with two conserved interacting domains named Kinase Interacting Sequence (KIS) and Association with SNF1 Complex (ASC), and a highly variable N-terminal domain. In this paper we describe the characterisation of AKINbeta3, a novel protein related to AKINbeta subunits of Arabidopsis thaliana, containing a truncated KIS domain and no N-terminal extension. Interestingly the missing region of the KIS domain corresponds to the glycogen-binding domain (beta-GBD) identified in the mammalian AMPKbeta1. In spite of its unusual features, AKINbeta3 complements the yeast sip1Deltasip2Deltagal83Delta mutant. Moreover, interactions between AKINbeta3 and other AKIN complex subunits from A. thaliana were detected by two-hybrid experiments and in vitro binding assays. Taken together these data demonstrate that AKINbeta3 is a beta-type subunit. A search for beta-type subunits revealed the existence of beta3-type proteins in other plant species. Furthermore, we suggest that the AKINbeta3-type subunits could be plant specific since no related sequences have been found in any of the other completely sequenced genomes. These data suggest the existence of novel SnRK1 complexes including AKINbeta3-type subunits, involved in several functions among which some could be plant specific.