Avec l’avènement des BioMEMS (Bio-MicroElectroMechanical Systems), ce sont toutes les pratiques médicales, biologiques, environnementales et agro-alimentaires qui entament une nouvelle ère. Les enjeux scientifique et industriel se rejoignent dans la miniaturisation des systèmes de détection, l’amélioration de leurs sensibilités et la simplification de leurs procédés de fabrication. Cette thèse hautement interdisciplinaire s’inscrit dans le cadre de ces enjeux. Elle expose la fabrication, la bio-fonctionnalisation et l’application d’un BioMEMS pour l’analyse de réactions enzymatiques en temps réel et à l’échelle micrométrique. Deux choix stratégiques ont été adoptés pour ce travail: le premier concerne l’utilisation de la technologie des plasmas froids ou polymérisation plasma pour la fonctionnalisation de surface à travers le dépôt de films polymères d’allylamine. Ce dépôt a permis ultérieurement l’immobilisation covalente de la trypsine (enzyme modèle protéolytique) au sein du BioMEMS. Cette technologie a été également utilisée pour développer une méthode simple de microfabrication des circuits microfluidiques compatible avec une production à grande échelle. Le second choix concerne l’utilisation de ce bioMEMS autour d’une transduction TeraHertz (THz) mise au point au sein de l’équipe. La spectroscopie THz vise à détecter les événements moléculaires à l’échelle de la picoseconde, sans marqueur et d’une manière non-invasive, en sondant directement les liaisons chimiques de faible énergie. Au cours de ce travail, nous avons donc développé un procédé de fonctionnalisation de surfaces par des amines, optimisé une méthode de greffage des enzymes, et étudié l’activité de la trypsine immobilisée. Nous avons ensuite intégré ces étapes dans le procédé de microfabrication du BioMEMS. Les mesures réalisées dans le domaine sub-THz (0,06-0,11 THz) sur une réaction de biocatalyse confirment la faisabilité d’une telle approche comme méthode analytique en biologie. Les résultats des différentes études montrent également que le mariage des plasmas froids avec les méthodes lithographiques représente une voie efficace, rapide et très compétitive pour le transfert de la technologie des BioMEMS à l’échelle industrielle. / The applications of miniaturized devices are no longer limited to electronic industry. Today, a new kind of microsystems called BioMEMS (Bio-MicroElectroMechanical Systems) are spreading in different fields, including biomedical, environmental and food industry applications. Recurring challenges are focusing on enabling processes for smaller, cost-effective, high-functionality devices, with more sensitivity and suitability for industrial scale development. This highly interdisciplinary thesis work attempts to provide new solutions to meet some of the needs mentioned above. It reports the fabrication, functionalization, and applications of a BioMEMS for enzyme reaction monitoring. First, we have developed a PECVD (plasma enhanced chemical vapor deposition) process for the surface functionalization by plasma polymerized allylamine. Films with high amine density and enhanced stability in aqueous environment were obtained. The amine functions were then used for enzymes immobilization. The covalently bonded trypsin molecules were extensively characterized and kinetic parameters determined using several microscopic and spectroscopic methods. Finally, both optimized processes were applied to the biofunctionalization of a TeraHertz (THz)-based BioMEMS. THz spectroscopy is the only non-invasive analytic method able to monitor molecular events at the picosecond timescale by probing low binding energies directly. It is used here for sensing a biocatalysis reaction inside the bioMEMS microchannels. Sub-THz measurements (0.06-0.11 THz) showed that combining microfluidic microsystems technology with THz detection could be a promising alternative for label-free real-time detection of biological interactions at the microscale. Additionally, we have developed a new microchannel fabrication process using direct plasma polymerization of TMDS (TetraMethylDiSiloxane) on micropatterned surfaces. This achievement demonstrates that cold plasma processes could be used not only for functionalization purposes or surface treatment but for the 3D microfabrication as well. This highly reduces processing time and manual handling steps, which is of a great importance for further industrial scale production.

Monodispersed nanoparticles of poly-N-isopropylacrylamide-co-allylamine (PNIPAM-co-allylamine) and PNIPAM-co-acrylic acid (AA) have been synthesized and used as building blocks for creating three-dimensional networks. The close-packed PNIPAM-co-allylamine and PNIPAM-co-AA nanoparticles were stabilized by covalently bonding neighboring particles at room temperature and at neutral pH; factors which make these networks amicable for drug loading and release. Controlled release studies have been performed on the networks using dextran markers of various molecular weights as model macromolecular drugs. Drug release was quantified under various physical conditions including a range of temperature and molecular weight. These nanoparticle networks have several advantages over the conventional bulk gels for controlling the release of biomolecules with large molecular weights. Monodispersed nanoparticles of poly-N-isopropylacrylamide-co-allylamine (PNIPAM-co-allylamine) can self-assemble into crystals with a lattice spacing on the order of the wavelength of visible light. By initiating the crystallization process near the colloidal crystal melting temperature, while subsequently bonding the PNIPAM-co-allylamine particles below the glass transition temperature, a nanostructured hydrogel has been created. The crystalline hydrogels exhibit iridescent patterns that are tunable by the change of temperature, pH value or even protein concentration. This kind of soft and wet hydrogel with periodic structures may lead to new sensors, devices, and displays operating in aqueous solutions, where most biological and biomedical systems reside. The volume-transition equilibrium and the interaction potential between neutral PINPAM particles dispersed in pure water were investigated by using static and dynamic light-scattering experiments. From the temperature-dependent size and energy parameters, the Sutherland-like potential provides a reasonable representation of the inter-particle potential for PNIPAM particles in swollen and in collapsed phases. An aqueous dispersion of PNIPAM particles can freeze at both high and low temperatures. At low temperatures, the freezing occurs at a large particle volume fraction, similar to that in a hard-sphere system; while at high temperature, the freezing occurs at low particle concentrations, driven by the strong van der Waals attraction due to the collapsed microgel particles. The calculated phase diagram has been confirmed semi-quantitatively by experiments.

NF-κB activity is critical in the regulation of atherosclerotic vascular smooth muscle cell (vSMC) phenotypes induced following oxidative injury by allylamine. The present studies were designed to detail dysregulation of NF-κB activity in these altered phenotypes, and to assess the importance of NF-κB in the regulation of osteopontin, a cytokine which modulates atherosclerosis. Increased degradation of IκBα was observed in allylamine-induced atherosclerotic vSMC phenotypes (henceforth referred to as allylamine cells). Enhanced phosphorylation of I-κ-kinases was observed by Western immunoblotting. NF-κB DNA binding activity as assessed by electrophoretic mobility shift assay demonstrated changes in the kinetics and magnitude of induction of binding. Enhancement of NF-κB binding activity was evident in allylamine cells compared to controls when seeded on plastic, fibronectin, and laminin, but not collagen I. Posttranscriptional alterations in Rel protein expression and nuclear localization partly account for changes in NF-κB DNA binding activity. Promoter-specific NF-κB binding profiles suggest altered dimer prevalence as a consequence of the changes in Rel protein expression. The expression of NF-κB regulated genes osteopontin and MMP-2 was enhanced in allylamine-treated aortas, while cyclin D1 and MMP-9 were unchanged. As the importance of osteopontin in atherosclerosis has been described in several models, subsequent studies were designed to assess osteopontin promoter activity. Activity of the osteopontin promoter was significantly reduced in allylamine cells compared to controls as assessed using a luciferase reporter. Deletion analysis suggested the presence of inhibitory cis-acting elements in the regulatory region of the gene. Mutation of these elements, including VDRE, AP-1, NF-κB, and USF1, indicated that NF-κB and USF1 mediate suppression of osteopontin promoter activity in allylamine cells. Decreased serine phosphorylation of immunoprecipitated RelA/p65 was observed in allylamine cells, indicating decreased ability of this protein to transactive gene promoters. NF-κB was found to play a role in suppression of osteopontin promoter activity by collagen I-mediated integrin signaling. These findings suggest that enhancements in NF-κB activity suppress osteopontin promoter activity in oxidant-activated vSMC cultures. Dysregulation of NF-κB activity occurs as a result of altered matrix and intracellular signaling upstream of the nucleus and possibly differential dimer assembly leading to cell-specific profiles of NF-κB-dependent gene regulation.

The present research involves two projects: a surface science study of the room-temperature adsorption and thermal evolution of allylamine and ethanolamine on Si(100)2×1, studied by using temperature-dependent X-ray photoelectron spectroscopy (XPS) and thermal desorption spectrometry (TDS), as well as Density Functional Theory (DFT) calculations; and a materials science study on the shape control of copper nanoparticles (Cu NPs) deposited on H-terminated Si(100) substrate with an extended size regime of 5-400 nm, by using a simple, one-step electrochemical method. The Cu NPs of three primary shapes were characterized with scanning-electron microscopy (SEM), glancing-incidence X-ray diffraction (GIXRD) and XPS. In the first surface science study, the presence of broad N 1s XPS features at 398.9-399.1 eV, corresponding to N–Si bonds, indicates N–H dissociative adsorption for both allylamine and ethanolamine on Si(100)2×1. For allylamine, the presence of C 1s features at 284.6 eV and 286.2 eV, corresponding to C=C and C−N, respectively, and the absence of the Si−C feature expected at 283.5 eV show that the reactions involving the ethenyl group such as the [2+2] C=C cycloaddition or those producing the [N, C, C] tridentate adstructures do not occur at room temperature. For ethanolamine, the O 1s feature at 533.1 eV indicates the formation of Si−O bond and O−H dissociation, which confirms an [O, N] bidentate adstructure and excludes the N−H and O−H dissociation unidentate structures. These XPS data are consistent with the N−H unidentate, and N−H and O−H double dissociation [O, N] bidentate adstructures for allylamine and ethanolamine, respectively, as predicted by the DFT calculations. TDS and temperature-dependent XPS data further show the desorption of propene and ethylene at 580 K and of acetylene at 700 K for allylamine and the desorption of ethylene at 615 K for ethanolamine, while the lack of N- or O-containing desorbates suggests that the dissociated N and O species are likely bonded to multiple surface Si atoms or diffused into the bulk at elevated temperatures (as confirmed by the corresponding temperature-dependent XPS spectra). Unlike the multidentate allyl alcohol and allylamine adstructures that have been found to be not favored kinetically, the present [O, N] bidentate ethanolamine adstructure appears to be kinetically favored on Si(100)2×1. In the second materials science study, Cu NPs of three primary shapes have been deposited on H-terminated Si(100) by a simple, one-step electrochemical method. By precisely manipulating the electrolyte concentration [CuSO4.5H2O] below their respective critical values, cubic, cuboctahedral, and octahedral Cu NPs of ranges of average sizes and number densities can be easily obtained by varying the deposition time. Combined GIXRD and depth-profiling XPS studies show that these Cu NPs have a crystalline core-shell structure, with a face-centered cubic metallic Cu core and a simple cubic Cu2O shell with a CuO outerlayer. The shape control of Cu NPs can be understood in terms of a progressive growth model under different kinetic conditions as dictated by different [CuSO4.5H2O] concentration regimes. The two studies in the present work lay the foundation for future investigation of surface biofunctionalization of these fascinating Cu NPs with different shapes and therefore different surface chemistries as controlled by the relative amounts of the (100) and (111) facets, and their boundaries.

The present research involves two projects: a surface science study of the room-temperature adsorption and thermal evolution of allylamine and ethanolamine on Si(100)2×1, studied by using temperature-dependent X-ray photoelectron spectroscopy (XPS) and thermal desorption spectrometry (TDS), as well as Density Functional Theory (DFT) calculations; and a materials science study on the shape control of copper nanoparticles (Cu NPs) deposited on H-terminated Si(100) substrate with an extended size regime of 5-400 nm, by using a simple, one-step electrochemical method. The Cu NPs of three primary shapes were characterized with scanning-electron microscopy (SEM), glancing-incidence X-ray diffraction (GIXRD) and XPS. In the first surface science study, the presence of broad N 1s XPS features at 398.9-399.1 eV, corresponding to N–Si bonds, indicates N–H dissociative adsorption for both allylamine and ethanolamine on Si(100)2×1. For allylamine, the presence of C 1s features at 284.6 eV and 286.2 eV, corresponding to C=C and C−N, respectively, and the absence of the Si−C feature expected at 283.5 eV show that the reactions involving the ethenyl group such as the [2+2] C=C cycloaddition or those producing the [N, C, C] tridentate adstructures do not occur at room temperature. For ethanolamine, the O 1s feature at 533.1 eV indicates the formation of Si−O bond and O−H dissociation, which confirms an [O, N] bidentate adstructure and excludes the N−H and O−H dissociation unidentate structures. These XPS data are consistent with the N−H unidentate, and N−H and O−H double dissociation [O, N] bidentate adstructures for allylamine and ethanolamine, respectively, as predicted by the DFT calculations. TDS and temperature-dependent XPS data further show the desorption of propene and ethylene at 580 K and of acetylene at 700 K for allylamine and the desorption of ethylene at 615 K for ethanolamine, while the lack of N- or O-containing desorbates suggests that the dissociated N and O species are likely bonded to multiple surface Si atoms or diffused into the bulk at elevated temperatures (as confirmed by the corresponding temperature-dependent XPS spectra). Unlike the multidentate allyl alcohol and allylamine adstructures that have been found to be not favored kinetically, the present [O, N] bidentate ethanolamine adstructure appears to be kinetically favored on Si(100)2×1. In the second materials science study, Cu NPs of three primary shapes have been deposited on H-terminated Si(100) by a simple, one-step electrochemical method. By precisely manipulating the electrolyte concentration [CuSO4.5H2O] below their respective critical values, cubic, cuboctahedral, and octahedral Cu NPs of ranges of average sizes and number densities can be easily obtained by varying the deposition time. Combined GIXRD and depth-profiling XPS studies show that these Cu NPs have a crystalline core-shell structure, with a face-centered cubic metallic Cu core and a simple cubic Cu2O shell with a CuO outerlayer. The shape control of Cu NPs can be understood in terms of a progressive growth model under different kinetic conditions as dictated by different [CuSO4.5H2O] concentration regimes. The two studies in the present work lay the foundation for future investigation of surface biofunctionalization of these fascinating Cu NPs with different shapes and therefore different surface chemistries as controlled by the relative amounts of the (100) and (111) facets, and their boundaries.

NF-κB activity is critical in the regulation of atherosclerotic vascular smooth muscle cell (vSMC) phenotypes induced following oxidative injury by allylamine. The present studies were designed to detail dysregulation of NF-κB activity in these altered phenotypes, and to assess the importance of NF-κB in the regulation of osteopontin, a cytokine which modulates atherosclerosis. Increased degradation of IκBα was observed in allylamine-induced atherosclerotic vSMC phenotypes (henceforth referred to as allylamine cells). Enhanced phosphorylation of I-κ-kinases was observed by Western immunoblotting. NF-κB DNA binding activity as assessed by electrophoretic mobility shift assay demonstrated changes in the kinetics and magnitude of induction of binding. Enhancement of NF-κB binding activity was evident in allylamine cells compared to controls when seeded on plastic, fibronectin, and laminin, but not collagen I. Posttranscriptional alterations in Rel protein expression and nuclear localization partly account for changes in NF-κB DNA binding activity. Promoter-specific NF-κB binding profiles suggest altered dimer prevalence as a consequence of the changes in Rel protein expression. The expression of NF-κB regulated genes osteopontin and MMP-2 was enhanced in allylamine-treated aortas, while cyclin D1 and MMP-9 were unchanged. As the importance of osteopontin in atherosclerosis has been described in several models, subsequent studies were designed to assess osteopontin promoter activity. Activity of the osteopontin promoter was significantly reduced in allylamine cells compared to controls as assessed using a luciferase reporter. Deletion analysis suggested the presence of inhibitory cis-acting elements in the regulatory region of the gene. Mutation of these elements, including VDRE, AP-1, NF-κB, and USF1, indicated that NF-κB and USF1 mediate suppression of osteopontin promoter activity in allylamine cells. Decreased serine phosphorylation of immunoprecipitated RelA/p65 was observed in allylamine cells, indicating decreased ability of this protein to transactive gene promoters. NF-κB was found to play a role in suppression of osteopontin promoter activity by collagen I-mediated integrin signaling. These findings suggest that enhancements in NF-κB activity suppress osteopontin promoter activity in oxidant-activated vSMC cultures. Dysregulation of NF-κB activity occurs as a result of altered matrix and intracellular signaling upstream of the nucleus and possibly differential dimer assembly leading to cell-specific profiles of NF-κB-dependent gene regulation.

Embryonic stem (ES) cells have the capacity to form any cell type. However, their propagation and differentiation is limited by current two dimensional (2D) culture techniques which offer little flexibility in terms of surface structure and functionalisation with bioactive molecules. The aim of the current work was to produce a novel scaffold that could manipulate ES cell behaviour using both architectural and biological cues. Electrospinning is a flexible technique that creates nonwoven meshes that mimic the fibrous architecture of the ECM. Initial work focused on investigating the suitability of electrospun poly(lactic-co-glycolic acid) (PLGA) meshes for 2D and three dimensional (3D) culture of mouse ES cells, with the hypothesis that the fibrous architecture would assist in maintaining pluripotency. The study also sought to functionalise the scaffolds with biologically active molecules. Heparan sulphate proteoglycans (HSPGs) reside at the cell surface and within the ECM where they mediate growth factor binding, assist cell attachment and stabilise the ECM. Furthermore, ES cells modulate their own microenvironment by controlling the composition of heparan sulphate (HS), regulating the binding of growth factors such as fibroblast growth factor (FGF) family members. Therefore, we aimed to immobilise HS and heparin (a highly sulphated structural analogue of HS) on the fibre surface in a form that was freely accessible for protein/cell interactions and that retained its biological activity. Electrospinning parameters were optimised to produce microfibre electrospun meshes with an average fibre diameter of 570nm. Cell morphology, proliferation and pluripotency were monitored using an Oct4-GFP reporter cell line and results compared with flat spin coated films. To investigate the potential for 3D culture, spinning parameters were altered to increase fibre diameter to >3micro metre with infiltration assessed using pro-migratory E-cadherin-/- ES cells. Scaffolds were coated with plasma polymerised allylamine (ppAm) to enable non-covalent immobilisation of HS/heparin. Ligand binding assays with the link module of TSG-6 and anti-heparin/HS antibodies were used to probe HS/heparin presentation on the fibre surface. The biological activity of the immobilised HS/heparin was analysed by testing the ability of coated scaffolds to rescue the neural differentiation capacity HS deficient EXT1-/- ES cells. Finally, human ES cells were cultured on the surface of ppAm scaffolds +/- HS in both unconditioned and mouse embryonic fibroblast (MEF) conditioned media for 5 days. Both microfibre meshes and flat spin coated films supported the attachment, growth and pluripotency of mouse ES cells. Cells adopted distinct morphologies, with mouse ES cells aggregating in rounded colonies on microfibre scaffolds and demonstrating increased spreading on spin coated films. Fibres >3micro metre created a thicker mesh with potential for 3D culture supporting the infiltration of E-cadherin-/- ES cells. ppAm enabled non-covalent immobilisation of HS/heparin in a form that was free to participate in protein interactions and which presented essential sulphation motifs within the HS/heparin chains. Bound HS was biologically active and functioned in synchrony with FGF4 to enhance neural differentiation of EXT1-/- ES cells. The constructs also supported the attachment and growth of human ES cells, with HS functionalised scaffolds demonstrating a slight increase in compatibility during culture in unconditioned media. The successful functionalisation of electrospun meshes with HS/heparin creates a highly versatile scaffold for ES cell culture and differentiation. The architecture of the meshes can be manipulated to either serve as a fibrous substrate for maintenance of pluripotency or support the formation of complex cell interactions present in vivo. The immobilisation of HS provides an extra dimension of versatility, as the scaffold can be tailored with specific HS species, potentially enabling the differential regulation of growth factor binding.

Etude et développement de dépôts d'allylamine assistés par plasma basse pression specifiques aux stents coronariens recouverts Les stents coronariens sont des dispositifs médicaux, généralement fabriqués en acier inoxydable 316L, utilisés pour traiter des maladies cardiovasculaires comme l'athérosclérose. Les stents recouverts ou à relargage contrôlé de médicaments sont des solutions prometteuses pour réduire les phénomènes de resténose. Ce travail a pour objectif le développement d'un procédé plasma basse pression capable de déposer une couche de polymère permettant de protéger la surface des stents contre l'agressivité du milieu physiologique. L'allylamine est choisie comme précurseur moléculaire pour assurer un taux élevé de fonctions amines primaires. Ces fonctions pourront être utilisées, successivement, pour l'immobilisation de molécules bioactives afin d'augmenter la biocompatibilité des stents. Les dépôts sont effectués sur des substrats d'acier inoxydable 316L en utilisant un réacteur plasma basse pression (70 kHz). Les différentes techniques d'analyse de surface utilisées (angle de contact, XPS, FTIR-ATR) montrent que les variations de puissance de la décharge et du temps de traitement ne modifient pas significativement la composition chimique de surface des dépôts. Cependant, grâce à une technique de dérivation chimique nous avons mis en évidence une meilleure sélectivité vis-à-vis des fonctions amines primaires pour les couches déposées à faibles valeurs de puissance. En effet, des analyses in-situ de la phase plasmagène (spectrométrie de masse, spectroscopie d'émission optique) révèlent qu'une augmentation de la puissance de la décharge conduit à l'augmentation de son caractère énergétique et, ainsi, à l'augmentation du taux de fragmentation du précurseur. La stabilité des revêtements au lavage dans l'eau de-ionisée a été aussi évaluée. Les dépôts obtenus pour une puissance de la décharge de 2W présentent le meilleur compromis entre rétention des fonctions amines primaires et stabilité. Enfin, nous avons évalué les propriétés d'adhérence des couches après déformation plastique en utilisant le " small punch test ", permettant de reproduire les conditions qu'on retrouve lorsque les stents sont déployés dans les artères. Les dépôts présentent des adéquates propriétés de cohésion et d'adhérence au substrat pour répondre à la déformation sans se fissurer et/ou délaminer. Ces résultats montrent que les couches d'allylamine déposées par procédé plasma basse pression présentent des caractéristiques prometteuses afin d'être utilisées comme revêtement performant pour les stents coronariens. Plasma Polymerized allylamine films deposited on 316L stainless steel

Polymers rich in primary amine groups are proposed to be effective adsorbents for the reversible adsorption of CO2 from moderately dilute gas streams (10% CO2) and ultra-dilute gas streams (e.g. ambient air, 400 ppm CO2), with their performance under ultra-dilute conditions being competitive with or exceeding the state-of-the-art adsorbents based on supported poly(ethyleneimine) (PEI). The CO2 adsorption capacity (mmol CO2/g sorbent) and amine efficiency (mmol CO2/mmol amine) of linear poly(allylamine) (PAA), cross-linked poly(allylamine) prepared by post-polymerization crosslinking with epichlorohydrin (PAAEPI), and branched poly(allylamine) prepared by branching of poly(allylamine) with divinylbenzene (PAADVB) are presented here and compared with state-of-the-art adsorbents based on supported PEI, specifically branched and linear, low molecular weight PEI. Silica mesocellular foam, MCF, serves as the support material for impregnation of the amine polymers. In general, branched polymers are found to yield more effective adsorbents materials. Overall, the results of this work show that linear PAA, cross-linked PAAEPI, and branched PAADVB are promising candidates for solid adsorbents with high capacity for CO2.

Flavin adenine dinucleotide (FAD) is a coenzyme that participates in the redox process of flavoenzymes. Attempts to characterize the catalytic pathways of these enzymes have relied in part on the use of FAD chemical models. The efforts described in this dissertation focus on the chemical model approach to investigate the mechanism of the monoamine oxidase (MAO) catalyzed oxidation of the cyclic tertiary allylamine 1-methyl-4-(2-methyl-1H-pyrrol-2-yl)-1,2,3,6-tetrahydropyridine (TMMP), which is a close analog of the parkinsonian-inducing designer drug 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). MAO-B catalyzes the conversion of MPTP and its derivatives into active neurotoxins in the brain that subsequently mediate neurogenerative processes that mimic the events leading to idiopathic Parkinson\'s disease. Monoamine oxidase inhibitors are currently used to treat early stages of Parkinson\'s disease. Two FAD chemical models are examined in this project: 5-ethyl-3-methyllumiflavinium perchlorate (5Et3MLF+ClO4-) and 3-methyllumiflavin (3MLF). The flavinium salt 5Et3MLF+ClO4- is an activated form of 3MLF. These FAD chemical models have been used to examine the MAO catalyzed oxidation. MAO-B is expressed in the brain and is known to be involved in the conversion of TMMP into the neurotoxic metabolite 1-methyl-4-phenyl pyridnium (MMP+). MAO-B is responsible for the alpha-carbon oxidation of TMMP to yield 1-methyl-4-(2-methylpyrrol-2-yl)-2,3-dihydropyridinium (DHP+), which then undergoes a second 2-electron oxidation to MMP+. Previous findings demonstrated that 3MLF and 5Et3MLF+ClO4- promoted the oxidation reaction of primary and secondary amines but not tertiary amines. However, the cyclic tertiary allylamine TMMP has not been examined experimentally. Therefore, the alpha-carbon oxidation of TMMP in the presence of the FAD chemical models is reported in this dissertation. The effect of dioxygen and water on the activity of these FAD models is also investigated. / Ph. D.