Le système endocannabinoïde dans la rétine du singe : expression, localisation et fonctions

Le cannabis produit de nombreux effets psychologiques et physiologiques sur le corps humain. Les molécules contenues dans cette plante, désignées comme « phytocannabinoïdes », activent un système endogène qu’on appelle le système endocannabinoïde (eCB). Les effets de la consommation de cannabis sur la vision ont déjà été décrits sans cependant de formulation sur les mécanismes sous-jacents. Ces résultats comportementaux suggèrent, malgré tout, la présence de ce système eCB dans le système visuel, et particulièrement dans la rétine. Cette thèse vise donc à caractériser l’expression, la localisation et le rôle du système eCB dans la rétine du singe vervet, une espèce animale ayant un système visuel semblable à celui de l’humain. Nous avons mis au point un protocole expérimental d’immunohistochimie décrit dans l’article apparaissant dans l’Annexe I que nous avons utilisé pour répondre à notre objectif principal. Dans une première série de quatre articles, nous avons ainsi caractérisé l’expression et la localisation de deux récepteurs eCBs reconnus, les récepteurs cannabinoïdes de type 1 (CB1R) et de type 2 (CB2R), et d’un 3e présumé récepteur aux cannabinoïdes, le récepteur GPR55. Dans l’article 1, nous avons démontré que CB1R et une enzyme clé de ce système, la fatty acid amide hydrolase (FAAH), sont exprimés dans les parties centrale et périphérique de la rétine, et abondamment présents dans la fovéa, une région où l’acuité visuelle est maximale. Dans l’article 2, nous avons localisé le CB2R dans des cellules gliales de la rétine : les cellules de Müller et nous avons proposé un modèle sur l’action de cette protéine dans la fonction rétinienne faisant appel à une cascade chimique impliquant les canaux potassiques. Dans l’article 3, nous avons observé le GPR55 exclusivement dans les bâtonnets qui sont responsables de la vision scotopique et nous avons soumis un deuxième modèle de fonctionnement de ce récepteur par le biais d'une modulation des canaux calciques et sodiques des bâtonnets. Vu que ces 3 récepteurs se retrouvent dans des cellules distinctes, nous avons suggéré leur rôle primordial dans l’analyse de l’information visuelle au niveau rétinien. Dans l’article 4, nous avons effectué une analyse comparative de l’expression du système eCB dans la rétine de souris, de toupayes (petits mammifères insectivores qui sont sont considérés comme l’étape intermédiaire entre les rongeurs et les primates) et de deux espèces de singe (le vervet et le rhésus). Ces résultats nous ont menés à présenter une hypothèse évolutionniste quant à l’apparition et à la fonction précise de ces récepteurs. Dans les articles subséquents, nous avons confirmé notre hypothèse sur le rôle spécifique de ces trois récepteurs par l’utilisation de l’électrorétinographie (ERG) après injection intravitréenne d’agonistes et d’antagonistes de ces récepteurs. Nous avons conclu sur leur influence indéniable dans le processus visuel rétinien chez le primate. Dans l’article 5, nous avons établi le protocole d’enregistrement ERG normalisé sur le singe vervet, et nous avons produit un atlas d’ondes ERG spécifique à cette espèce, selon les règles de l’International Society for Clinical Electrophysiology of Vision (ISCEV). Les patrons électrorétinographiques se sont avérés semblables à ceux de l’humain et ont confirmé la similarité entre ces deux espèces. Dans l’article 6, nous avons démontré que le blocage de CB1R ou CB2R entraine une modification de l’électrorétinogramme, tant au niveau photopique que scotopique, ce qui supporte l’implication de ces récepteurs dans la modulation des ondes de l’ERG. Finalement, dans l’article 7, nous avons confirmé le modèle neurochimique proposé dans l’article 3 pour expliquer le rôle fonctionnel de GPR55, en montrant que l’activation ou le blocage de ce récepteur, respectivement par un agoniste (lysophosphatidylglucoside, LPG) ou un antagoniste (CID16020046), entraine soit une augmentation ou une baisse significative de l’ERG scotopique seulement. Ces données, prises ensemble, démontrent que les récepteurs CB1R, CB2R et GPR55 sont exprimés dans des types cellulaires bien distincts de la rétine du singe et ont chacun un rôle spécifique. L’importance de notre travail se manifeste aussi par des applications cliniques en permettant le développement de cibles pharmacologiques potentielles dans le traitement des maladies de la rétine.

Copper Doped Methylene Blue Sensitized Poly(vinyl alcohol)–Acrylamide Films for Stable Diffraction Efficiency

Copper doped methylene blue sensitized poly(vinyl alcohol) (MBPVA)–acrylamide films were fabricated to improve the storage life of recorded gratings. The films were fabricated using gravity settling method and the copper chloride concentration was optimized as 3:18 10 3 mol/l for a dye concentration of 6:2 10 4 mol/l. The gratings recorded on the optimized film constitution could be stored for months with stable diffraction efficiency (24%) without any chemical or thermal fixing techniques. The resolution of the material is found to be unaffected with the addition of copper chloride.

Copper Doped Methylene Blue Sensitized Poly(vinyl alcohol)–Acrylamide Films for Stable Diffraction Efficiency

Copper doped methylene blue sensitized poly(vinyl alcohol) (MBPVA)–acrylamide films were fabricated to improve the storage life of recorded gratings. The films were fabricated using gravity settling method and the copper chloride concentration was optimized as 3:18 10 3 mol/l for a dye concentration of 6:2 10 4 mol/l. The gratings recorded on the optimized film constitution could be stored for months with stable diffraction efficiency (24%) without any chemical or thermal fixing techniques. The resolution of the material is found to be unaffected with the addition of copper chloride.

New Simulated Corrugated Scaltering Surface Giving Widebandcharacter I Sti Cs

Simultaneous elimination of specular reflection and backscattered power from a plane metallic surface by simulated corrugated surfaces of constant period and variable strip width for TM polarisation is reported. This new configuration offers almost a ten-fold frequency bandwidth compared with a regularly spaced strip grating of the same size.

Polarisation agile active microstrip patch arrays

Two three-clement polarisation-agile active microstrip patch arrays have been developed . The radiating elements are square patches each with two transistors mounted on adjacent edges. The patches radiate orthogonal modes , the relative phase of which can be varied. Radiation patterns show good agreement with predictions from theory, in both linear and circular polarization, and no grating lobes were observed

Analysis and Testing of a Novel Oadm Based on FBG and Machzehnder Interferometer

A novel optical add-drop multiplexer (OADM) based on the Mach-Zelauler interferometer (MZI) and the fiber Bragg grating (FBG) is proposed for the first tittle to the authors ' knowledge. In the structure, the Mach-Zehnder interferometer acts as an optical switch. The principle of the OADM is analyzed in this paper. The OADM can add/drop one of the multi-input channels or pass the channel directly by adjusting the difference of the two arms of the interferometer. The channel isolation is more than 20 dB

Design. Fabrication and Characterization of Fiber Optic Sensors for Physical and Chemical Applications

Advent of lasers together with the advancement in fiber optics technology has revolutionized the sensor technology. Advancement in the telemetric applications of optical fiber based measurements is an added bonus. The present thesis describes variety of fiber based sensors using techniques like micro bending, long period grating and evanescent waves. Sensors to measure various physical and chemical parameters are described in this thesis.

Fabrication and characterisation of polymer optical fibers for smart sensing and optical amplification

The thesis presented the fabrication and characterisation of polymer optical fibers in their applications as optical amplifier and smart sensors.Optical polymers such as PMMA are found to be a very good host material due to their ability to incorporate very high concentration of optical gain media like fluorescent dyes and rare earth compounds. High power and high gain optical amplification in organic dye-doped polymer optical fibers is possible due to extremely large emission cross sections of oyes. Dye doped (Rhodamine 6G) optical fibers were fabricated by using indigenously developed polymer optical fiber drawing tower. Loss characterization of drawn dye doped fibers was carried out using side illumination technique. The advantage of the above technique is that it is a nondestructive method and can also be used for studying the uniformity in fiber diameter and doping. Sensitivity of the undoped polymer fibers to temperature and microbending were also studied in its application in smart sensors.Optical amplification studies using the dye doped polymer optical fibers were carried out and found that an amplification of l8dB could be achieved using a very short fiber of length lOcm. Studies were carried out in fibers with different dye concentrations and diameter and it was observed that gain stability was achieved at relatively high dye concentrations irrespective of the fiber diameter.Due to their large diameter, large numerical aperture, flexibility and geometrical versatility of polymer optical fibers it has a wide range of applications in the field of optical sensing. Just as in the case of conventional silica based fiber optic sensors, sensing techniques like evanescent wave, grating and other intensity modulation schemes can also be efficiently utilized in the case of POF based sensors. Since polymer optical fibers have very low Young's modulus when compared to glass fibers, it can be utilized for sensing mechanical stress and strain efficiently in comparison with its counterpart. Fiber optic sensors have proved themselves as efficient and reliable devices to sense various parameters like aging, crack formation, weathering in civil structures. A similar type of study was carried out to find the setting characteristics of cement paste used for constructing civil structures. It was found that the measurements made by using fiber optic sensors are far more superior than that carried out by conventional methods. More over,POF based sensors were found to have more sensitivity as well.

Long period gratings in multimode optical fibers: application in chemical sensing

We propose and demonstrate a new technique for evanescent wave chemical sensing by writing long period gratings in a bare multimode plastic clad silica fiber. The sensing length of the present sensor is only 10 mm, but is as sensitive as a conventional unclad evanescent wave sensor having about 100 mm sensing length. The minimum measurable concentration of the sensor reported here is 10 nmol/l and the operating range is more than 4 orders of magnitude. Moreover, the detection is carried out in two independent detection configurations viz., bright field detection scheme that detects the core-mode power and dark field detection scheme that detects the cladding mode power. The use of such a double detection scheme definitely enhances the reliability and accuracy of the results. Furthermore, the cladding of the present fiber need not be removed as done in conventional evanescent wave fiber sensors.

Rotational analysis of the C1Π-X1 Σ+ system of indium monofluoride

Indium monofluoride was excited in a high-frequency discharge and the C-X system was photographed at a reciprocal dispersion of 0.3 AA mm-1 using a plane-grating spectrograph. Rotational analyses of the 0,0 1,0 2,2 3,3 4,4 2,4 3,5 4,6 and 5,7 bands have been carried out and the following molecular constants have been evaluated. Be'=0.2670(+or-3) cm-1, Be"=0.2628(+or-4) cm-1, alpha e'=0.0050(+or-4) cm-1, alpha e"=0.0020(+or-1) cm-1, De'=3.65(+or-5)*10-7 cm-1, De"=2.5(+or-3)*10-7 cm-1, beta e'=0.5(+or-2)*10-7 cm-1, beta e"=0.2(+or-1)*10-7 cm-1, re'=1.9672(+or-3) AA, re"=1.9853(+or-2) AA. The re" value agrees with the microwave absorption value 1.9854 AA.

Fabrication And Parametric Measurements Of N2 Laser Pumped Dye Laser And Its Application To Energy Transfer Studies In Mixed Dyes

Eventhough a large number of schemes have been proposed and develoned for N9 laser ouined dye lasers the relatively low efficiency compelled the scientists to device new methods to improve the system efficiencs. Energy transfer mechanism has been shown to he a convenien tool for the enhancement of efficiency of dye lasers. Th p resent work covers a detailed study of the performance characteristics of a N2 laser pumped dye laser in the con— ventional mode and also, when pumped by the energy transfer mechanism. For .th.e present investigations a dye laser pumped by a'N2 laser (A4200 kw peak power) was fabricated. The grating at grazing incidence was used as the beam expanding device; A t its best performance the system was giving an output peak power of l5 kW for a 5 X lC"3H/l Rh—€ solution in methanol. T he conversion efficiency was 7.5; The output beam was having 3 divergence of 2 mrad and bandwidth o.9 A. Suitable modifications were suggested for obtaining better conversion efficiency and bandwidth.

Investigation on the panchromaticity of silver-doped poly(vinyl alcohol)/acrylamide photopolymer

An investigation on the panchromaticity of a silver-doped poly(vinyl alcohol)/acrylamide photopolymer system is presented in this paper. Frequency-doubled Nd:YAG (532 nm) and Arþ (488 nm) lasers were used for the characterization of the films. Previous studies using an He–Ne laser (632:8 nm) showed that plane-wave transmission grating with a high diffraction efficiency of 75% could be stored in the optimized film. From the present study, it was noted that transmission gratings with 70% diffraction efficiency could be recorded using Arþ and Nd:YAG lasers, thereby elucidating the possibility of using the developed photopolymer system as a competent panchromatic recording medium

Effect of Concentration of DYE on the Storage Life of Plane Wave Gratings on Photopolymer Film

Holographic grating with good storage life in poly(vinyl alcohol) based photopolymer film, prepared by gravity settling method, with reduced concentration of the dye was found to give good diffraction efficiency without crosslinking. The material was found to show good diffraction efficiency and sensitivity (75% diffraction efficiency at exposure energy of 80 mJ/cm2). The shelf life of the photopolymer solution could be improved by storage at a temperature 4 C in refrigerator

Development of directly modulated high speed telecommunication lasers based on surface defined feedback gratings

High-speed semiconductor lasers are an integral part in the implemen- tation of high-bit-rate optical communications systems. They are com- pact, rugged, reliable, long-lived, and relatively inexpensive sources of coherent light. Due to the very low attenuation window that exists in the silica based optical fiber at 1.55 μm and the zero dispersion point at 1.3 μm, they have become the mainstay of optical fiber com- munication systems. For the fabrication of lasers with gratings such as, distributed bragg reflector or distributed feedback lasers, etching is the most critical step. Etching defines the lateral dimmensions of the structure which determines the performance of optoelectronic devices. In this thesis studies and experiments were carried out about the exist- ing etching processes for InP and a novel dry etching process was de- veloped. The newly developed process was based on Cl2/CH4/H2/Ar chemistry and resulted in very smooth surfaces and vertical side walls. With this process the grating definition was significantly improved as compared to other technological developments in the respective field. A surface defined grating definition approach is used in this thesis work which does not require any re-growth steps and makes the whole fabrication process simpler and cost effective. Moreover, this grating fabrication process is fully compatible with nano-imprint lithography and can be used for high throughput low-cost manufacturing. With usual etching techniques reported before it is not possible to etch very deep because of aspect ratio dependent etching phenomenon where with increasing etch depth the etch rate slows down resulting in non-vertical side walls and footing effects. Although with our de- veloped process quite vertical side walls were achieved but footing was still a problem. To overcome the challenges related to grating defini- tion and deep etching, a completely new three step gas chopping dry etching process was developed. This was the very first time that a time multiplexed etching process for an InP based material system was demonstrated. The developed gas chopping process showed extra ordinary results including high mask selectivity of 15, moderate etch- ing rate, very vertical side walls and a record high aspect ratio of 41. Both the developed etching processes are completely compatible with nano imprint lithography and can be used for low-cost high-throughput fabrication. A large number of broad area laser, ridge waveguide laser, distributed feedback laser, distributed bragg reflector laser and coupled cavity in- jection grating lasers were fabricated using the developed one step etch- ing process. Very extensive characterization was done to optimize all the important design and fabrication parameters. The devices devel- oped have shown excellent performance with a very high side mode suppression ratio of more than 52 dB, an output power of 17 mW per facet, high efficiency of 0.15 W/A, stable operation over temperature and injected currents and a threshold current as low as 30 mA for almost 1 mm long device. A record high modulation bandwidth of 15 GHz with electron-photon resonance and open eye diagrams for 10 Gbps data transmission were also shown.

Análisis comparativo de remoción de la membrana limitante interna en agujeros maculares con y sin azul tripan

Objetivo: comparar los resultados anatómicos y funcionales de los pacientes con Agujero Macular idiopático estadíos II, III y IV de Gass, quienes fueron sometidos a cirugía vitreoretiniana, con y sin uso de Azul Tripan, para la remoción de la Membrana Limitante Interna, en la Fundación Oftalmológica Nacional, Bogotá, Colombia, a partir de la revisión de historias clínicas de pacientes operados entre Enero de 2006 y Noviembre de 2009 Métodos: se realizó un estudio de cohorte retrospectivo, Los registros de historias clínicas de 32 pacientes operados de agujero macular idiopático fueron recolectados retrospectivamente a partir de 3 cirujanos. A todos los pacientes se les había realizado vitrectomía vía pars plana, se dividieron en 2 grupos aquellos con remoción de la MLI asistida con azul tripan y sin ésta. Se evaluaron variables, preoperatorias, intraoperatorias y postoperatorias. Resultados: la edad media de presentación del agujero macular fue de 61,33 años, la agudeza visual postoperatoria presentó diferencia entre los 2 grupos (p=0,008), la obtención del cierre del agujero macular utilizando azul tripan para la remoción de la membrana limitante interna no fue estadísticamente significativa (p=0,383) Conclusiones: la agudeza visual postoperatoria mejora, en los pacientes en quienes se utiliza el azul tripan para asistir la remoción de la MLI, en pacientes con agujeros maculares, en nuestro estudio la mejoría de la agudeza visual fue en promedio de 0,143. 2. No hubo diferencias entre los grupos estudiados con respecto al cierre anatómico primario del agujero macular. Se recomienda realizar estudios prospectivos, longitudinales para evaluar la asociación entre agujeros maculares, cierre anatómico primario y agudeza visual con el uso de azul tripan.