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PhD subjects

9 sujets IRAMIS

Dernière mise à jour : 17-09-2019


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• Soft matter and complex fluids

 

Lithium metal polymer" Batteries: towards operation at ambient temperature

SL-DRF-19-0563

Research field : Soft matter and complex fluids
Location :

Laboratoire Léon Brillouin

Groupe Biologie et Systèmes Désordonnés

Saclay

Contact :

Jean-Marc ZANOTTI

Starting date : 01-10-2019

Contact :

Jean-Marc ZANOTTI
CEA - DRF/IRAMIS/LLB/GBSD

+33(0)476207582

Thesis supervisor :

Jean-Marc ZANOTTI
CEA - DRF/IRAMIS/LLB/GBSD

+33(0)476207582

Personal web page : http://iramis.cea.fr/Pisp/jean-marc.zanotti/

Laboratory link : http://www-llb.cea.fr/

More : http://liten.cea.fr/cea-tech/liten/Pages/Accueil.aspx

This PhD subject proposes an original way to allow the use of "lithium metal polymer" batteries at room temperature. This objective will be achieved by combining three effects:

i) The nanometric confinement of the electrolyte in membranes based on vertically aligned Carbon NanoTubes (VA-CNT),

ii) The use of low molecular weight polymer (here Poly(Ethylene Oxide (PEO))

iii) One-dimensional ionic conduction.



The conduction of the CNT confined Lithium will be driven by two characteristic distances: the pore diameter (1-4 nm) and the total VA-CNT length (from 10 to 500 micrometers). Rational modeling of the transport properties over distances differing by orders of magnitude naturally calls for a multiscale approach.

Therefore, as for its fundamental Science aspect, the primary goal is to develop an experimental multi-scale approach to bridge the broad time and spatial scales (eight orders of magnitude) relevant to the high-mobility-in-tight-1D-spaces we are seeking.

The applied research fold of the study is to prove the validity of the concept.
Reaction-ready cubosomes: a self-assembled nano-platform for multifunctional delivery

SL-DRF-19-0936

Research field : Soft matter and complex fluids
Location :

Service Nanosciences et Innovation pour les Materiaux, la Biomédecine et l'Energie

Laboratoire Interdisciplinaire sur l'Organisation Nanométrique et Supramoléculaire

Saclay

Contact :

Christophe FAJOLLES

Patrick GUENOUN

Starting date : 01-11-2019

Contact :

Christophe FAJOLLES
CEA - DSM/IRAMIS/NIMBE/LIONS

01 69 08 99 60

Thesis supervisor :

Patrick GUENOUN
CEA - DRF/IRAMIS/NIMBE/LIONS

01-69-08-74-33

Personal web page : http://iramis.cea.fr/Pisp/patrick.guenoun/index.html

Laboratory link : http://iramis.cea.fr/nimbe/lions/

Cubosomes are nanometric particles formulated from lipid bicontinuous cubic phases. Water-lipid bicontinuous cubic phases [Luzzati, 1962] are made from a single continuous lipid bilayer, where the bilayer midplane is coincident with an infinite periodic minimal surface and subdivides the space into two interpenetrating, but not connected water channels [Scriven, 1976]. The large surface area of the lipid-water interface (400 m2g-1) makes these nano-objects suitable for entrapment of proteins, peptides, and other biomolecules. Also their polar-apolar continuous domains allow for the encapsulation of a broad range of hydrophilic-hydrophobic molecules and for a slow release of cargo, maintaining the therapeutic concentration range over a longer period of time. Cubosomes suffer however from limited possibilities to control load and delivery of potential cargo. The size of the water channels, one of the few available structural parameters, can only be varied in a relatively narrow size range and the chemical nature of the lipids(here monoolein named MO), defining also the lipid-water interface, cannot be significantly modified without compromising the integrity of the structure.



In this PhD project we aim at bringing the control of uptake and release in cubosomes to new possibilities by engineering reaction-ready cubosomes and achieving flexibility in cubosome functionalization by allowing for in-situ multi-dimensional orthogonal reactions combined with the power of inclusion complexation provided by cyclodextrins (CDs). In practice, standard structure-forming lipids will be functionalized to allow for orthogonal click-chemistry reactions to occur at the lipidwater interface in the channels of these lipid mazes. This will open the pathway for devising sophisticated strategies for the kinetics of charging and discharging the nanoparticles with a variety of relevant payloads.

The structure of cubosomes will be characterized by several techniques, such as small angle X-ray scattering (SAXS), cryo-electron microscopy (cryo-EM), NMR and differential scanning calorimetry (DSC) [Angelova, 2015; Li, 2015].

Despite extensive work on cubosomes, outstanding questions still hamper progress in the understanding and control of the encapsulation and release mechanisms. How many of the channels actually communicate with the solution, are all cubosomes open, what is the role of exact shape and size on the exchange processes, why are cubosomes most often decorated with single bilayer handles and what role do they play … these questions require designing advanced experimental geometries to inter-relate structure and properties. We will address them by performing kinetic experiments on single cubosomes under a fluorescence microscope. Although for many therapeutic applications cubosomes are brought into contact with cell cultures, experiments on model systems are required to assess the fundamental parameters controlling how cubosomes behave at oil/water or lipid bilayer interfaces [Falchi, 2015]. We will apply synthetic chemistry following robust pathways [Osornio, 2012] to achieve in-situ functionalization summarised. Pathways are flexible enough to accommodate efficient labelling requirements (deuterium or fluorescence)or modifications required during the course of the project such as adjusting a spacer between lipids and the desired functional groups. Functional groups are chosen to be consistent with modern ligation methods in order to allow further access to large libraries of compounds and high throughput methods. They have also to allow selective orthogonal reactions according to Sharpless click chemistry paradigm. Of particular interest, the Huisgen reaction AAC (Alkyne-azidecycloaddition), usually referred as the first click reaction, will be performed using different sets of parameters (Strain, Copper catalysis, Ruthenium catalysis…). Functional lipids are designed to react after cubosome formation but could also enter the preparation of cubosomes in a one-pot process. Multicompartment nanocarrier cubosomes were recently demonstrated through incorporation of amphiphilic cyclodextrins (CD) that could carry water insoluble substances [Zerkoune, 2016]. In this project, the channels surfaces will be functionalised with CDs by in-situ reactions. Cyclodextrins will be considered as:



i) model molecules to link to the internal cubosome membrane in the aqueous channel (e.g. common Azido-cyclodextrin)

ii) carrier of hidden molecules to the membrane

iii)cages to control release and modulate affinity constants, typically of moderately hydrophilic compound with known affinity toward cyclodextrin cavity.



Importantly, according to their ring size, Cds and interaction capability, CDs can offer multiple specific behaviour with pluronic stabilizers. With little chemical effort, inclusion compounds could be favoured or hampered. Structure determination. In addition to X-ray and other standard techniques, deuterated species will be synthesized for structure determination by contrast matching in neutron scattering. Deuterated MO will be prepared in a multistep yet simple synthesis from deuterated oleic acid [Darwish, 2013]. Mixtures of deuterated unmodified MO and chemically modified MO will allow controlling the degree of mixing of the modified molecules with bare ones. Deuterated compounds will also allow NMR experiments to inspect diffusion phenomena, reaction kinetics, and membrane organization.





[Angelova, 2015] Angelov, B., Angelova, A., Drechsler, M., Garamus, V.M., Mutafchieva, R. and Lesieur, S. (2015) Identification of large channels in cationic PEGylated cu- bosome nanoparticles by synchrotron radiation SAXS and Cryo-TEM imaging. Soft Matter, 11, 3686–3692, 2015.

[Astolfi, 2017] Astolfi, P.; Giorgini, E.; Gambin, V.; … Vita, F.; Francescangeli, O.; Marchini, C. & Pisani, M. (2017), Lyotropic Liquid- Crystalline Nanosystems as Drug Delivery Agents for 5-Fluorouracil: Structure and Cytotoxicity, Langmuir 33, 12369-12378.

[Barriga, 2018] Barriga H.M.G., Holme M.N. and Stevens M.M. (2018), Cubosomes: the next generation of smart lipid nanoparticles, Angew. Chemie Int. Ed. 57, 2.

[Chong, 2011] Chong, Y.T.J., Mulet, X., Waddington, L.J., Boyd, B.J. and Drummond, C.J. (2011), Steric stabilisation of selfassembled cubic lyotropic liquid crystalline nanoparticles: high throughput evaluation of triblock polyethylene oxide-polypropylene

oxide-polyethylene oxide copolymers. Soft Matter, 7:4768.

[Darwish, 2013] Darwish, T.A., Luks, E., Moraes, G., Yepuri, N.R., Holden, P.J., James, M., (2013), Synthesis of deuterated oleic acid and its phospholipid derivative [D64]dioleoyl-sn-glycero-3-phosphocholine. J. Label. Compd. Radiopharm. 56, 520–529.

[Elamari, 2010] Elamari, H., Jlalia, I., Louet, C., Herscovici, J., Meganem, F., Girard, C., (2010), On the reactivity of activated alkynes

in copper and solvent-free Huisgen’s reaction. Tetrahedron Asymmetry, Henri Kagan: An 80th Birthday Celebration Special Issue – Part 1 21, 1179–1183.

[Falchi, 2015] Falchi, A.M., Rosa, A., Atzeri, A., Incani, A., Lampis, S., Meli, V., Caltagirone, C. and Murgia, S., (2015), Effects of

monoolein-based cubosome formulations on lipid droplets and mitochondria of HeLa cells. Toxicology Research, 4, 1025-1036.

[Garg, 2006] Garg, G.; Singh, D.; Saraf, S. & Saraf, S. (2006), Management of benign prostate hyperplasia: An overview of alphaadrenergic

antagonist, Biological & Pharmaceutical Bulletin. 29, 1554-1558.

[Hyde, 1984] Hyde, S.T. and Andersson S.A (1984) Cubic structure consisting of a lipid bilayer forming an infinite periodic minimum

surface of the gyroid type in the glycerol monooleate-water system. Crystalline Materials, 1984.

[Kluzek, 2017] Kluzek, M., Tyler, A.I., Wang, S., Chen, R., Marques, C.M., Thalmann, F., Seddon, J.M. and Schmutz, M., 2017. Influence of a pH-sensitive polymer on the structure of monoolein cubosomes. Soft Matter, 13, 7571-7577.

[La, 2014] La, Y.; Park, C.; Shin, T. J.; Joo, S. H.; Kang, S. & Kim, K. T. (2014), Colloidal inverse bicontinuous cubic membranes of block copolymers with tunable surface functional groups, Nat. Chem. 6(6), 534-541.

[Landh, 1994] Landh, T. (1994), Phase behavior in the system pine needle oil Monoglycerides-Poloxamer 407-Water at 20ºC. J. Phys. Chem., 98, 8453.

[Li, 2015] Li, J.C., Zhu, N., Zhu, J.X., Zhang, W.J, Zhang, H.M., Wang, Q.Q, Wu, X.X., Wang, X., Zhang, J. and Hao, J.F. (2015) Self- Assembled cubic liquid crystalline nanoparticles for transdermal delivery of paeonol. Med Sci Monit., 21, 3298–3310.

[Luzzati, 1962] Luzzati V . and Husson F. (1962) The structure of the liquid-crystalline phases of lipid-water systems., J Cell Biol, 12(2):207–219.

[Osornio, 2012] Osornio, Y. M.; Uebelhart, P.; Bosshard, S.; Konrad, F.; Siegel, J. S. & Landau, E. M. (2012), Design and Synthesis

of Lipids for the Fabrication of Functional Lipidic Cubic-Phase Biomaterials, JOC 77, 10583-10595.

[Scriven, 1976] Scriven, L.E.(1976) Equilibrium bicontinuous structure. Nature, 263(5573):123–125

[Zerkoune, 2016] Zerkoune, L., Lesieur, S., Putaux, J.-L., Choisnard, L., Gèze, A., Wouessidjewe, D., Angelov, B., Vebert-Nardin, C., Doutch, J., Angelova, A. (2016), Mesoporous self-assembled nanoparticles of biotransesterified cyclodextrins and nonlamellar lipids as carriers of water-insoluble substances. Soft Matter 12, 7539–7550.
Fabrication of omniphobic surfaces

SL-DRF-19-0656

Research field : Soft matter and complex fluids
Location :

Service Nanosciences et Innovation pour les Materiaux, la Biomédecine et l'Energie

Laboratoire Interdisciplinaire sur l'Organisation Nanométrique et Supramoléculaire

Saclay

Contact :

Christophe FAJOLLES

Patrick GUENOUN

Starting date : 01-11-2019

Contact :

Christophe FAJOLLES
CEA - DSM/IRAMIS/NIMBE/LIONS

01 69 08 99 60

Thesis supervisor :

Patrick GUENOUN
CEA - DRF/IRAMIS/NIMBE/LIONS

01-69-08-74-33

Personal web page : http://iramis.cea.fr/Pisp/patrick.guenoun/index.html

Laboratory link : http://iramis.cea.fr/nimbe/lions/

For many applications (defrosting, anti-adhesion, cleaning) one aims at surfaces where consensed droplets can be easily removed. The usual method for water is to create hydrophobic surface coatings to create water droplets with a high contact angle and a low hysteresis force: the drops are then easily evacuated under the influence of gravity for example. However, these coatings are generally not very effective for oil drops and are often fragile in the long term. In addition, the coatings may consist of chemical species that will be soon regulated or banned or, in the case of nanostructured coatings, are too sensitive to the pressure that makes them ineffective (impalment transition).



In this PhD project, in collaboration with an industrial partner, we propose to explore a new strategy recently developed in our laboratory and in the literature and which consists in creating a liquid-like coating : as nanostructured surfaces mimick lotus leaves, this strategy is inspired from pitcher plants [1]. A first method consists to infusing a liquid in a porous layer [2] but the stability of the liquid can be problematic. More recently, polymers of poly (dimethylsiloxane) kind, grafted or adsorbed on glass surfaces have shown such liquid-like behaviors such as condensed oil drops could slide very easily [3,4]. Yet convincing similar properties for drops of water have not been shown yet since the remnant hysteresis is still quite large.



We therefore propose a training and doctoral project that will consist of optimizing truly omniphobic surfaces. To do this, we will explore different types of polymers and different methods of adsorpton or grafting by controlling especially the chemical nature of surface groups and the associated surface energy as well as the behavior of drops on the surfaces. Particular attention will be paid to the thermal annealing of the layers as well as to their aging over time upon pH variations notably.





[1] Bohn H.F., Federle W., 14138–14143 PNAS September 28, 2004 vol. 101 no. 39

[2] Wong, T. S.; Kang, S. H.; Tang, S. K.; Smythe, E. J.; Hatton, B.

D.; Grinthal, A.; Aizenberg, J. Bioinspired Self-Repairing Slippery

Surfaces with Pressure-Stable Omniphobicity. Nature 2011, 477, 443-

447.

[3] Wang, L.; McCarthy, T. J. Covalently Attached Liquids: Instant

Omniphobic Surfaces with Unprecedented Repellency. Angew. C hem.,

Int. Ed. 2016, 55, 244-248.

[4] Liu, P.; Zhang, H.; He, W .; Li, H.; Jiang, J.; Liu, M.; Sun, H.; He,

M.; Cui, J.; Jiang, L.; Yao, X. Development of “Liquid-Like”

Copolymer Nanocoatings for Reactive Oil-Repellent Surfac e. ACS

Nano 2017, 11 (2), 2248-2256.

“Smart” Composite Membranes for Lithium-Metal-Polymer Batteries.

SL-DRF-19-0850

Research field : Soft matter and complex fluids
Location :

Laboratoire Léon Brillouin

Groupe Biologie et Systèmes Désordonnés

Saclay

Contact :

Quentin BERROD

Jean-Marc ZANOTTI

Starting date : 01-10-2019

Contact :

Quentin BERROD
CNRS - DRF/INAC/SyMMES/STEP

(+33)(0)438786425

Thesis supervisor :

Jean-Marc ZANOTTI
CEA - DRF/IRAMIS/LLB/GBSD

+33(0)476207582

Personal web page : http://iramis.cea.fr/Pisp/jean-marc.zanotti/

Laboratory link : http://www-llb.cea.fr/

More : https://icr-amu.cnrs.fr/

At the present stage, in the electrochemical device landscape, solid-state polymer lithium batteries offer an interesting compromise in terms of specific stored energy and power. Nevertheless, to achieve practical conduction they need to operate at relatively high temperature (80°C). This condition significantly hampers the performances of the system. The top-one priority of manufacturers in the field is to decrease the working temperature of their products. This project proposes a fundamental science approach targeting the delivery of a “proof of concept” polymer based lithium metal battery working at room temperature.



This ambitious goal will be achieved by taking advantage of i) the confinement of the electrolyte within composite Carbon NanoTube (CNT) membranes (Gibbs-Thomson effect), ii) one-dimensional (1D) ionic conductivity, and iii) the use of low molecular mass PEO (high mobility). The reduction of dimensionality will be obtained by using the quasi-perfect 1D topology offered by vertically aligned CNT forests.



The suppression of the electrical conductivity of the CNT is a critical aspect to use 1D CNT membranes as battery separators. Short PEO chains will be therefore grafted onto the CNT caps to achieve at once good ionic conduction at the CNT pore entrance and ensure electrical insulation of the CNT/electrode contact. Depending on the physico-chemical conditions on one side of the membrane (pH, temperature…), one can expect drastic changes in the conformation of the CNT-tips-grafted-polymer layer: from extended to mushroom conformation. Therefore, beyond the present project, such smart membranes could be turned into “nano-valves”, able to gate the flow between different media.

Irreversibility and out-of-equilibrium in turbulence

SL-DRF-19-0825

Research field : Soft matter and complex fluids
Location :

Service de Physique de l'Etat Condensé

Systèmes Physiques Hors-équilibre, hYdrodynamique, éNergie et compleXes

Saclay

Contact :

François DAVIAUD

Starting date : 01-10-2019

Contact :

François DAVIAUD
CEA - DRF/IRAMIS/SPEC/SPHYNX

01 69 08 72 40

Thesis supervisor :

François DAVIAUD
CEA - DRF/IRAMIS/SPEC/SPHYNX

01 69 08 72 40

Personal web page : http://iramis.cea.fr/Pisp/francois.daviaud

Laboratory link : http://iramis.cea.fr/spec/SPHYNX

Turbulent flows are fundamentally out of equilibrium. They are characterized by fluxes of quantity of movement, heat or energy over a range of scales covering several orders of magnitude. The corresponding number of degrees of freedom is very large and does not allow their direct implementation on a computer. To progress, it is therefore necessary to be able to model these flows taking into account the out of equilibrium nature of turbulence.



The purpose of this thesis is to investigate the validity of the formalism of out of equilibrium physics (fluctuation-dissipation theorems, function of large deviations) in a turbulent von Karman model flow. We will try to obtain direct measurements of the velocity fields, to characterize, in the scale space, the different flows and their relationship with the forcing constraints. We will use 2D and 3D Particle Image Velocimetry measurements and a new multi-scale analysis method similar to the wavelet transform.

Self-assembled metamaterials made by block copolymers

SL-DRF-19-0901

Research field : Soft matter and complex fluids
Location :

Service Nanosciences et Innovation pour les Materiaux, la Biomédecine et l'Energie

Laboratoire Interdisciplinaire sur l'Organisation Nanométrique et Supramoléculaire

Saclay

Contact :

Patrick GUENOUN

Starting date : 01-10-2018

Contact :

Patrick GUENOUN
CEA - DRF/IRAMIS/NIMBE/LIONS

01-69-08-74-33

Thesis supervisor :

Patrick GUENOUN
CEA - DRF/IRAMIS/NIMBE/LIONS

01-69-08-74-33

Personal web page : http://iramis.cea.fr/Pisp/patrick.guenoun/index.html

Laboratory link : http://iramis.cea.fr/nimbe/lions/

Metamaterials are "artificial" materials which are created to reach properties inaccessible to natural homogeneous materials. Optical properties like negative refractive indices could be achieved by an adequate structuring of materials at a scale lower than the wavelength of the light. In this PhD work, we shall obtain such a structuration by combining the self-assembly of copolymers on surfaces and the insertion of gold nanoparticles in the copolymer matrix. The copolymer matrix of copolymers provides the nanostructuration and the desired geometry thanks to microphase separation on top of the substrate whereas the gold nanoparticles presence confers the expected optical properties. This PhD thesis project in collaboration between LIONS at CEA Saclay (U. P. Saclay) and the Paul Pascal Research Center (CRPP) in Bordeaux will benefit from both environments to lead an experimental study which will consist in preparing surfaces where cylindrical or bicontinuous phases of copolymers will be directed perpendicularly to the substrate. After synthesis in the laboratory and insertion of gold nanoparticles in the structures, the optical properties of the obtained material will be measured and analyzed for modeling.

Single-ion hybrid polymer electrolytes for Li-metal battery

SL-DRF-19-0554

Research field : Soft matter and complex fluids
Location :

Laboratoire Léon Brillouin

Groupe de Diffusion Neutron Petits Angles

Saclay

Contact :

Jacques JESTIN

Starting date : 01-10-2019

Contact :

Jacques JESTIN
CNRS - LLB01/Laboratoire de Diffusion Neutronique

0661476825

Thesis supervisor :

Jacques JESTIN
CNRS - LLB01/Laboratoire de Diffusion Neutronique

0661476825

Biophysical and dynamical study of chromatin conformation during genome replication

SL-DRF-19-0435

Research field : Soft matter and complex fluids
Location :

Service Nanosciences et Innovation pour les Materiaux, la Biomédecine et l'Energie

Laboratoire Interdisciplinaire sur l'Organisation Nanométrique et Supramoléculaire

Saclay

Contact :

Frédéric GOBEAUX

Patrick GUENOUN

Starting date : 01-10-2018

Contact :

Frédéric GOBEAUX
CEA - DRF/IRAMIS/NIMBE/LIONS

01 69 08 24 74

Thesis supervisor :

Patrick GUENOUN
CEA - DRF/IRAMIS/NIMBE/LIONS

01-69-08-74-33

Personal web page : http://iramis.cea.fr/Pisp/frederic.gobeaux/

Laboratory link : http://iramis.cea.fr/nimbe/lions/

The tridimensional organization of the genome and its dynamics in live cells are decisive to perform its functions. It is crucial to understand them and to identify the parameters controlling them. Current state of the art allows describing the short range (<10 nm) and long range (>250 nm) organization of chromatin conformation in the nucleus. However, there is an intermediate range (10-250 nm) where chromatin organization is difficult to apprehend. This range corresponds to the size of protein complexes that modify chromatin and harness genome replication.



We propose to monitor cell cultures during genome replication and other cellular events using small angle x-ray scattering. Thanks to a dedicated experimental set-up we will study chromatin conformation dynamics during genome duplication and complement this analysis with numerical simulations (molecular dynamics) so as to correlate chromatin dynamics with that of genome duplication. We will use different cell mutants and the addition of drugs to perturb the system and modify the observed structures.



This project is a collaboration between two teams of physicists and biologists and will consist for the student to reach a dual expertise in both disciplines.
Effect of structure on the digestion of proteins: from animal to plant

SL-DRF-19-1093

Research field : Soft matter and complex fluids
Location :

Laboratoire Léon Brillouin

Groupe de Diffusion Neutron Petits Angles

Saclay

Contact :

Fabrice COUSIN

Annie BRULET

Starting date : 01-11-2019

Contact :

Fabrice COUSIN
CEA - DRF/IRAMIS/LLB/MMB

01 69 08 67 73

Thesis supervisor :

Annie BRULET
CNRS - DRF/IRAMIS/LLB/MMB

0169086669

Personal web page : http://iramis.cea.fr/llb/Phocea/Pisp/index.php?nom=fabrice.cousin

Laboratory link : http://iramis.cea.fr/llb/Phocea/Pisp/index.php?nom=francois.boue

More : http://www-llb.cea.fr/

The thesis will be focused on the physiological process of digestion, studied from a purely physico-chemical point de view (asides of the one of physiologists and nutritionists). It aims at monitoring and modelising unraveling of model food, more or less micro-structured at the smallest scales, comparing animal and plant products like dairy gels and plant proteins.



The experimental part will consist in following the effect of digestive enzymes (gastric and intestinal) on model food, solutions or gels:

- by Small Angle Neutron Scattering and X-ray scattering (SANS & SAXS): in volume on homogeneous samples (neutrons, using contrast variation on mixtures of proteins, lipids or glucids, which have different scattering length density), or at interfaces of films (RX, narrow beam) ;

- by transmission imaging, technique mastered by the Paul Scherrer Institut team: permeation of water in the food piece, evolution of this piece by swelling/deswelling and vanishing of the eddible material;

- by scattering imaging: more difficult but promising, in which PSI is strongly implicated.



A modelisation of all steps, difficult using differential equations, will use agent-based simulations (“distributed artificial intelligence”) in collaboration with an Institut National de la Recherche Agronomique team.

 

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