The EU Water Framework Directive recently encouraged the development of innovative ecotoxicological tools. However and due to the lack of reference genome in most environmentally relevant invertebrates species, as Gammarus fossarum, the biomarkers currently used are indirect assays previously developed and validated in vertebrates. Nevertheless molecular divergence and diversity of protein functions acquired through animal evolution rules out the possibility and the relevance to directly use many vertebrates-derived biomarkers. Moreover, the fact that each assay is specific to only each biomarker makes difficult their routinely use. At last, biomarker measurements are currently performed on whole organism (or pools of organisms), leading to a decrease in its sensitivity due to biological dilution and/or specific tissue responses and making impracticable to predict and provide information on impaired physiological functions (organ). Based on a multidisciplinarity and complementary consortium (high-throughput proteomics, ecotoxicology, TK/TD modelling, analytical chemistry and regulatory), the objective of the APPROve project is to define, through a case-study with metals, a framework to formalize the fate and effects of contaminants in a sentinel species, Gammarus fossarum, in order to improve water quality monitoring with an effective transfer of new tools to the operational sphere. The APPROve project has two mains objectives: 1) producing knowledge on accumulation and fate of metals in G. fossarum and their associated responses at organs level, in order to propose specific and relevant biomarkers and, 2) improving diagnostic tools currently available with caged G. fossarum for the assessment of contamination and toxicity in aquatic systems through TK/TD models. Based on advanced analytical approaches (radioelements and mass spectrometry), critical lack of knowledge will be addressed about i) bioaccumulation and fate (organotropism) of metals in an invertebrate species and the identification of the organ playing a major role in the underlying processes, ii) the molecular physiology in G. fossarum to better describe and understand the impact of metals, in order to propose relevant biomarkers and to establish links with life trait impairments. The bioaccumulation dynamic of Ag, Cd, and Zn within the different organs of G. fossarum will be formalized by a TK model for each metal that will take into account the contributions of the different bioaccumulation pathways and the concentration-dependence. Then, the effects measured on biological endpoints will be linked to internal concentrations through a TD model (DEBtox model). Based on outputs of modeling and proteomic approaches, diagnostic tools currently available with caged G. fossarum for the assessment of contamination levels and toxicity impacts of aquatic systems will be improved, and at last transferred to the operational sphere. On one hand, based on TK/TD models, metal contamination levels in caged G. fossarum could be also used i) to check the compliance to water-EQS and ii) to predict a toxic risk. On the other hand, based on field-expertise and experiment capacity of the SME BIOMAE, threshold values will be defined for new molecular biomarkers, in order to validate their use as diagnostic tools for field biomonitoring. APPROve will (i) develop and calibrate models, allowing to better formalize accumulation and fate of metals into organisms, (ii) bridge the gap in molecular ecophysiology of a sentinel species, giving strong information and data on key organs involved in the toxicity of metallic contaminants, (iii) propose benchmark values for each new targeted-organ biomarker validated and at last (iv) highlight the originality of coupling proposed methodologies to develop TK/TD models and of their used to propose integrated diagnostic tools (molecular and physiological) for environmental monitoring.

MALDI-TOF mass spectrometry has revolutionized pathogen identification by shortening this step by 24h compared to traditional biochemical tests. In contrast, the antibiotic therapy choice remains largely driven by conventional antibiotic susceptibility testing, hence with a delay of 18-24h after ID. Commercially available products exist today for quicker determination of resistance determinants, either based on imaging, molecular biology tools, or on colorimetric and immuno-chromatographic testing. There is however no universal technology available for rapid (t<6hrs), cost effective, concomitant ID, AST characterization and virulence evaluation. By leveraging on an innovative in-real-time adaptive targeted mass spectrometry technology, we propose to overcome this limitation, directly from a positive blood culture, and with a time-to-result of less than an hour.

The aim of the OxCyCat-CO2 project is to use CO2 as both a reagent and a solvent (supercritical conditions) to prepare cyclic carbonates of interest starting from the corresponding olefins via a one-pot reaction with hydrogen peroxide or molecular oxygen. To achieve this, new multifunctional heterogeneous catalysts will be developed by combining, on the same mesoporous support, active phases that are able to catalyze the epoxidation of alkenes and the further cycloaddition of CO2 under the same experimental conditions. Two ligand libraries, lacunary polyoxometalates (W-POMs) and nitrogen Schiff bases (BS-4Ns) are available within the consortium. W-POM species will be used to catalyze the epoxidation of alkenes in the presence of H2O2 and will be compared to Mn(III) complexes involving BS-4N ligands also able to catalyze epoxidations in the presence of dioxygen and of an aldehyde. The same nitrogen Schiff bases associated with zinc (II) will also be exploited for the cycloaddition reaction of CO2 onto epoxides. These different active phases will then be combined, thus allowing the definition of two classes of bifunctional catalytic systems according to whether they involve i) both polyoxometalates and complexes with nitrogen Schiff bases ("Mix POM / BS-4N ") intended for operations with H2O2 / CO2, or (ii) only complexes derived from Schiff bases (" all BS-4Ns ") intended for operations with O2 / CO2. In a first approach, these combinations will be tested in solution. In particular, we shall ensure that the two active phases can work in synergy under comparable solvent, temperature and pressure conditions without interfering with one another. An important challenge of the project is to lower the optimum temperature of the CO2 cycloaddition reaction via the design of suitable BS-4N ligands with the help of theoretical chemistry colleagues involved in the consortium. The reactions will first be carried out in organic solvent, a priori in acetonitrile, and then transposed into supercritical CO2. This last solvent will be preferred, especially for combinations of CO2 with O2 in order to limit the risks of explosiveness. Moreover, this association suggests the possibility of using CO2 emissions that are not completely purified. Some terminal and internal alkenes representative of industrial requirements will be considered. Chiral versions of the catalytic "all BS-4Ns" system will be developed and then tested in the context of the enantioselective epoxidation of alkenes and then in the overall process. Polyoxometalates will then be grafted by amide bonds to the surface of mesoporous silicas and complexes with BS-4N ligands by Si-O-Si bonds. For the sake of simplification, and ultimately, in the case of "all BS-4Ns", the use of a single metal / BS-4N complex to perform both reactions (epoxidation and cycloaddition) will be preferred. In the case of "Mix POM / BS-4N", the pre-association of the two entities in one edifice that will be grafted later through amide linkages will be encouraged.

This program is dedicated to a new type of resistance to thyroid hormone (RTHa), which is due to a mutation of the TRa1 receptor, encoded by the THRA gene .This rare genetic disease, for which the clinical features are surprisingly variable, was discovered in 2012. The patients are heterozygous and carry mutations transforming the receptor into a dominant negative. The pleiotropic consequences of the mutation usually include impaired growth and ossification, mental retardation and constipation. However TH levels in serum remain within the normal range. As most clinicians are not aware of the disease, and since the symptoms are not highly specific, it is likely that the number of patients is highly underestimated, despite its debilitating consequences. We gathered 4 partners with complementary skills, ranging from biochemistry, nuclear magnetic resonance analysis and mouse genetics, to clinical studies to shed new light on all aspects of this disease: diagnosis, etiology and therapy. Our first goal is to generate knowledge to facilitate RTHa diagnosis. For this purpose, our program will use available animal and cellular models of RTHa, and generate new ones. The molecular basis underlying the phenotype variability associated with the different THRA mutations will be deciphered. Finally we will provide proof of principle for treatment. To reach these objectives we have divided the program into the following tasks. Task1 is dedicated to the dissemination of RTHa knowledge and the discovery of new patients. An international network of hospitals, in France and China, will help to identify putative RTHa patients for which DNA sequencing of THRA will be performed. Task 2 goal is to generate new mouse models. Although we already possess a large collection of mouse strains with THRA mutations, we will generate other “tailor-made’” models, using the CRISPR/Cas9 editing method. We will copy mutations found in the ligand binding or DNA binding domain of TRa1, present in patients or only in anonymous exome databases. In Task 3, we will use the full collection of mouse models to look for a discriminant metabonomic signature of RTHa in urine or serum. This approach will then transpose to patient samples in order to provide new diagnostic tools. Task 4 is conceived to develop a pipeline for a full structural and functional characterization of THRA mutations. This will include biochemical studies of the mutant receptors in vitro, transcriptional analyses of cells co-expressing mutant and wild-type TR and phenotyping mice carrying these mutations. Our preliminary data encourages us to focus our analysis on 2 cell types, of high clinical relevance: GABAergic neurons and macrophages. Task 5 intends to promote further clinical exploration in patients and to provide a proof of principle for therapeutic intervention. We will test in our mouse and cellular models the possibility of using approved drugs, which have the property of inhibiting the activity of the co-repressors that mutant TRa1 recruits in a TH-independent manner on the chromatin. Such compounds should thus antagonize the detrimental effect exerted by the mutant TRa1 receptor and provide some improvement of the RTHa symptoms.

The BIOPOLIOL project proposes a new way to manufacture bio-based polyhydroxyurethane (PHU) foams, derived from technical lignins for the insulation industry. Lignin is a natural resource with a renewable aromatic structure that is co-produced in the pulp industry in large quantities (up to 100 million tons/year) and that has been under-used until now. An innovative valorization of lignin consists in using it as a macromonomer for the preparation of new biosourced polymers. Continuously growing, due to the high demand in many applications of everyday life, the polymer industry is now facing global warming and the depletion of fossil fuels by using sustainable resources. According to recent reports, of the 360 Mt of polymers produced worldwide, the share of bio-based polymers is only 1%. The incorporation of lignin or modified lignin into industrial polymers can lead to foams with new properties such as mechanical strength and fire resistance. Polyurethane foams (PUR) are widely used in the construction industry as insulating materials. However, the synthesis of PUR relies on the use of toxic isocyanates. The most promising alternative to isocyanate-free polyurethanes is the polyaddition of cyclic carbonate and amine groups to produce UHP, which are good candidates to replace conventional PUR. The main objective of the BIOPOLIOL project is to provide access to new biobased polyhydroxyurethane (PHU) polymers for the industry through the incorporation of functionalized lignin derivatives. Oligomers enriched in OH functions will be targeted by a controlled depolymerization of lignin in the presence of catalysts promoting the breaking of ether bonds and the demethylation of aromatic methoxy functions (P1, IRCELYON). The characterization of lignins and converted fractions will be performed by classical methods well known by the partners such as GPC, NMR and GCXGC. However, these methods do not allow to characterize precisely the produced oligomers and dedicated analytical methods must be implemented. In particular, we propose the development of a complete LC x SFC-QToF method to perform the characterization of OH oligomers and its subsequent use to monitor lignin depolymerization (P3, ISA). The separation of OH-enriched oligomers is also a challenge. Purification will be implemented using membranes either as an innovative approach in combination with liquid/liquid extraction, with a hollow fiber membrane contactor, or as a more conventional approach in the form of a membrane cascade by P2 (ABI). Then, the functionalization of the oligomeric fragments will be carried out in particular by using OH groups. Two types of modifications will be developed, to obtain lignins or oligomers with cyclic carbonate or primary amine groups by P4 (ICPEES). The modified lignins or oligomers will then be reacted with bifunctional molecules presenting complementary chemical functions (amines or cyclic carbonates). Their incorporation in formulations containing surfactants and blowing agents will finally allow the production of NIPU foams (Soprema, P5). The comparison between the reactivity and the final properties of the material obtained with the various lignin oligomers will allow to identify the structure-properties relationships according to the main characteristics of the oligomers. Finally, Soprema will carry out a technico-economic study to evaluate the interest of this approach for an industrial valorization on the basis of the partners' data.