El estudio de la lágrima como ejemplo de traslacionalidad: del laboratorio a la clínica / The study of the tear as example of translation: From the bench to the clinic

No dsiponible

Distribution of Neurotensin and Somatostatin-28 (1-12) in the Minipig Brainstem.

Using an indirect immunoperoxidase technique, an in depth study has been carried out for the first time on the distribution of fibres and cell bodies containing neurotensin and somatostatin-28 (1-12) (SOM) in the minipig brainstem. The animals used were not treated with colchicine. The distribution of neurotensin- and SOM-immunoreactive fibres was seen to be quite similar and was moderate in the minipig brainstem: a close anatomical relationship between both neuropeptides was observed. The distribution of cell bodies containing neurotensin or SOM was quite different and restricted. Cell bodies containing neurotensin were found in four brainstem nuclei: nucleus centralis raphae, nucleus dorsalis raphae, in the pars centralis of the nucleus tractus spinalis nervi trigemini and in the nucleus ventralis raphae. Cell bodies containing SOM were found in six nuclei/regions of the brainstem: nucleus ambiguus, nucleus dorsalis motorius nervi vagus, formatio reticularis, nucleus parabrachialis medialis, nucleus reticularis lateralis and nucleus ventralis raphae. According to the observed anatomical distribution of the immunoreactive structures containing neurotensin or SOM, the peptides could be involved in sleep-waking, nociceptive, gustatory, motor, respiratory and autonomic mechanisms.

Efecto de las células de Müller en la supervivencia y neuritogénesis de las células ganglionares de la retina / Effect of Müller cells on the survival and neuritogenesis in retinal ganglion cells

OBJETIVO: Las células ganglionares de la retina (RGC) son las células que se afectan primero en neuropatologías como el glaucoma, por eso es importante investigar nuevas estrategias neuroprotectoras. Las células de Müller son células gliales que proporcionan a las neuronas factores tróficos y soporte físico. El propósito del presente estudio fue analizar el efecto de las células de Müller en la supervivencia y la formación de neuritas en las RGC. MÉTODO: Se cultivaron células de Müller de rata hasta obtener un cultivo confluente sobre el que se añadieron RGC. Cultivos puros de RGC fueron usados como control. Se marcaron las RGC con anticuerpos anti-βIII-tubulina y las células de Müller con antiglutamina sintetasa, los núcleos se tiñeron con DAPI. Se cuantificó el número de RGC y número y longitud de las neuritas. RESULTADOS: No hay diferencias en el número de RGC entre el control y las células crecidas sobre el sustrato de Müller. Aumenta la proporción de neuritas en RGC que crecieron sobre células de Müller, el número RGC con 1-3 neuritas pasa de un 19 a un 43%. También aumenta la longitud de las neuritas, el número de RGC con neuritas de entre 50-200 μm aumenta de un 21 en control a un 41% en cocultivo y con más de 200 μm pasa de un 6 a un 20%. CONCLUSIONES: Las células de Müller mantienen la supervivencia de las RGC e inducen un aumento tanto del número como de la longitud de las neuritas de las RGC

OBJECTIVE: The purpose of this study is to assess the effectiveness of the topical application of cacicol regenerating agent (RGTA) in an experimental model of corneal ulcer after photorefractive keratectomy (PRK) in mice. METHODS: Mice were subjected to PRK surgery with a 2.0 mm ablation zone on the central cornea and 45 mm of depth on a VISX Star S2 excimer laser. Corneas were treated topically with cacicol drops 1 hour and 48 hours after injury. Control groups received balanced salt solution (BSS) in the same dosage. Clinical and histopathological events were evaluated at 1, 2, 3 and 7 days after surgery. Sections obtained through the central region of the corneas were used to analyze the histopathological events of injured and healed corneas. αSMA (myofibroblast transformation), E cadherin (assembly of epithelial cells) and neuronal class III β-tubulin (innervation) were performed. RESULTS: Corneas treated topically with cacicol for 7 days showed a greater degree of transparency compared to controls. cacicol treated corneas showed improved epithelial cytoarchitecture. Analysis of αSMA profiles in the stroma showed that cacicol reduced or delayed the presence of myofibroblasts in the stroma compared to BSS (P<0.001). Finally, a putative neuroregenerative effect of cacicol was found in corneas subjected to an experimental PRK lesion. In some cases some interindividual variability could be observed due to the design of the experimental model. This is a limitation to consider, despite the statistical significance of the data. CONCLUSIONS: In a model of laser induced surgical lesions in the cornea, topical application of an RGTA (i.e. cacicol) could be involved in avoiding myofibroblast scarring formation and promoting nerve regeneration

Effect of Müller cells on the survival and neuritogenesis in retinal ganglion cells.

OBJECTIVE: Retinal ganglion cells (RGCs) are the first affected cells in neuropathies like glaucoma, for that reason it is very important to explore new methods to neuroprotect these neurons. Müller cells are glial cells that provide the neurons with trophic factors and scaffold. The purpose of this study was to analyze the effect of Müller cells on survival and neurite formation in RGCs. METHOD: Rat Müller cells were grown until a confluent culture on which rat RGCs were added, using pure culture of rat RGCs as controls. RGCs were labeled with ßIII-tubulin, and Müller cells with glutamine synthetase antibodies. In addition, nuclei were stained with DAPI. The number of RGCs and number and neurite length were measured. RESULTS: No differences were found in the number of RGCs between control and cells grown on the substrate of Müller cells. The proportion of RGCs with neurites increased when they grew on Müller (RGCs with 1-3 neurites increased from 19% to 43%. The length of neurites also increased in RGCs grown on Müller cells, with the number of RGCs with neurites from 50 to 200µm increasing from 21% to 41%, and with neurites of more than 200µm the increase was from 6% to 20%. CONCLUSIONS: Müller cells support the survival of RGCs and induced an increase in the number and length of neurites of RGCs.

Changes in tear protein profile in keratoconus disease.

PURPOSE: To analyze tear protein profile variations in patients with keratoconus (KC) and to compare them with those of control subjects. SUBJECTS AND METHODS: Tears from 12 normal subjects and 12 patients with KC were analyzed by two-dimensional gel electrophoresis (2-DE) and liquid chromatography-mass spectrometry (LC-MS). Analysis of the 2-DE gels was performed using Progenesis SameSpots software (Nonlinear Dynamics). Proteins exhibiting high variation in expression levels (P-value <0.05) were identified using matrix-assisted laser desorption/ionization-TOF spectrometry. For LC-MS analysis, a label-free quantification approach was used. Tears were digested with trypsin, subjected to data-independent acquisition (MS(E)) analysis, and identified proteins were relatively quantified using ProteinLynx Global Server software (Waters). RESULTS: The 2-DE and LC-MS analyses revealed a significant decrease in the levels of members of the cystatin family and an increase in lipocalin-1 in KC patients. A 1.43-fold decrease was observed for cystatin-S by 2-DE, and 1.69- and 1.56-fold for cystatin-SN and cystatin-SA by LC-MS, respectively. The increase in lipocalin-1 was observed by both methods with fold changes of 1.26 in the 2-DE approach and 1.31 according to LC-MS. Significant protein upregulation was also observed for Ig-κ chain C and Ig J chain proteins by 2-DE. Levels of lipophilin-C, lipophilin-A, and phospholipase A2 were decreased in tears from KC patients according to LC-MS. Serum albumin was found to be increased in KC patients according to LC-MS. CONCLUSION: The results show differences in the tear protein profile of KC and control subjects. These changes are indicative of alterations in tear film stability and in interactions with the corneal surface in KC patients.

[Cannabinoid applications in glaucoma]. / Aplicaciones de los cannabinoides en glaucoma.

INTRODUCTION: Glaucoma is a slowly progressive optic neuropathy that is one of the leading causes of legal blindness throughout the world. Currently there is a limited group of topical drugs for the medical treatment of glaucoma is currently limited, and research needs to be focused on new therapeutic horizons, such as the potential usefulness of the cannabinoid agonists for the treatment of glaucoma. AIM: To review the current scientific literature related to the beneficial effects derived from the different ways of administration of cannabinoids indicated for the glaucomatous optic neuropathy. DEVELOPMENT: Cannabinoid receptors have shown an intense expression in ocular tissues implicated in the regulation of the intraocular pressure, as well as inner layers of the retina. Through activation of CB1 and CB1 specific receptors and through other still unknown pathways, the cannabinoid agonists have shown both a clear hypotensive, as well as an experimentally proved neuroprotective effect on retinal ganglion cells. CONCLUSIONS: Some cannabinoid agonists (WIN 55212-2, anandamide) have demonstrated, in experimental studies, to act as «ideal drugs¼ in the management of glaucoma, as they have been shown to have good tolerability after topical application, efficiently reduce intraocular pressure, and behave as neuroprotectors on retinal ganglion cells. Further studies as regards the safety and clinical assays must be carried out in order to examine the effectiveness of these drugs for the treatment of glaucoma in our daily clinical practice.

Aplicaciones de los cannabinoides en glaucoma / Cannabinoid applications in glaucoma

IntroducciónEl glaucoma es una neuropatía óptica lentamente progresiva que constituye una de las principales causas de ceguera legal en el mundo. Actualmente existe un limitado grupo de fármacos tópicos para su manejo médico, siendo necesario enfocar la investigación hacia nuevos horizontes terapéuticos como el potencialmente útil grupo de los agonistas de cannabinoides.ObjetivoRevisar a través de la literatura científica actual, los efectos beneficiosos a través de distintas vías de administración de los cannabinoides para la neuropatía óptica glaucomatosa.DesarrolloLos receptores de cannabinoides han demostrado una amplia expresión en los tejidos oculares implicados en la regulación de la tensión ocular, así como en las capas internas de la retina. Mediante la activación de receptores específicos CB1, CB2 y vías aún no bien conocidas, los agonistas de cannabinoides han demostrado un claro efecto hipotensor ocular, así como un probado efecto neuroprotector sobre las células ganglionares de la retina en estudios experimentales.ConclusionesAlgunos cannabinoides (WIN 55212-2, anandamida) han demostrado a nivel experimental actuar como «fármacos ideales» en el manejo del glaucoma, al presentar buena tolerancia tras su aplicación tópica, reducir de forma eficaz la presión intraocular, y presentar un probado carácter neuroprotector sobre las células ganglionares de la retina.Se deben realizar más estudios sobre su seguridad y ensayos clínicos para poder examinar la utilidad de estos fármacos en el tratamiento del glaucoma en nuestra clínica diaria(AU)

Introduction: Glaucoma is a slowly progressive optic neuropathy that is one of the leadingcauses of legal blindness throughout the world. Currently there is a limited group of topical drugs for the medical treatment of glaucoma is currently limited, and research needs to befocused on new therapeutic horizons, such as the potential usefulness of the cannabinoidagonists for the treatment of glaucoma.Aim: To review the current scientific literature related to the beneficial effects derived fromthe different ways of administration of cannabinoids indicated for the glaucomatous opticneuropathy.Development: Cannabinoid receptors have shown an intense expression in ocular tissuesimplicated in the regulation of the intraocular pressure, as well as inner layers of the retina.Through activation of CB1 and CB1 specific receptors and through other still unknownpathways, the cannabinoid agonists have shown both a clear hypotensive, as well as anexperimentally proved neuroprotective effect on retinal ganglion cells.Conclusions: Some cannabinoid agonists (WIN 55212-2, anandamide) have demonstrated, inexperimental studies, to act as «ideal drugs» in the management of glaucoma, as they havebeen shown to have good tolerability after topical application, efficiently reduce intraocularpressure, and behave as neuroprotectors on retinal ganglion cells.Further studies as regards the safety and clinical assays must be carried out in order toexamine the effectiveness of these drugs for the treatment of glaucoma in our daily clinicalpractice(AU)

Modelos animales en el estudio del glaucoma: pasado, presente y futuro / Animal models in the study of the glaucoma; past, present and future

No disponible

Terapia génica contra la retinosis pigmentaria / Gene therapy against retinosis pigmentary

No disponible