Depuis plusieurs années, il est devenu plus aisé de détecter et d’étudier les mécanismes biologiques in cellulo grâce aux techniques d’imagerie optique que sont la fluorescence, la bioluminescence et la chimiluminescence. Bien que la fluorescence soit la technique la plus employée de nos jours, la bioluminescence et la chimiluminescence, qui sont la conséquence d’une cascade de réactions (bio)chimiques, sont très étudiées depuis quelques décennies. En effet, elles permettent de contourner la source des problèmes rencontrés dans l’utilisation d’un fluorophore : la lumière excitatrice. La chimiluminescence est par conséquent une méthode de choix pour s’affranchir de l’auto-fluorescence tissulaire. Le 1,2-dioxétane est un des motifs chimiluminescent qui, par décomposition, peut générer un état excité sur un fluorophore auquel il est connecté, autorisant ainsi sa luminescence intrinsèque. Ainsi, le but de ce projet de thèse a été de développer et d’étudier des nouvelles sondes chémiluminescentes à motif 1,2-dioxétane à coeurs naphtolique et phénolique, compatibles avec le vivant pour une détection ciblée de complexes enzymatiques. La stratégie envisagée passait par la préparation d'une plateforme chimiluminescente comportant un motif 1,2-dioxétane thermiquement stable, sur laquelle il est possible de faire varier le déclencheur (i.e. possibilité d'adapter cette plateforme à l'analyte ou événement que l'on souhaite détecter) et d'accrocher un fluorophore NIR. Deux méthodes ont été tentées : d’une part une greffe d’une version hydrosoluble d’un fluorophore connu (i.e. le Nile red) pour réaliser un transfert d’énergie à travers les liaisons (i.e. TBET), et d’autre part un couplage à un complexe de lanthanide permettant un transfert d’énergie à travers l’espace (i.e. CRET). / Since several years, it is easier to exploit biological phenomena in cellulo through technologies dealing with bioimaging. This method gathers fluorescence, bioluminescence and chemiluminescence. Even though fluorescence is the most employed technic, bioluminescence and chemiluminescence, being the consequence of (bio)chemical reaction, have been widely studied for decades. In fact, they can avoid the main problems encountered in fluorophore use: exciting light. Chemiluminescence is thus the appropriate approach to avoid biological autofluorescence. 1,2-dioxetane is one of the moieties that, upon decomposition, can generate an excited state on a connected fluorophore, giving rise to its intrinsic luminescence. The aim of our project was to develop and study new 1,2-dioxetan chemiluminescent probes based on phenol or naphthol moieties, for targeted enzymatic complexes detection with in cellulo and in vivo bioimaging. The strategy relied on the synthesis of chemiluminescent scaffolds comprising thermally stable 1,2-dioxetan, on which the trigger and the connected NIR fluorophore can be easily diversified. Two methods have been attempted: 1) coupling to a water-soluble version of a known fluorophore (i.e. Nile red) allowing an energy transfer through bonds (i.e. TBET) 2) connection with a lanthanide complex giving rise to an energy transfer through space (i.e. CRET).

La valorisation des effluents gazeux à faible pouvoir calorifique, sous-produits de différents procédés industriels (gazéification du charbon ou de la biomasse, rejets industriels) apparait aujourd’hui comme une solution alternative pour accroître l’efficacité globale des systèmes de combustion par réduction des coûts énergétiques et contrôle des rejets dans l’environnement. Dans ce contexte, une étude expérimentale d'oxyflammes d'un gaz à très bas pouvoir calorifique, le gaz de haut fourneau (BFG), est réalisée pour évaluer l'effet conjoint de l'oxycombustion et du préchauffage des réactifs pour la stabilisation des flammes turbulentes. La configuration du brûleur consiste en un jet annulaire de gaz de haut fourneau (BFG), entouré de deux injections d’oxygène (interne ‘O2i’ et externe ‘O2e’). Son dimensionnement s’appuie sur une méthodologie originale basée sur la détermination d’une vitesse de convection critique UC* à l’extinction de flamme, dérivée d’une valeur expérimentale d'un nombre de Damköhler critique Dac*. Les structures de flammes sont caractérisées par imagerie de chimiluminescence OH* et les propriétés thermiques et chimiques sont évaluées par mesures de température et flux thermique à la paroi et de la composition des fumées. Les champs aérodynamiques 2D des écoulements réactifs sont mesurés par PIV. Quatre principales topologies de flamme sont observées avec cette configuration de brûleur et classées suivant leur mode de combustion. Sans préchauffage, les deux flammes concentriques, interne ‘O2i-BFG’ et externe ‘BFG-O2e’, sont attachées au brûleur à basse puissance (Type A) ; la flamme BFG-O2e peut présenter une stabilité intermittente (Type B), ou se décrocher du brûleur (Type C) à la puissance nominale de dimensionnement. Avec préchauffage des réactifs, la flamme annulaire BFG-O2e reste toujours accrochée au brûleur et la flamme centrale O2i-BFG présente une zone d’extinction locale pour des fortes valeurs de vitesse d’O2i (Type D). L’ensemble des résultats a permis de mettre en avant un bon accord entre les prédictions théoriques et les valeurs expérimentales de UC* avec un élargissement des domaines de stabilité de flamme avec le préchauffage. L'analyse aérodynamique permet de caractériser les transitions entre les structures de flammes. Une validation du critère de dimensionnement de l’oxy-brûleur est aussi effectuée par changement d’échelle, grâce à des mesures réalisées sur une installation semi-industrielle de 180 kW. / The effective utilisation of low calorific value fuel, as gaseous by-products of coal/biomass or industrial residual gases, provides not only excellent opportunities for low cost power generation but also for the reduction of environmental impact of combustion. The present work aims to consider the combination of oxyfuel combustion with fuel and/or oxygen preheating in order to increase thermal efficiency by heat recovery and enhance oxyfuel flame stabilization of blast furnace gas (BFG). This experimental study is performed with a burner consisting in an annular jet of BFG surrounded by two injections of oxygen (internal 'O2i' and external 'O2e'). Its dimensions are determined from an original design strategy based on an experimentally critical Damköhler number Dac*, which represents the theoretical limit of stabilisation of a turbulent diffusion BFG-O2 flame with preheated reactants. Flames structures are characterized by OH* chemiluminescence imaging. Thermal and chemical flame properties are evaluated by temperature and radiative flux analysis and pollutant emissions measurements. The 2D aerodynamic fields of reactive flows are determined by velocity measurements by PIV. Four LCV flames structures are resulting from this burner configuration. Without preheating, two concentric flames, internal 'O2i-BFG' and external 'BFG-O2e', are anchored at the burner (Type A) at low thermal power. When increasing the latter, the external flame BFG-O2e manifests some local fluctuations (Type B) or is lifted-off (Type C). With reactants preheating, the BFG-O2e flame is always anchored at the burner tip and the O2i-BFG flame could have local extinction zone for very high values of internal oxygen velocity (Type D). The results highlight a good agreement between theoretical and experimental critical velocity UC* which significantly increases with preheating. The aerodynamics study points out the transitions between the different flames structures. At semi-industrial scale, flames show similar structures to those obtained at laboratory scale. This validates the burner design strategy of preheated oxyfuel combustion adapted to LCV fuels, as well as the scale up criteria used.

E-learning has surged in popularity due to technological advancements and the COVID-19 pandemic, proving effective across various fields, including the healthcare sector. DiaSorin AB, a subsidiary of DiaSorin, a multinational biotechnology company, is exploring e-learning as an alternative to the on-site courses they are currently offering their customers, who mainly consist of laboratory personnel. One of DiaSorin AB’s long-term goals is to provide an e-learning platform for their chemiluminescence analyzer LIAISON XL, potentially reducing costs, commute time, and emissions while achieving positive learning outcomes. This thesis investigates the needs and challenges faced by laboratory personnel using chemiluminescence analysers, specifically LIAISON XL, and supports DiaSorin AB in developing an e-learning platform for LIAISON XL. This was accomplished through a user-centred design process consisting of six main components: 1) conducting a field study, 2) carrying out an interview study, 3) performing thematic analysis on the collected data, 4) creating personas, 5) establishing requirements, and 6) developing a prototype for an e-learning platform. By considering the established requirements, a potential e-learning platform for LIAISON XL can better support laboratory personnel and enhance their proficiency in using LIAISON XL. Future work should prioritise usability testing and refinement of both the prototype and the requirements. / E-lärande har blivit mycket populärt på grund av teknologiska framsteg och COVID-19-pandemin, och har visat sig vara effektivt inom olika områden, inklusive hälso- och sjukvårdssektorn. DiaSorin AB, ett dotterbolag till DiaSorin, en global ledare inom laboratoriediagnostik, utforskar e-lärande som ett alternativ till de kurser som de för närvarande erbjuder sina kunder, som huvudsakligen består av laboratoriepersonal. Ett av DiaSorin AB:s långsiktiga mål är att kunna erbjuda en e-lärandeplattform för sin kemiluminiscensanlaysator LIAISON XL, vilket potentiellt kan minska kostnader, restider och utsläpp samtidigt som positiva inlärningsresultat uppnås. Denna avhandling uppsats behoven och utmaningarna som laboratoriepersonal står inför när de använder kemiluminiscensanlaysatorn, särskilt LIAISON XL, och stöder DiaSorin AB i utvecklingen av en e-lärandeplattform för LIAISON XL. Detta åstadkoms genom en användarcentrerad designprocess bestående av sex huvudkomponenter: 1) genomförandet av en fältstudie, 2) genomförandet av en intervjustudie, 3) utförandet av tematisk analys av insamlad data, 4) skapandet av personas, 5) etablerandet av krav och 6) utvecklingen av en prototyp för en e-lärandeplattform. Genom att beakta de etablerade kraven kan en potentiell e-inlärningsplattform för LIAISON XL bättre stödja laboratoriepersonal och förbättra deras kompetens i att använda LIAISON XL. Framtida arbete bör prioritera användbarhetstester och förbättring av både prototypen och kraven.

In der Konservierungsforschung werden unterschiedlichste Materialien analysiert und auf ihre Stabilität hin geprüft, um gezielte Konservierungs- oder Restaurierungsmassnahmen durchzuführen und deren Nachhaltigkeit besser einschätzen zu können. Die Chemilumineszenz - im Folgenden kurz CL - ist ein Phänomen, bei dem ein Elektron aus einem energetisch höher gelegenen Orbital in einen tiefer liegenden Zustand relaxiert und dabei seine Energie in Form von Photonen abgibt. Der angeregte Zustand wird durch eine chemische Reaktion verursacht. Dieser Vorgang ist sehr spezifisch und tritt u.a. bei Oxidationsreaktionen auf. Durch Rekombination zweier Peroxide wird z.B. eine angeregte Carbonylverbindung erzeugt, die ihre Energie als Licht emittiert. Dieser Zusammenhang zwischen der Oxidation organischer Substanzen und die Emission von Photonen bedingt die hier zentral gestellte Frage, ob die Chemilumineszenz als Methode zur Charakterisierung von Materialien, die in der Konservierungsforschung relevant sind, eingesetzt werden kann. Innerhalb einer Forschungskooperation zwischen Departementen der Berner Fachhochschule BFH konnte ein Chemilumineszenz-Messgerät konstruiert, modifiziert und für diese Arbeit eingesetzt werden. In der Arbeit wurde ein standardisiertes Messverfahren an verschiedenen Substanzen, wie z.B. Adamantylidenadamantan-1.2-dioxetan geprüft und die verschiedenen Einflüsse auf das Chemilumineszenz-Signal untersucht. Dabei konnten für das Messprinzip wichtige Daten, wie z.B. Präzision, Nachweis- und Bestimmungsgrenze erhoben werden. Anhand der Stabilität von Dammar, einem Triterpen-Naturharz, konnte das Verfahren innerhalb der Arbeit, an einem komplexen Beispiel aus der Konservierungsforschung experimentell erprobt werden. Dabei wurden publizierte Resultate zum Oxidationsverhalten von Dammarharz teilweise verifiziert. Durch das spezifische Signal konnten die thermisch initiierten Reaktionen bei inerten und oxidativen Gas-Atmosphären dokumentiert werden. Die Ergebnisse wurden mit etablierten Methoden, wie z.B. DSC, TGA und ESR- Spektroskopie verglichen und Konsistenzen zu den Signalen erfasst. Der in der Literatur diskutierte Einfluss von Stabilisatoren auf Dammarharz-Filme wurde an ungealterten und künstlich gealterten Proben mit der Chemilumineszenz untersucht. Dabei konnten ebenfalls in der Literatur empfohlene Formulierungen und Wirkungen bestimmter synergetischer Mischungen nachgewiesen und bestätigt werden. Aus den ermittelten Daten der CL-Messungen liessen sich in einem Ausblick kinetische Berechnungen zu Aktivierungsparametern erstellen. Die ermittelten Parameter wurden zur Prognose von Reaktionsverläufen bei unterschiedlich simulierten Temperaturprofilen genutzt und machten eine Vorhersage zur Lebensdauer der Materialien möglich. Neben der hohen Leistungsfähigkeit der Methode zeigte sich v.a. das hohe Potential der ermittelten Messdaten, v.a. in Bezug auf die Anwendung in der Modellfreien Kinetik. Neben diesen Möglichkeiten wurden aber auch die Grenzen der Methode aufgezeigt. Diese sind einerseits durch die Besonderheit des Signals gegeben, welches mechanistisch bis heute noch nicht vollständig aufgeklärt ist, andererseits liegen sie bei der Weiterverarbeitung der Messdaten durch thermokinetische Software. Hier wird deutlich, dass die Qualität der Berechnungen zu Reaktionsverläufen nur so gut ist, wie die zuvor erhobenen Daten und, dass erstellte Prognosen zur Stabilität eines Materials lediglich eine Vorstellung vermitteln können. / During conservation studies different materials are analysed and tested for their solidity in order to be able to carry out precise conservation and restoration measures and to be able to assess their sustainability. Chemiluminescence is a phenomenon in which an electron is relaxed from an energetically higher orbital into a lower situated condition while simultaneously releasing energy in the shape of photons. As a consequence the stimulated state is caused by a chemical reaction. This process is very specific and occurs during an oxidation reaction among other things. Through the recombination of two peroxides for example a stimulated carbonyl compound is generated which emits its energy as light. This connection between the oxidation of organic substances and the emission of photons determines the main question, whether chemiluminescence can be used as a method for characterisation of materials which are relevant to the research of conservation. As a result of a research project between departments of the Bern University of Applied Sciences BFH it was possible to construct a chemiluminescence measuring device and modify it in a way that it could be used for this work. Within this experiment a standardised method of measurement was tested on different substances, for example adamantylidenadamantan-1.2-dioxetan, and the different impacts on the chemiluminescence-signal investigated. Through this process important facts were determined for the concept of measurement, for example precision, proof- and definition perimeters. Based on the solidity of dammar, a triterpene resin, the method could be experimentally verified within the test using a complex example from conservation research. Within that process published results about the oxidation properties of dammar resin were verified. Due to the specific signal, the thermally initiated responses in inert as well as oxidative gas atmospheres could be documented. The results were compared with established methods like DSC, TGA and ESR spectroscopy and the consistencies to the signals logged. The already much discussed influence of stabilisers on dammar resin was tested on nonaged and artificially aged specimens using chemiluminescence. Through these previously suggested formulations, effects of certain synergistic composites were detected and confirmed. Using the data from the chemiluminescence measurements kinetic calculations about activation parameters were compiled. The identified parameters were used for the prognosis of the course of reactions during differently simulated temperature profiles, making a prediction of the materials’ life span possible. Through these results not only the high performance ability of the method became apparent but also the potential of the determined measurement data, mainly in connection with the application within the model-free kinetic. However, alongside these possibilities limitations of the method were revealed. These are partly caused by the characteristics of the signal which has yet to be mechanistically fully clarified. Simultaneously, further processing of the measurement data via thermokinetic software makes clear that the quality of the calculations of the reaction process is only as good as the quality of the initial data. This suggests the prognoses about the stability of the material only conveys a concept of their qualities.

The main objective of this Thesis is the study of the autoignition phenomenon of reactive mixtures from a theoretical and experimental point of view. A wide parametric study has been carried out in a Rapid Compression-Expansion Machine (RCEM) for different initial temperatures, compression ratios, equivalence ratios and molar fractions of oxygen (by using synthetic EGR) for different fuels. The ignition delay referred to cool flames (if it can be identified), as well as the ignition delay referred to the high-temperature stage of the ignition, have been experimentally obtained and their trends have been explained regarding the chemical kinetics of each fuel. The different effects of the species that compose the synthetic EGR on the ignition delay have been studied, decoupling the thermodynamic effects from the chemical ones. Different compositions have been taken into account to generate the synthetic EGR, and validation limits have been obtained for each mixture. The thermodynamic and the chemical effects have shown to be opposed, while the dominant one is different depending on the working temperature. Several chemical kinetic mechanisms have been validated by comparison to the experimental results. A detailed mechanism for iso-octane and n-heptane blends and a reduced mechanisms for n-dodecane have been analyzed. Moreover, a sub-model for the generation and decay of excited OH* has been validated by comparison to chemiluminescence and spectroscopy results. The different radiation sources have been studied for iso-octane and n-heptane by means of spectroscopy techniques. Besides, chemiluminescence measurements filtered at 310nm (OH* emission wavelength) have been performed in order to analyze the generalization and propagation velocity of the autoignition front. The ignition propagation has shown to depend on the thermodynamic conditions reached in the combustion chamber when the first ignition spot occurs and not on the global reactivity of the mixture. Furthermore, two different radiation sources have been found at 310nm in the spectroscopic analysis depending on the ignition intensity: the decay of the OH* radical from excited to ground state and the oxidation of CO to CO2 (CO continuum). However, these optical techniques have been applied only in the experiments carried out with iso-octane and n-heptane due to technical limitations. Finally, a new predictive model has been theoretically developed starting from the Glassman's model for autoignition. This method is based on modeling the accumulation rate of chain carriers up to reach their critical concentration (obtaining the ignition delay referred to cool flames) and, afterwards, modeling the disappearance rate of such chain carriers up to their consumption (when the maximum heat release rate is reached, obtaining the ignition delay referred to the high-temperature stage of the process). The predictive capability of the model has been compared to the ability of other methods that can be found in the literature, such as the Livengood & Wu integral method. The validity of each method has been tested, defining a working methodology to obtain reasonable predictions for the ignition delay. / El objetivo de esta Tesis Doctoral es el estudio del fenómeno de autoencendido de mezclas reactivas desde un punto de vista teórico y experimental. Se ha realizado un amplio estudio paramétrico en una Máquina de Compresión-Expansión Rápida (RCEM por sus siglas en inglés) barriendo diversas temperaturas iniciales, relaciones de compresión, dosados relativos y fracciones molares de oxígeno (mediante el uso de EGR sintético) para distintos combustibles. El tiempo de retraso del fenómeno de llamas frías (en el caso de existir), así como el tiempo de retraso de la etapa de alta temperatura, han sido obtenidos experimentalmente y sus tendencias explicadas mediante cinética química. Se han estudiado los diferentes efectos de las distintas especies involucradas en el EGR sintético sobre el tiempo de retraso, desligando aquellos de carácter termodinámico de los efectos puramente químicos. Se han tenido en cuenta distintas composiciones para definir dicho EGR, estableciendo límites de validez para cada una de las mezclas propuestas. Los efectos termodinámicos y químicos resultaron ser opuestos, siendo dominante uno u otro a distintos rangos de temperatura de trabajo. Varios mecanismos de cinética química han sido validados gracias a los resultados experimentales obtenidos. Además de un mecanismo detallado para mezclas PRF de iso-octano y n-heptano, se ha llevado a cabo la validación de otro mecanismo simplificado para el n-dodecano. Por otro lado, un submodelo de formación y decaimiento de OH* excitado ha sido validado contra resultados de quimioluminiscencia y espectroscopía. Se han estudiado las diferentes fuentes de radiación del proceso de autoencendido para el iso-octano y el n-heptano mediante técnicas de espectroscopía. Además, se han realizado medidas de quimioluminiscencia filtrada a 310nm (longitud de onda de emisión del radical OH*) para el análisis de la generalización y velocidad de propagación del frente de autoencendido. La propagación del encendido ha mostrado ser dependiente de las condiciones termodinámicas alcanzadas en la cámara de combustión en el instante de ignición más que de la reactividad de la mezcla. Se han encontrado dos fuentes de radiación distintas a 310nm mediante espectroscopía, dependiendo de la intensidad del encendido: el decaimiento del radical OH* de estado excitado a estado natural y la oxidación del CO a CO2 (continuo del CO). No obstante, estas técnicas han sido utilizadas solamente para los dos combustibles de referencia de la escala de octanaje debido a limitaciones técnicas. Finalmente, se ha desarrollado un nuevo modelo predictivo de manera teórica partiendo del modelo de Glassman para el autoencendido. Este método se basa en modelar primero la tasa de acumulación de portadores de cadena hasta su concentración crítica (obteniendo así el tiempo de retraso referido a la etapa de llamas frías) y, tras dicho instante, modelar la tasa de consumo de dichos portadores de cadena hasta su completa desaparición (instante en el cual se produce la máxima exotermia del proceso, prediciendo el tiempo de retraso referido a la etapa de alta temperatura del encendido). La capacidad predictiva del modelo ha sido comprobada para cada uno de los seis combustibles ensayados. Además, dicha capacidad predictiva ha sido comparada con la de otros métodos existentes en la literatura, como la integral de Livengood & Wu. La validez de cada uno de los métodos ha sido analizada, definiendo una metodología de uso para obtener predicciones razonables del tiempo de retraso. / L'objectiu d'aquesta Tesi Doctoral és l'estudi del fenomen d'autoencesa de mescles reactives des d'un punt de vista teòric i experimental. S'ha realitzat un ampli estudi paramètric en una Màquina de Compressió-Expansió Ràpida (RCEM per les seues sigles en anglès) cobrint diverses temperatures inicials, relacions de compressió, dosatges relatius i fraccions molars d'oxigen (mitjançant l'ús de EGR sintètic) per a diferents combustibles. El temps de retard del fenomen de flames fredes (en el cas d'existir), així com el temps de retard de l'etapa d'alta temperatura, han sigut obtinguts experimentalment i les seues tendències explicades mitjançant cinètica química. S'han estudiat els diferents efectes de les diferents espècies involucrades en l'EGR sintètic sobre el temps de retard, deslligant aquells de caràcter termodinàmic dels efectes purament químics. S'han tingut en compte diferents composicions per a definir aquest EGR, establint límits de validesa per a cadascuna de les mescles proposades. Els efectes termodinàmics i químics van resultar ser oposats, sent dominant un o un altre a diferents rangs de temperatura de treball. Diversos mecanismes de cinètica química han sigut validats gràcies als resultats experimentals obtinguts. A més d'un mecanisme detallat per a mescles PRF d'iso-octà i n-heptà, s'ha dut a terme la validació d'un altre mecanisme simplificat per al n-dodecà. D'altra banda, un submodel de formació i decaïment d'OH* excitat ha sigut validat contra resultats de quimioluminescència i espectroscopía. S'han estudiat les diferents fonts de radiació del procés d'autoencesa per a l'iso-octà i l'n-heptà mitjançant tècniques d'espectroscopía. A més, s'han realitzat mesures de quimioluminescència filtrada a 310nm (longitud d'ona d'emissió del radical OH*) per a l'anàlisi de la generalització i velocitat de propagació del front d'autoencesa. La propagació de l'encesa ha mostrat ser depenent de les condicions termodinàmiques aconseguides en la cambra de combustió en l'instant d'ignició més que de la reactivitat de la mescla. S'han trobat dues fonts de radiació diferents a 310nm mitjançant espectroscopía, depenent de la intensitat de l'encesa: el decaïment del radical OH* d'estat excitat a estat natural i l'oxidació del CO a CO2 (continu del CO). No obstant açò, aquestes tècniques han sigut utilitzades solament per als dos combustibles de referència de l'escala de octanaje a causa de limitacions tècniques. Finalment, s'ha desenvolupat un nou model predictiu de manera teòrica partint del model de Glassman per a l'autoencesa. Aquest mètode es basa a modelar primer la taxa d'acumulació de portadors de cadena fins a la seua concentració crítica (obtenint així el temps de retard referit a l'etapa de flames fredes) i, després d'aquest instant, modelar la taxa de consum d'aquests portadors de cadena fins a la seua completa desaparició (instant en el qual es produeix la màxima exotermia del procés, predient el temps de retard referit a l'etapa d'alta temperatura de l'encesa). La capacitat predictiva del model ha sigut comprovada per a cadascun dels sis combustibles assajats. A més, aquesta capacitat predictiva ha sigut comparada amb la d'altres mètodes existents en la literatura, com la integral de Livengood & Wu. La validesa de cadascun dels mètodes ha sigut analitzada, definint una metodologia d'ús per a obtenir prediccions raonables del temps de retard. / López Pintor, D. (2017). Theoretical and experimental study on the autoignition phenomena of homogeneous reactive mixtures [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/90642

Le présent travail a porté sur l’évaluation des propriétés anti-inflammatoires et anticancéreuses de Agelanthus dodoneifolius (Loranthaceae), communément appelée «gui africain». Cette plante hémiparasite est utilisée en médecine traditionnelle africaine pour le traitement de pathologies chroniques telles que l’asthme, l’hypertension, des gastroentérites et le cancer. Actuellement, les maladies chroniques représentent un problème mondial de santé publique. En effet, elles constituent la première cause de mortalité dans le monde surtout dans les pays à revenu faible ou intermédiaire. <p><p>Cette étude a été réalisée dans le but d’apporter une validation scientifique quant à certaines utilisations traditionnelles de Agelanthus dodoneifolius. <p>Pour évaluer l’effet anti-inflammatoire de Agelanthus dodoneifolius, nous avons testé les différentes fractions de la plante sur la production des espèces réactives de l’oxygène, la libération et l’activité spécifique de la myéloperoxydase (MPO), enzyme libérée par le neutrophile au cours de la phagocytose pour détruire les microorganismes. L’identification et la quantification des composés a été faite grâce à une combinaison des méthodes chromatographiques, spectrophotométriques et spectrométriques. L’activité anticancéreuse de Agelanthus dodoneifolius a consisté, d’abord, à déterminer l’effet d’inhibition de croissance de diverses fractions de la plante, de la quercétine ainsi que de ses dérivés sur des lignées cellulaires cancéreuses. Nous avons ensuite déterminé les effets de la quercétine sur l’activité de plus de 300 kinases. <p><p>Les résultats obtenus montrent qu’Agelanthus dodoneifolius est capable de moduler les activités biologiques des neutrophiles. En effet, le décocté aqueux et les fractions organiques de la plante inhibent de manière dose-dépendante la production des espèces réactives de l’oxygène, la dégranulation du neutrophile et l’activité spécifique de la myéloperoxydase. Nous avons pu identifier et quantifier dix composés polyphénoliques dont quatre acides phénoliques :l’acide gallique, l’acide coumarique, l’acide chlorogénique et l’acide ellagique et six flavonoïdes :la quercétine, le kaempférol, la catéchine, l’isoquercitrine ou quercétine 3-O-glucoside, la rutine et la miquelianine ou quercétine-3-O-glucuronide. <p>Concernant l’activité anticancéreuse, les résultats montrent que seules les fractions à l’éther diéthylique et à l’acétate d’éthyle ont une activité antiproliférative. La quercétine a des effets inhibiteurs de croissance, cytostatiques et présente un large spectre d’activité sur plusieurs kinases surexprimées dans certains cancers. <p><p>En conclusion, l’ensemble de ces résultats constitue des bases scientifiques qui pourraient justifier certaines utilisations traditionnelles de Agelanthus dodoneifolius. <p>À notre connaissance, cette étude est la première à évaluer d’une part l’effet, in vitro, des différentes fractions de Agelanthus dodoneifolius sur des neutrophiles stimulés et sur la MPO et d’autre part l’effet inhibiteur de croissance de lignées cellulaires cancéreuses par certaines fractions de la plante. En outre, cette étude a permis pour la première fois d’identifier et de quantifier des composés polyphénoliques dans Agelanthus dodoneifolius. Les nombreuses propriétés de ces composés, notamment celles anti-inflammatoires et anticancéreuses, peuvent expliquer en partie les résultats reportés dans ce travail.<p><p>This work focused on evaluating anti-inflammatory and anticancer activities of Agelanthus dodoneifolius (Loranthaceae), commonly called "African mistletoe". This plant is used in African traditional medicine for the treatment of chronic conditions such as asthma, hypertension, gastroenteritis and cancer. Currently, chronic diseases are a global public health problem. Indeed, they are the leading cause of death worldwide, especially in countries with low and middle income.<p>The study was conducted to provide scientific validation for some traditional uses of Agelanthus dodoneifolius.<p><p>To characterize the anti-inflammatory activity of Agelanthus dodoneifolius, we tested the different fractions of the plant on reactive oxygen species production, release and the specific activity of myeloperoxidase, an enzyme released by neutrophils during phagocytosis to destroy microorganisms. The identification and quantification of compounds were made through a combination of chromatographic, spectrophotometric and spectrometric techniques. The anticancer activity of Agelanthus dodoneifolius consisted, first, to determine, the antiproliferative effect of fractions of the plant, quercetin and its derivatives on cancer cell lines. Then, we determined the effects of quercetin on the activity of more than 300 kinases.<p><p>The results show that Agelanthus dodoneifolius is capable of modulating the biological activities of neutrophils. In fact, the decoction aqueous and organic fractions of the plant inhibited in a dose-dependent manner the production of reactive oxygen species, degranulation of neutrophils and specific activity of myeloperoxidase. <p>We were able to identify and quantify ten polyphenolic compounds including four phenolic acids: gallic acid, coumaric acid, chlorogenic acid and ellagic acid and six flavonoids: quercetin, kaempferol, catechin, isoquercitrin or quercetin 3-O-glucoside, rutin and miquelianin or quercetin-3-O-glucuronide.<p>Regarding the anticancer activity, the results show that only fractions with diethyl ether and ethyl acetate have antiproliferative activity. Quercetin has antiproliferative and cytostatic effects and presents a broad spectrum of activity on several kinases overexpressed in certain cancers.<p><p>In conclusion, all these findings are scientific basis that could justify some traditional uses of Agelanthus dodoneifolius. To our knowledge, this is the first study to evaluate the effect firstly, by in vitro tests, of the different fractions of Agelanthus dodoneifolius on stimulated neutrophils and the MPO and secondly the growth inhibitory effect of cancer cell lines by certain fractions. Also, this study is the first to identify and quantify the phenolic compounds in Agelanthus dodoneifolius. The many properties of these compounds, including anti-inflammatory and anticancer, may partly explain the results reported in the present work. / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

The cyclic peroxides: diphenoyl peroxide (1), spiro-adamantyl-1,2-dioxetanone (2) and 4-(3-tert-butyldimethylsilyloxyphenyl)-4-methoxyspiro[1.2-dioxetane-3.2\'-adamantane] (3) synthesized, purified and characterized, also their kinetic and chemiluminescence (CL) properties were determined. The influence of solvent polarity on the chemiexcitation parameters of the catalyzed decomposition of 1 and 2, as well as the induced decomposition of 3 were examined in several binary solvent mixtures with different polarity and polarizability parameters, but similar viscosities. For solvents with low polarity, singlet quantum yields for the intermolecular catalyzed decomposition of diphenoyl peroxide (1,) and 1,2-dioxetanone (2) increase in function of medium polarity, showing maximum values in mediums of intermediate polarity, and decreasing in highly polar mixtures. For the induced decomposition of 3, the quantum yield showed to increase with increasing solvent polarity, including high polar solvent systems. In the binary solvent systems studied, the polarizability parameter showed to be contrary to the polarity values, therefore, it might appear that an increase of polarizability leads to a decrease in the singlet quantum yields for catalyzed decomposition of diphenoyl peroxide (1) and the induced decomposition of 3, however an increase in the singlet quantum yields for 1,2-dioxetanone 2. The three CL systems were also studied in binary solvent mixtures with different viscosities but similar polarity and polarizability parameters and singlet quantum yields showed an increase with increasing medium viscosity. Data were analyzed by using the collisional and the free volume models. The highly efficient intramolecular system induced 1,2-dioxetane 3 decomposition showed to be much more sensible to viscosity effects than the inefficient intermolecular systems. This surprising fact indicates that, even showing a significant solvent-cage effect, the induced 1,2-dioxetane should occur by an intramolecular electron back transfer. Moreover, chemiluminescence parameters of these systems were studied in several pure solvents. The obtained singlet quantum yields were correlated with viscosity, polarizability and polarity parameters by using multiple linear regression analysis. / Os peróxidos cíclicos: peróxido de difenoila (1), spiro-adamantil-1,2-dioxetanona (2) e 4-(3-terc-butildimetilsililoxifenil)-4-metoxispiro[1,2-dioxetano-3.2\'-adamantano] (3) foram sintetizados, purificados e caracterizados e as suas propriedades cinéticas e de quimiluminescência (CL) determinadas. A influência da polaridade do solvente sobre os parâmetros de quimi-excitação da decomposição catalisada de 1 e 2, e a decomposição induzida de 3 foi examinada em diversas misturas binárias de solventes com parâmetros de polaridade e polarizabilidade diferente, mas viscosidades semelhantes. Para solventes com baixa polaridade, os rendimentos quânticos singlete para a decomposição catalisada intermolecular do peróxido de difenoila (1) e 1,2-dioxetanona (2) aumentam em função da polaridade do meio, mostrando valores máximos em meios com polaridade intermediária, e diminuim para misturas altamente polares. Para a decomposição induzida de 3, o rendimento quântico mostrou aumentar com o aumento da polaridade do solvente, inclusive para sistemas com alta polaridade. Nos sistemas binários de solventes estudados, os parâmetros de polarizabilidade mostraram-se contrários aos valores de polaridade, portanto, parece que um aumento da polarizability leva a uma diminuição nos rendimentos quânticos singlete para a decomposição catalisada do peróxido de difenoila (1) e a decomposição induzida de 3, no entanto, um aumento nos rendimentos quânticos singlete para a 1,2-dioxetanona 2. Os três sistemas de CL também foram estudados em misturas binárias de solventes com diferentes viscosidades, mas com parâmetros de polaridade e polarizabilidade semelhantes e os rendimentos quânticos singlete mostraram aumentar com o aumento da viscosidade do meio. Os dados foram analisados usando tanto o modelos colisional quando o modelo de volume livre. Surpreendentemente, o sistema altamente eficiente decomposição induzida intramolecular do 1,2-dioxetano 3 mostrou-se muito mais sensível aos efeitos da viscosidade do que os sistemas intermoleculares ineficientes, o que indica claramente que a decomposição induzida do 1,2-dioxetano deve ocorrer por um processo de retro-transferência de elétron intramolecular. Além disso, os parâmetros de quimiluminescência destes sistemas foram estudados em vários solventes puros. Os rendimentos quânticos singlete obtidos foram correlacionados com parâmetros de viscosidade, polarizabilidade e polaridade usando análise de regressão linear múltipla.

Estudos envolvendo o oxigênio molecular singlete (1O2) tem uma relevância biológica, uma vez que esta espécie, devido ao caráter eletrofílico, reage com moléculas ricas em elétrons como proteínas, lipídeos e DNA provocando danos que resultam em perdas de função e integridade celular. Em sistemas biológicos, a presença de outras espécies reativas de oxigênio e nitrogênio, dificultam a identificação de lesões específicas causadas por 1O2 .Neste contexto, este trabalho foi desenvolvido objetivando a síntese de endoperóxidos isotopicamente marcados com 17O para serem utilizados como fonte geradora limpa de 17[1O2] em estudos mecanísticos. A capacidade de geração de 17[1O2] pelo endoperóxido N,N\'-di(2,3-dihidroxipropil)-3,3\'-(1,4 naftilideno) dipropanamida 17O (DHPN17O2) foi confirmada utilizando o captador químico sulfato mono-{2-[10-(2-sulfoxi-etil)-antracen-9-il]-etil}éster de sódio e o nucleosídeo 2\'- desoxiguanosina. Os produtos isotopicamente marcados com 17O formados foram analisados por espectrometria de ressonância magnética nuclear e cromatografia líquida de alta eficiência acoplado ao espectrômetro de massa. Os lipídeos, em especial o colesterol ao reagir com o 1O2 geram hidroperóxidos de colesterol como produtos de oxidação primária e na presença de metais resulta em compostos de maior reatividade e toxicidade, como os radicais peroxila, que contribuem para a propagação da peroxidação lipídica. Neste trabalho, demonstramos que os hidroperóxidos de colesterol são capazes de gerar 1O2 na presença de metal através de medidas de luminescência, utilização de supressores e captador químico de 1O2. Os mecanismos de reação envolvidos foram estudados e determinados por espectrometria de massa acoplada a cromatografia líquida de alta eficiência. Por fim, a caracterização detalhada dos produtos formados por espectrometria de ressonância magnética nuclear e massa na reação do colesterol com 1O2 mostrou que além dos hidroperóxidos a reação também produz um aldeído, o 3&#946; -hidroxi-5&#946;-hidroxi-B-norcolestano-6&#946;-carboxialdeído. Até o momento, este composto havia sido identificado como um produto específico da ozonização do colesterol. Neste estudo, baseado nos estudos por reações de quimiluminescência, é proposto o mecanismo de formação deste aldeído em reações de oxidação de colesterol por 1O2 envolvendo intermediário dioxetano. / Studies involving singlet molecular oxygen (1O2) has biological relevance, once this species, due to its eletrophylic character, reacts with rich electron molecules such as proteins, lipids and DNA causing damages that result in loss of function and cellular integrity. In biological system, the presence of other reactive species of oxygen and nitrogen impair the identification of lesions caused by 1O2. In this context, this work was developed with the aim of synthesizing <SUP17O-labeleded endoperoxides to be used as a clean source of (1O2) in mechanistic studies. The ability of 17[1O2]generation by N,N\'-di(2,4-dihydroxypropyl)-1,4-naphthalene-dipropanamide labeled with 17O(DHNP17O2) was observed using the disodium salt of anthracene-9,10-diyldiethyl disulfate as a chemical trap and the nucleoside 2\'-deoxyguanosine. The products isotopically labeled with 17O were analyzed by nuclear magnetic resonance spectroscopy and high performance liquid chromatography coupled to a mass spectrometer. Lipds, in special the cholesterol, when reacting with singlet molecular oxygen generate cholesterol hydroperoxides as primary products and in the presence of metals result in compounds of higher reactivity and toxicity, such as peroxyl radicals which contribute to the propagation of lipid peroxidation. In this work, we demonstrated that cholesterol hydroperoxides are able to generate singlet molecular oxygen in the presence of metal by chemiluminescence measurements by testing the effect of singlet molecular oxygen quencher and by chemical trap. The involved reaction mechanisms were studied and determined by mass spectrometry coupled to the high performance liquid chromatography. Finally, we detailed characterization of the products formed in the reaction of cholesterol with 1O2 by nuclear magnetic resonance and mass spectroscopy showed that besides cholesterol hydroperoxides, the reaction also produces an aldehyde, 3&#946;hydroxy-5&#946;-hydroxy-B-norcholestan-6&#946;-carboxyaldehyde which had been identified as a specific product of cholesterol ozonization. In this study, based on the studies of chemiluminescence reactions, the mechanism of formation of this aldehyde in reaction of oxidation of cholesterol by 1O2 involving a dioxetane intermediate has been proposed.

Shelf seas represent an important source of iron (Fe) to the open ocean. Additionally, shelf seas are highly productive environments which contribute to atmospheric carbon dioxide drawdown and support large fisheries. The work presented in this thesis describes the seasonal cycle of Fe in the Celtic and Hebridean Shelf Seas, and determines the physico-chemical speciation of Fe supplied from oxic margins. The results from repeated field surveys of the central Celtic Sea showed a nutrient type seasonal cycling of dissolved Fe (< 0.2 µm; dFe), which is surprising in a particle rich shelf system, suggesting a balance of scavenging and remineralisation processes. Coincident drawdown of dFe and nitrate (NO3-) was observed during the phytoplankton spring bloom. During the bloom, preferential drawdown of soluble Fe (< 0.02 µm; sFe) over colloidal Fe (0.02-0.2 µm; cFe) indicated greater bioavailability of the soluble fraction. Throughout summer stratification, it is known that NO3- is drawn down to < 0.02 µM in surface waters. This study revealed that both dFe and labile particulate Fe (LpFe) were also seasonally drawn down to < 0.2 nM. Consequently, it is hypothesised that the availability of Fe seasonally co-limits primary production in this region. At depth both dFe and NO3- concentrations increased from spring to autumn, indicating that remineralisation is an important process governing the seasonal cycling of dFe in the central Celtic Sea. In spring, summer and autumn, distinctive intermediate nepheloid layers (INL) were observed emanating from the Celtic Sea shelf slope. The INLs were associated with elevated concentrations of dFe (up to 3.25 ± 0.16 nM) and particulate Fe (up to 315 ± 1.8 nM) indicating that they are a persistent conduit for the supply of Fe to the open ocean. Typically > 15% of particulate Fe was labile and 60-90% of dFe was in the colloidal fraction. Despite being < 50 km from the 200 m isobath, the concentration of dFe was < 0.1 nM in surface waters at several stations. Broadly, the concentration of nutrients in surface waters described an oligotrophic environment where co-limitation between multiple nutrients, including Fe, appears likely. Over the Hebridean shelf break, residual surface NO3- concentrations (5.27 ± 0.79 µM) and very low concentrations of dFe (0.09 ± 0.04 nM) were observed during autumn, implying seasonal Fe limitation. The dFe:NO3- ratio observed is attributed to sub-optimal vertical supply of Fe relative to NO3- from sub-surface waters. In contrast to the shelf break, surface water in coastal regions contained elevated dFe concentrations (1.73 ± 1.16 nM) alongside low NO3-. Seasonal Fe limitation is known to occur in the Irminger and Iceland Basins; therefore, the Hebridean shelf break likely represents the eastern extent of sub-Arctic Atlantic seasonal Fe limitation, thus indicating that the associated weakening of the biological carbon pump exists over a wider region of the sub-Arctic Atlantic than previously recognised. These key findings demonstrate that the availability of Fe to phytoplankton may seasonally reach limiting levels in temperate shelf waters and that oxic margins persistently supply Fe dominated by colloidal and particulate fractions to the ocean.

Au cours de cette thèse, deux sujets distincts ont été étudiés : d’une part la synthèse d’analogues de la coelentérazine, substrat de différentes luciférases, pour une application en imagerie in vivo, et d’autre part la synthèse d’analogues du (-)-EGCG, inhibiteur de la kinase Dyrk1a, impliquée notamment dans les troubles cognitifs rencontrés dans la trisomie 21.La problématique du premier sujet consistait à obtenir des analogues de la coelentérazine conservant leur activité sur deux luciférases, la luciférase Renilla et l’aequorine, tout en induisant un déplacement vers le rouge de la bioluminescence produite par ces enzymes. L’aequorine, sensible au calcium, représentait la cible biologique principale du projet.Sept analogues dont six originaux ont été obtenues par des méthodes de synthèse classiques, et leurs activités ont pu être testées sur les deux luciférases choisies, avec des résultats probants : malgré des émissions de lumière moins intenses que celles obtenues avec la coelentérazine native, plusieurs molécules ont entrainé un déplacement vers le rouge de la longueur d’onde d’émission de lumière par bioluminescence allant jusqu’à 27 nm pour l’aequorine et plus de 120 nm pour la luciférase Renilla.Le second sujet, de chimie médicinale classique, a principalement consisté à la synthèse d’analogues du gallate d’épigallocatéchine (EGCG) au squelette simplifié, et au sein desquels le cycle pyranique caractéristique des catéchines a été remplacé par un carbocycle. Plusieurs molécules ont pu être synthétisées, dont deux présentant le motif hexaphénol. Leur activité inhibitrice de Dyrk1a a pu être testée in vitro et l’une d’entre elle s’est déjà révélée plus active que l’EGCG. / During this thesis, two distinct projects were studied: on the one hand, the synthesis of coelenterazine analogs, substrate of several luciferases, in the purpose of using them for in vivo imaging, and on the other, the synthesis of (-)-EGCG analogs, inhibitor of the Dyrk1a kinase, which interests us for the role it plays in the mental retardation existing in the Down Syndrome disease.The problematic of the first project consisted in obtaining coelenterazine analogs that would not only maintain their activity on two luciferases, the Renilla luciferase and aequorine, but they should also induce a red-shift of the bioluminescence produced by these enzymes. Because of its sensitivity to calcium, aequorine was the main biologic target of this project.Seven analogs, of which six had an original structure, were synthesized through usual synthetic methodologies and their activities on both aequorine and Renilla luciferase were tested in vitro, with interesting results: even if the intensities of light emission were weaker than those obtained with native coelenterazine, several molecules produced a red-shift of the emission wavelength of bioluminescence, up to 27nm for aequorine and more than 120nm for the Renilla luciferase. The second project, of classical medicinal chemistry, mainly consisted in the synthesis of epigallocatechin gallate analogs (EGCG) with a simplified backbone and in which the pyranic ring typical of catechins was replaced by a carbocycle. Several molecules were synthesized, two of them possessing the hexaphenol motif. Their inhibiting activity of Dyrk1a was tested in vitro and one already showed a better activity than natural EGCG.