Role of Reactive Gliosis and Neuroinflammation in Experimental Glaucoma

Le glaucome est la principale cause de cécité irréversible dans le monde. Chez les patients atteints de cette pathologie, la perte de la vue résulte de la mort sélective des cellules ganglionnaires (CGR) de la rétine ainsi que de la dégénérescence axonale. La pression intraoculaire élevée est considérée le facteur de risque majeur pour le développement de cette maladie. Les thérapies actuelles emploient des traitements pharmacologiques et/ou chirurgicaux pour diminuer la pression oculaire. Néanmoins, la perte du champ visuel continue à progresser, impliquant des mécanismes indépendants de la pression intraoculaire dans la progression de la maladie. Il a été récemment démontré que des facteurs neuroinflammatoires pourraient être impliqués dans le développement du glaucome. Cette réponse est caractérisée par une régulation positive des cytokines pro-inflammatoires, en particulier du facteur de nécrose tumorale alpha (TNFα). Cependant, le mécanisme par lequel le processus neuroinflammatoire agit sur la mort neuronale reste à clarifier. L’hypothèse principale de ce doctorat propose que les facteurs pro-inflammatoires comme le TNFα et la phosphodiestérase 4 (PDE4) interagissent avec les mécanismes moléculaires de la mort neuronale, favorisant ainsi la survie et la protection des CGRs au cours du glaucome. Dans la première partie de ma thèse, J’ai utilisé un modèle in vivo de glaucome chez des rats Brown Norway pour montrer que l’expression du TNFα est augmentée après l'induction de l'hypertension oculaire. L'hypothèse spécifique de cette étude suggère que les niveaux élevés de TNFα provoquent la mort des CGRs en favorisant l'insertion de récepteurs AMPA perméables au calcium (CP-AMPAR) à la membrane cytoplasmique. Pour tester cette hypothèse, j’ai utilisé un inhibiteur sélectif de la forme soluble du TNFα, le XPro1595. L'administration de cet agent pharmacologique a induit une protection significative des somas et des axones des neurones rétiniens. L'évaluation de la perméabilité au cobalt a montré que le TNFα soluble est impliqué dans l'insertion de CP-AMPAR à la membrane des CGRs lors du glaucome. L’exposition des neurones à une pression oculaire élevée est à l’origine de la hausse de la densité membranaire des CP-AMPARs, grâce à une diminution de l’expression de la sous-unité GluA2. La présence de GluA2 au sein du récepteur ne permet pas l’entrée du calcium à l’intérieur de la cellule. L'administration intraoculaire d’antagonistes spécifiques des CP-AMPARs promeut la protection des somas et des axones des CGRs. Ces résultats montrent que les CP-AMPARs jouent un rôle important dans la pathologie du glaucome. Dans la deuxième partie de ma thèse, j’ai caractérisé l'effet neuroprotecteur d’un inhibiteur de la PDE4, l’ibudilast, dans notre modèle de glaucome. L'hypothèse spécifique s’oriente vers une atténuation de la réponse neuroinflammatoire et de la gliose par l’administration d’ibudilast, favorisant ainsi la protection neuronale. Les résultats montrent que dans les rétines glaucomateuses, l’ibudilast diminue la gliose et l'expression de plusieurs facteurs tels que le TNFα, l'interleukine-1β (IL-1β), l’interleukine-6 (IL-6) et le facteur inhibiteur de la migration des macrophages (MIF). Chez les rats glaucomateux, nous avons observé une expression notable de PDE4A dans les cellules de Müller, qui est en corrélation avec l'accumulation de l’AMP cyclique (AMPc) dans ces cellules après un traitement d’ibudilast. Finalement, nous avons démontré que la protection des CGRs via l’administration d’ibudilast est un mécanisme dépendent de l’AMPc et de la protéine kinase A (PKA). En conclusion, les résultats présentés dans cette thèse identifient deux mécanismes différents impliqués dans la perte des CGRs au cours du glaucome. Ces mécanismes pourraient fournir des perspectives potentielles pour le développement de nouvelles stratégies de traitement du glaucome.

Optimization of pH and Direct Imaging Conditions of Complexed Methylene Blue Sensitized Poly(vinyl chloride) Films

Significant results of our experimental investigations on the dependence of pH on real time transmission characteristics on recording media fabricated by doping PVC with complexed methylene blue are presented. The optimum pH value for faster bleaching was found to be 4×5. In typical applications, the illumination from one side, normal to the surface of this material, initiates a chemical sequence that records the incident light pattern in the polymer. Thus direct imaging can be successfully done on this sample. The recorded letters were very legible with good contrast and no scattering centres. Diffraction efficiency measurements were also carried out on this material.

Reusable Recording Medium Based on MBPVA and Vinyl Acetate

A new photopolymerizable recording media is introduced based on poly (vinyl alcohol) and vinyl acetate sensitized with methylene blue. It is observed that this MBPVA/VAc system can be reused a number of times without significant decrease in diffraction efficiency. The PVA-VAc ratio was optimized at 2:1. Diffraction efficiency of 6.3% was obtained without any fixing at a dye concentration of 9.3 10 4 mol/l at an exposure of 750 mJ/cm2. The material is attractive on account of its reusability.

Photosensitivity of Laser Dye Mixtures in Polymer Matrix – A Photoacoustic Study

Laser induced photoacoustic (PA) technique is used in the study of photostability of polymethyl methacrylate (PMMA) films doped with Rhodamine 6G -Rhodamine B dye system. Energy transfer from a donor molecule to an acceptor molecule in a dye mixture affects the output of the dye system. Details of investigations on the role of laser power, modulation frequency and the irradiation wavelength on the photosensitivity of the dye mixture doped PMMA films are presented.

Effect of Chromium Doping on the Diffraction Efficiency of Methylene Blue Sensitized PVA/Acrylamide Films

The effect of chromium doping on methylene blue sensitized Poly (vinyl alcohol)/Acrylamide was carried out by varying the ratios of Ammonium dichromate and methylene blue. In the case of films without ammonium dichromate, the diffraction efficiency was found to decrease on storage. On chromium doping the storage life was improved. Interestingly, a self-enhancement in efficiency was observed for a particular ratio of methylene blue and ammonium dichromate.

Optimization of pH and Direct Imaging Conditions of Complexed Methylene Blue Sensitized Poly(vinyl chloride) Films

Significant results of our experimental investigations on the dependence of pH on real time transmission characteristics on recording media fabricated by doping PVC with complexed methylene blue are presented. The optimum pH value for faster bleaching was found to be 4×5. In typical applications, the illumination from one side, normal to the surface of this material, initiates a chemical sequence that records the incident light pattern in the polymer. Thus direct imaging can be successfully done on this sample. The recorded letters were very legible with good contrast and no scattering centres. Diffraction efficiency measurements were also carried out on this material.

Reusable Recording Medium Based on MBPVA and vinyl Acetate

A new photopolymerizable recording media is introduced based on poly (vinyl alcohol) and vinyl acetate sensitized with methylene blue. It is observed that this MBPVA/VAc system can be reused a number of times without significant decrease in diffraction . The PVA-VAc ratio was optimized at 2:1. Diffraction efficiency of 6.3% was obtained without any fixing at a dye concentration of 9.3 10 4 mol/l at an exposure of 750 mJ/cm2. The material is attractive on account of its reusability.

Reusable recording medium based on MBPVA and vinyl acetate

A new photopolymerizable recording media is introduced based on poly (vinyl alcohol) and vinyl acetate sensitized with methylene blue. It is observed that this MBPVA/VAc system can be reused a number of times without significant decrease in diffraction efficiency. The PVA-VAc ratio was optimized at 2:1. Diffraction efcienc4y of 6.3% was obtained without any fixing at a dye concentration of 9.3 x 10- mol/l at an exposure of 750mJ/cm2. The material is attractive on account of its reusability.

Complex methylene-blue-sensitized polyvinyl chloride: a polymer matrix for hologram recording

A new polymer matrix sensitized with methylene blue for use as an optical recording material is described here. The characterization is done to determine the optimal recording conditions. These films need no chemical development and are found to be stable for several months. The matrix has excellent shelf life and needs an exposure only as short as 20 s. Direct imaging was done on this material.

Optimization of pH and direct imaging conditions of complexed methylene blue sensitized poly(vinyl chloride) films

Significant results of our experimental investigations on the dependence of pH on real time transmission characteristics on recording media fabricated by doping PVC with complexed methylene blue are presented. The optimum pH value for faster bleaching was found to be 4 . 5. In typical applications, the illumination from one side, normal to the surface of this material, initiates a chemical sequence that records the incident light pattern in the polymer. Thus direct imaging can be successfully done on this sample. The recorded letters were very legible with good contrast and no scattering centres. Diffraction efficiency measurements were also carried out on this material.

Reusable Recording Medium Based on MBPVA and Vinyl Acetate

A new photopolymerizable recording media is introduced based on poly (vinyl alcohol) and vinyl acetate sensitized with methylene blue. It is observed that this MBPVA/VAc system can be reused a number of times without significant decrease in diffraction efficiency. The PVA-VAc ratio was optimized at 2:1. Diffraction efcienc4y of 6.3% was obtained without any fixing at a dye concentration of 9.3 x 10- mol/l at an exposure of 750mJ/cm2. The material is attractive on account of its reusability.

Reusable Recording Medium based on MBPVA and Vinyl Acetate

A new photopolymerizable recording media is introduced based on poly (vinyl alcohol) and vinyl acetate sensitized with methylene blue. It is observed that this MBPVA/VAc system can be reused a number of times without significant decrease in diffraction efficiency. The PVA-VAc ratio was optimized at 2:1. Diffraction efcienc4y of 6.3% was obtained without any fixing at a dye concentration of 9.3 x 10- mol/l at an exposure of 750mJ/cm2. The material is attractive on account of its reusability.

Laser induced fluorescence based optical fiber probe for analyzing bacteria

Optical fiber based laser induced fluorescence (LIF) measurements were carried out using Rhodamine B to analyze two different species of bacteria , a Gram-positive bacteria namely Bacillus .cmithii , and fibrin alginolvticus, a Gram-' negative bacteria . The fiber sensor was clearly able to distinguish between the two species of bacteria . Quenching effect of the dye Rhodamine B by Bacillus smitltii was observed . The effect of dye on the samples was also studied in detail.