Dissertations / Theses on the topic 'Physico-Chemical interactions'

In this thesis, several physico-chemical phenomena occurring at the two-dimensional environment of a Langmuir monolayer are studied. In the first section, the characterization of Langmuir monolayers of an azobenzene derivative is presented, together with the induction of chirality by the action of a chemical and a physical force, inside labile condensed domains formed by the trans isomer. This has been compared with results of another study done in the group for a homologue azobenzene molecule. In addition, we have studied a heterochiral recognition phenomenon in monolayers by means of the interaction of two enantiomeric chiral azobenzene derivative surfactants. The following section focuses on the field of two-dimensional microfluidics, and encompasses the analysis of different transport processes occurring at the interface between two monolayers that are coflowing through a channel. In this context, studies of the diffusion between co-flowing monolayers have resulted in a slight increase in the diffusion coefficient value for experiments with curvilinear channels, and when vortices are introduced in the system. In contrast, increasing the concentration gradient between co-flowing monolayers has not been successful at producing a noticeable impact in the measured diffusion coefficient values. On the other hand, a two-dimensional dissolution process has been visualized, with a rate that follows a classic dissolution model. Different unsuccessful attempts to realize a chemical reaction between coflowing monolayers are presented. Moreover, we have addressed the flow of monolayers in open microchannels driven by surface tension gradients, determining that drag at the interface and lateral confinement result in the formation of backflow patterns that carry the subphase liquid upstream through self-organized micro-ducts. This backflow patterns depend on channel geometry and are bound by velocity stagnation surfaces, whose shape and location has been related to the interfacial velocity profiles by means of the combination of a simple model and particle image velocimetry measurements. In the last section, we adapt the protocols used in two-dimensional microfluidics to study the insertion and diffusion of a protein in a phospholipid monolayer.
En aquesta tesi s’han estudiat diferents fenòmens físico-químics en l’àmbit de les monocapes de Langmuir. En una primera secció, s’han caracteritzat monocapes de Langmuir d’un derivat d’azobenzè que presenta una separació de fases entre els seus dos possibles isòmers. S’ha explorat la inducció de quiralitat en dominis condensats aïllats, rics en isòmer trans, mitjançant l’acció d’una força quiral química (un dopant quiral), i una de física (un vòrtex). Els resultats obtinguts s’han comparat amb els d’un altre estudi realitzat prèviament en el grup, fet amb un derivat d’azobenzè homòleg. A part, s’ha estudiat un fenomen de reconeixement quiral en monocapes, degut a la interacció de dos enantiòmers quirals d’un altre derivat azobenzènic. Una segona secció de la tesi es centra en el camp de la microfluídica bidimensional, i mostra l’estudi de diferents fenòmens de transport que ocorren en la interfase entre dues monocapes que flueixen al llarg d’un canal. En concret, s’han fet experiments de difusió entre monocapes que co-flueixen per un canal, per tal de trobar les condicions en les que s’obté un increment de la difusió, traduït en un augment del valor del coeficient de difusió mesurat. D’aquesta manera, els experiments realitzats en circuits amb forma de serpentí, o en els que s’hi ha creat vòrtexs, presenten un augment en la difusió. En canvi, l’efecte de l’augment del gradient de concentració existent entre les dues monocapes no ha resultat en canvis apreciables en els coeficients de difusió mesurats. D’altra banda, s’ha visualitzat un procés de dissolució en dues dimensions que segueix el model de dissolució clàssic, i s’han fet experiments per tal d’aconseguir visualitzar una reacció en la línia de contacte entre dues monocapes. A més, s’ha estudiat la formació d’un flux de retorn en la subfase com a resultat del flux d’una monocapa en la interfase, i s’ha pogut relacionar amb perfils de velocitat interficials anòmals obtinguts. En l’última secció, s’han adaptat els protocols experimentals utilitzats en la microfluídica bidimensional, per tal d’estudiar la inserció i difusió d’una proteïna en una monocapa de fosfolípids.

In this work, in-situ measurements of aerosol chemical composition, particle number size distribution, cloud-relevant properties and ground-based cloud observations were combined with high-resolution satellite sea surface chlorophyll-a concentration and air mass back-trajectory data to investigate the impact of the marine biota on aerosol physico-chemical and cloud properties. Studies were performed over the North-Eastern Atlantic Ocean, the central Mediterranean Sea, and the Arctic Ocean, by deploying both multi-year datasets and short-time scale observations. All the data were chosen to be representative of the marine atmosphere, reducing to a minimum any anthropogenic input. A relationship between the patterns of marine biological activity and the time evolution of marine aerosol properties was observed, under a variety of aspects, from chemical composition to number concentration and size distribution, up to the most cloud‐relevant properties. At short-time scales (1-2 months), the aerosol properties tend to respond to biological activity variations with a delay of about one to three weeks. This delay should be considered in model applications that make use of Chlorophyll-a to predict marine aerosol properties at high temporal resolution. The impact of oceanic biological activity on the microphysical properties of marine stratiform clouds is also evidenced by our analysis, over the Eastern North Atlantic Ocean. Such clouds tend to have a higher number of smaller cloud droplets in periods of high biological activity with respect to quiescent periods. This confirms the possibility of feedback interactions within the biota-aerosol-cloud climate system. Achieving a better characterization of the time and space relationships linking oceanic biological activity to marine aerosol composition and properties may significantly impact our future capability of predicting the chemical composition of the marine atmosphere, potentially contributing to reducing the uncertainty of future climate predictions, through a better understanding of the natural climate system.

Pendant longtemps, les microorganismes présents dans l’atmosphère n’ont été considérés qu’en tant que particules inertes subissant les conditions hostiles de cet environnement. Cependant, de récentes études mettant en évidence la présence de microorganismes métaboliquement actifs dans la phase aqueuse des nuages incitent à s’interroger sur le rôle que ces organismes pourraient avoir sur les processus physiques et chimiques des nuages. En effet, la formation de gouttelettes de nuage ou de cristaux de glace à des températures supérieures à -36°C nécessite la présence de particules dites « noyaux de condensation » ou « noyaux glaçogènes », dont les bactéries pourraient être des représentantes. De plus, plusieurs travaux ont révélé une importance potentielle des microorganismes dans la transformation de la matière organique dans les nuages. L’objectif de ces travaux de thèse a donc été d’étudier les interactions réciproques entre les microorganismes et les conditions physico-chimiques des nuages. Dans un premier temps, les composantes physico-chimiques et microbiologiques ont été caractérisées au moyen de prélèvements nuageux au sommet du puy de Dôme (1465 m, France) et des études statistiques ont permis de mettre en avant des corrélations entre les différents paramètres physico-chimiques et/ou biologiques. Puis, cinq souches microbiennes appartenant à des genres microbiens cultivables majeurs dans les nuages ont été soumises à quatre stress rencontrés dans les nuages : la lumière solaire, la présence de peroxyde d’hydrogène, les variations de chocs osmotiques intervenant lors de la formation et de la dissipation des gouttelettes d’eau et les cycles de gel et de dégel. Il a ainsi été mis en évidence que la lumière solaire et le peroxyde d’hydrogène dans des conditions nuageuses n’ont que peu ou pas d’impact sur la viabilité des cellules. A l’inverse, les chocs osmotiques et le gel-dégel peuvent être hautement délétères selon les souches considérées. La troisième partie de ce travail s’est focalisé à mettre en évidence la présence de souches bactériennes glaçogènes dans l’eau de nuage. Sept souches ont ainsi ont été identifiées et décrites, et l’une d’entre elles a été choisie comme modèle pour étudier le comportement de bactéries (survie et activité glaçogène) dans une chambre de simulaion de nuage (AIDA, Allemagne). En parallèle, l’activité glaçogène biologique de l’eau de nuage a été mesurée à partir de prélèvements au puy de Dôme et l’activité glaçogène bactérienne a été estimée. L’ensemble de ces travaux met en avant une sous-estimation jusqu’alors des proportions de bactéries glaçogènes dans les modèles numériques simulant les processus microphysiques d’initiation de la glace et des précipitations dans les nuages. Ces données vont désormais pouvoir être considérées dans de tels modèles. Enfin, afin d’estimer l’étendue de l’importance des microorganismes dans la chimie atmosphérique, il est nécessaire d’avoir recours à des modèles numériques. La dernière étude de cette thèse s’est consacrée à déterminer des constantes cinétiques de biodégradation de trois composés organiques majeurs des nuages par trois souches bactériennes isolées de cet environnement qui pourront servir à paramétrer des modèles numériques. Une première approche simple a permis de confirmer les résultats précédents de l’équipe en mettant en avant une contribution non négligeable des microorganismes dans leur dégradation
Airborne microorganisms have long been considered as inert, passive particles dealing with hostile conditions. Recent studies highlighting metabolic activity in cloud water raised questions about the role these organisms may play on physical and chemical processes in clouds. Indeed, cloud droplets and ice crystals formation at temperature warmer than -36°C need the presence of particles called “cloud condensation nuclei” or “ice nuclei”. Bacteria could be one of them. In addition, several works revealed a potential importance of microorganisms in organic matter transformation in clouds. The objective of this thesis was to study the reciprocal interactions between microorganisms and physico-chemical conditions in clouds. First, cloud physico-chemical and microbiological compositions were described by cloud sampling at the puy de Dôme station (1465 m, France) and statistical analyses were performed to highlight correlations between physico-chemical and/or biological parameters. Secondly, five microbial strains belonging to genera frequently isolated from cloud water were subjected to four atmospheric stresses: sunlight, hydrogen peroxide, osmotic shocks occurring when water droplets condensate or evaporate and freeze-thaw cycles. Thus, it was pointed that sunlight and hydrogen peroxide at cloud concentration have no or little impact on cell viability. On the opposite, osmotic shocks and freeze-thaw can be highly deleterious depending on the considered strain. The third part of this thesis focused on the detection of ice nucleating bacterial strains in cloud water. Seven strains were thus identified and described, and one of them was selected as a model to study its behavior (survival and ice nucleation activity, INA) in a cloud simulation chamber (AIDA, Germany). In parallel, biological ice nucleation activity was measured directly on cloud samples and bacterial INA was estimated. All these experiments highlighted underestimations of ice nucleation active bacteria in models simulating microphysical processes in clouds. This new dataset may be used as new parameterization in this kind of models. Finally, in order to estimate the bacterial contribution in cloud chemistry, numerical means are needed. Therefore, the last study of this thesis focused on the determination of biological kinetic constants that may be implemented in atmospheric chemistry models. The biodegradation of three major organic compounds encountered in cloud water by three bacterial strains isolated from clouds was measured. A first approach confirmed precedent team results highlighting a considerable contribution of microorganisms on the transformation of these compounds

Le béton de chanvre est l'un des matériaux de construction biosourcés le plus en vogue en France. Il est souvent utilisé comme matériau de remplissage et/ou d'isolation, dans un système éco-constructif. Compte tenu de sa forte porosité, ce béton présente des performances thermiques et hygriques particulièrement intéressantes. Toutefois, alors que son niveau d'isolation thermique s'améliore lorsqu'on diminue sa densité, ce béton de chanvre voit parallèlement ses propriétés mécaniques diminuer. C'est la raison pour laquelle, il est nécessaire d'optimiser sa formulation, afin d'obtenir le meilleur compromis entre ses performances mécaniques, hygriques et thermiques, selon le domaine d'emploi (porteur et/ou isolant). La première piste d'optimisation envisagée consiste à faire varier le taux de granulats végétaux dans la composition du béton, mais aussi la nature et la quantité de liant utilisé. Le travail de cette thèse a pour objectif l'étude des propriétés mécaniques, thermiques et hygriques d'un béton de chanvre en fonction de sa formulation. Dans ce mémoire, on s'intéresse à la faisabilité de matériaux de construction légers à base de particules de chanvre et d'un liant à base de chaux, qui présentent de bonnes performances mécaniques. L'objectif principal ici est de réduire la concentration des ions Ca2+ libres responsables de la dégradation des particules végétales par minéralisation, et qui, par conséquent, induisent la perte de performance mécanique du matériau composite final. Pour y parvenir, une fraction du liant de base a été remplacée par du métakaolin qui interagit avec ces ions Ca2+ suivant une réaction pouzzolanique, afin d'empêcher leur migration vers les particules végétale. Dans un premier temps, nous avons étudié l'influence de différents pourcentages de métakaolin sur les liants témoins à base de chaux: chaux NHL5 et chaux préformulée Tradical PF70. Une étude préliminaire de caractérisation mécanique, en compression et en flexion, a permis suivant une approche "performantielle" de définir une formulation optimale des liants utilisés pour la fabrication des bétons étudiés. Ainsi, les matériaux composites finis sont testés pour différents pourcentages en volume (2V et 3V) de particules de chanvre. Les résultats de la caractérisation des propriétés physico-mécaniques de ces matériaux sont présentés et argumentés. Il en ressort une relation étroite entre la teneur en métakaolin et les propriétés physico-mécaniques du béton de chanvre, qui résulte de la réaction pouzzolanique. Des analyses microscopiques au MEB couplées à l'EDX ont montré une diminution de la zone de transition à l'interface particules de chanvre-liant, mais aussi une faible migration des ions Ca2+ responsables de la minéralisation des particules végétales. Puis, nous avons comparé les performances hygro-thermiques des bétons de chanvre formulés comprenant respectivement 2 volumes et 3 volumes de particules végétales pour un volume de liant, sans ou avec 20% de métakaolin, pourcentage optimal défini précédemment. Cette étude est particulièrement utile pour établir les proportions adéquates d'adjuvants à utiliser dans les bétons destinés aux travaux de rénovation ou aux constructions neuves. D'une manière générale et en fonction de la formulation, on obtient un matériau bon, voire excellent régulateur de l'humidité ambiante, avec des performances thermiques intéressantes pour l'isolation des bâtiments. Ce travail de caractérisation complète les nombreux travaux déjà réalisés sur les bétons biosourcés avec différents types de végétaux, et doit permettre d'alimenter les bases de données indispensables pour la simulation du comportement d'une paroi ou de l'enveloppe d'un bâtiment sous différents climats et garantir le confort des occupants
Hemp concrete is one of the most popular bio-based building materials in France. It is often used as a filling and / or insulation material, in an eco-construction system. Given its high porosity, this concrete has particularly advantageous thermal and hygric performance. However, the lighter this hemp concrete, the higher its level of thermal insulation, while its mechanical properties decrease. This is why it is necessary to optimize its formulation, in order to obtain the best compromise between its mechanical, hygric and thermal performance, depending on the field of use (carrier and / or insulator). The first optimization approach considered consists in varying the rate of plant aggregates in the composition of the concrete, but also the nature and quantity of binder used. The objective of this thesis is to study the mechanical, thermal and hygric properties of a hemp concrete according to its formulation. In this thesis, we are interested in the feasibility of lightweight building materials based on hemp particles and a lime-based binder, which present good mechanical performances. The main objective is to overcome the migration process of free Ca2+ to the lumen of hemp particles that are responsible for their degradation and which, consequently, induce the loss of mechanical performance of the final composite material. To achieve this, a fraction of the base binder is replaced by metakaolin. First, we studied the influence of different percentages of metakaolin on the carbonation of two lime-based control mortars: NHL5 lime and Tradical PF70 pre-formulated lime. Then, we measured their mechanical resistances in compression and flexion in order to define an optimal formulation. Thus, the finished composite materials are tested for different volume percentages (2v and 3v) of hemp particles. The results of the characterization of the physico-mechanical properties of the specimens are presented and argued. A close relationship between the metakaolin content and the physico-mechanical properties of the hemp concrete has been observed, due to the additional hydration products derived from pozzolanic reaction mechanism. The MEB and EDX analyses have shown the enhancement of hemp particles-binder Interfacial Zone Transition, while the pozzolanic reaction leads to reduce the migration process of free Ca2+ to lumen of vegetable particles thus reducing their mineralization. Then, we compared the hygro-thermal performances of hemp concretes formulated with respectively 2 volumes and 3 volumes of plant particles for one volume of binder, without or with 20% metakaolin, the optimal percentage defined previously. This study is particularly useful to establish the adequate proportions of admixtures to be used in concretes intended for renovation works or new constructions. Generally speaking, and depending on the formulation, we obtain a material that is a good or even excellent regulator of ambient humidity, with interesting thermal performances for the insulation of buildings. This characterization work completes the many works already carried out on biobased concrete with different types of plants, and should make it possible to feed the databases essential for the simulation of the behavior of a wall or of the envelope of a building under different climates and guarantee the comfort of the occupants

Sizing (hydrophobization) is one of the most important process steps within the added-value chain of about 1/3rd of the worldwide produced paper & board products. Even though sizing with so-called reactive sizing agents, such as alkenyl succinic anhydride (ASA) was implemented in the paper industry decades ago, there is no total clarity yet about the detailed chemical and physical mechanisms that lead to their performance. Previous research was carried out on the role of different factors influencing the sizing performance, such as bonding between ASA and cellulose, ASA hydrolysis, size revision as well as the most important interactions with stock components, process parameters and additives during the paper making process. However, it was not yet possible to develop a holistic model for the explanation of the sizing performance given in real life application. This thesis describes a novel physico-chemical approach to this problem by including results from previous research and combining these with a wide field of own basic research and a newly developed method that allows tracing back the actual localization of ASA within the sheet structure. The carried out measurements and trial sets for the basic field of research served to evaluate the stock and process parameters that most dominantly influence the sizing performance of ASA. Interactions with additives other than retention aids were not taken into account. The results show that parameters, such as the content of secondary fibers, the degree of refining, the water hardness as well as the suspension conductivity, are of highest significance. The sample sets of the trials with the major impacting parameters were additionally analyzed by a newly developed localization method in order to better understand the main influencing factors. This method is based on optical localization of ASA within the sheet structure by confocal white light microscopy. In order to fulfill the requirements at magnification rates of factor 100 optical zoom, it was necessary to improve the contrast between ASA and cellulose. Therefore, ASA was pretreated with an inert red diazo dye, which does not have any impact on neither the sizing nor the handling properties of ASA. Laboratory hand sheets that were sized with dyed ASA, were analyzed by means of their sizing performance in correlation to measurable ASA agglomerations in the sheet structure. The sizing performance was measured by ultrasonic penetration analysis. The agglomeration behavior of ASA was analyzed automatically by multiple random imaging of a sample area of approx. 8650 µm² with a minimum resolution for particles of 500 nm in size. The gained results were interpreted by full factorial design of experiments (DOE). The trials were carried out with ASA dosages between 0% and 0.8% on laboratory hand sheets, made of 80% bleached eucalyptus short fiber kraft pulp and 20% northern bleached softwood kraft pulp, beaten to SR° 30, produced with a RDA sheet former at a base weight of 100 g/m² oven dry. The results show that there is a defined correlation between the ASA dosage, the sizing performance and the number and area of ASA agglomerates to be found in the sheet structure. It was also possible to show that the agglomeration behavior is highly influenced by external factors like furnish composition and process parameters. This enables a new approach to the explanation of sizing performance, by making it possible to not only examine the performance of the sizing agent, but to closely look at the predominant position where it is located in the sheet structure. These results lead to the explanation that the phenomenon of sizing is by far not a pure chemical process but rather a more physical one. Based on the gained findings it was possible so far to optimize the ASA sizing process in industrial-scale by means of ~ 50% less ASA consumption at a steady degree of sizing and improved physical sheet properties.

La protéine majeure du lactosérum, la β-lactoglobuline (βlg) est bien reconnue pour ses propriétés structurales intéressantes lui permettant d’établir des interactions avec des ligands de taille et de caractéristiques différentes. La riboflavine (RF) et la vitamine D3 (D3) ont été sélectionnées comme modèles de petits nutraceutiques amphiphiles et hydrophobes, respectivement, et le lysozyme (Lyso), comme ligand protéique de plus grande taille. La capacité de la βlg à lier la RF a été étudiée par des méthodes spectroscopiques. La βlg et la RF forment le complexe βlg-RF dont la photoactivation génère une activité antiproliférative contre les cellules cancéreuses de la peau, démontrée en utilisant le protocole du NCI/NIH Developmental Therapeutics Program. La cytotoxicité serait probablement due à la génération d’espèces oxydatives réactives résultant de l’interaction entre la RF et la βlg. L’impact de la formation du βlg-D3 sur la solubilité et stabilité de la D3 a été étudié en utilisant des méthodes spectroscopiques et de chromatographie. Les résultats ont démontré que le complexe βlg-D3 était stable aux pHs gastrique et intestinal et augmentait la solubilité de la D3. De plus, une matrice protéique appelée coagulum enrichie en D3 (94.5 ± 1.8 % de taux d’encapsulation) été formée à partir du complexe βlg-D3 grâce à l’aptitude de la βlg à s’auto-associer. Les images de microscopie électronique ont montré que les interactions électrostatiques entre la βlg et Lyso ont pour leur part, abouti à la formation de microsphères pouvant encapsuler la D3 à un taux élevé (90.8 ± 4.8 %). La capacité des matrices à base de βlg à transporter, protéger et à améliorer la solubilité, la stabilité et la biodisponibilité de la D3 a été évaluée en effectuant des expériences in vitro et in vivo chez des modèles animaux. Les matrices protéiques à base de βlg ont significativement augmenté la solubilité, stabilité et biodisponibilité de la D3 (p < 0.001). Ces études prouvent que la βlg, grâce à ses caractéristiques suturales, pourrait former des matrices protéiques compatibles avec une administration orale et les aliments, tout en préservant l’activité biologique de la RF, de la D3 et donc possiblement d’autres molécules bioactives
The major whey protein, β-lactoglobulin (βlg) is well recognized for its interesting structural properties and ability to interact with ligands with varying size and characteristics. Riboflavin (RF) and vitamin D3 (D3) were selected as small amphiphilic and hydrophobic nutraceutical models, respectively, and Lysozyme, as a larger size ligand model. Spectroscopic methods were used to demonstrate interaction between βlg and RF. βlg and RF form a complex, which was irradiated according to the NCI/NIH Developmental Therapeutics Program. The βlg-RF complex exhibited an important anti-proliferative activity against skin melanoma cancer cell lines, probably due to the generation of reactive oxygen species as the result of the interaction between RF and βlg. The impact of the βlg-D3 complex on the solubility and stability of the D3 was studied using spectroscopic methods and chromatography. The findings indicate that the βlg-D3 complex is stable at the gastric and intestinal pHs and increases the solubility of the vitamin. A βlg-based scaffold, named coagulum, enriched with D3 (94.5 ± 1.8 % of encapsulation efficiency) was prepared by using the capacity of βlg to self-aggregate. Electronic microscopy images showed that microspheres, with high D3 encapsulation efficiency (90.8 ± 4.8 %), were formed as the result of electrostatic interactions between βlg and Lyso. The efficiency of βlg-based scaffolds to improve the solubility, stability and bioavailability of the D3 was evaluated by performing in vitro and in vivo experiments using animal model. The βlg-based scaffolds significantly increased the solubility, stability bioavailability of D3 (p < 0.001). Overall, the present study showed that βlg, due to its structural properties, can be used to form protein-based matrices compatible with a food and an oral administration while preserving the biological activity of RF, D3 and possibly other bioactive molecules.

La protéine majeure du lactosérum, la β-lactoglobuline (βlg) est bien reconnue pour ses propriétés structurales intéressantes lui permettant d’établir des interactions avec des ligands de taille et de caractéristiques différentes. La riboflavine (RF) et la vitamine D3 (D3) ont été sélectionnées comme modèles de petits nutraceutiques amphiphiles et hydrophobes, respectivement, et le lysozyme (Lyso), comme ligand protéique de plus grande taille. La capacité de la βlg à lier la RF a été étudiée par des méthodes spectroscopiques. La βlg et la RF forment le complexe βlg-RF dont la photoactivation génère une activité antiproliférative contre les cellules cancéreuses de la peau, démontrée en utilisant le protocole du NCI/NIH Developmental Therapeutics Program. La cytotoxicité serait probablement due à la génération d’espèces oxydatives réactives résultant de l’interaction entre la RF et la βlg. L’impact de la formation du βlg-D3 sur la solubilité et stabilité de la D3 a été étudié en utilisant des méthodes spectroscopiques et de chromatographie. Les résultats ont démontré que le complexe βlg-D3 était stable aux pHs gastrique et intestinal et augmentait la solubilité de la D3. De plus, une matrice protéique appelée coagulum enrichie en D3 (94.5 ± 1.8 % de taux d’encapsulation) été formée à partir du complexe βlg-D3 grâce à l’aptitude de la βlg à s’auto-associer. Les images de microscopie électronique ont montré que les interactions électrostatiques entre la βlg et Lyso ont pour leur part, abouti à la formation de microsphères pouvant encapsuler la D3 à un taux élevé (90.8 ± 4.8 %). La capacité des matrices à base de βlg à transporter, protéger et à améliorer la solubilité, la stabilité et la biodisponibilité de la D3 a été évaluée en effectuant des expériences in vitro et in vivo chez des modèles animaux. Les matrices protéiques à base de βlg ont significativement augmenté la solubilité, stabilité et biodisponibilité de la D3 (p < 0.001). Ces études prouvent que la βlg, grâce à ses caractéristiques suturales, pourrait former des matrices protéiques compatibles avec une administration orale et les aliments, tout en préservant l’activité biologique de la RF, de la D3 et donc possiblement d’autres molécules bioactives
The major whey protein, β-lactoglobulin (βlg) is well recognized for its interesting structural properties and ability to interact with ligands with varying size and characteristics. Riboflavin (RF) and vitamin D3 (D3) were selected as small amphiphilic and hydrophobic nutraceutical models, respectively, and Lysozyme, as a larger size ligand model. Spectroscopic methods were used to demonstrate interaction between βlg and RF. βlg and RF form a complex, which was irradiated according to the NCI/NIH Developmental Therapeutics Program. The βlg-RF complex exhibited an important anti-proliferative activity against skin melanoma cancer cell lines, probably due to the generation of reactive oxygen species as the result of the interaction between RF and βlg. The impact of the βlg-D3 complex on the solubility and stability of the D3 was studied using spectroscopic methods and chromatography. The findings indicate that the βlg-D3 complex is stable at the gastric and intestinal pHs and increases the solubility of the vitamin. A βlg-based scaffold, named coagulum, enriched with D3 (94.5 ± 1.8 % of encapsulation efficiency) was prepared by using the capacity of βlg to self-aggregate. Electronic microscopy images showed that microspheres, with high D3 encapsulation efficiency (90.8 ± 4.8 %), were formed as the result of electrostatic interactions between βlg and Lyso. The efficiency of βlg-based scaffolds to improve the solubility, stability and bioavailability of the D3 was evaluated by performing in vitro and in vivo experiments using animal model. The βlg-based scaffolds significantly increased the solubility, stability bioavailability of D3 (p < 0.001). Overall, the present study showed that βlg, due to its structural properties, can be used to form protein-based matrices compatible with a food and an oral administration while preserving the biological activity of RF, D3 and possibly other bioactive molecules.

La curcumine est un polyphénol de couleur jaune extrait du rhizome de la plante (Curcuma longa). Identifié comme un principe actif du curcuma, cette épice est largement consommée compte tenu de ses propriétés antioxydantes, antimicrobiennes et antitumorales. Cependant, sa solubilité et sa biodisponibilité restent limitées en raison de son hydrophobicité et diminuent de ce fait les applications potentielles dans les domaines nutraceutique et pharmaceutique. L’objectif de ce travail s’est focalisé sur l’étude des propriétés physico-chimiques de cette molécule vectorisée sous forme liposomale. Différentes formulations de nanovecteurs préparées à partir de lipides polaires plus ou moins riches en acides gras polyinsaturés à longue chaîne variables, issus de source marine ou végétale, ont été utilisées pour modifier la fluidité membranaire des vecteurs. Ces travaux ont permis d’optimiser le pourcentage d’encapsulation de cette biomolécule et d’étudier les effets de cytotoxicité et de sa biodisponibilité sur différentes lignées cellulaires (MCF7, cellules embryonnaires de neurones corticaux). Les résultats montrent des effets significatifs lors de l’utilisation de nanoliposomes formulés avec de la lécithine marine, protégés par un enrobage de polymère de chitosane. Les analyses physicochimiques de taille par diffraction de la lumière (zetasizer, nanosight), de stabilité (mesure de la taille et la mobilité électrophorétique), de structure par microscopie électronique en transmission et la libération de la molécule vectorisée, ont permis de mieux comprendre les effets des interactions polymère-phospholipides et le relargage de la curcumine encapsulée dans les conditions environnementales drastiques de digestion gastrique et intestinale. Une caractérisation multi-échelle est proposée afin d’améliorer la compréhension des différentes propriétés du nanovecteur et du principe actif qu’il vectorise, ainsi que les interactions possibles entre eux. Les techniques utilisées sont la spectroscopie infrarouge à transformée de Fourier (FTIR), les rayons X, ainsi que la microscopie électronique en transmission
Curcumin, a yellow lipid-soluble natural pigment, a hydrophobic polyphenol derived from the rhizome of Curcuma longa, has been identified as the active principle of turmeric. Curcumin has already become a research focus due to its numerous biological and pharmacological benefits such as antioxidant, antitumor, anti-inflammatory, antimicrobial properties and other desirable medical benefits. However, due to its low absorption, the poor bioavailability of curcumin limits its clinical use. The objective of this work has focused on the study of the physicochemical properties of this molecule encapsulated in form of liposome. Different formulations of nanocarriers prepared from polar lipids more or less rich in polyunsaturated fatty acids with variable long chain, derived from vegetable or marine source, have been used to modify liposomes membrane fluidity. These works have allowed to optimize the encapsulation efficiency of the curcumin and to study its bioavailability and cytotoxicity effects on different cell lines (MCF7, embryonic cortical neurons). The results show significant effects when using nanoliposomes formulated with marine lecithin, protected by chitosan as the coating polymer. The physico-chemical analyses of the size by light scattering (Zetasizer, NanoSight), the stability (measurement of the size and the electrophoretic mobility), the structure by transmission electron microscopy and the release of the encapsulated molecule, allowed to better understand the effects of polymer-phospholipid interactions and the release of encapsulated curcumin in drastic environmental conditions of gastric and intestinal digestion. A multiscale characterization is proposed in order to improve the understanding of the various properties of the active agent (curcumin) and the nanovector, as well as the possible interactions occuring between them. The techniques used are Fourier Transform Infrared Spectroscopy (FTIR), X-rays, Rheometer and Transmission Electron Microscopy

Ce manuscrit est consacré à l'étude et au développement de microsystèmes capteurs de gaz sélectifs au dioxyde d'azote, destinés au contrôle de la qualité de l'air atmosphérique. La stratégie que nous avons développée consiste à associer une structure sensible à base de matériaux semi-conducteurs partiellement sélectifs aux gaz oxydants et des filtres sélectifs à l'ozone. L'objectif premier est la mise en oeuvre et la caractérisation de matériaux chimiques strictement imperméables à l'ozone (O3) et non-réactifs vis-à-vis du dioxyde d'azote (NO2). Notre choix s'est focalisé sur un matériau moléculaire, l'indigo, connu pour sa réactivité vis-à-vis de O3, et plusieurs nanomatériaux carbonés. Pour ces derniers, la possibilité de conformer leurs textures, leurs morphologies et leurs chimies de surface par traitements thermiques, chimiques et mécaniques, permet d'étendre le panel de matériaux potentiels et d'identifier les facteurs d'influence de leur réactivité avec les espèces gazeuses. La caractérisation de l'ensemble de ces matériaux a nécessité l'utilisation de techniques adaptées et complémentaires (adsorption de N2 à 77 K, spectroscopies Raman, XPS, IR en mode ATR, RPE et NEXAFS). Les filtres chimiques les plus efficaces (hauts rendements de filtration et grande durabilité) ont été sélectionnés d'après des tests de soumission aux gaz selon une méthodologie adaptée. Enfin, l'association de ces meilleurs filtres et de la structure capteur a conduit à l'élaboration de prototypes microsystèmes capteurs de gaz optimisés. De plus, une contribution à la compréhension des mécanismes d'interaction de l'indigo et de certains nanocarbones avec O3 et NO2 a aussi permis d'améliorer le microsystème en développant des méthodologies pertinentes et innovantes mais également en réalisant la synthèse de nouveaux filtres indigo / nanocarbone.

Dans les écosystèmes marins des zones tempérées, la majeure partie de la production primaire annuelle est générée au printemps lors de phénomènes d’accumulation rapide de biomasse phytoplanctonique, appelés efflorescences, supportant la diversité et le fonctionnement de ces systèmes. Plusieurs mécanismes physico-chimiques et biologiques expliquant l’initiation des efflorescences phytoplanctoniques sont évoqués pour ces écosystèmes. En revanche pour les zones côtières peu profondes sous influence de forçages complexes, les mécanismes à la base de ce phénomène restent encore mal connus. L’objectif de cette thèse était donc d’identifier les facteurs contribuant à l’initiation des efflorescences phytoplanctoniques dans ces zones, notamment le rôle des forçages physico-chimiques et des interactions biologiques au sein du réseau microbien, mais également de tenter de comprendre les conséquences de l’élévation de la température sur ce fonctionnement dans le contexte du réchauffement climatique. Dans cette optique, un suivi à deux approches a été réalisé dans la lagune de Thau : un suivi in situ à haute fréquence (15 min) des paramètres hydrologiques, météorologiques et biologiques ; et un suivi hebdomadaire de l’abondance et la diversité de la communauté microbienne de l’hiver au printemps sur deux années consécutives, 2015 et 2016. En plus de ces suivis, une expérimentation en mésocosmes in situ a été réalisée au printemps 2018, simulant l’élévation de la température selon le scénario du réchauffement climatique attendu dans le futur, et en présence et absence du mésozooplancton afin d’identifier les effets directs du réchauffement et ceux indirects dus au zooplancton sur la dynamique, la composition pigmentaire et la succession du phytoplancton, avant, pendant et après une efflorescence phytoplanctonique. Une analyse basée sur les réseaux de corrélations entre 110 différents groupes/taxon/espèces observés a permis de mettre en évidence les interactions majeures au sein du réseau planctonique microbien. Pendant les périodes d’efflorescences les interactions de compétition intraguilde au sein du phytoplancton dominaient, tout comme les interactions mutualistes entre le phytoplancton et les bactéries hétérotrophes, suggérant un transfert de l’énergie basée à la fois sur la biomasse phytoplanctonique et bactérienne effectué par la prédation du microzooplancton. Pendant les épisodes sans efflorescence, les interactions entre les ciliés et les bactéries (bactérivorie) dominaient, suggérant un transfert de l’énergie basée essentiellement sur la biomasse bactérienne. Dans le même temps, les résultats obtenus par le suivi à haute fréquence ont permis de mettre en évidence le rôle prépondérant de l’augmentation de la température de l’eau, notamment à la sortie de l’hiver, dans l’initiation des efflorescences phytoplanctoniques. La combinaison entre le métabolisme du phytoplancton stimulé par l’augmentation de la température et la faible pression de broutage, permettrait l’accumulation de la biomasse phytoplanctonique à l’origine des efflorescences phytoplanctoniques. De plus, l’année 2016, avec l’hiver le plus chaud jamais enregistré, était caractérisée par une plus faible accumulation de la biomasse phytoplanctonique, une dominance du phytoplancton de plus petite taille au détriment des diatomées, et une dominance des interactions entre microorganismes de petite taille. Les résultats de l’expérimentation en mésocosmes in situ ont confirmé l’effet de l’élévation de la température dans la réduction de l’amplitude des efflorescences et de la favorisation du petit phytoplancton comme les petites algues vertes ainsi que des dinoflagellés au détriment des diatomées. De plus, ils ont permis de mettre en évidence que cette modification de l’amplitude et de la composition des efflorescences était principalement liée à un effet indirect sur le phytoplancton due à l’augmentation de la pression de broutage exercée par le zooplancton
In temperate marine ecosystems, the major part of the annual primary production is generated in spring during rapid phytoplankton biomass accumulation periods, called ‘blooms’, supporting the diversity and the functioning of these ecosystems. Several physical, chemical and biological mechanisms triggering the bloom initiation were evocated for these ecosystems. However, for shallow coastal zones, under the influence of complex environmental forcing factors, mechanisms triggering blooms are not well known. The objective of the present thesis was to identify and classify the forcing factors contributing to the bloom initiation in these zones, especially the role of physical and chemical forcing factors and biological interactions in the microbial network, but also to understand the consequences of the temperature elevation on this functioning in the global warming context.In this frame, a monitoring with a dual approach was carried out in Thau lagoon: a high frequency (15 min) in situ monitoring of hydrological , meteorological and biological parameters; and a weekly monitoring of the abundance of the microbial community (virus, bacteria, phytoplankton, heterotrophic flagellates and ciliates), and its diversity, with a particular look at phytoplankton. These monitoring were carried out from winter to spring in two consecutive years, 2015 and 2016. Besides these monitoring, an in situ mesocosm experiment was carried out during the 2018 spring to simulate the temperature elevation according to the global warming scenario, in the presence and the absence of mesozooplankton. The objective of this experiment was to identify the direct effect of warming and the indirect effect of the zooplankton on the phytoplankton dynamic, the pigment composition and succession, during the pre-bloom, bloom and post-bloom periods. A correlation network analysis between 110 various groups/taxa/species highlighted the major interactions characterizing the microbial interaction network during the bloom and the non-bloom periods and the differences between these two years. During the bloom periods, intraguild phytoplankton competition and mutualism between phytoplankton and heterotrophic bacteria dominated the microbial food web. This suggested an energy transfer based on both bacterial and phytoplanktonic biomass, through the microzooplankton predation. During the non-bloom periods, interaction between ciliates and heterotrophic bacteria (bacterivory) dominated, suggesting an energy transfer mainly based on bacterial biomass. Besides, the high frequency monitoring highlighted the predominant role of the water temperature increase, especially during the early spring, in the initiation of the phytoplankton blooms. The combination between the phytoplankton metabolism stimulated by the temperature increase and the low grazing pressure triggered the phytoplankton biomass accumulation starting the blooms. Furthermore, 2016 year, with the warmer winter recorded in France (Meteofrance), was characterized by a weaker phytoplankton biomass accumulation during the early spring, a dominance of the small phytoplankton at the expanse of diatoms, and a dominance of interactions between small size microorganisms. The mesocosm experiment confirmed the role of the temperature elevation on the bloom amplitude reduction (diminution of 50% of the chlorophyll a concentration) and the promotion of small phytoplankton such as small green algae and dinoflagellates, at the expanse of diatoms. This amplitude and composition modification of phytoplankton blooms was mainly due to the indirect effect of the zooplankton grazing increase under warming. Furthermore, the results underlined that it was microzooplankton which mainly controlled the phytoplankton dynamic and biomass and the mesozooplankton was mainly accomplished the role of the secondary consumer in this system

L'objectif de ce travail était d'étudier les interactions entre une plante « phytoremédiatrice », Lantana camara (Verbenaceae), le ver de terre, Pontoscolex corethrurus (Glossoscolecidae) et les microorganismes telluriques d'un sol pollué au plomb. Dans un premier temps, il apparaît que dans les sols contaminés, la présence de ver conduit à un accroissement de la biomasse des parties aériennes et racinaires des plantes ainsi qu'à une augmentation de l'absorption de plomb. La caractérisation physico-chimique des agrégats racinaires a montré que l'activité des vers augmente le taux de matière organique, la capacité d'échange cationique ainsi que l'azote total, le potassium total et disponible. De plus, la présence des vers augmente certaines activités enzymatiques de la rhizosphère. La croissance accrue de L. camara pourrait résulter de ces différentes actions. L'action des vers de terre sur les plantes se ferait via les communautés microbiennes telluriques. Ainsi, la biomasse des microorganismes, bactéries et champignons, des agrégats racinaires augmente en présence de vers. La PCR-DGGE n'a pas permis de mettre en évidence de modifications de la structure taxonomique des communautés bactériennes sous l'influence du Pb et/ou du vers, par contre l'analyse des profils physiologiques par plaques Biolog montre clairement une diversification fonctionnelle bactérienne. Les communautés fongiques voient, elles, leur diversité taxonomique, augmenter sous l'action des vers. La restructuration des populations microbiennes, en présence de vers, des agrégats racinaires élaborés par les plantes en milieu pollué au plomb est l'élément déterminant pour la compréhension de l'impact de P. corethrurus sur la croissance et la phytoremédiation de L. camara. L'association de ces deux organismes aurait donc un potentiel considérable pour le traitement de sites industriels pollués au plomb

La thèse a pour but d’étudier les effets de changements de mouillabilité de roches dans des conditions d’injections d’eau douce en tant que méthode de récupération d’hydrocarbures. Afin d’identifier le ou les mécanismes à l’origine du gain additionnel de récupération nous avons utilisé un microtomographe RX. Nous avons ainsi imagé les états de saturations finales d’un milieu poreux rempli de saumures et d’huiles. Une fois le drainage primaire réalisé nous avons effectué deux phases d’imbibitions : avec une saumure (récupération secondaire) puis une imbibition d’eau douce (récupération tertiaire). L’analyse de la mouillabilité à l’échelle du pore a permis de mettre en évidence l’effet de la température et de la salinité sur la mouillabilité. Nous avons montré que les changements de mouillages des roches n’étaient pas occasionnées par la seule expansion de la couche électrique en revanche des changements de mouillabilité ont été montrés. Ces changements s’expliquant par des transitions de mouillages de second ordre observées non seulement pour des gouttes d’huiles sur de l’eau mais également sur un substrat en verre. Au final, la mouillabilité en milieux poreux doit être mise en évidence à une échelle sous-Micrométrique ce qui est relativement nouveau dans le domaine pétrolier
The aim of the thesis is to study rock wettability change effects caused by Low Salinity brine injection as tertiary recovery method. To identify the underlying mechanism or mechanisms of additional oil recovery X-Ray imaging technology was applied. We have also imaged the end-Point saturations of filled by brine and water core samples. Once the primary drainage is realized we carried out two phases imbibitions: with high salinity brine (waterflooding) and with low salinity brine (tertiary recovery mode). The wettability analysis at pore scale permitted to put in evidence the thermal and saline effects playing a decisive role in rock wettability. We have showed wettability changes are not caused by only electrical double layer expansion, however wettability changes was shown. These changes are explained by wettability transition of second order and observed not only for oil droplet on brine, but also for oil deposited on glass substrate. Finally, the pore space wettability needs to be evidenced at sub-Micrometric scale that is new for the petroleum domain

L’objectif principal de cette thèse était double : (i) mesurer l’impact d’une espèce de fourmi sur son écosystème, afin (ii) d’en déduire des applications potentielles dans le domaine de la restauration écologique.Les fourmis sont parmi les organismes les plus abondants des écosystèmes terrestres et occupent des zones géographiques très variées. Elles jouent des rôles écologiques clés dans de nombreux écosystèmes comme ingénieurs du sol, prédateurs ou régulateurs de la croissance et de la reproduction des plantes. Cependant les données collectées localement sont souvent parcellaires et ne permettent pas d’avoir une vision complète de l’impact d’une espèce sur son milieu.Messor barbarus (L.), connue pour redistribuer les graines et pour modifier les propriétés physico-chimiques du sol, est largement répandue dans le Sud-Ouest de l’Europe notamment au sein des pelouses méditerranéennes. Elle pourrait donc jouer un rôle majeur dans la composition et structuration de ces pelouses caractérisées par une forte biodiversité mais dont le nombre et la superficie ont drastiquement diminué ces dernières décennies.Dans un premier temps, par une étude multi-compartiments, nous avons confirmé l’hypothèse selon laquelle M. barbarus est une ingénieure de l’écosystème au sein des pelouses méditerranéennes. Elle transforme cet habitat en modifiant, comme attendu, les propriétés physico-chimiques du sol. Ces modifications sont associées à une augmentation de la biomasse et de l’hétérogénéité des communautés végétales ainsi qu’à des changements dans les faunes épigée et endogée (abondance, occurrence et structure des communautés). De plus, M. barbarus modifie profondément les relations trophiques et non trophiques interspécifiques et entre les espèces et leur habitat. L’hétérogénéité créée à l’échelle locale par l’activité de cette fourmi, entraine une diversification des niches écologiques au sein de ces pelouses.Malgré leur rôle souvent majeur sur le fonctionnement des écosystèmes, les fourmis ne sont que très rarement considérées en restauration écologique. Sur notre site d’étude, un chantier de réhabilitation d’une pelouse sèche après une fuite d’hydrocarbures et un transfert de sol, M. barbarus a permis d’accélérer la restauration des propriétés physico-chimiques du sol mais aussi de la banque de graines à moyen terme - sept ans après la réhabilitation du site. Ces résultats font donc de cette espèce une bonne candidate en ingénierie écologique.Afin de généraliser l’utilisation des fourmis en restauration écologique, nous proposons une méthodologie à destination des gestionnaires basée sur l’utilisation de traits fonctionnels et d’histoire de vie. Pour cela nous avons évalué le potentiel des fourmis en écologie de la restauration, puis nous avons listé l’ensemble des traits connus pour affecter les compartiments abiotiques et biotiques et/ou pertinent pour effectuer un suivi du succès de la phase de restauration. La méthodologie proposée permet une première sélection des espèces potentiellement utilisables en fonction des objectifs de restauration
The main objective of this thesis was double: (i) to assess the impact of an ant species on its ecosystem, in order to (ii) deduce potential applications in the field of ecological restoration.Ants are among the most abundant organisms in terrestrial ecosystems and occupy a wide range of geographical areas. They play key ecological roles in many ecosystems as soil engineers, predators or regulators of plant growth and reproduction. However, the information collected locally is often fragmented and does not provide a complete overview of the impact of a species on its environment.Messor barbarus (L.), known to redistribute seeds and to modify the soil physico-chemical properties, is widespread in South-Western Europe, particularly in Mediterranean grasslands. Therefore, it may play a major role in the composition and structuring of these ecosystems, which are characterised by high biodiversity but whose abundance and surface area have decreased drastically in recent decades.Through a multi-compartment study, we confirmed the hypothesis that M. barbarus is an ecological engineer in Mediterranean grasslands. This species changes this habitat by modifying, as expected, soil physico-chemical properties. These modifications are associated with an increase in both biomass and heterogeneity of plant communities, as well as changes in above- and belowground fauna (abundance, occurrence and structure of communities). Messor barbarus profoundly changes trophic and non-trophic relationships within and between species and their habitat. The heterogeneity created locally by the activity of M. barbarus leads to a diversification of ecological niches within these grasslands.Despite their major role in the functioning of ecosystems, ants are rarely considered in restoration ecology. In our study site, corresponding to a dry grassland rehabilited after an oil leak and a soil transfer, M. barbarus contributed to accelerate the restoration of the soil physico-chemical properties but also of the seed bank in the medium term - seven years after the rehabilitation. These results make this species a good candidate for ecological engineering.In order to generalise the use of ants in restoration ecology, we propose a trait-based methodology for stakeholders. We evaluated the potential of ants in restoration ecology, then listed all the traits known to affect abiotic and biotic compartments and/or relevant to monitor the success of the restoration phase. The proposed methodology provides a first selection of potentially relevant species according to the restoration objectives

Upon ageing, a progressive disruption of protein homeostasis often leads to extensive protein aggregation and neurodegeneration. It is therefore important to study at the proteome level the origins and consequences of such disruption, which so far have remained elusive. Addressing this problem has recently become possible by major advances in mass spectrometry-based (MS) proteomics, which allows the identifications and quantification of thousands of proteins in a variety of biological samples. In the first part of this thesis, I analyse proteome-wide MS data for the nematode worm C. elegans upon ageing, in wild type (WT), long-lived and short-lived mutant strains. By comparing the total abundance and the soluble abundance for nearly 4000 proteins, I provide extensive evidence that proteins are expressed in adult worms at levels close to their solubility limits. With the use of sequence-based prediction tools, I then identify specific physico-chemical properties associated with this age-related protein homeostasis impairment. The results that I obtained reveal that the total intracellular protein content remains constant, in spite of the fact that the proteome undergoes wide remodeling upon ageing, resulting into severe protein homeostasis disruption and widespread protein aggregation. These results suggest a protein-dependent decrease in solubility associated with the protein homeostasis failure. In the second part of the thesis, I determine and classify potential interactions of misfolded protein oligomers with other proteins. This phenomenon is widely believed to give rise to cytotoxicity, although the mechanisms by which this happens are not fully understood. To address this question, I process and analyse MS data from structurally different oligomers (toxic type A and nontoxic type B) of the protein HypF-N, incubated in vitro with proteins extracted from murine cell cultures. I find that more than 2500 proteins are pulled down with the misfolded oligomers. These results indicate that the two types of oligomers interact with the same pool of proteins and differ only in the degree of binding. Functional annotation analysis on the groups reveals a preference of the oligomers to bind proteins in specific biological pathways and categories, including in particular mitochondrial membrane proteins, RNA-binding proteins and molecular chaperones. Overall, in this study I complement the powerful and high-throughput experimental approach of MS proteomics with bioinformatics analyses and prediction algorithms to define the physical, chemical and biological features of protein homeostasis disruption upon ageing and the interactome of misfolded oligomers.

La calcite (carbonate de calcium) est un matériau extrêmement répandu dans les roches telles que le marbre et comme constituant dans des domaines variés (bâtiment, pharmacie, papier, art). La compréhension des propriétés mécaniques des suspensions de calcite constitue une étape importante pour améliorer à la fois leur maniabilité ainsi que les propriétés finales du matériau fabriqué. Cette étude relie les propriétés rhéologiques (élasticité, écoulement) de ces suspensions à leurs interactions microscopiques. Les interactions attractives entre particules de calcite confèrent aux pâtes les propriétés de gels colloidaux caractérisés par un module élastique et une déformation critique, et ce pour une large gamme de concentrations. L'étude de ces grandeurs en fonction de la concentration a permis de mettre en évidence pour la première fois l'existence de deux régimes de déformation (liens forts et faibles) prédits théoriquement et de caractériser la dimension fractale. L'étude des interactions a été réalisée grâce à la mesure du potentiel Zeta des pâtes, du pH et au calcul de la longueur de Debye résultant de la force ionique. L'ajout d'additifs tels que la chaux ou la soude modifie les interactions. La chaux réduit fortement l'élasticité initiale des pâtes, facilitant leur maniabilité et renforçant la réactivité de la pâte en présence de CO2. L'ajout de soude augmente fortement l'attraction entre les particules ce qui se traduit à l'échelle macroscopique par l'existence de bandes de cisaillement. Cette manifestation de l'attraction entre colloïdes à l'échelle macroscopique avait été observée dans des émulsions concentrées mais encore jamais dans les gels colloïdaux
Calcite (calcium carbonate) is an extremely widespread material that can be found naturally in rocks (i.e. marble, limestone) and is employed in many industrial fields such as paper filling, pharmaceutical, art or construction. Understanding the mechanical properties of calcite suspensions is a first step to improve the workability of the paste as well as the final properties of solid mineral materials. Macroscopic characterization of calcite suspensions via rheological measurements are linked to microscopic interactions, via DLVO analysis. Our calcite pastes are weakly attractive systems showing a typical colloidal gel behavior and characterized by an elastic shear modulus and a critical strain. The elastic domain of pure calcite suspensions is characterized for a wide range of volume concentrations. The deformation at the end of linearity exhibits a minimum versus concentration, a major prediction of colloidal gel theory, never verified so far. The interaction forces between particles are tuned by addition of simple ionic species. Rheological measurements are analyzed through DLVO calculations, obtained by chemical speciations and ζ potential measurements on dense suspensions. Addition of calcium hydroxide improves initially the workability of the paste, enhancing the reactivity when in contact with CO2. The role of interaction forces is also evaluated with flow measurements. The addition of sodium hydroxide increases strongly the attraction between particles, inducing shear bands at the macroscopic scale. This correlation is well known for emulsions but never verified so far for colloidal gels

Les tentatives menées pour détruire des cellules cancéreuses avec les agents radiothérapeutiques à base de 211 At qui ont été synthétisés jusqu’à présent ne sont pas encore pleinement satisfaisantes car elles sont entachées par une deastatination in vivo. Étant donné que ce problème est lié aux connaissances actuelles qui sont limitées concernant la chimie de base de l’astate et de ses espèces, des recherches fondamentales combinant des expériences à l’échelle des ultra-traces et des études théoriques ont été lancées. Dans cette thèse, une étude théorique de plusieurs espèces de l’astate est réalisée au moyen de méthodes relativistes basées sur la théorie de la fonctionnelle de la densité ou des méthodes à basées sur la fonction d’onde. Tout d'abord, les méthodes qui peuvent être utilisées pour faire des prédictions pertinentes sont établies. A l’aide de ces approches, nous rationaliserons les structures électroniques, géométries et propriétés physicochimiques des différents systèmes d'intérêt théorique ou expérimental, en particulier les espèces AtF3 et AtO+. Finalement, nous identifierons formellement une nouvelle espèce de l’astate à l’aide de résultats expérimentaux et de calculs, ce qui non seulement complète le diagramme de Pourbaix de l’astate en milieu aqueux non complexant, mais aussi donne des informations cruciales pour identifier des conditions expérimentales pour rendre le plus « réactif » possible le précurseur At−, qui est de nos jours impliqué dans la synthèse d’agents radiothérapeutiques innovants
Trials to destroy cancer cells with currently synthesized 211 At-based radiotherapeutic agents are not yet fully satisfactorily since they resume to in vivo deastatination. Since this issue is related to the limited knowledge of the basic chemistry of At and its species, fundamental researches combining ultra-trace experiments and computational studies have been initiated. In this thesis, a computational study of several At species is performed, by means of relativistic density functional theory and wave-function-based calculations. First, the quantum mechanical approaches that can safely be used to make adequate predictions are established. Using these approaches, we attempt to rationalize the electronic structures, geometries, and physico-chemical properties of various systems of theoretical and/or experimental interest, in particular the AtF3 and AtO+ ones. By the end, we firmly identify a new At species by combining outcomes of experiments and calculations. This new species not only completes the Pourbaix diagram of At in aqueous and non-complexing media, but also gives clues of identifying experimental conditions to make best reactive the At– precursor, which is currently involved in the synthesis of promising radiotherapeutic agents

Shale is often the most difficult of all formations to maintain a stable wellbore in when drillincr ::> for oil and gas. Time and money spent overcoming this problem during drilling, together with overall reduced profit margins. has led the oil industry to devote considerable time and effort to solve the problem of unstable boreholes in shales. It has long been established that the moisture adsorption (or desorption) of shale rocks can be controlled by the salinity of drilling fluid. When compacted shale (under constant compaction stress) adsorbs moisture, its total volume increases and swelling strains develop. Developed swelling strains then become an integral part of the effective radial stress acting on the shale formation contributing to borehole failure. A mathematical model has been developed for predicting the swelling behaviour of shale when placed in contact with water under moderate pressures and the effect of the swelling on borehole (in)stability. The model is based on thermodynamic theory which suggests that fluid movement into or out of a shale is driven by an imbalance in the partial molar free energy of the shale and the contacting fluid. Conversion of the free energy of each system (fluid and shale) into "total swelling pressure" made it possible to model transient pressures and strains generated in shale. The analytical solution of the radial diffusivity equation is reduced to a simpler form for the model. The model was validated using equipment and experimental techniques which allow continuous monitoring of shale swelling as function of time and distance from the wetting end. It was found that increasing the compaction stress acting on the shale reduced the rate of swelling, and increasing the hydraulic pressure of the fluid on the shale's wetted surface increased the rate of swelling. This behaviour was adequately described by the model which therefore represents a new method for predicting shale swelling as function of time and radial distance under different environments. Swelling strains are then used to predict related changes in shale mechanical properties (failure criteria) and well (in)stability. Several well-site index tests have been developed to study shale-drilling fluid interaction at wellsite. These index tests can provide input data for the mathematical model. Drilling fluids can be screened for their ability to control shale swelling, thus minimising the risk of well bore instability.

Polymer-bioglass composites are favourable materials for bone repair. However, early failure in the interface of components is a common problem in physical mixtures. The aim of this project was to address the issue of phase separation by creating a hybrid material in which the polymer is bonded to bioglass. Synthetic non-biodegradable poly(methyl methacrylate) (PMMA) and natural biodegradable chitosan were selected as two types of polymers for fabrication of hybrid with bioglass. PMMA and chitosan were functionalised with appropriate silane coupling agents and covalently bonded to bioglass. The polymer and bioglass were then co-condensed during sol-gel method to form hybrid. The results of molecular scale analyses demonstrated that at optimum condition (0.1 coupling agent:MMA mol ratio, 60:40 vol% polymer:bioglass), the covalent bond between PMMA and bioglass occurred and resulted in the fabrication of hybrid. The presence of nano-scale interaction resulted in improved physico-chemical and biological properties compared with physical mixtures and bioglass. Furthermore, by manipulating process parameters such as replacing tetrahydrofuran with ethanol, increasing the temperature to 70 °C and adding sodium bicarbonate as catalyst, the gelation time was reduced and a more condensed structure was produced. The chitosan-bioglass hybrid was optimized for the volume ratio of chitosan:bioglass and coupling agent. Furthermore, a new method was developed for the creation of porosity in polymer-bioglass composites in which sodium bicarbonate was used as a gas foaming agent and a biocompatible alternative for the commonly used hydrofluoric acid during sol-gel method. In conclusion, the presence of nano-scale interaction significantly improved the physico-chemical properties of polymer-bioglass hybrids via promoting the homogenous distribution of phases. These hybrids open an avenue for the applications of polymers-bioglass composites for bone replacement and tissue engineering.

2006 - 2007
Negli ultimi anni è stato dimostrato che sequenze di DNA (o RNA) ricche in guanine sono in grado di formare una nuova classe di strutture: le quadruple eliche. Importante e' la presenza di quadruple eliche in regioni promotrici e regolatrici di molti geni e nella parte terminale dei cromosomi, i telomeri. Recentemente, lo studio di queste molecole ha avuto un notevole sviluppo grazie alle diverse applicazioni che esse potrebbero avere in campo medico e farmaceutico. In particolare, le quadruple eliche hanno una potenziale applicazione nella terapia anticancro quali inibitori della telomerasi, un enzima la cui iperattività è sicuramente collegata allo sviluppo del cancro. Un promettente approccio per inibire l’attività della telomerasi riguarda l’utilizzo di agenti che possano legare e stabilizzare le quadruple eliche presenti ai telomeri bloccando, in tal modo, l’attività catalitica dalla telomerasi ed agendo da antitumorali. Inoltre, è stato scoperto che oligonucleotidi con struttura a quadrupla elica possono agire da aptameri, cioè hanno la capacità di legare specificamente delle proteine bersaglio, inibendole. L’aptamero denominato TBA (Thrombin Binding Aptamer) è un oligomero a DNA con struttura a quadrupla elica, scoperto in vitro, che è in grado di legare la trombina inibendone la funzione. Il TBA per le sue proprietà anticoagulanti è stato oggetto di numerosi studi sia di tipo strutturale sia di natura farmacologica. L’attività anticoagulante del TBA in vitro è tale da giustificare un suo eventuale impiego in terapia, ma purtroppo tale attività risulta piuttosto bassa in vivo poichè il DNA è rapidamente degradato dalle nucleasi. Uno degli obiettivi di questa tesi di dottorato è stato quello di affrontare uno studio chimico-fisico di composti che sono in grado di legarsi in maniera specifica alle quadruple eliche e dei fattori che regolano gli equilibri in gioco; tali composti sono, infatti, in grado di incrementare la stabilità di questi sistemi e di conseguenza ne possono aumentare l’effetto terapeutico. Per questo motivo è stata studiata la termodinamica dell’interazione di alcune molecole di interesse farmacologico, quali la distamicina, due suoi derivati e la porfirina cationica, con diverse quadruple eliche. Le quadruple eliche prese in esame sono state la d[AG3(TTAG3)3] e la [d(TAGGGTTAGGG)]2 che rappresentano due quadruple eliche formate da un diverso troncamento dal DNA telomerico umano e la d[(TGGGGT)]4 formata da una sequenza troncata del DNA telomerico di Oxytricha e Tetrahymena. Un altro obiettivo è stato quello di caratterizzare la stabilità termodinamica del TBA e di un aptamero modificato (mTBA) ed il processo di binding di questi aptameri con la trombina. L’mTBA presenta una modifica chimica in grado di rendere resistente l’aptamero all’azione delle nucleasi che potrebbe rendere concreto un suo eventuale utilizzo come principio attivo di un farmaco anticoagulante. Inoltre, è stato anche affrontato uno studio computazionale di due quadruple eliche bimolecolari formate da sequenze analoghe di RNA e DNA allo scopo di chiarire i motivi per cui si formano due diverse strutture a quadrupla elica ed i fattori che le stabilizzano. Lo studio è stata condotto principalmente attraverso tecniche calorimetriche quali la calorimetria isoterma (ITC), la microcalorimetria differenziale a scansione (DSC), il dicroismo circolare (CD), e metodi computazionali quali meccanica e dinamica molecolare e docking. I risultati ottenuti per i sistemi quadrupla elica-ligando dimostrano che la distamicina ed un suo derivato contenente un anello metil pirrolico in più ed un gruppo ammidico terminale, legano le quadruple eliche in esame, al contrario un secondo derivato della distamicina contenente due anelli metil pirrolici in più ed un gruppo ammidico terminale non ha mostrato avere interazioni specifiche. I parametri termodinamici ottenuti indicano che le interazioni quadrupla elica-ligando sono fortemente influenzate dallo ione presente in soluzione e che anche la stechiometria di legame è dipendente dal tipo di soluzione. Il calcolo dei parametri termodinamici ha mostrato che, sia in sodio che in potassio, il legame della distamicina e del composto I alle quadruple eliche in esame è entropicamente guidato. Questo dato in particolare suggerisce che il binding di queste molecole potrebbe avvenire nei solchi delle quadruple eliche. Inoltre, un risultato particolarmente interessante è che il composto I ha mostrato, in entrambe le soluzioni, una maggiore affinità della distamicina per le quadruple eliche. I risultati ottenuti per l’interazione porfirina-[d(TAGGGTTAGGG)]2 sono in generale accordo con la struttura cristallografica, recentemente riportata, del complesso ed indicano un binding entalpicamente guidato con una stechiometria 2:1. Lo studio della stabilità termodinamica del TBA e del TBA modificato ha rivelato che l’introduzione della modifica aumenta sia la stabilità termica che termodinamica dell’aptamero. Inoltre, i valori delle variazioni di entalpia ed entropia per la dissociazione del TBA modificato risultano più elevati rispetto al TBA, suggerendo una struttura più rigida e la presenza di ulteriori interazioni intramolecolari nell’aptamero modificato. Questi risultati sono stati confermati ed interpretati sulla base dei risultati delle dinamiche molecolari dei due aptameri. I parametri termodinamici del processo di binding del TBA e del TBA modificato con la trombina mostrano che l’interazione è esotermica e che la stechiometria del complesso che si forma è 1:2 (aptamero:proteina). Il TBA modificato risulta però avere una più alta affinità per la trombina e la sua interazione è associata ad una variazione di entalpia maggiormente favorevole. I risultati delle dinamiche molecolari dei complessi suggeriscono che la maggior affinità dell’aptamero modificato sia dovuta essenzialmente ad una migliore interazione con l’esosito II della trombina. Lo studio di dinamica molecolare delle quadruple eliche di RNA e DNA ha evidenziato che la maggiore differenza tra le due molecole deriva dalla presenza del gruppo 2’-OH del ribosio, che contribuisce considerevolmente al numero di legami idrogeno formando più interazioni intramolecolari nella molecola di RNA. I legami che hanno una maggiore persistenza sono quelli formati con l’ossigeno del gruppo fosfato. Questo tipo di legame idrogeno conferisce rigidità alla struttura ed è in parte responsabile della stabilità delle esadi presenti nella quadrupla elica di RNA. Inoltre, durante le simulazioni, gli ioni Na+, inizialmente non direttamente coordinati alle quadruple eliche, vanno a coordinare in modo stabile le macromolecole indicando un possibile meccanismo di coordinazione non ancora osservato sperimentalmente. Parte della ricerca è stata svolta in collaborazione con la Prof. Concetta Giancola del Dipartimento di Chimica dell’Università Federico II di Napoli. Inoltre, una parte dello studio è stato realizzato a Londra presso i laboratori del Prof. Stephen Neidle della School of Pharmacy dell’University of London, e presso il gruppo della Dott. Franca Fraternali, della Randall Division del King’s College London. [a cura dell'autore]
VI n.s.

La carbonatation atmosphérique et la pénétration des ions chlorures sont les principales causes du déclenchement de la corrosion des armatures dans les bétons armés. Chacun de ses mécanismes a fait l'objet de nombreuses études dans la bibliographie. Pourtant les études prenant en compte en même temps les deux types d'agression sont rares. Le couplage des deux phénomènes existe cependant. La carbonatation intervient en effet dans tout ouvrage dès le décoffrage et il suffit que cet ouvrage soit aussi par exemple situé à proximité d'eau de mer pour que les embruns apportent des ions chlorures dans le matériau. L'objectif de cette thèse a été de quantifier les différentes phases de la matrice cimentaire et ses capacités de fixation des ions chlorures par le biais d'isothermes d'interactions pour des matériaux sains et carbonatés. Ces isothermes d'interactions obtenues notamment par la méthode des équilibres ont permis de voir que les matrices partiellement ou totalement carbonatées fixaient moins d'ions chlorures que les matrices saines. L'utilisation de moyens complémentaires d'analyses tels que l'analyse thermogravimétrique (ATG), la diffraction des rayons X (DRX) et la résonance magnétique nucléaire (RMN) a permis de mieux comprendre cette perte de fixation. Une diminution des sels de Friedel et des chlorures fixés par les C-S-H a été ainsi mise en évidence dans les matériaux carbonatés. Ces diminutions qui on été quantifiées sont attribuées à une modification sous carbonatation des phases capables de fixer les ions chlorures (phases aluminates, sulfo-aluminates et C-S-H). Des différences au niveau du réseau poreux ont été également mises en évidence par la porosimétrie par intrusion de mercure (diminution de la macroporosité et de la connectivité en général pour les matrices carbonatées par formation de CaCO3). L'objectif a été aussi de corréler toutes ces modifications observées avec les propriétés de transport des ions qui ont été obtenues par des tests de diffusion et migration des ions chlorures (et/ou sulfates) ainsi que des mesures de résistivité électrique. Les résultats expérimentaux ont montré que le coefficient de diffusion apparent est plus grand dans les matériaux carbonatés. Certaines expériences de diffusion faites en présence à la fois d'ions chlorures et sulfates ont aussi montré la concurrence de ces deux ions pour se fixer à la matrice cimentaire. Les matériaux cimentaires testés (bétons et pâtes) ont été formulés sans ou avec additions minérales (métakaolin, cendres volantes, laitier) et les propriétés de ces matériaux ont été observées à différents âges

L'émission dans l'atmosphère de nanoparticules issues des transports préoccupe la communauté scientifique à cause de leurs impacts probables sur le changement climatique. La compréhension de l'effet de ces émissions sur l'environnement reste faible principalement à cause du manque de données expérimentales sur la caractérisation de ces nanoparticules. Ce travail est axé sur les propriétés physico-chimiques des nanoparticules de combustion et sur leur interaction avec l'eau. L'hydroscopicité est l'un des paramètres déterminants liés à la formation des noyaux de condensation des nuages (NCN). Les données expérimentales montrent des différences dans la microstructure, la composition chimique et la morphologie des suies de laboratoire et des résidus émis par les transports. Les quantités d'eau absorbées sur la suie de chambre de combustion de moteur d'avion et sur les résidus issus de la combustion de fioul lourd et de diesel sont bien plus importantes que sur les suies de laboratoire. Nous pouvons ainsi supposer que ces particules agissent comme des noyaux actifs dans la formation des NCN
Transport emission of nanoparticles into atmosphere is of major interest because of its possible effect on climate changes. The understanding of the potential environmental effect of the aviation and ship emission is still poor maintly because of the lack in the experimental characterization of these nanoparticules. The present work focuses on physico-chemical properties of combustion nanoparticles and their interaction with water. Hygroscopicity is on of the key parameters that are related to could condensation nuclei (CCN) activity and the environnemental effect. Experimental data show differences in the microstructure, elemental composition residuals coming from marine transport emitted residuals. Water uptake on combustion residuals coming from marine transport and aviation is higher than for laboratory-produced samples. We can therfore guess that these particules act as active CCN in the atmosphere

La synthèse de polymères hydrosolubles présentant un caractère associatif a été effectuée, par réaction de diverses alkylamines à chaine longue (Cn-NH2, 8≤n≤16) sur l'alginate de propylène glycol (PGA), un dérivé partiellement estérifié d'un polysaccharide extrait des algues. Les propriétés physico-chimiques des dérivés hydrophobisés résultants ont été étudiées, comparativement à celles des polymères parents, en rhéologie, en spectrophotométrie de fluorescence et par des mesures de tension superficielle, dans l'eau pure ou en présence de sels. De façon générale, on observe que le caractère associatif des dérivés synthétisés augmente avec le taux et la longueur des segments hydrophobes immobilisés sur le PGA. Ainsi, si les effets d'interactions hydrophobes ne sont pas mis en évidence pour le dérivé en C8, les dérivés hydrophobisés par des chaines alkyles longues (C12 et C14) présentent, quant à eux, d'importantes variations de comportement par rapport aux polymères parents. Celles-ci résultent de la mise en place d'interactions hydrophobes entre les chaines alkyles immobilisées sur le squelette polysaccharidique. Ces interactions sont essentiellement de nature intramoléculaire en solution diluée tandis que les associations intermoléculaires deviennent prépondérantes en régime semi-dilué. Des entités de volume hydrodynamique élevé apparaissent alors et conduisent à une augmentation spectaculaire de la viscosité, pouvant aller jusqu'à la formation d'un réseau tridimensionnel ayant une structure de gel physique

Tese de doutoramento em Engenharia Têxtil
O uso de toalhetes à base de fibras impregnados com desinfetantes são o método mais comum para a desinfeção de superfícies em contexto hospitalar, indústrias de processamento e aplicações domésticas. Este trabalho estuda a interação entre o desinfetante cloreto de alquildimetilbenzilamónio (ADBAC) e toalhetes comerciais compostos por três materiais distintos, W1 – 100% poliéster, W2 – 55% celulose/45% poliéster e W3 – 100% celulose, a sua eficácia antimicrobiana e o efeito do tratamento com descarga plasmática de barreira dielétrica (DBD). Demonstrou-se que a adsorção de ADBAC é significativamente influenciada pela composição do substrato têxtil em termos de teor de celulose, razão de banho e tempo de imersão. Os substratos com maior teor de celulose apresentam maior adsorção de ADBAC. No entanto, na análise por espectroscopia de raio-X (XPS), o substrato de poliéster foi o que apresentou a maior concentração de ADBAC na superfície. Os resultados de XPS também demonstraram a geração de novas espécies de oxigénio na sua superfície após o tratamento DBD, o que aumentou de forma substancial a concentração de ADBAC. Na celulose, o plasma apenas promoveu um efeito de erosão. No estudo de envelhecimento não se observaram diferenças significativas na força e alongamento à rotura durante o armazenamento de W1 e W2. No entanto, a força à rotura no sentido da trama do substrato W3 tratado com plasma diminuiu. Os resultados das análises mecânicas dinâmicas demonstraram que a imersão em ADBAC e o tratamento DBD tiveram uma influência significativa nas propriedades viscoelásticas de W1, pelo melhoramento da resposta elástica devido à diminuição da mobilidade das cadeias poliméricas, e de W3, pelo melhoramento da resposta não elástica devido à erosão plasmática. O substrato W1 foi o que demonstrou maior eficácia antimicrobiana, sendo este efeito mais evidenciado e mantido ao longo do tempo nas amostras tratadas com plasma. Por outro lado, o W2 não tratado com plasma exibiu maior efeito antimicrobiano em bactérias Gram-positivas, mas com o tratamento DBD, a atividade antimicrobiana foi maior nas Gram-negativas. O substrato W3 apresenta baixa atividade antimicrobiana, confirmando os resultados apresentados na literatura para a celulose. Em geral, este projeto permitiu o desenvolvimento de uma nova classe de toalhetes à base de poliéster de custo acessível e ambientalmente sustentável. Estes possuem uma eficácia antimicrobiana superior, melhorada pelo tratamento DBD, devido à maior concentração de ADBAC na superfície do toalhete, podendo ser armazenados por período de tempo mais longos.
Surface disinfection by disinfectant-impregnated wipes is the most prevalent disinfection method used in nosocomial environment, food processing industry and other domestic situations. This work studies the interaction between the alkyldimethylbenzylammonium chloride (ADBAC) disinfectant and three untreated and dielectric barrier discharge (DBD) plasma-treated commercial wiping materials of W1 – 100% polyester, W2 – 55% cellulose/45% polyester and W3 – 100% cellulose (cotton) affecting the antimicrobial efficacy. Wipe material type in terms of cellulose content, liquor ratio and immersion time demonstrated a significant influence on the adsorption of ADBAC in both untreated and plasma-treated samples. The higher the content of cellulose in the material, the higher is the adsorption of ADBAC active ingredient. Nevertheless, X-ray photoelectron spectroscopy (XPS) analysis found higher ADBAC concentration on the surface of polyester wipe than the other two. Also, it confirmed the interaction of the ADBAC with the newly generated oxygen species on the polyester surface when plasma treatment was applied, drastically increasing the ADBAC concentration on the surface. Whereas, plasma treatment only resulted in etching effects on cellulose. In the ageing study, no significant changes in breaking force and elongation during storage for untreated and plasma-treated W1 and W2 were observed. However, plasma treatment affects W3 in weft direction reducing the force at break. Dynamic mechanical analysis results showed that ADBAC immersion and plasma treatment have a significant influence in viscoelastic properties of W1, by improving its elastic response limiting the polymeric chains mobility and of W3, by enhancing the non-elastic response due to the etching effect. W1 displayed the highest antimicrobial efficacy, with more enhanced and prolonged performance in plasma-treated samples. W2 showed high antimicrobial effect but plasma treatment caused an inversion in its performance against Gram-positive and Gram-negative bacteria. W3 exhibited low antimicrobial activity, confirming the negative result presented in literature. Overall, plasma treatment allows a new class of low-cost and environmental-friendly polyester-based wiping materials with superior antimicrobial efficacy due to improved ADBAC concentration on the wipe surface, which is maintained during a longer storage period.
I am greatly indebted to the Fundação para a Ciência e a Tecnologia for its financial aid of the PhD Project (SDRH/BD/130028/2017), and to the Centro de Ciência e Tecnologia Têxtil (2C2T) – Universidade do Minho for the support of the PhD work with all the available research facilities. I appreciate very much of the organisation European Cooperation in Science & Technology (COST) for its financial support under COST action CA15114 of the ICPIC 2019 conference and STSM in the University Medical Centre of Amsterdam.

The fidelity of biological processes and reactions, inspite of the widespread diversity, is programmed by highly specific physico-chemical principles. This underlines our basic understanding of different interesting phenomena of biological relevance, ranging from enzyme specificity to allosteric communication, from selection of fold to structural organization / states of oligomerization, from half-sites-reactivity to reshuffling of the conformational free energy landscape, encompassing the dogma of sequence-structure dynamics-function of macromolecules. The role of striking an optimal balance between rigidity and flexibility in macromolecular 3D structural organisation is yet another concept that needs attention from the functional perspective. Needless to say that the variety of protein structures and conformations naturally leads to the diversity of their function and consequently many other biological functions in general. Classical models of allostery like the ‘MWC model’ or the ‘KNF model’ and the more recently proposed ‘population shift model’ have advanced our understanding of the underlying principles of long range signal transfer in macromolecules. Extensive studies have also reported the importance of the fold selection and 3D structural organisation in the context of macromolecular function. Also ligand induced conformational changes in macromolecules, both subtle and drastic, forms the basis for controlling several biological processes in an ordered manner by re-organizing the free energy landscape. The above mentioned biological phenomena have been observed from several different biochemical and biophysical approaches. Although these processes may often seem independent of each other and are associated with regulation of specialized functions in macromolecules, it is worthwhile to investigate if they share any commonality or interdependence at the detailed atomic level of the 3D structural organisation. So the nagging question is, do these diverse biological processes have a unifying theme, when probed at a level that takes into account even subtle re-orchestrations of the interactions and energetics at the protein/nucleic acid side-chain level. This is a complex problem to address and here we have made attempts to examine this problem using computational tools. Two methods have been extensively applied: Molecular Dynamics (MD) simulations and network theory and related parameters. Network theory has been extensively used in the past in several studies, ranging from analysis of social networks to systems level networks in biology (e.g., metabolic networks) and have also found applications in the varied fields of physics, economics, cartography and psychology. More recently, this concept has been applied to study the intricate details of the structural organisation in proteins, providing a local view of molecular interactions from a global perspective. On the other hand, MD simulations capture the dynamics of interactions and the conformational space associated with a given state (e.g., different ligand-bound states) of the macromolecule. The unison of these two methods enables the detection and investigation of the energetic and geometric re-arrangements of the 3D structural organisation of macromolecule/macromolecular complexes from a dynamical or ensemble perspective and this has been one of the thrust areas of the current study. So we not only correlate structure and functions in terms of subtle changes in interactions but also bring in conformational dynamics into the picture by studying such changes along the MD ensemble. The focus was to identify the subtle rearrangements of interactions between non-covalently interacting partners in proteins and the interacting nucleic acids. We propose that these rearrangements in interactions between residues (amino acids in proteins, nucleic acids in RNA/DNA) form the common basis for different biological phenomena which regulates several apparently unrelated processes in biology. Broadly, the major goal of this work is to elucidate the physico-chemical principles underlying some of the important biological phenomena, such as allosteric communication, ligand induced modulation of rigidity/flexibility, half-sites-reactivity and so on, in molecular details. We have investigated several proteins, protein-RNA/DNA complexes to formulate general methodologies to address these questions from a molecular perspective. In the process we have also specifically illuminated upon the mechanistic aspects of the aminoacylation reaction by aminoacyl-tRNA synthetases like tryptophanyl and pyrrolysyl tRNA synthetase, structural details related to an enzyme catalyzed reaction that influences the process of quorum sensing in bacteria. Further, we have also examined the ‘dynamic allosterism’ that manipulates the activity of MutS, a prominent component of the DNA bp ‘mismatch repair’ machinery. Additionally, our protein structure network (PSN) based studies on a dataset of Rossmann fold containing proteins have provided insights into the structural signatures that drive the adoption of a fold from a repertoire of diverse sequences. Ligand induced percolations distant from the active sites, which may be of functional relevance have also been probed, in the context of the S1A family of serine proteases. In the course of our investigation, we have borrowed several concepts of network parameters from social network analysis and have developed new concepts. The Introduction (Chapter-1) summarizes the relevant literature and lays down a suitable background for the subsequent chapters in the thesis. The major questions addressed and the main goal of this thesis are described to set an appropriate stage for the detailed discussions. The methodologies involved are discussed in Chapter-2. Chapter-3 deals with a protein, LuxS that is involved in the bacterial quorum sensing; the first part of the chapter describes the application of network analysis on the static structures of several LuxS proteins from different organisms and the second part of this chapter describes the application of a dynamic network approach to analyze the MD trajectories of H.pylori LuxS. Chapter-4 focuses on the investigation of human tryptophanyl-tRNA synthetase (hTrpRS), with an emphasis to identify ligand induced subtle conformational changes in terms of the alternation of rigidity/flexibility at different sites and the re-organisation of the free energy landscape. Chapter-5 presents a novel application of a quantum clustering (QC) technique, popular in the fields of pattern recognition, to objectively cluster the conformations, sampled by molecular dynamics simulations performed on different ligand bound structures of the protein. The protein structure network (PSN) in the earlier studies were constituted on the basis of geometric interactions. In Chapters 6 and 7, we describe the networks (proteins+nucleic acids) using interaction energy as edges, thus incorporating the detailed chemistry in terms of an energy-weighted complex network. Chapter-6 describes an application of the energy weighted network formalism to probe allosteric communication in D.hafniense pyrrolysyl-tRNA synthetase. The methodology developed for in-depth study of ligand induced changes in DhPylRS has been adopted to the protein MutS, the first ‘check-point protein’ for DNA base pair (bp) mismatch repair. In Chapter-7, we describe the network analysis and the biological insights derived from this study (the work is done in collaboration with Prof. David Beveridge and Dr. Susan Pieniazek). Chapter-8 describes the application of a network approach to capture the ligand-induced subtle global changes in protein structures, using a dataset of high resolution structures from the S1A family of serine proteases. Chapter-9 deals with probing the structural rationale behind diverse sequences adopting the same fold with the NAD(P)-binding Rossmann fold as a case study. Future directions are discussed in the final chapter of the thesis (Chapter-10).

The fidelity of biological processes and reactions, inspite of the widespread diversity, is programmed by highly specific physico-chemical principles. This underlines our basic understanding of different interesting phenomena of biological relevance, ranging from enzyme specificity to allosteric communication, from selection of fold to structural organization / states of oligomerization, from half-sites-reactivity to reshuffling of the conformational free energy landscape, encompassing the dogma of sequence-structure dynamics-function of macromolecules. The role of striking an optimal balance between rigidity and flexibility in macromolecular 3D structural organisation is yet another concept that needs attention from the functional perspective. Needless to say that the variety of protein structures and conformations naturally leads to the diversity of their function and consequently many other biological functions in general. Classical models of allostery like the ‘MWC model’ or the ‘KNF model’ and the more recently proposed ‘population shift model’ have advanced our understanding of the underlying principles of long range signal transfer in macromolecules. Extensive studies have also reported the importance of the fold selection and 3D structural organisation in the context of macromolecular function. Also ligand induced conformational changes in macromolecules, both subtle and drastic, forms the basis for controlling several biological processes in an ordered manner by re-organizing the free energy landscape. The above mentioned biological phenomena have been observed from several different biochemical and biophysical approaches. Although these processes may often seem independent of each other and are associated with regulation of specialized functions in macromolecules, it is worthwhile to investigate if they share any commonality or interdependence at the detailed atomic level of the 3D structural organisation. So the nagging question is, do these diverse biological processes have a unifying theme, when probed at a level that takes into account even subtle re-orchestrations of the interactions and energetics at the protein/nucleic acid side-chain level. This is a complex problem to address and here we have made attempts to examine this problem using computational tools. Two methods have been extensively applied: Molecular Dynamics (MD) simulations and network theory and related parameters. Network theory has been extensively used in the past in several studies, ranging from analysis of social networks to systems level networks in biology (e.g., metabolic networks) and have also found applications in the varied fields of physics, economics, cartography and psychology. More recently, this concept has been applied to study the intricate details of the structural organisation in proteins, providing a local view of molecular interactions from a global perspective. On the other hand, MD simulations capture the dynamics of interactions and the conformational space associated with a given state (e.g., different ligand-bound states) of the macromolecule. The unison of these two methods enables the detection and investigation of the energetic and geometric re-arrangements of the 3D structural organisation of macromolecule/macromolecular complexes from a dynamical or ensemble perspective and this has been one of the thrust areas of the current study. So we not only correlate structure and functions in terms of subtle changes in interactions but also bring in conformational dynamics into the picture by studying such changes along the MD ensemble. The focus was to identify the subtle rearrangements of interactions between non-covalently interacting partners in proteins and the interacting nucleic acids. We propose that these rearrangements in interactions between residues (amino acids in proteins, nucleic acids in RNA/DNA) form the common basis for different biological phenomena which regulates several apparently unrelated processes in biology. Broadly, the major goal of this work is to elucidate the physico-chemical principles underlying some of the important biological phenomena, such as allosteric communication, ligand induced modulation of rigidity/flexibility, half-sites-reactivity and so on, in molecular details. We have investigated several proteins, protein-RNA/DNA complexes to formulate general methodologies to address these questions from a molecular perspective. In the process we have also specifically illuminated upon the mechanistic aspects of the aminoacylation reaction by aminoacyl-tRNA synthetases like tryptophanyl and pyrrolysyl tRNA synthetase, structural details related to an enzyme catalyzed reaction that influences the process of quorum sensing in bacteria. Further, we have also examined the ‘dynamic allosterism’ that manipulates the activity of MutS, a prominent component of the DNA bp ‘mismatch repair’ machinery. Additionally, our protein structure network (PSN) based studies on a dataset of Rossmann fold containing proteins have provided insights into the structural signatures that drive the adoption of a fold from a repertoire of diverse sequences. Ligand induced percolations distant from the active sites, which may be of functional relevance have also been probed, in the context of the S1A family of serine proteases. In the course of our investigation, we have borrowed several concepts of network parameters from social network analysis and have developed new concepts. The Introduction (Chapter-1) summarizes the relevant literature and lays down a suitable background for the subsequent chapters in the thesis. The major questions addressed and the main goal of this thesis are described to set an appropriate stage for the detailed discussions. The methodologies involved are discussed in Chapter-2. Chapter-3 deals with a protein, LuxS that is involved in the bacterial quorum sensing; the first part of the chapter describes the application of network analysis on the static structures of several LuxS proteins from different organisms and the second part of this chapter describes the application of a dynamic network approach to analyze the MD trajectories of H.pylori LuxS. Chapter-4 focuses on the investigation of human tryptophanyl-tRNA synthetase (hTrpRS), with an emphasis to identify ligand induced subtle conformational changes in terms of the alternation of rigidity/flexibility at different sites and the re-organisation of the free energy landscape. Chapter-5 presents a novel application of a quantum clustering (QC) technique, popular in the fields of pattern recognition, to objectively cluster the conformations, sampled by molecular dynamics simulations performed on different ligand bound structures of the protein. The protein structure network (PSN) in the earlier studies were constituted on the basis of geometric interactions. In Chapters 6 and 7, we describe the networks (proteins+nucleic acids) using interaction energy as edges, thus incorporating the detailed chemistry in terms of an energy-weighted complex network. Chapter-6 describes an application of the energy weighted network formalism to probe allosteric communication in D.hafniense pyrrolysyl-tRNA synthetase. The methodology developed for in-depth study of ligand induced changes in DhPylRS has been adopted to the protein MutS, the first ‘check-point protein’ for DNA base pair (bp) mismatch repair. In Chapter-7, we describe the network analysis and the biological insights derived from this study (the work is done in collaboration with Prof. David Beveridge and Dr. Susan Pieniazek). Chapter-8 describes the application of a network approach to capture the ligand-induced subtle global changes in protein structures, using a dataset of high resolution structures from the S1A family of serine proteases. Chapter-9 deals with probing the structural rationale behind diverse sequences adopting the same fold with the NAD(P)-binding Rossmann fold as a case study. Future directions are discussed in the final chapter of the thesis (Chapter-10).

Sizing (hydrophobization) is one of the most important process steps within the added-value chain of about 1/3rd of the worldwide produced paper & board products. Even though sizing with so-called reactive sizing agents, such as alkenyl succinic anhydride (ASA) was implemented in the paper industry decades ago, there is no total clarity yet about the detailed chemical and physical mechanisms that lead to their performance. Previous research was carried out on the role of different factors influencing the sizing performance, such as bonding between ASA and cellulose, ASA hydrolysis, size revision as well as the most important interactions with stock components, process parameters and additives during the paper making process. However, it was not yet possible to develop a holistic model for the explanation of the sizing performance given in real life application. This thesis describes a novel physico-chemical approach to this problem by including results from previous research and combining these with a wide field of own basic research and a newly developed method that allows tracing back the actual localization of ASA within the sheet structure. The carried out measurements and trial sets for the basic field of research served to evaluate the stock and process parameters that most dominantly influence the sizing performance of ASA. Interactions with additives other than retention aids were not taken into account. The results show that parameters, such as the content of secondary fibers, the degree of refining, the water hardness as well as the suspension conductivity, are of highest significance. The sample sets of the trials with the major impacting parameters were additionally analyzed by a newly developed localization method in order to better understand the main influencing factors. This method is based on optical localization of ASA within the sheet structure by confocal white light microscopy. In order to fulfill the requirements at magnification rates of factor 100 optical zoom, it was necessary to improve the contrast between ASA and cellulose. Therefore, ASA was pretreated with an inert red diazo dye, which does not have any impact on neither the sizing nor the handling properties of ASA. Laboratory hand sheets that were sized with dyed ASA, were analyzed by means of their sizing performance in correlation to measurable ASA agglomerations in the sheet structure. The sizing performance was measured by ultrasonic penetration analysis. The agglomeration behavior of ASA was analyzed automatically by multiple random imaging of a sample area of approx. 8650 µm² with a minimum resolution for particles of 500 nm in size. The gained results were interpreted by full factorial design of experiments (DOE). The trials were carried out with ASA dosages between 0% and 0.8% on laboratory hand sheets, made of 80% bleached eucalyptus short fiber kraft pulp and 20% northern bleached softwood kraft pulp, beaten to SR° 30, produced with a RDA sheet former at a base weight of 100 g/m² oven dry. The results show that there is a defined correlation between the ASA dosage, the sizing performance and the number and area of ASA agglomerates to be found in the sheet structure. It was also possible to show that the agglomeration behavior is highly influenced by external factors like furnish composition and process parameters. This enables a new approach to the explanation of sizing performance, by making it possible to not only examine the performance of the sizing agent, but to closely look at the predominant position where it is located in the sheet structure. These results lead to the explanation that the phenomenon of sizing is by far not a pure chemical process but rather a more physical one. Based on the gained findings it was possible so far to optimize the ASA sizing process in industrial-scale by means of ~ 50% less ASA consumption at a steady degree of sizing and improved physical sheet properties.:Acknowledgment I Abstract III Table of Content V List of Illustrations XI List of Tables XVI List of Formulas XVII List of Abbreviations XVIII 1 Introduction and Problem Description 1 1.1 Initial Situation 1 1.2 Objective 2 2 Theoretical Approach 3 2.1 The Modern Paper & Board Industry on the Example of Germany 3 2.1.1 Raw Materials for the Production of Paper & Board 5 2.2 The Sizing of Paper & Board 8 2.2.1 Introduction to Paper & Board Sizing 8 2.2.2 The Definition of Paper & Board Sizing 10 2.2.3 The Global Markets for Sized Paper & Board Products and Sizing Agents 11 2.2.4 Physical and Chemical Background to the Mechanisms of Surface-Wetting and Penetration 13 2.2.4.1 Surface Wetting 14 2.2.4.2 Liquid Penetration 15 2.2.5 Surface and Internal Sizing 17 2.2.6 Sizing Agents 18 2.2.6.1 Alkenyl Succinic Anhydride (ASA) 19 2.2.6.2 Rosin Sizes 19 2.2.6.3 Alkylketen Dimer (AKD) 23 2.2.6.4 Polymeric Sizing Agents (PSA) 26 2.2.7 Determination of the Sizing Degree (Performance Analysis) 28 2.2.7.1 Cobb Water Absorption 29 2.2.7.2 Contact Angle Measurement 30 2.2.7.3 Penetration Dynamics Analysis 31 2.2.7.4 Further Qualitative Analysis Methods 33 2.2.7.4.1 Ink Stroke 33 2.2.7.4.2 Immersion Test 33 2.2.7.4.3 Floating Test 34 2.2.7.4.4 Hercules Sizing Tester (HST) 34 2.2.8 Sizing Agent Detection (Qualitative Analysis) and Determination of the Sizing Agent Content (Quantitative Analysis) 35 2.2.8.1 Destructive Methods 35 2.2.8.2 Non Destructive Methods 36 2.3 Alkenyl Succinic Anhydride (ASA) 36 2.3.1.1 Chemical Composition and Production of ASA 37 2.3.1.2 Mechanistic Reaction Models 39 2.3.1.3 ASA Application 42 2.3.1.3.1 Emulsification 42 2.3.1.3.2 Dosing 44 2.3.1.4 Mechanistic Steps of ASA Sizing 46 2.3.2 Physico-Chemical Aspects during ASA Sizing 48 2.3.2.1 Reaction Plausibility 48 2.3.2.1.1 Educt-Product Balance / Kinetics 48 2.3.2.1.2 Energetics 51 2.3.2.1.3 Sterics 52 2.3.2.2 Phenomena based on Sizing Agent Mobility 53 2.3.2.2.1 Sizing Agent Orientation 54 2.3.2.2.2 Intra-Molecular Orientation 55 2.3.2.2.3 Sizing Agent Agglomeration 55 2.3.2.2.4 Fugitive Sizing / Sizing Loss / Size Reversion 56 2.3.2.2.5 Sizing Agent Migration 58 2.3.2.2.6 Sizing Reactivation / Sizing Agent Reorientation 59 2.3.3 Causes for Interactions during ASA Sizing 60 2.3.3.1 Process Parameters 61 2.3.3.1.1 Temperature 61 2.3.3.1.2 pH-Value 62 2.3.3.1.3 Water Hardness 63 2.3.3.2 Fiber Types 64 2.3.3.3 Filler Types 65 2.3.3.4 Cationic Additives 66 2.3.3.5 Anionic Additives 67 2.3.3.6 Surface-Active Additives 68 2.4 Limitations of State-of-the-Art ASA-Sizing Analysis 69 2.5 Optical ASA Localization 71 2.5.1 General Background 71 2.5.2 Confocal Microscopy 72 2.5.2.1 Principle 72 2.5.2.2 Features, Advantage and Applicability for Paper-Component Analysis 74 2.5.3 Dying / Staining 75 3 Discussion of Results 77 3.1 Localization of ASA within the Sheet Structure 77 3.1.1 Choice of Dyes 77 3.1.1.1 Dye Type 78 3.1.1.2 Evaluation of Dye/ASA Mixtures 80 3.1.1.2.1 Maximum Soluble Dye Concentration 80 3.1.1.2.2 Thin Layer Chromatography 81 3.1.1.2.3 FTIR-Spectroscopy 82 3.1.1.3 Evaluation of the D-ASA Emulsion 84 3.1.1.4 Paper Chromatography with D-ASA & F-ASA Emulsions 85 3.1.1.5 Evaluation of the D-ASA Emulsion’s Sizing Efficiency 86 3.1.2 The Localization Method 87 3.1.2.1 The Correlation between ASA Distribution and Agglomeration 88 3.1.2.2 Measurement Settings 89 3.1.2.3 Manual Analysis 90 3.1.2.4 Automated Analysis 92 3.1.2.4.1 Automated Localization / Microscopy Measurement 92 3.1.2.4.2 Automated Analysis / Image-Processing 93 3.1.2.5 Result Interpretation and Example Results 96 3.1.2.6 Reproducibility 97 3.1.2.7 Sample Mapping 98 3.1.3 Approaches to Localization-Method Validation 102 3.1.3.1 Raman Spectroscopy 102 3.1.3.2 Confocal Laser Scanning Fluorescent Microscopy 102 3.1.3.3 Decolorization 103 3.2 Factors Impacting the Sizing Behavior of ASA 104 3.2.1 ASA Type 105 3.2.2 Emulsion Parameters 107 3.2.2.1 Hydrolyzed ASA Content 107 3.2.2.2 ASA/Starch Ratio 109 3.2.2.3 Emulsion Age 110 3.2.3 Stock Parameters 111 3.2.3.1 Long Fiber/Short Fiber Ratio 111 3.2.3.2 Furnish Type 112 3.2.3.3 Degree of Refining 114 3.2.3.4 Filler Type/Content 116 3.2.4 Process Parameters 119 3.2.4.1 Temperature 119 3.2.4.2 pH-Value 120 3.2.4.3 Conductivity 122 3.2.4.4 Water Hardness 123 3.2.4.5 Shear Rate 125 3.2.4.6 Dwell Time 127 3.2.4.7 Dosing Position & Dosing Order 128 3.2.4.8 Drying 130 3.2.4.9 Aging 131 3.3 Factors Impacting the Localization Behavior of ASA 132 3.3.1 Degree of Refining 132 3.3.2 Sheet Forming Conductivity 135 3.3.3 Water Hardness 136 3.3.4 Retention Aid (PAM) 137 3.3.5 Contact Curing 138 3.3.6 Accelerated Aging 139 3.4 Main Optimization Approach 141 3.4.1 Optimization of ASA Sizing Performance Characteristics 142 3.4.2 Emulsion Modification 144 3.4.2.1 Lab Trials / RDA Sheet Forming 146 3.4.2.2 TPM Trials 147 3.4.2.3 Industrial-Scale Trials 149 3.4.2.4 Correlation between Sizing Performance Optimization and Agglomeration Behavior on the Example of PAAE 152 3.5 Holistic Approach to Sizing Performance Explanation 154 4 Experimental Approach 157 4.1 Characterization of Methods, Measurements and Chemicals used for the Optical Localization-Analysis of ASA 157 4.1.1 Characterization of used Chemicals 157 4.1.1.1 Preparation of Dyed-ASA Solutions 157 4.1.1.2 Thin Layer Chromatography 157 4.1.1.3 Fourier Transformed Infrared Spectroscopy 157 4.1.1.4 Emulsification of ASA 158 4.1.1.5 Paper Chromatography 159 4.1.1.6 Particle Size Measurement 159 4.1.2 Optical Analysis of ASA Agglomerates 160 4.1.2.1 Microscopy 160 4.1.2.2 Automated Analysis 163 4.1.2.2.1 Adobe Photoshop 163 4.1.2.2.2 Adobe Illustrator 164 4.1.2.3 Confocal Laser Scanning Fluorescent Microscopy 166 4.2 Characterization of Used Standard Methods and Measurements 166 4.2.1 Stock and Paper Properties 166 4.2.1.1 Stock pH, Conductivity and Temperature Measurement 166 4.2.1.2 Dry Content / Consistency Measurement 167 4.2.1.3 Drainability (Schopper-Riegler) Measurement 167 4.2.1.4 Base Weight Measurement 168 4.2.1.5 Ultrasonic Penetration Measurement 168 4.2.1.6 Contact Angle Measurement 169 4.2.1.1 Cobb Measurement 169 4.2.1.2 Air Permeability Measurements 170 4.2.1.3 Tensile Strength Measurements 170 4.2.2 Preparation of Sample Sheets 171 4.2.2.1 Stock Preparation 171 4.2.2.2 Laboratory Refining (Valley Beater) 171 4.2.2.3 RDA Sheet Forming 171 4.2.2.4 Additive Dosing 173 4.2.2.5 Contact Curing 174 4.2.2.6 Hot Air Curing 174 4.2.2.7 Sample Aging 174 4.2.2.8 Preparation of Hydrolyzed ASA 175 4.2.2.9 Trial Paper Machine 175 4.2.2.10 Industrial-Scale Board Machine 177 4.3 Characterization of used Materials 178 4.3.1 Fibers 178 4.3.1.1 Reference Stock System 178 4.3.1.2 OCC Fibers 179 4.3.1.3 DIP Fibers 179 4.3.2 Fillers 180 4.3.3 Chemical Additives 180 4.3.3.1 ASA 180 4.3.3.2 Starches 181 4.3.3.3 Retention Aids 181 4.3.3.4 Poly Aluminum Compounds 181 4.3.3.5 Wet Strength Resin 181 4.3.4 Characterization of used Additives 182 4.3.4.1 Solids Content 182 4.4 Description of Implemented Advanced Data Analysis- and Visualization Methods 183 4.4.1 Design of Experiments (DOE183 4.4.2 Contour Plots 184 4.4.3 Box-Whisker Graphs 185 5 Conclusion 186 6 Outlook for Further Work 191 7 Bibliography 192 Appendix 207 7.1 Localization Method Reproducibility 207 7.2 DOE - Coefficient Lists 208 7.2.1 Trial 3.3.4 – Impact of Retention Aid (PAM) on Agglomeration Behavior and Sizing Performance 208 7.2.2 Trial 3.3.5 – Impact of Contact Curing on Agglomeration Behavior and Sizing Performance 208 7.2.3 Trial 3.3.6 – Impact of Accelerated Aging on Agglomeration Behavior and Sizing Performance 209

Flaxseed (Linum usitatissimum L.) gum (FG) is a material with many potential food and non-food applications. Consistent performance is critical for FG utilization and this is possible through selection of genotype, characterization and optimization of constituents, and chemical modification. Physico-chemical and functional properties of FG aqueous solutions from six Canadian flaxseed cultivars were investigated. FG yield, carbohydrate composition, protein content, and zeta potential (ζ) varied among these cultivars. FG solution properties were also affected by temperature, solution pH, NaCl concentration, and sucrose concentration. Detailed studies were conducted on CDC Bethune FG (FGB) proteins that were separated by 2D-gel electrophoresis. Conlinin was identified as the major protein. Protease treatment decreased FGB solution emulsification properties suggesting that conlinin might enhance emulsification. Formation of BSA-FGB coacervates was monitored by turbidimetric analysis as a function of solution pH, biopolymer mixing ratio, NaCl and urea. Coacervates were stabilized primarily by attractive electrostatic forces and secondarily by hydrogen bonds. Further, anionic carboxymethyl ether moieties were introduced to FGB structure through ether forming reactions using monochloroacetic acid (MCA) to produce products with uniform properties. The highest degree of substitution (DS) was obtained at 70 °C, 7.0 M NaOH, and a molar ratio of MCA to FGB of 10:1 over 3 h. Carboxymethylated FGB (CMFG) exhibited both modified surface morphology and thermal behaviour. Solutions of CMFG demonstrated shear-thinning behaviour and apparent viscosity decreased with increased DS. A more liquid-like flow behaviour was observed for CMFG as DS increased. Findings here will introduce and expand FG applications in food or related fields with targeted performance.

In the present Ph.D. thesis an extensive structural and functional study on model bio-membranes is presented. The first part of my research has been focused to understand how the lipid composition of the bio-membranes affects their biophysical properties and modulates their interactions with peptides. One of the biological processes involving lipid composition is the interaction between antimicrobial peptides (AMPs) and biological membranes. In fact, the selective interaction of AMPs with prokaryotic cells arises from the difference in the chemical composition between prokaryotic and eukaryotic membranes. Different mechanisms of membrane destruction have been proposed, depending on physico-chemical properties of AMPs and of the target bio-membranes. Among the large number of AMPs present in nature, Myxinidin, from hagfish (Myxine glutinosa L.), is a promising antimicrobial candidate due to its antibacterial activity against different pathogenic Gram negative and Gram positive bacteria. This thesis reports a comparative study of the interaction between Myxinidin and its mutant WMR with two model bio-membranes at different composition and complexity. In particular, in order to understand the role of lipid composition in the peptide-membrane interaction, two different models of bio-membranes have been studied mimicking P. aeruginosa and E. coli cell cytoplasmic membranes. The final goal was to elucidate the effect of amino acid residues substitutions of the peptides and the role of lipid composition on the antibacterial activity of Myxinidin and WMR against these two model bio-membranes.The collected data have allowed to recognize the AMPs specificity for a particular lipid composition and to propose a mechanism of membrane destabilization. In order to study the role of lipid composition in biological processes, another important model bacterial bio-membrane has been studied. In particular the work has been focused on a particular model of bio-membrane representative of Bradyrhizobium BTAi1 Gram negative bacterium, containing an unusual lipopolysaccharide (LPS) in which the lipid A is covalently linked to a hopanoid moiety The aim of this study was to understand the effect of this unique lipid A in modulating the stability and rigidity of the outer membrane of Bradyrhizobium BTAi1 strain. To obtain a wide physico-chemical characterization of the analyzed systems, a combined experimental strategy has been adopted, including spectroscopic and calorimetric techniques such as Circular Dichroism (CD) to study the secondary structure of peptides and its changes in lipid environment; Fluorescence to estimate the microenvironment of the peptides in the vesicles; Dynamic Light Scattering (DLS) to estimate the size and distribution of the liposomes in the absence and in the presence of peptides; (NMR) to obtain information about the conformation of the peptides in membrane environment; Electron Paramagnetic Resonance (EPR) to investigate the dynamics of the lipid hydrophobic tails in the bilayer; Differential Scanning Calorimetry (DSC) to understand the thermotropic behavior of liposomes and the effect of peptides on their phase transition; Isothermal Titration Calorimetry (ITC) to study the energetic of the interaction process between peptides and liposomes.

The thyroid transcription factor 1 (TTF-1) is a tissue-specific transcription factor involved in the development of thyroid and lung. It is a protein of 371 amino acids that possesses two independent transcriptional activation domains, one located in the N-terminal region of homeodomain and the other in the C-terminal region. The homeodomain is a highly conserved DNA-binding motif that is found in numerous transcription factors throughout a large variety of species from yeasts to human. These gene-specific transcription factors play critical roles in development and adult homeostasis, and therefore, any germline mutations associated with these proteins can lead to a number of congenital abnormalities. Remarkable features of structural and functional conservation have been observed within homeodomain family members. This high degree of conservation between the sequence and structure makes the homeodomain an ideal model for studying protein-DNA interactions and gene regulation. In this PhD thesis, the study on the conformational stability and DNA binding energetics of the rat thyroid transcription factor 1 homeodomain (TTF-1HD) is reported. Furthermore, to gain a molecular-level understanding of the interactions determining the association of TTF-1HD to the target DNA sequence, Some key point mutants (W48, Q50 and Y54) and a deletion mutant (desTTF-1HD), lacking five residues from the N-terminal region have been produced and studied. Since it has been demonstrated that N-terminal region of TTF-1 also interact with Pax-8 (transcription factor belonging to the PAX genes superfamily) in the regulation of thyroid-specific gene expression, the study of the thermal stability and DNA binding energetics of a fusion protein, containing the N-terminal region and the homeodomain of TTF-1 (TTF-1NH2HD), and a recombinant protein containing C-terminal region of Pax-8 (Pax-8 Prd) has been reported.