Les PLLs sont une famille d’enzyme dont l’activité catalytique est double. L’activité lactonase permet entre autre de détruire les AHLs, molécules médiatrices de la communication chez les bactéries Gram négatives, empêchant ainsi la synchronisation à l’échelle de la population de la sécrétion de facteurs de virulence ou la formation de biofilm. Elles sont non seulement capables d’hydrolyser les molécules possédant un noyau lactone, mais aussi les phosphotriésters par promiscuité de substrat. L’activité phosphotriestérase permet de dégrader les composés organophosphorés (OPs) hautement toxiques, que ce soit les insecticides (Paraoxon) ou les agents neurotoxiques de guerre (Sarin, VX). Les travaux effectués ont consisté à évaluer l’efficacité de la PLL SsoPox, provenant de l’archée extrêmophile Sulfolobus solfataricus, pour empêcher la mise en place de la virulence et du biofilm chez Pseudomonas aeruginosa PAO1. Cette évaluation a été effectuée in vitro et in vivo via un modèle d’infection pulmonaire chez le rat. D’autre part, une étude in vitro a été réalisée sur 73 souches cliniques de P. aeruginosa isolés sur des patients atteints de pied diabétique, en évaluant la quantité de pyocyanine et l’activité protéolytique. Enfin, une phase prospective pour identifier de nouvelles PLLs a permis la caractérisation enzymatique et structurale de 2 nouvelles enzymes, SacPox et VmoLac, contribuant ainsi à affiner la connaissance sur cette famille et leur potentiel d’amélioration par ingénierie protéique. Au final, les PLLs offrent un intérêt biotechnologique majeur et peuvent mener à une valorisation concrète pour la santé humaine mais également pour la bioremédiation des OPs. / Phosphotriesterase-Like Lactonases (PLLs) are a family of enzyme displaying a dual catalytic activity. Lactonase activity allows for, among others, the destruction of Acyl Homoserine Lactones (AHLs). These molecules mediate the communication in Gram negative bacteria, allowing them to synchronize group behavior like virulence factor secretion or biofilm formation. Beside their ability to hydrolyze molecules with a lactones moiety, PLL also show substrate promiscuity for hydrolyzing highly toxic organophosphate compounds (OPs), such as pesticides (i.e. Paraoxon) or Chemical Warfare Nerve Agents (CWNAs) (e.g. Sarin & VX). The work described here consisted in evaluating the efficacy of the PLL SsoPox, originating from the extremophile Archaea Sulfolobus solfataricus, to prevent virulence and biofilm formation on the model strain Pseudomonas aeruginosa PAO1. This evaluation was performed both in vitro and in vivo, by using a rat pulmonary infection model. A study has been performed on a strain collection of 73 clinical strains of P. aeruginosa isolated from diabetic foot, to assess the enzyme effects on pyocyanin secretion and protease activity. Finally, a prospective phase to identify new PLLs allowed for the enzymatic and structural characterization of two new enzymes, SacPox and VmoLac, thus contributing to refine the knowledge about this enzyme family and their engineering potential. In conclusion, PLLs are of prime biotechnological interest and could lead to developments in human health, but also for OPs bioremediation with a non aggressive decontamination solution.

Les matrices d’excitation et d’émission de fluorescence (MEEF) sont utilisées pour caractériser la matière organique naturelle (MON). Afin de mieux exploiter ces informations, un algorithme trilinéaire, PARAFAC, est employé. Après l’élimination des diffusions Rayleigh et Raman et la correction de l’effet d’écran, cette méthode permet de séparer les composants spectraux présents dans les MEEF.Ce travail présente deux études : la qualification et la quantification de la MON selon son origine environnementale et le calcul des constantes de complexation de la MON et du cuivre sous forme ionique.Les composants spectraux et leurs intensités relatives sont calculés par PARAFAC à partir 1146 échantillons regroupés suivant les missions, leur type de milieu, ou le niveau de salinité. Pour étudier ces composants, une nouvelle représentation spectrale est proposée afin de mettre en évidence leur variabilité spectrale. Les résultats montrent que le regroupement d’échantillons d’origine diverse conserve le recouvrement spectral global et les intensités relatives. Sur l’ensemble du domaine spectral, les zones correspondant aux substances humiques sont peu variables, comparées à la zone protéinique.La complexation des métaux par la MON est analysée par une technique combinant quatre outils : l’ajout logarithmique d’ions métalliques, la mesure de MEEF, la méthode PARAFAC et l’algorithme PROSECE. La mesure du quenching de fluorescence ne se limite pas seulement à la modélisation d’une intensité de fluorescence mais à celle de l’intensité relative de chaque composant PARAFAC surpassant ainsi les méthodes utilisées jusqu’à présent. Finalement, l’application de cette technique originale permet de quantifier les propriétés de complexation de la MON à l'aide d'un modèle de complexation utilisant 2 sites de complexation par composant en utilisant la totalité du signal de fluorescence. / Fluorescence excitation and emission matrices (EEM) are used to characterize natural organic matter (NOM). To make best use of this information, PARAFAC, a trilinear algorithm is employed. After removing Rayleigh and Raman scattering and correction of the inner filter effect, this method allows separating the spectral components present in MEEF.This work presents two studies: the characterization and quantification of NOM according to its origin and calculation of environmental complexation constants of NOM towards copper as ionic form.Spectral components and their relative intensities are calculated by PARAFAC from 1146 samples gathered according to the missions, the medium type, or the salinity level. To study these components, a new spectral representation is proposed in order to highlight their spectral variability. The results show that even when samples spectra of various origins are clustered, the overall spectral overlap and the relative intensities remain almost similar. On the whole spectral range, areas corresponding to humic substances are quite variable, compared to the protein zone.Metal complexation by NOM is analyzed by combination of four tools: metal ions logarithmic addition, MEEF measurement, the PARAFAC dissociation method and the PROSECE modelling algorithm. Fluorescence quenching measurement is not only limited to the modelling of fluorescence intensity but also to the relative intensity of each PARAFAC-dissociated component though surpassing the methods used so far. Finally, the application of this improving technique leads to quantify NOM complexation properties using a two-complexing sites complexation model for each PARAFAC-dissociated component by using the whole fluorescence signal.

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Detection, identification and separation of polycyclic aromatic compounds in environmental samples are of extreme importance since many of these compounds are well known for their potential carcinogenic and/or mutagenic activities. Selective quenching of molecular fluorescence can be utilized effectively to analyze mixtures containing different polycyclic aromatic hydrocarbons. Molecularly organized assemblies are used widely in detection and separation of these compounds mainly because of less toxicity and enhanced solubilization capabilities associated with these media. Feasibility of using nitromethane and the alkylpyridinium cation as selective fluorescence quenching agents for discriminating between alternant versus nonalternant polycyclic aromatic hydrocarbons (PAHs) is critically examined in several molecularly organized micellar solvent media. Fluorescence quenching is used to probe the structural features in mixed micelles containing the various combinations of anionic, cationic, nonionic and zwitterionic surfactants. Experimental results provide valuable information regarding molecular interactions between the dissimilar surfactants.

Cette thèse porte sur la modélisation de l'ébullition qui joue un rôle important dans les vitessesde refroidissement des pièces, elle possède un volet numérique et un volet expérimental. Lessimulations et les expériences envisagées se situent à deux échelles. A l'échelle d'une ou quelquesbulles de vapeur, il s'agit de faire des simulations multiphasiques très précises en prenant encompte, la tension de surface, les calculs directs d'écoulement à grand nombre de Reynolds, etrendant compte du détachement et de la coalescences des bulles. Des observations expérimentalessont réalisées à la même échelle en contrôlant en surface la nucléation d'une bulle de vapeur àl'aide d'une caméra rapide. Des mesures de champs de vitesse par PIV et de température par twocolor LIF thermometry sont réalisées dans les mêmes conditions. Ceci a permit de confronter lacroissance, la dynamique et les formes des bulles observées et calculés. Les techniques numériquesles plus avancées sont utilisées : Eléments finis stabilisé VMS, level set, adaptation anisotropeet calcul intensif. Les modéles numériques proposés dans cette thèse permettent de passer àl'échelle macroscopique des pièces industrielles en considérant un film de vapeur (ou une phasede mélange liquide vapeur). L'enjeu supplémentaire étant de modéliser la turbulence induite parl'ébullition dans une approche de type CFD. / This thesis focuses on the modelling of boiling that plays an important role on the coolingand heat treatment in quenching processes, it has two components: numerical simulations andexperimental measurements. Both simulations and experiments are envisaged for two scales. Thefirst one concerns small scales: the scale of one or few bubbles. In this case, the focus is put onvery precise numerical simulations for multiphase flows taking into account the surface tension,the direct computations of flows at high Reynolds number, and on reflecting the detachmentand coalescence of bubbles. On that same scale, experimental observations are performed tocontrol, in the volume or at surface, the nucleation of a vapour bubble using a high speedcamera. Measurements of velocity fields by PIV and the temperature by PLIF are realizedunder the same conditions. This will allow us to compare the growth dynamics and shapesof bubbles observed and calculated. Advanced numerical methods are used to fulfil this task:VMS stabilized finite elements, level set, anisotropic adaptation and parallel computing. Thenumerical models proposed in this work are extended and also used to deal with macroscopicscales: at the level of industrial parts considering the vapour films (or a phase of liquid vapourmixture). The additional challenge resides in the modelling of turbulence induced by boiling ina CFD approach.

Um novo ciclo de tratamento térmico denominado como têmpera e partição vem sendo desenvolvido em aços com elevados teores de silício, como rota para obtenção de estruturas com frações consideráveis de austenia retida. Essa rota de tratamento m térmico consiste em realizar uma têmpera temperaturas intermediárias entre Ms e Mf, seguido de um reaquecimento com manutenção em patamares isotérmicos por certos intervalos de tempo, objetivando estabilizar a austenita remanescente através da partição do carbono a partir da martensita supersaturada. No presente trabalho, duas ligas de ferros fundidos nodulares convencionais, com diferentes teores de silício e manganês, foram submetidas a ciclos de têmpera e partição. As amostras foram austenitizadas a 900°C por duas horas. Uma das ligas foi temperada em óleo a 160°C e a outra a 170°C por 2 minutos. Imediatamente após a têmpera as amostras foram reaquecidas em temperaturas entre 300 e 450°C por intervalos de tempo que variaram entre 2 e 180 minutos. A caracterização microestrutural foi realizada através de microscopia eletrônica de varredura (MEV) e difração de raios x. A caracterização mecânica foi feita através de ensaios de energia absorvida ao impacto, dureza HRC e ensaios de tração. A caracterização microestrutural evidenciou que os ciclos de têmpera e partição são viáveis na obtenção de frações consideráveis de austenita retida nos ferros fundidos nodulares. A caracterização mecânica evidenciou que foi possível obter boas combinações de energia absorvida ao impacto, resistência à tração e alongamento. Em todas as condições testadas é possível perceber uma janela de processo bem definida caracterizada por valores crescentes das propriedades mecânicas nos primeiros minutos do ciclo de partição e que decrescem após certo intervalo de tempo. O conjunto de propriedades mecânicas obtidas através dessa rota de tratamentos térmicos indica que os ferros fundidos nodulares submetidos ao ciclo de têmpera e partição podem se constituir como alternativa tecnológica para aplicações comerciais nas quais os ferros fundidos nodulares austemperados já são materiais consolidados. / A new heat treatment cycle known as quenching and partitioning has been developed in commercial steel alloys containing silicon as a way to obtain structures with controlled fractions of retained austenite. This heat treatment cycle consists in performing a quenching in temperatures between Ms and Mf, followed by a reheating with isothermal holding by different time intervals. The aim of this cycle is to achieve the austenite stabilization by diffusion of carbon from the supersaturated plates of martensite. In this work, two conventional ductile cast iron alloys, with two different contents of silicon and manganese were heat-treated in quenching and partitioning cycle. The samples were austenitized at 900°C for two hours, followed by quenching in oil at 160° C and 170° C for two minutes. Immediately after quenching, the samples were reheated at temperatures between 300 and 450°C for time intervals between 2 and 180 minutes. The microstructural characterization was performed using electronic microscopy (SEM) and x-ray diffraction. The mechanical characterization was performed using impact tests, hardness and tensile strength tests. The microstructural characterization showed that the cycles of quenching and partitioning are viable to obtain considerable fractions of retained austenite in nodular cast by this heat treatment route. The mechanical characterization showed that it was possible to obtain good combinations of energy absorbed in the impact, tensile strength and elongation. In all tested conditions was possible to perceive a well-defined process window characterized by increasing values of mechanical properties in the first minutes of the partitioning step, and decrease after certain time intervals. The set of mechanical properties obtained by this route of heat treatments indicates that nodular cast iron subjected to tempering and partitioning cycle can be constituted as an alternative technology for commercial applications in which austempered ductile irons are already consolidated materials.

The finite difference method was used to calculate the variable heat transfer coefficient required to maximize mechanical properties of heat treated wrought 7075 aluminum alloy without causing residual stress. Quench simulation enabled determination of maximum surface heat flux bordering on inducing plastic flow in the work piece. Quench Factor Analysis was used to correlate cylinder diameter to yield strength in the T73 condition. It was found that the maximum bar diameter capable of being quenched without residual stress while meeting military mechanical design minimums is 2". It was also found that the cooling rate must increase exponentially and that the maximum cooling rate needed to achieve minimum mechanical properties is well within the capability of metals heat treatment industry.

Photosystem II (PSII) is the primary generator of electrons required for photosynthesis. The reaction center protein of PSII (RCII) is the most susceptible component of the photosynthetic machinery to damage. Photodamage can lead to long-term downregulation of photosynthesis. This occurs because plants are exposed to rapid light fluctuations and high light conditions, leading to the over accumulation of excess energy around PSII. Plants have developed a mechanism to dissipate this excess energy called nonphotochemical quenching (NPQ). In order to quantify the protectiveness of NPQ (pNPQ), a novel methodology was developed and employed. During methodology development, development, it is shown that a variable PSI fluorescence should be taken into account, and how it can be calculated. Application of the procedure assessed the contribution of xanthophylls lutein, violaxanthin, zeaxanthin, and the PsbS protein to pNPQ. Results show that the most important factors governing photoprotection are the PsbS protein and the correct xanthophyll composition in their natural binding sites. The more xanthophyll variation, the greater the photodamage at the end of the pNPQ assessment procedure. PsbS is essential to achieve the maximum pNPQ. PsbS increases the aggregation of LHCII. Arabidopsis with excess PsbS has three-times more aggregated LHCII than wild type levels of PsbS. The phototolerance and pNPQ required for Arabidopsis grown under different conditions and for leaves of different ages was also calculated. Plants grown under low light conditions accumulate disconnect antenna (LHCII), which is inefficient at protecting RCII, despite the high NPQ levels. Investigating plants of different ages, it was found that eight-week old Arabidopsis are the optimum age for pNPQ effectiveness. Younger and older leaves suffer photodamage at lower light intensities and form less pNPQ. This thesis demonstrates the novelty and adaptability of the pNPQ assessment procedure, and offers a sound case for its use in acclimation and photoinhibition experiments.

What drives the transition of galaxies from the disc dominated, star forming blue cloud to the elliptical dominated, quiescent red sequence? What role does the morphology, central supermassive black hole and galaxy environment play in this transition? I have attempted to answer these questions by using Bayesian statistics to infer a simple star formation history (SFH) describing the time, t_q, and exponential rate, τ, that quenching occurs in a galaxy. I use both the optical and NUV photometry of a galaxy in order to infer the posterior distribution of its SFH across the two dimensional [t_q, τ] parameter space. I then utilise the Galaxy Zoo 2 morphological classifications to obtain a morphology weighted, combined population distribution across each quenching parameter for a sample of galaxies. I apply this method across the blue cloud, green valley and red sequence of a sample of 126,316 galaxies and find a clear difference between the quenching timescales preferred by smooth and disc weighted populations, with three major routes through the green valley dominated by smooth (rapid rates, attributed to major mergers), intermediately classified (intermediate rates, attributed to galaxy interactions) and disc morphologies (slow rates, attributed to secular evolution). I hypothesise that morphological changes occur in systems which have undergone quenching with an exponential rate, τ < 1.5 Gyr, in order for the evolution of galaxies in the green valley to match the ratio of smooth to disc galaxies observed in the red sequence. I repeat this SFH analysis for a sample of 1,244 Type 2 AGN host galaxies and find statistical evidence for recent, rapid quenching, suggesting that this may be caused by AGN feedback. However I find that rapid quenching rates cannot account for all the quenching across the AGN host population; slow quenching rates, attributed to secular evolution, are also significant in the evolution of AGN host galaxies. I investigate this possible secular co-evolution of galaxies and black holes further by measuring the black hole masses of a sample of 101 bulgeless AGN host galaxies and compare them to typical black hole-galaxy scaling relations. I find that the measured black holes of the bulgeless galaxies are ~1-2 dex more massive than they should be, given their lack of bulges. This suggests that black hole-galaxy scaling relations may arise due to mutual correlations to the overall gravitational potential of the dark matter halo of the galaxy. I also considered the effect of the group environment on the time and rate that quenching occurs, with respect to the group-centric radius, for 4,629 satellite galaxies. I find that although mergers, mass quenching and morphological quenching are all occurring in groups, environmentally driven quenching mechanisms are also prevalent. However, I find that these environmentally driven quenching processes are not correlated with the velocity of a satellite within a group, ruling out ram pressure stripping as a possible mechanism. I discuss how all of these quenching mechanisms are likely to affect a galaxy across its lifetime, acting in concert to reduce the SFR, which in turn produces the wide distribution of quenching timescales seen across the colour-magnitude diagram. I discuss ideas for future work using the method employed in this work, including applying it to forthcoming data from large integral field unit surveys.