Among the myraid energy storage technologies, polymer electrolytes have been widely employed in diverse applications such as fuel cell membranes, battery separators, mechanical actuators, reverse-osmosis membranes and solar cells. The polymer electrolytes used for these applications usually require a combination of properties, including anisotropic orientation, tunable modulus, high ionic conductivity, light weight, high thermal stability and low cost. These critical properties have motivated researchers to find next-generation polymer electrolytes, for example ion gels. This dissertation aims to develop and characterize a new class of ion gel electrolytes based on ionic liquids and a rigid-rod polyelectrolyte. The rigid-rod polyelectrolyte poly (2,2'-disulfonyl-4,4'-benzidine terephthalamide) (PBDT) is a water-miscible system and forms a liquid crystal phase above a critical concentration. The diverse properties and broad applications of this rigid-rod polyelectrolyte may originate from the double helical conformation of PBDT molecular chains. We primarily develop an ionic liquid-based polymer gel electrolyte that possesses the following exceptional combination of properties: transport anisotropy up to 3.5×, high ionic conductivity (up to 8 mS cm⁻¹), widely tunable modulus (0.03 – 3 GPa) and high thermal stability (up to 300°C). This unique platform that combines ionic liquid and polyelectrolyte is essential to develop more advanced materials for broader applications. After we obtain the ion gels, we then mainly focus on modifying and then applying them in Li-metal batteries. As a next generation of Li batteries, the Li-metal battery offers higher energy capacity compared to the current Li-ion battery, thus satisfying our requirements in developing longer-lasting batteries for portable devices and even electric vehicles. However, Li dendrite growth on the Li metal anode has limited the pratical application of Li-metal batteries. This unexpected Li dendrite growth can be suppressed by developing polymer separators with high modulus (~ Gpa), while maintaining enough ionic conductivity (~ 1 mS/cm). Here, we describe an advanced solid-state electrolyte based on a sulfonated aramid rigid-rod polymer, an ionic liquid (IL), and a lithium salt, showing promise to make a breakthrough. This unique fabrication platform can be a milestone in discovering next-generation electrolyte materials. / Ph. D. / Among the myraid energy storage technologies, polymer-based electrolytes have been widely employed in diverse applications such as fuel cell membranes, battery electrolytes, “artificial muscle” mechanical actuators, reverse-osmosis membranes and solar cells. The materials used for each of these applications usually require a specific combination of properties, which include anisotropic orientation, tunable mechanical stiffness (modulus), high ionic conductivity, light weight, high thermal stability and low cost. These critical properties have motivated researchers to find next-generation polymer-based electrolytes, for example “ion gels” that consist of a polymer combined with ionic liquids or salts. This thesis describes development of an ion gel that possesses the following exceptional combination of properties: high ionic conductivity (up to 8 mS cm^-1), widely tunable modulus (0.03 ‒ 3 GPa), ion transport anisotropy up to 3.5×, and high thermal stability (up to 300°C). Thus, this unprecedented material shows liquid-like ion motions inside a matrix with solid-like stiffness, and in a material that can withstand extreme temperatures and will not burn. After obtaining these ion gels, we are then mainly focusing on modifying them for application in safe and high density Li-metal batteries. As a next generation of Li batteries, the Li-metal battery offers higher energy capacity compared to the current Liion battery, thus satisfying our requirements in developing longer-lasting batteries for portable devices and even electric vehicles. However, Li dendrite growth on the Li metal anode has limited the pratical application of Li-metal batteries. This unexpected Li dendrite growth can be supressed by developing polymer electrolytes with high modulus (~ GPa), while maintaining sufficient ionic conductivity (~ 1 mS/cm) for efficient battery operation. In short, this thesis describes an advanced solid-state electrolyte based on a kevlar-like (sulfonated aramid) rigid-rod polymer, an ionic liquid (IL), and a lithium salt, showing promise to make a breakthrough and enable practical Li-metal batteries. Furthermore, the unique fabrication platform for these ion gels represents a new paradigm for discovering next-generation electrolyte materials for a wide variety of applications.

Ces travaux de thèse s'inscrivent dans le cadre d'un vaste projet qui vise à construire des membranes hybrides constituée d'un support solide nanoporeux et de protéines canal-ionique biologiques. Nous nous intéressons ici à un film polymère nanoporeux en polycarbonate et à la Gramicidine-A. La membrane ainsi réalisée est étudiée par des mesures expérimentales. Ce travail peut être divisé en deux parties. Dans la première, nous rapportons l'étude du confinement de la protéine canal ionique, au sein des nanopores du film « track-etched » en polycarbonate. Après imprégnation de gA, la membrane est étudiée par Spectroscopie de Fluorescence Confocale. Les premiers résultats expérimentaux particulièrement encourageants montrent que la gA est localisée dans les nanopores et non pas à la surface de la membrane. Dans la deuxième, les propriétés de transport ionique de la membrane hybride sont caractérisées par le biais de deux grandeurs : d'une part les coefficients de diffusion mesurés à partir d'une cellule et d'autre part les conductivités via la Spectroscopie d'Impédance Complexe (S.I.C). Les électrolytes aqueux étudiées sont : XCl(2) où X=Na, K, Mg et Ca à des concentrations comprises entre 5.10-3 à 1M. Une étude statistique approfondie des données obtenues par la méthode de la variance permet de déterminer les effets relatifs des différentes variables : nature et concentration du sel, présence de la Gramicidine A et traitement à l'éthanol de la membrane. Cette analyse révèle clairement que la présence de Gramicidine A au sein des nanopores de 15nm modifie de façon positive le transport ionique. Il est, par contre, difficile de conclure sur la nature sélective du transport ionique en présence de cette protéine. Ce travail de thèse ouvre un champ de recherche très prometteur dans le domaine de la nanofiltration. / This project of thesis is to build of a bio-inspired hybrid membrane made of a thin nanoporous polymer film in which a biological ionic channel is confined. Thus, this work may be divided in two parts. First, we report the confinement of the biological ionic channel, i.e. Gramicidin A, inside the nanopore of nanoporous thin film, i.e. a track etched polycarbonate film (Whatman NucleoporeTM). After impregnation with Gramicidine-A, the membrane is studied by means of confocal fluorescence spectroscopy. The results show the ionic channel is well located into the nanopores and not at the surface of the membrane. Secondly, ionic transport properties are measured by means of two experiments: on the one hand, ionic diffusion coefficients are measured using a cell and on the other hand, ionic dc conductivity is measured via Complex Impedance Spectroscopy (SIC). Various aqueous electrolytes (XCl(2) where X=Na,K, Mg et Ca) at different concentrations ranging from 5.10-3 à 1M are carried out. A statistical analysis of the data so-obtained allows to determine the relative effects of the different parameters: the nature and concentration of the electrolytes, the presence of Gramicidine A and the membrane pre-treatment with ethanol treatment. It is thus clearly pointed out that the presence of Gramicidine A inside the 15nm nanopores improves ion permeability. However, it is difficult to conclude about ionic selectivity of the hybrid membrane. Nevertheless, this work which is the first attempt ever to build such a bio-inspired system opens an extremely promising field of research in the domain of nanofiltration.

Différents travaux de recherche ont été décrits dans cette thèse. Les travaux de recherche peuvent être résumés comme suit. Le premier chapitre a porté sur l'identification d’inhibiteurs puissants efficaces vis-à-vis de de l'isoenzyme anhydrase carbonique humaine I (hCAI). Considérant l'importance pharmacologique de trouver des inhibiteurs (CAIs) et des activateurs (AACs) sélectifs aux isoformes de l’anhydrase carbonique ), l'anhydrase carbonique humaine I (hCAI) a été confrontée en parallèle à diverses bibliothèques dynamiques constitutionnelles (CDL). Dans le deuxième chapitre, des réseaux constitutionnels dynamiques ont été préparés sous forme de systèmes membranaires liquides et solides agissant comme un réseau pour le transport spécifique des ions lanthanides. Le transport est basé sur la capacité de complexation des lanthanides (La + 3, Lu + 3, Eu + 3) avec les groupes polyéther fonctionnels situés dans les matériaux membranaires. Dans le troisième chapitre, l'approche proposée consiste en l'utilisation de membranes liquides ioniques supportées (SILMs) comprenant deux enzymes différentes de l'anhydrase carbonique, l’enzyme thermo-résistante SspCA et l'enzyme bovine-CA, qui catalysent la réaction de conversion réversible du CO2 en bicarbonate en favorisant la force motrice vers le transport de CO2. La stabilité des membrane, leur perméabilité vis-à-vis de CO2 et de N2 ainsi que la sélectivité idéale (CO2 / N2) ont été déterminées pour les membranes développées. Le quatrième chapitre porte sur la synthèse et la caractérisation de membranes polymères denses pour une application en séparation de gaz. Les mesures de perméabilité aux gaz des membranes polymères synthétisées ont montré que la perméabilité de CO2 est supérieure à celle des autres gaz testés (CH4 et N2). Dans le dernier chapitre, des membranes de PVDF ont été fonctionnalisées avec une enzyme, la phosphotriestérase (PTE), selon deux méthodes différentes pour construire un réacteur à membrane biocatalytique (BMR) avec pour finalité la bioconversion et la séparation sélective du substrat paraoxon. La première méthode met en œuvre une dispersion réversible de nanoparticules magnétiques de PTE qui est immobilisée à la surface de la membrane de PVDF sous l’effet d'un champ magnétique externe. A l’inverse, la seconde méthode porte sur le greffage chimique de l'enzyme PTE, après modification de la surface de la membrane de PVDF native (DAMP-GA-enzymatique). Les deux techniques d'immobilisation d'enzymes ont montré une bonne efficacité et une sensibilité à l'égard de la bioconversion du paraoxon dans les différentes conditions appliquées dans un réacteur à membrane biocatalytique (BMR).De façon globale, les concepts développés dans ce travail de thèse permettront d’ouvrir de nouvelles pistes de recherche allant vers le développement d'une membrane polymère sélective au transport d’ions, de gaz mais aussi active dans les réactions catalytiques enzymatiques grâce à un contrôle bio-moléculaire au niveau des matériaux membranaires. / Different research works have been described in this thesis. The research works can be summarized as the following. The first chapter deals with the identification of effective potent inhibitors for the human carbonic anhydrase I (hCAI) isozyme. Considering the pharmacological importance to find selective CA inhibitors (CAIs) and CA activators (CAAs), human carbonic anhydrase I (hCAI) has been subjected to a parallel screening of various constitutional dynamic libraries (CDL). In the second chapter, constitutional dynamic networks have been used in liquid and solid membrane systems as a carrier network for transporting lanthanides. The transport is based on the complexing ability of lanthanides metals (La+3, Lu+3, and Eu+3) with the functional polyether groups in the membrane materials. In the third chapter, the proposed approach consists in using supported ionic liquid membranes (SILMs) comprising two different carbonic anhydrase enzymes, the thermo-resistant SspCA enzyme and the Bovine-CA enzyme, which catalyze the reaction of reversible conversion of CO2 to bicarbonate, enhancing the driving force for CO2 transport. Membrane stability, CO2 and N2 permeability and (CO2/N2) ideal selectivity were determined for the membranes developed. In the fourth chapter, the research work consists in the synthesis and characterization of dense polymeric membranes for gas separation application. The gas permeability measurements for the synthesized polymeric membranes showed that the permeability of CO2 is higher than other used gases (N2 and CH4). In the last chapter, two different methods of PVDF membrane functionalization with a phosphotriesterase (PTE) enzyme have been developed to construct biocatalytic membrane reactor (BMR) for bioconversion and selective separation of paraoxon substrate. The first method employs reversible dispersion of magnetic nanoparticle immobilized with PTE using an external magnetic field on the surface of native PVDF membrane. On the contrary, the second method comprises chemical grafting of the PTE enzyme, after surface modification of the native PVDF membrane (DAMP-GA-Enzyme). Both methods of enzyme immobilization showed good efficiency and sensitivity towards the bioconversion of paraoxon substrate at different conditions applied in a biocatalytic membrane reactor (BMR).In general, the concepts developed in this thesis research work will help bring new tracks on the way to the development of a polymeric membrane for selective ion and gas separation but also for selective catalytic reaction under bio(molecular) control.

The Two-pore channel (TPC1-3) family represents a recently identified class of endolysosomal ion channels. TPCs were originally proposed to be promising candidate channels for NAADP-induced Ca²⁺ release. However, subsequent studies have emerged to propose an alternative view where TPCs may be Na+-selective channels regulated by the lysosome-specific phosphoinositide PI(3,5)P2 or voltage in an isoform-dependent manner. This thesis asks the question of whether pharmacological and ion permeation properties of TPCs, in particular the human TPC2, may satisfy or may be consistent with the requirement of a potential NAADP-sensitive Ca²⁺-release channel. These fundamental properties of hTPC2 were approached using patch-clamp electrophysiology and confocal fluorescence microscopy, and were analysed quantitatively to extract relevant physical parameters important to our understanding of their physiological and functional significance. Chapter 2 presents the basic electrophysiological characterisation of hTPC2. It follows a logical way by first determining the ion permeation properties, followed by the investigation of its physical relation with fractional Ca²⁺ current and Ca²⁺ nanodomains to rigorously prove that this Na⁺ selectivity is sufficient to ensure negligible Ca²⁺ leakage both experimentally and theoretically. This follows the logic that matter must not be created nor destroyed so that a Na+-selective channel that poses a physiologically significant energy barrier to Ca²⁺ permeation from one side would not lead to the creation of Ca²⁺ on the other side. Chapter 3 represents a natural progression from Chapter 2 and is aimed at investigating the underlying mechanisms responsible for the electrophysiological ion selectivity observed. This chapter also follows a logical way by first identifying spermine as a high valence intracellular blocker, its mutual antagonism with different external ionic species that allows the determination of ion-binding affinity, followed by the determination of the concentration dependence of ion conduction to identify possible lower affinity binding. By considering all the above qualities, the outcome is a coherent description and connection of ion binding selectivity, kinetic selectivity and ion binding configuration with the observed electrophysiological selectivity. Chapter 4 discusses the missing puzzles and how these questions might be addressed.

In der vorliegenden Arbeit habe ich wichtige Teilmechanismen der Erregungs-Sekretionskopplung in der Speicheldrüse der Schabe Periplaneta americana (L.) untersucht. Die Speicheldrüse ist von dopaminergen und serotonergen Fasern innerviert (Baumann et al., 2002). Beide Transmitter stimulieren eine unterschiedliche Reaktion der Drüse: Dopamin (DA) stimuliert die P-Zellen der Acini und die Ausführgangzellen, während Serotonin (5-HT) die P- und C-Zellen der Acini stimuliert, nicht jedoch die Ausführgangzellen. Der Endspeichel ist nach einer DA-Stimulierung proteinfrei. Dagegen enthält er nach einer 5-HT-Stimulierung Proteine, die von den C-Zellen sezerniert werden (Just & Walz, 1996). Im ersten Teil meiner Arbeit habe ich mittels Kapillarelektrophoretischer Analyse (CE-Analyse) die Elektrolytkonzentrationen im Endspeichel untersucht sowie die Raten der Flüssigkeitssekretion gemessen. Damit wollte ich klären, welche Transporter an der Sekretion des Primärspeichels und an dessen Modifikation beteiligt sind. Ausserdem wollte ich die Rolle der transportaktiven Epithelzellen der Ausführgänge für die Modifikation des Primärspeichels untersuchen. Dafür habe ich einen Vergleich der Elektrolytkonzentrationen im DA- und 5-HT-stimulierten Endspeichel durchgeführt. Der Elektrolytgehalt des DA- und 5-HT-stimulierten Endspeichels unterscheidet sich nicht signifikant voneinander. Er ist nach beiden Stimulierungen hypoosmotisch zum verwendeten Ringer. Die Ausführgangzellen werden durch DA stimuliert und modifizieren den Primärspeichel durch eine netto-Ionenreabsorption. Meine Versuche zeigen jedoch, dass auch die während einer 5-HT-Stimulierung der Drüse unstimulierten Ausführgangzellen den Primärspeichel modifizieren. In einer nachfolgenden Versuchsreihe habe ich den Einfluss von Ouabain, einem Hemmstoff der Na+-K+-ATPase, und Bumetanid, einem Hemmstoff des NKCC, auf die Raten der Flüssigkeitssekretion sowie den Elektrolytgehalt des Endspeichels untersucht. Ich habe gefunden, dass die Aktivität der Na+-K+-ATPase wichtig für die Modifikation des DA-stimulierten Primärspeichels ist. Im Gegensatz dazu ist sie für die Modifikation des 5-HT-stimulierten Primärspeichels nicht von Bedeutung. Bezüglich der Flüssigkeitssekretion habe ich keinen Einfluss der Na+-K+-ATPase-Aktivität auf die DA-stimulierten Sekretionsraten gefunden, dagegen ist die 5-HT-stimulierte Sekretionsrate in Anwesenheit von Ouabain gesteigert. Die Aktivität des NKCC ist für beide sekretorische Prozesse, die Ionen- und die Flüssigkeitssekretion, wichtig. Eine Hemmung des NKCC bewirkt eine signifikante Verringerung der Raten der Flüssigkeitssekretion nach DA- und 5-HT-Stimulierung sowie in beiden Fällen einen signifikanten Abfall der Ionenkonzentrationen im Endspeichel. Im zweiten Teil meiner Arbeit habe ich versucht, Änderungen der intrazellulären Ionenkonzentrationen in den Acinuszellen während einer DA- oder 5-HT-Stimulierung zu messen. Diese Experimente sollten mit der Methode des "ratiometric imaging" durchgeführt werden. Messungen mit dem Ca2+-sensitiven Fluoreszenzfarbstoff Fura-2 zeigten keinen globalen Anstieg in der intrazellulären Ca2+-Konzentration der P-Zellen. Aufgrund von Problemen mit einer schlechten Beladung der Zellen, einer starken und sich während der Stimulierung ändernden Autofluoreszenz der Zellen sowie Änderungen im Zellvolumen wurden keine Messungen mit Na+- und K+-sensitiven Fluoreszenzfarbstoffen durchgeführt. Im dritten Teil dieser Arbeit habe ich die intrazellulären Signalwege untersucht, die zwischen einer 5-HT-Stimulierung der Drüse und der Proteinsekretion vermitteln. Dazu wurde der Proteingehalt im Endspeichel biochemisch mittels eines modifizierten Bradford Assay gemessen. Eine erstellte Dosis-Wirkungskurve zeigt, dass die Rate der Proteinsekretion von der zur Stimulierung verwendeten 5-HT-Konzentration abhängt. In einer Serie von Experimenten habe ich die intrazellulären Konzentrationen von Ca2+, cAMP und / oder cGMP erhöht und anschließend den Proteingehalt im Endspeichel gemessen. Ein Anstieg der intrazellulären Ca2+-Konzentration aktiviert nur eine geringe Rate der Proteinsekretion. Dagegen kann die Steigerung der intrazellulären cAMP-Konzentration eine stärkere Proteinsekretion aktivieren, die sich nicht signifikant von der nach 5-HT-Stimulierung unterscheidet. Die cAMP-stimulierte Proteinsekretion kann durch gleichzeitige Erhöhung der intrazellulären Ca2+-Konzentration weiter gesteigert werden. Dagegen aktivierte eine Erhöhung der intrazellulären cGMP-Konzentration die Proteinsekretion nicht. Aufgrund dieser Ergebnisse postuliere ich die Existenz eines die Adenylatcyclase aktivierenden 5-HT-Rezeptors in der Basolateralmembran der C-Zellen. / The aim of this PhD-work was to investigate major mechanisms of excitation-secretion coupling in the salivary gland of the cockroach Periplaneta americana (L.). This salivary gland is innervated by dopaminergic and serotonergic fibres (Baumann et al., 2002). The two transmitters stimulate different processes in the gland: Dopamine (DA) stimulates the p-cells of the acini and the salivary duct cells, whereas 5-HT (serotonin) activates the p- and the c-cells of the acini, but not the salivary duct cells. Final saliva is completely protein-free after dopamine stimulation. It contains proteins, which are secreted by the c-cells of the acini, after a 5-HT-stimulation (Just & Walz, 1996). In the first part of my work I measured the electrolytic composition of the final saliva by capillary electrophoretic analysis and measured the rates of fluid secretion, in order to answer the following questions: 1.) Which transporters affect the production of primary saliva and its modification? 2.) What is the function of the transport-active salivary duct cells for the modification of the primary saliva? Electrolytic composition of the DA- and 5-HT-stimulated final saliva is not significantly different from each other, and is hypoosmotic to the Ringer used. Salivary duct cells are stimulated by DA and modify the primary saliva by a netto ion-reabsorption. My experiments also show that the duct cells, which are unstimulated during a 5-HT-stimulation of the gland, modify the primary saliva. In the next series of experiments I investigated the effects of ouabain, an inhibitor of the Na+-K+-ATPase, and bumetanide, an inhibitor of the NKCC on the rates of fluid secretion and the electrolytic composition of the final saliva. I found, that the activity of the Na+-K+-ATPase is important for the modification of DA-stimulated primary saliva during its flow through the stimulated duct system. In contrast, it is not important for modification of the 5-HT-stimulated primary saliva. Inhibition of the Na+-K+-ATPase does not affect rates of DA-stimulated fluid secretion, but it increases the rates of 5-HT-stimulated fluid secretion. Activity of the NKCC is important for both secretory processes: the ion and the fluid secretion. Inhibition of the NKCC results in a significant drop in the rates of fluid secretion after DA- and 5-HT-stimulation, as well as a drop in electrolytic concentrations in the saliva. In the second part of my work, I tried to measure changes in the intracellular ionic concentrations (Ca2+, Na+, and K+) within the acinar cells during a DA- or 5-HT-stimulation. The experiments should be performed by ratiometric imaging. Measurements with the Ca2+-sensitive dye Fura-2 did not show any global increase in the intracellular Ca2+-concentration in the p-cells of the acini. Problems concerning a bad loading of the cells, a strong autofluorescence which changed during the time course of the stimulation, as well as changes in the cell volume were the reason, that no measurements using Na+- or K+-sensitive dyes were performed. In the third part of my work I investigated the intracellular signalling pathways, which activate protein secretion after 5-HT-stimulation of the gland. A modified Bradford Assay was used for measuring the protein content in the final saliva. In a dose-response curve I showed that rates of protein secretion are dependent on the 5-HT-concentrations used to stimulate the glands. In another set of experiments I increased the intracellular concentrations of Ca2+, cAMP and / or cGMP, and measured the protein content in the final saliva. An increase in the intracellular Ca2+-concentration activates only a low rate of protein secretion. After an increase in the intracellular cAMP-concentration a much higher rate of protein secretion can be activated, which is not significantly different from the 5-HT stimulated rate of protein secretion. The cAMP-stimulated protein secretion can be further increased by a simultaneous rise in the intracellular Ca2+-concentration. In contrast, cGMP does not activate protein secretion. Therefore I propose the expression of an adenylyl cyclase activating 5-HT-receptor in the basolateral membrane of the protein secreting c-cells.

[Truncated abstract] This study describes the results of a four and a half year investigation examining local regulation of ion transport through pumping epithelial cells. The study focussed on the standard isolated toad skin preparation, made famous by Hans Ussing. Originally, the objective was to perform some simple manipulations on the isolated toad skin, a standard and well-tested epithelial layer, which, according to the literature, was a well-behaved and stable preparation. The purpose of doing these toad skin experiments was to gain familiarity with the experimental techniques, such as measuring the open-circuit voltage (Voc) and the short-circuit current (Isc) across an epithelium. In the process, the experimental information that was obtained was to assist in the development and refinement of a mathematical model of a single pumping epithelial cell . . . Finally, it should be emphasised the toad skin was a convenient tissue model for exploring more general issues such as: (i) how pumping epithelial cells may adjust to changes in the extracellular environment by locally regulating their membrane conductances; (2) how the topology of a cell can influence its function (i.e. the topology can determine whether a cell is optimised for salt transport or water transport). (3) how different cells, with different functions, may be positioned in apposition in a pumping epithelial tissue so that gradients generated by one cell type can be utilised by another. From a broader perspective, it is likely that such issues are also applicable to other pumping epithelia, and ultimately, may assist in understanding how these epithelia function.

Thesis--University of Oklahoma. / Bibliography: leaves 90-92.

L’Hypertension Hyperkaliémique Familiale (HHF) est une forme rare d’hypertension associée à une hyperkaliémie et une acidose métabolique hyperchlorémique, très sensible aux diurétiques thiazidiques. Les premières analyses génétiques ont permis d’identifier deux gènes responsables, WNK1 et WNK4, mais qui n’expliquaient que 8 % de notre cohorte. L’objectif de ma thèse était de rechercher de nouveaux gènes ou variants à l’origine de l’HHF. Notre stratégie initiale a été de combiner une analyse de liaison à un séquençage d’exome entier sur trois grandes familles atteintes. Nous avons ainsi identifié un nouveau gène responsable de la maladie codant pour un acteur jusque là insoupçonné, KLHL3 (Kelch-like 3), responsable de 39% des cas de notre cohorte. La majorité des mutations sont présentes à l’état hétérozygote entrainant un phénotype modéré, alors que les rares patients porteurs d’une mutation homozygote, tous issus de familles consanguines, présentent un phénotype plus marqué. Le spectre des mutations a montré l’importance des structures en boucles de cette protéine qui joue un rôle d’adaptateur de substrat dans un complexe d’ubiquitination (publié dans Nature Genetics, 2012). La découverte d’un type unique de mutations sur le gène CUL-3 par une équipe concurrente a été confirmée dans notre cohorte, entrainant un phénotype plus précoce et plus sévère. Ces mutations ont mis en lumière l’importance de ces deux protéines dans la constitution du complexe E3 ubiquitine-ligase et la dégradation des WNKs dans le néphron, par le protéasome après ubiquitination. Nous avons aussi identifié des mutations faux-sens dans le domaine acide de WNK1 très conservé chez des patients ayant un phénotype HHF mais sans hypertension artérielle. Ce motif, similaire à celui porteur de mutations sur WNK4 est responsable de la liaison à l’adaptateur de substrat KLHL3. Les sujets atteints présentent un âge plus précoce d’apparition de la maladie avec des valeurs de pression artérielle normales. La comparaison phénotypique avec les cas porteurs d’une mutation WNK4 et d’une délétion de l’intron 1 de WNK1 a montré des différences de pression artérielle significatives. La transfection d’ARNc mutés dans les œufs de Xénope, effectuées en collaboration, a permis de démontrer que ces nouvelles mutations faux-sens de WNK1 entrainent une accumulation de son isoforme rénale KS-WNK1 (soumis à J Am Soc Nephrol). L’ensemble de ces résultats ouvre une nouvelle voie de compréhension moléculaire de la régulation du transport des ions sodium, potassium et chlore au niveau du rein et par conséquence de la pression artérielle. / Familial Hyperkalemia Hypertension (FHHt), also known as Gordon syndrome is a rare form of hypertension associated with hyperkalemia and hyperchloremic metabolic acidosis, very sensitive to thiazide diuretics. In 2001, the first genetic analysis identified two genes, coding for two serine/threonine kinases WNK1 and WNK4, which explained only 8% of our cohort. The aim of my thesis was to search new genes or variants responsible for FHHt. We decided to combine a linkage analysis and a whole exome sequencing in three affected families. We identified a new gene responsible for the disease coding for an unsuspected actor KLHL3 (Kelch-like 3), responsible for 39% of our cohort. The majority of the mutations are present at a heterozygous state leading to a moderate phenotype, whereas patients with homozygous mutation, all from consanguineous families, displayed a stronger phenotype. The spectrum of mutations showed the importance of the loop structures of this protein playing an adaptor role in an ubiquitination complex (published in Nature Genetics, 2012). The discovery of a particular type of mutations in CUL-3 by another team was confirmed in our cohort, leading to an earlier and more severe phenotype. These changes have highlighted the importance of these two proteins in the formation of the E3 ubiquitin-ligase-complex and in the WNKs degradation in the nephron by the proteasome after ubiquitination. We have also identified missense mutations in the acidic motif of WNK1, highly conserved in patients with FHHt without hypertension. This pattern is similar to the WNK4 mutations and is responsible for the binding of the substrate adaptor KLHL3. Affected individuals have an earlier age of onset with normal blood pressure values for most of them. Phenotypic comparison with cases carrying WNK4 mutations and deletion of the intron 1 of WNK1 showed significant differences in blood pressure values. Transfection of mutated cRNA in Xenopus laevis oocyte demonstrated that these new WNK1 missense mutations result in the accumulation of the renal isoform KS-WNK1 (submitted to J Am Soc Nephrol). Taken together, these results open a new pathway for understanding the molecular regulation of ion transport and WNK kinases in the kidney and consequently the regulation of blood pressure.

Die vorliegende Dissertation beschäftigt sich mit der Synthese, den Li+-Transporteigenschaften und der Thermodynamik von Olivin-Typ LiFePO4. Es werden verschiedene Solvothermalsynthesen untersucht. Neben der Einstellung von Partikelgröße und Partikelmorphologie steht die Analyse der Hydrothermalsynthese aus Li3PO4 und Vivianit durch in situ Messung der elektrolytischen Leitfähigkeit im Vordergrund. Die Untersuchung des Li+-Transportes geschieht auf Basis von Redoxreaktionen. Die formalkinetische Auswertung von Lithiierungs- und Delithiierungsreaktionen und eine Nukleationsanalyse wird durch ein Modell zur Auswirkung von antisite-Defekten auf die Kapazität des Elektrodenmaterials ergänzt. Die Ramanspektroskopie wird in Verbindung mit Lösungsenthalpien zur Identifizierung reaktiver Spezies herangezogen. Schwerpunkt der thermodynamischen Charakterisierung ist die experimentelle Ermittlung der Wärmekapazität. Diese wurde unter Berücksichtigung einer magnetischen Phasenumwandlung im Bereich von 2 K bis 773 K ermittelt. Die Daten erlauben die Berechnung wichtiger thermodynamischer Funktionen.

Two-dimensional polymers (2DPs) and their layer-stacked 2D covalent organic frameworks (2D COFs) are classes of structurally defined crystalline polymeric materials with exotic physical and chemical properties. Yet, synthesizing 2DP and 2D COF single crystals via irreversible reactions remains challenging. Here we report the synthesis of charged 2DP (C2DP) single crystals through an irreversible Katritzky reaction, under pH control, on a water surface. The periodically ordered 2DPs comprise aromatic pyridinium cations and counter BF4− anions. The C2DP crystals, which are composed of linked porphyrin and pyrylium monomers (C2DP-Por), have a tunable thickness of 2–30 nm and a lateral domain size up to 120 μm2. Single crystals with a square lattice (a = b = 30.5 Å) are resolved by imaging and diffraction methods with near-atomic precision. Furthermore, the integration of C2DP-Por crystals in an osmotic power generator device shows an excellent chloride ion selectivity with a coefficient value reaching ~0.9 and an output power density of 4 W m−2, superior to those of graphene and boron nitride.