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Solar damage due to ultraviolet radiation (UVR) is implicated in the development of two proliferative lesions of the ocular surface: pterygium and pinguecula. Pterygium and pinguecula specimens were collected, along with adjacent healthy conjunctiva specimens. RNA was extracted and sequenced. Pairwise comparisons were made of differentially expressed genes (DEGs). Computational methods were used for analysis. Transcripts from 18,630 genes were identified. Comparison of two subgroups of pterygium specimens uncovered evidence of genomic instability associated with inflammation and the immune response, these changes were also observed in pinguecula, but to a lesser extent. Among the top DEGs were four genes encoding tumor suppressors that were downregulated in pterygium: C10orf90, RARRES1, DMBT1 and SCGB3A1, C10orf90 and RARRES1 were also downregulated in pinguecula. Ingenuity Pathway Analysis overwhelmingly linked DEGs to cancer for both lesions, however, both lesions are clearly still benign, as evidenced by the expression of other genes indicating their well-differentiated and non-invasive character. Pathways for epithelial cell proliferation were identified that distinguish the two lesions, as well as genes encoding specific pathway components. Upregulated DEGs common to both lesions, including KRT9 and TRPV3, provide a further insight into pathophysiology. Our findings suggest that pterygium and pinguecula, while benign lesions, are both on the pathological pathway towards neoplastic transformation.

Introduction The intestinal microbiota plays an important role in modulating host immune responses. Oral Toxoplasma gondii infection can promote intestinal inflammation in certain mice strains. The IDO-AhR axis may control tryptophan (Trp) metabolism constituting an important immune regulatory mechanism in inflammatory settings. Aims In the present study, we investigated the role of the intestinal microbiota on Trp metabolism during oral infection with T gondii. Methods and results Mice were treated with antibiotics for four weeks and then infected with T gondii by gavage. Histopathology and immune responses were evaluated 8 days after infection. We found that depletion of intestinal microbiota by antibiotics contributed to resistance against T gondii infection and led to reduced expression of AhR on dendritic and Treg cells. Mice depleted of Gram-negative bacteria presented higher levels of systemic Trp, downregulation of AhR expression and increased resistance to infection whereas depletion of Gram-positive bacteria did not affect susceptibility or expression of AhR on immune cells. Conclusion Our findings indicate that the intestinal microbiota can control Trp availability and provide a link between the AhR pathway and host-microbiota interaction in acute infection with T gondii.

The aim of this mini review is to describe the prevalence of Environmental Proteinaceous Corneal Degeneration (EPCD), pinguecula and pterygium and the association among them. We performed a transversal/observational study in individuals living all their lives in the El Cuy Department, province of Río Negro in the Argentinean Patagonia. The patients investigated were consecutive non-probabilistic. The prevalence of ophthalmoheliosis was 34% of pinguecula, followed by 30% of EPCD and 12% of pterygium. A statistically significant association was found between EPCD and male patients (p Fil: Suarez, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Serra, Horacio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina

Resistance to Trypanosoma cruzi infection is dependent on a rapid induction of Th1-type and CD8+ T cell responses that should be promptly balanced to prevent immunopathology. T. cruzi-infected B6 mice are able to control parasite replication but show a limited expansion of Foxp3+regulatory T (Treg) cells that results in the accumulation of effector immune cells and the development of acute liver pathology. AhR is a ligand-activated transcription factor that promotes Treg cell development and suppression of pro-inflammatory cytokine production in dendritic cells, altering the course of adaptive immune response and the development of immunopathology. Here, we used different AhR-dependent activation strategies aiming to improve the Treg response, and B6 congenic mice carrying a mutant AhR variant with low affinity for its ligands (AhRd) to evaluate the role of AhR activation by natural ligands during experimental T. cruzi infection. The outcome of TCDD or 3-HK plus ITE treatments indicated that strong or weak AhR activation before or during T. cruzi infection was effective to regulate inflammation improving the Treg cell response and regularizing the ratio between CD4+ CD25- to Treg cells. However, AhR activation shifted the host-parasite balance to the parasite replication. Weak AhR activation resulted in Treg promotion while strong activation differentially modulated the susceptibility and resistance of cell death in activated T and Treg cells and the increase in TGF-β-producing Treg cells. Of note, T. cruzi-infected AhRd mice showed low levels of Treg cells associated with strong Th1-type response, low parasite burden and absence of liver pathology. These mice developed a Treg- and Tr1-independent mechanism of Th1 constriction showing increased levels of systemic IL-10 and IL-10-secreting CD4+ splenocytes. In addition, AhR activation induced by exogenous ligands had negative effects on the development of memory CD8+ T cell subsets while the lack/very weak activation in AhRd mice showed opposite results, suggesting that AhR ligation restricts the differentiation of memory CD8+T cell subsets. We propose a model in which a threshold of AhR activation exists and may explain how activation or inhibition of AhR-derived signals by infection/inflammation-induced ligands, therapeutic interventions or exposure to pollutants can modulate infections/diseases outcomes or vaccination efficacy. Fil: Ambrosio, Laura Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba; Argentina Fil: Insfran, Constanza. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Volpini, Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina Fil: Acosta Rodriguez, Eva Virginia. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Serra, Horacio Marcelo. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Quintana, Francisco Javier. The Broad Institute of MIT and Harvard; Estados Unidos. Harvard Medical School; Estados Unidos Fil: Cervi, Laura Alejandra. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Motran, Claudia Cristina. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina

As metabolism is controlled by the input of genes and the environment, metabolic disorders result from some disturbance in the interaction between genes and environmental factors. Many metabolic disorders consist in congenital enzyme deficiencies, also known as "inborn errors of metabolism," that may be disabling or cause severe illness and death and are predominantly inherited in an autosomal recessive fashion. The deposit in cells and tissues of storage substances from errors in metabolic processes may produce a wide variety of disorders affecting different organs and functions, with different degrees of severity, and often present around the time of birth or early childhood. Distinctive ocular and skin manifestations accompany many metabolic diseases and may provide clues for their diagnosis and evolution.

La Queratopatía Climática Esferoidea (QCE) es una enfermedad degenerativa de la córnea humana caracterizada por la agregación de proteínas bajo el epitelio y una progresiva opacidad corneal, causada por condiciones ambientales desfavorables. Si bien esta patología fue descripta por primera vez hace más de cien años, en la actualidad no existe un modelo experimental de QCE que permita avanzar en el conocimiento de la etiopatogenia de la misma para plantear posibles tratamientos. En el intento por desarrollar este modelo en cobayos y debido a la escasa bibliografía disponible que describa la anatomía, fisiología y superficie ocular de estos animales, realizamos diferentes tests fisiológicos de superficie ocular en cobayos y lo compararemos con el humano. Métodos: Para estos estudios se utilizaron 15 cobayos. Se realizaron videograbaciones de los mismos en un ambiente de tranquilidad y bajo ciertos estímulos para estudiar la dinámica de parpadeo y sueño. Con el fin de analizar la producción y la estabilidad del film lagrimal, se realizaron test de Schirmer, medición de la altura del menisco lagrimal inferior por OCT visante y tiempo de ruptura precorneal (TRP) respectivamente. Resultados: El cobayo parpadea de forma completa y parcial, siendo más frecuente ésta última modalidad. La frecuencia y tipo de parpadeos/ minuto fue: Total= 2.35± 0.87; completos= 0.68±0.43; parciales= 1.49±0.75. Su frecuencia de parpadeo es 5 veces menor que en humanos. La respuesta palpebral frente a diferentes estímulos externos fue escasa. Los cobayos no poseen hábitos nocturnos como muchos roedores y duermen por cortos períodos. A menudo no cierran completamente los ojos. Test de Schirmer cobayos: 8.14±1.86 mm/5 min (Humanos: 11.9 ± 6.79 mm/5 min). Altura menisco inferior cobayos: 0,206 ± 0,059 mm (Humanos: 0,277 ± 0,033 mm). TRP cobayos: 2-3 min. (Humanos: 10-15 seg). Conclusiones: Los resultados obtenidos muestran que el cobayo, a diferencia de humanos, tiene una frecuencia de parpadeo menor. A su vez, esto se puede correlacionar con una producción de lágrima disminuida (objetivada con Test de Schirmer) y una mayor estabilidad de la película lagrimal, como lo evidencia el prolongado TRP.

Evaluar los resultados del implante de válvula de Ahmed en pacientes con diferentes tipos de glaucoma refractario. Pacientes, material y métodos: 14 ojos de 13 pacientes mayores de 18 años con glaucoma refractario al tratamiento médico máximo y operados mediante la implantación de válvula de Ahmed (Modelo AGV-FP7) entreAgosto de 2007 y Abril de 2015, con un seguimiento mínimo de 24 meses,fueron evaluados retrospectivamente. Se consideró éxito absoluto cuando la presión intraocular (PIO) postoperatoria se hallaba entre 5 y 22 mmHgsin tratamiento médico antiglaucomatoso, éxito relativo cuando la PIO postoperatoria se hallaba dentro de ese rango pero con la ayuda de uno o más colirios antiglaucomatosos, y fracaso cuando la PIO postoperatoria era menor de 5 o mayor de 22 mmHg, o cuando se debió proceder al explante de la válvula. En los pacientes con glaucoma crónico de ángulo abierto(GCAA), el 14,29% (n=1) presentó éxito absoluto, el 57,14% (n=4) éxito relativo, y en el 28,57% (n=2) la cirugía valvular fracasó. En todos los pacientes con glaucoma pseufoexfoliativo, glaucoma asociado a uveítis yen glaucoma congénito se obtuvo un éxito relativo. El implante valvular de Ahmed fue eficaz para reducir la PIO en glaucomas refractarios al tratamiento médico en un número importante de casos en esta serie, llegando a un éxito relativo en la mayoría de ellos. To evaluate the results of Ahmed valve implantation in refractory glaucoma patients. Patients, Material and Methods: 14 eyes in 13 patients with refractory glaucoma to medical treatment, which underwent an implan-tation of an Ahmed valve between August 2007 and April 2015, with a minimum follow up of 24 months were retrospectively reviewed. Success was defined as a postoperative intraocular pressure (IOP) between 5 and 22 mmHg without antiglauco-matous medications, relative success when postoperative IOP was within that range but with one or more glaucoma drops, and failure when postoperative IOP was < 5 or > 22 mmHg or when the valve was explanted. An absolute success was obtained in 14.29% (n = 1) of patients with primary open angle glaucoma a relative success in 57.14% (n = 4), and a failure in 28.57% (n = 2). Patients with pseudoexfoliative glaucoma, uveitic glaucoma and congenital glaucoma, showed 100% relative success. Ahmed valve implantation was effective in reducing IOP in glaucoma refractory to medical treatment in a significant number of cases in this series, reaching a relative success in most of them. Fil: Monti, Jose Rodolfo. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina Fil: Esposito, Evangelina. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina Fil: Fornies Paz, Maria Elena. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina Fil: Crim, Nicolás. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina Fil: Dalmagro, Juan. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina Fil: Serra, Horacio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Urrets Zavalía, Julio Alberto. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina

The cornea plays important roles in the eye, since it maintains the ocular pressure, bears up different aggressions and forces, and constitutes the primary lens in many animals’ eyes. In order to fulfill these extremely crucial functions, the cornea requires sophisticated structures called epithelium, Bowman's membrane, stroma, Descement's layer, and endothelium. The complexity of this tissue is such that despite many efforts, it has not been possible to obtain clinically viable corneas in the lab yet. Vitamin C or l -ascorbic acid is an essential vitamin and a water-soluble important nutrient, which many animals chemically synthesize from the glucose by means of enzymatic reactions, the l -gulono-γ-lactone oxidase being the last enzyme. Among the multiple roles played by this vitamin, its primary functions are to act as a cofactor for reactions requiring a reduced iron or copper metalloenzyme and as a protective antioxidant that operates in the aqueous phases, both intra- and extracellularly. Ocular tissues, particularly the cornea, concentrate vitamin C, which might suggest that the different structures of the cornea need this vitamin to fulfill their roles. In this chapter, we present scientific evidences of ascorbic acid critical roles in maintaining the functions and ultrastructures of the corneal epithelium, the stroma, and the endothelium.

Purpose Climatic droplets keratopathy (CDK) is closely associated with superficial corneal erosions and lack of protective mechanisms against the harmful effects of ultraviolet radiation (UVR) during a prolonged period of time. One of the difficulties in studying the pathogenic mechanisms involved in this human disease is the lack of an experimental animal model. In this paper, a study is conducted on the effects of 4 types of lasers at various powers and time conditions on the normal guinea pig corneas in order to select only one laser condition that reversibly injures the epithelium and superficial stroma, without leaving scarring. Methods Damage was induced in the cornea of guinea pigs using different powers and exposure times of 4 types of laser: argon, CO2, diode and Nd–Yag, and any injuries were evaluated by biomicroscopy (BM) and optical microscopy. Corneas from other normal animals were exposed to argon laser (350 mW, 0.3 s, 50 μm of diameter), and the induced alterations were studied at different times using BM, optical coherence tomography (OCT) and transmission electron microscopy (TEM). Results Only argon laser at 350 mW, 0.3 s and 50 μm of diameter produced epithelium and superficial stroma lesions. Some leukomas were observed by BM, and they disappeared by day 15. Corneal thickness measured by OCT decreased in the eyes treated with argon laser during the first week. Using TEM, different ultrastructural alterations in corneal epithelium and stroma were observed during the early days, which disappeared by day 15. Conclusions It was possible to develop reproducible corneal epithelium and anterior stroma injuries using argon laser at 350 mW, 0.3 s and 50 μm of diameter. In vivo and in vitro studies showed that injured corneas with these laser conditions did not leave irreversible microscopic or ultrastructural alterations. This protocol of corneal erosion combined with exposure to UVR and partial deficiency of ascorbate in the diets of the animals for an extended period of time has been used to try to develop an experimental model of CDK.

Resumen Objetivo La queratopatia climatica esferoidea (QCE) esta intimamente asociada a erosiones corneales superficiales y a carencia de mecanismos protectores contra los efectos nocivos de la radiacion ultravioleta (RUV) durante muchos anos. Debido a que una de las dificultades en el estudio de los mecanismos patogenicos de esta enfermedad humana es la ausencia de un modelo experimental, en este trabajo quisimos identificar cual es el mejor metodo para estudiar una de las variables involucradas en su genesis (erosiones superficiales de la cornea). A tal fin investigamos los efectos producidos por 4 tipos de laser con diferentes condiciones de potencia y tiempo en la cornea de cobayos normales con el fin de seleccionar un laser y condicion que lesione solamente el epitelio y estroma superficial, de manera reversible, sin dejar cicatrices. Metodos Se indujeron danos en la cornea de cobayos utilizando distintas potencias y tiempos con 4 tipos de laser: argon, CO 2 , diodo y Nd-Yag en distintos grupos de animales y se evaluaron dichas lesiones por biomicroscopia (BM) y microscopia optica. Corneas de otros animales normales fueron expuestas a laser de argon (350 mW, 0,3 s, 50 μm de diametro) y las alteraciones inducidas se estudiaron en diferentes tiempos utilizando BM, tomografia de coherencia optica (TCO) y microscopia electronica (ME). Resultados Solo el laser argon a una potencia de 350 mW, 0,3 s, 50 μm de diametro produjo lesiones de epitelio y estroma superficial, manteniendose indemne el endotelio. Por BM se observaron algunos leucomas que desaparecieron hacia el dia 15. Mediante TCO se observo un adelgazamiento del espesor corneal en los ojos tratados con esas condiciones de laser argon durante la primera semana. Mediante ME, se observaron diferentes alteraciones ultraestructurales en epitelio y estroma corneal durante los primeros dias, las cuales desaparecieron hacia el dia 15. Conclusiones Fue posible desarrollar lesiones corneales en epitelio y estroma anterior de cobayos de manera reproducible mediante el uso de laser argon. Los estudios in vivo e in vitro demostraron que las corneas lesionadas con este laser y en esas condiciones no dejaron alteraciones microscopicas ni ultraestructurales irreversibles. Este modo de erosion corneal combinado con exposicion a RUV y deficit parcial de ascorbato en la dieta de los animales durante un periodo prolongado de tiempo esta siendo utilizado con el fin de intentar desarrollar un modelo experimental de QCE.

Astrocytes respond to environmental cues, including changes in temperatures. Increased deimination, observed in many progressive neurological diseases, is thought to be contributed by astrocytes. We determined the level of deimination and expression of peptidyl arginine deiminase 2 (PAD2) in isolated primary astrocytes in response to changes on either side (31°C and 41°C) of the optimal temperature (37°C). We investigated changes in the astrocytes by using a number of established markers and accounted for cell death with the CellTiter-Blue assay. We found increased expression of glial fibrillary acidic protein, ALDH1L1, and J1-31, resulting from increased incubation temperature and increased expression of TSP1, S100β, and AQP4, resulting from decreased incubation temperature vs. optimal temperature, suggesting activation of different biochemical pathways in astrocytes associated with different incubation temperatures. Mass spectrometric analyses support such trends. The PAD2 level was increased only as a result of increased incubation temperature with a commensurate increased level of deimination. Actin cytoskeleton and iso[4]LGE, a lipid peroxidase modification, also showed an increase with higher incubation temperature. Altogether, these results suggest that temperature, as an environmental cue, activates astrocytes in a different manner on either side of the optimal temperature and that increase in deimination is associated only with the higher temperature side of the spectrum.

Climatic droplet keratopathy (CDK) is a degenerative corneal disease of unknown etiology. We described CDK for the rst time in Latin America in the Argentinean Patagonia (El Cuy). A deeper knowledge of CDK pathogenic mechanisms will provide new therapeutic strategies. For that reason we investigated the prevalence of CDK in El Cuy and its existence in other 3 provinces with similar climate. Patients eyes were examined, habits throughout lives were inquired about, and serum ascorbate (sAA) was determined. All individuals work outdoors for most of the day. All regions had normal O3 levels. Individuals from regions 1, 2, and 3 had very low consumption of vegetables/fruits and low sAA levels. Conversely, region 4 individuals had balanced diet and higher sAA concentrations. CDK was only found in region 3 where individuals had partial de ciency of sAA and did not use eye protection. No CDK was found in regions 1 and 2 where individuals had similar work activities and dietary habits to those in region 3 but wear eye protection. No disease was found in region 4 where individuals work outdoors, have balanced diet, and use eye protection. To summarize, the CDK existence was related not only to climate but also to the dietary habits and lack of protection from sunlight. Fil: Suarez, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Correa, Leandro Javier. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Crim, Nicolás. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Esposito, Evangelina. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Monti, Jose Rodolfo. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Urrets Zavalía, Julio Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Serra, Horacio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina

Climatic droplet keratopathy (CDK) is an acquired degenerative disease predominantly affecting males over 40 years old. It results in progressive corneal opacities usually affecting both eyes. CDK is multifactorial and its etiology remains unknown. Our recent findings are consistent with CDK pathology being driven by environmental factors with oxidative stress playing an important role (e.g.,, contributing to lipid peroxidation) rather than climate factors. The changes in corneal lipid composition affected by environmental factors remain understudied. The purpose of this study was to systematically investigate phospholipids profile (phosphatidylcholine [PC] and phosphatidylserine [PS]) in corneas from CDK patients using tandem mass spectrometry. Samples from CDK areas and from non-affected areas were obtained from patients diagnosed with CDK who underwent cataract surgery, were subjected to lipid extraction using a modified Bligh and Dyer method; protein concentrations were determined using the Bradford's method. Lipids were identified and subjected to ratiometric quantification using TSQ Quantum Access Max triple quadrupole mass spectrometer, using appropriate class specific lipid standards. All phospholipid classes showed lower total amounts in affected areas compared to control areas from CDK's corneas. Comparative profiles of two phospholipid classes (PC, PS) between CDK areas and control areas showed several common species between them. We also found a few unique lipids that were absent in CDK areas compared to controls and vice versa. Lower amount of phospholipids in CDK areas compared to control areas could be attributed to the lipid peroxidation in the affected corneal regions as a consequence of increased oxidative stress. Fil: Suarez, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina Fil: Piqueras, Maria Carmen. University of Miami; Estados Unidos Fil: Correa, Leandro Javier. Universidad Católica de Córdoba. Facultad de Medicina. Clínica Universitaria Reina Fabiola; Argentina Fil: Esposito, Evangelina. Universidad Católica de Córdoba. Facultad de Medicina. Clínica Universitaria Reina Fabiola; Argentina Fil: Barros Centeno, María Fernanda. Universidad Católica de Córdoba. Facultad de Medicina. Clínica Universitaria Reina Fabiola; Argentina Fil: Bhattacharya, Sanjoy K.. University of Miami; Estados Unidos Fil: Urrets Zavalía, Julio Alberto. Universidad Católica de Córdoba. Facultad de Medicina. Clínica Universitaria Reina Fabiola; Argentina Fil: Serra, Horacio Marcelo. Universidad Nacional de Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina

Summary Our study performed qualitative and quantitative studies on the corneal ultrastructure of healthy female Merino sheep of ages 4 months and 6 years old from the Argentinean Pampa. The corneas were evaluated using ex vivo laserscanning confocal microscopy, light microscopy and transmission electron microscopy. Those studies allowed us to obtain detailed images of the corneal layers as well as quantitative data of the cellular and sub-basal nerve densities in the cornea from sheep of different ages. The density of the corneal cells was significantly different in the anterior versus the posterior epithelium and stroma. Moreover, the density of the epithelial, stromal cells and endothelial cells, as well as the sub-basal nerve density were significantly lower in adult than in young animals. Our work provided a wide-ranging description of the corneal ultrastructure of healthy female Merino sheep, which adds to the current knowledge about the ophthalmological aspects of this species and undoubtedly benefits veterinarians.

This paper shows that the population studied in the most populated departments of Paraguay does not present significant population substructure among the different geographical locations sampled, allowing the use of these genetic frequencies for forensic genetics purposes in Paraguay. Furthermore, information on 22 aSTR presented in this study increases the resolution in database searching, it is useful for resolution of deficient paternity cases or paternity cases with inconsistencies, increasing both, the Power of Discrimination and the Power of Exclusion. Fil: Vullo, Carlos. Laboratorio de Inmunogenetica y Diagnostico Molecular; Argentina Fil: Rotondo, Martina. Laboratorio de Genética Forense; Argentina Fil: Daguerre, Natalia. Laboratorio de Inmunogenetica y Diagnostico Molecular; Argentina Fil: Quiroz, Alfredo. Hospital Regional, Encarnación, Paraguay; Paraguay Fil: Velásquez, Vanesa. Hospital Regional, Encarnación, Paraguay; Paraguay Fil: Machado, Patricia. Hospital Regional, Encarnación, Paraguay; Paraguay Fil: Serra, Horacio Marcelo. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Borosky, Alicia. Laboratorio de Inmunogenetica y Diagnostico Molecular; Argentina

To present external eye findings and the observation of iris atrophy in patients with climatic droplet keratopathy (CDK).Twenty-three patients with CDK and 13 controls living in a semideserted plain of the Argentine Patagonia were studied. Besides a comprehensive eye examination, Cochet-Bonnet aesthesiometry, Schirmer II test, breakup time (BUT), and surface staining were performed. According to corneal findings, eyes were grouped as grade 1 (confluent translucent microdroplets localized in the limbic region of the horizontal quadrants); grade 2 (band-shaped subepithelial haziness compromising the central cornea); and grade 3 (previously described lesions with yellow subepithelial droplets). Results were analyzed with the Fisher, Mann-Whitney, and Spearman tests, which were considered significant at P0.05.Nineteen of 23 patients with CDK had bilateral disease, which was asymmetric in 2 of them. Sixteen eyes had grade 1, 21 eyes had grade 2, and 5 eyes had grade 3 disease. Aesthesiometry showed that the more advanced the disease, the more profound the corneal hypoesthesia (P = 0.0008). BUT and ocular surface staining significantly differed between eyes with grade 3 and grade 1, grade 3 and grade 2, and grade 3 and controls. In 38.09% of eyes with CDK and in none of the controls, sectorial depigmentation and atrophy of the inferior iris were observed.A severe decrease of corneal sensitivity was observed in advanced stages of CDK. Some degree of dry eye was present in most patients, but severe disease was infrequent at any stage of CDK. Inferior iris depigmentation and atrophy was frequently observed among patients with CDK.

Objective: The purpose of our work was to study and compare the cornea and tear film of people and sheep inhabiting in a region of Argentinean Patagonia (PATG), and in a geographically and climatically different region in the Argentinean Pampa (CAPT).Methods: Using cross sectional study, corneal examinations were performed in people and Merino sheep inhabiting PATG and CAPT regions, respectively. All people completed a questionnaire related to work activity, diet, and the use of hats or sunglasses during their life. Eyes from all participants were examined with a portable hand- held slit-lamp biomicroscope (BM) to evaluate corneal appearance, epithelium integrity, and transparency. Later on, in a subgroup of participants we studied eye blinking frequency (EBF), ocular surface staining (FS), breakup time (BUT), Schirmer Tear Test (STT), corneal structure using confocal laser scanning microscopy (CLSM) and light microscopy (LM), and concentrations of serum ascorbate (sAA).Results: BM studies revealed numerous cases of climatic droplet keratopathy (CDK) only in people inhabiting the PATG region. CLSM studies confirmed typical punctate deposits at the Bowman ́s layer in CDK patients living in that region. CLSM images from sheep did not show any abnormalities at the Bowman ́s layer but exhibited small hyper reflective dots at the epithelium only in animals pasturing in the PATG region. FS and EBF mean values were significant higher in sheep grazing in the PATG region (p

Background Conjunctival amyloidosis is a very rare condition, generally unilateral, and presents mostly as an isolated condition without systemic compromise. Our purpose is to present a new case of systemic amyloidosis with a bilateral conjunctival involvement. Case Presentation A 66-years-old caucasian female complaining of conjunctival hemorrhage and chemosis in both eyes for the last five years had been discontinuously treated with topical antibiotics and corticosteroids without any evident improvement. She presented with a pink-yellow infiltration in the inferior conjunctiva of both eyes. Conjunctival biopsy under optical microscopy revealed amyloid deposit, confirmed by Congo red staining. Mucosal biopsy from esophagus and rectus confirmed amyloidosis by Congo red stain. Immunohistochemistry of bone marrow biopsy showed an increased number of plasma cells and an over-expression of light chain kappa subunit. She was treated with corticosteroids and lubrication with an improvement of symptoms. Ocular lesions remained stable after a follow-up of 3 years. Conclusions Conjunctival amyloidosis is a rare entity that may be overlooked, and should be differentiated from chronic conjunctivitis and conjunctival malignancies. Although it presents most frequently as a local process, a systemic involvement should always be ruled out.

The aim of this study was to assess whether tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta1 (TGF-beta1) and interleukin-10 (IL-10) polymorphisms are among the factors influencing the development of pemphigus. Whole blood from 20 patients with pemphigus and 24 control subjects was taken. Genomic DNA was obtained and cytokine genotyping for IL-10 (-1082 G/A; -819 C/T), TGFB1 (codon 10 C/T, codon 25 G/C) and TNFA (-308 G/A) was performed using the ARMS-PCR method. The distribution of IL-10 (-819) alleles was significantly different between the pemphigus and control groups (P=0.009). In particular, allele T was associated with the disease (OR 3.291, 95% CI 1.350-8.020). Similar results were observed when only pemphigus vulgaris (PV) patients were analyzed (P=0.012, OR 3.410, 95% CI 1.346-8.639). An increased frequency of the low producer IL-10 haplotype (-1082/-819 A/T) in patients with pemphigus compared with controls was observed (OR 2.714, 95% CI 1.102-6.685) and this association was also significant when only PV patients were considered (OR 2.667, 95% CI 1.043-6.816). There were no differences between patients and controls in the frequency of any other gene polymorphism analyzed. The increased frequency of the low producer IL-10 haplotype (-1082 /-819 A/T) suggest that the carriage of this haplotype might predispose to pemphigus or the high and intermediate producer haplotypes may be protective factors. The prevalence of the allele IL-10 (-819 T) in pemphigus patients cannot be explained by the current hypothesis, according to which a particular allele of the gene is associated with a different level of cytokine production and therefore affects the predisposition to a particular disease. However, this cytokine polymorphism might be linked to an unknown susceptibility factor.

Chemokines are a superfamily of small cytokines with activities ranging from leukocyte traffick to hematopoiesis, angiogenesis, and tissue organogenesis. Secondary lymphoid-organ chemokine (SLC/CCL21) was originally reported as a chemokine constitutively expressed by stromal cells and high endothelial venules in secondary lymphoid tissues and endothelium of afferent lymphatics, directing CCR7+ cells. More recently, others and we have demonstrated that SLC/CCL21 is up-regulated in different skin inflammatory conditions. Thereafter, this molecule is much more than a constitutive chemokine, which could play a role in effector and regulatory immune functions.

Summary Background Leucocyte migration within inflammatory skin compartments in allergic contact dermatitis (ACD) is the result of a sophisticated multi-step event where multiple molecules are involved. Objective Since non-antigen-specific mechanisms have been described as an early participant in elicitation of ACD, we investigated the kinetics of the expression of monocyte chemoattractant protein-1 (MCP-1/CCL2) and the type of infiltrating cells. We compared the time course production of MCP-1/CCL2 with connecting segment-1 (CS-1) fibronectin and thymus and activation-regulated chemokine (TARC/ CCL17) expression. Methods Biopsies from 10 individuals challenged in their back with the antigen responsible for their contact dermatitis and an irrelevant antigen were taken at different times and histology, immunohistochemistry for CS-1 fibronectin, TARC/CCL17, CD3, CD68, CXCR3, CCR4 and in situ hybridization for MCP-1/CCL2 were performed. Results At positive antigen stimulated sites expression of MCP-1/CCL2 by basal keratinocytes and isolated cells in dermis started at 10 h. CS-1 fibronectin and TARC/CCL17 expression by blood endothelial cells was found at 2 and 10 h, respectively. This was followed by dermal accumulation of mononuclear cells with a significant increase of CD3 1 and CD68 1 cells. At 48 h, approximately 58% of infiltrating cells were CXCR3 1 , and 35% CCR4 1 . Conclusions We showed evidence of the fact that CS-1 fibronectin expression precedes the production of MCP-1/CCL2 and TARC/CCL17 in the skin of patients with ACD, suggesting that these molecules participate in the early complex process of migrating mononuclear cells during elicitation of ACD.

Background: Allergic contact dermatitis (ACD) is a common human dermatosis in which not all the mechanisms involved in its pathogenesis have been elucidated. Objective: To study the expression of CS-1 fibronectin, TARC and Th1-associated chemokine receptors in biopsies from allergic patch test reactions. Material and methods: Thirteen patients already diagnosed with ACD were challenged on the back with the antigen responsible of the disease and macroscopic responses and biopsies taken after 48 h. Skin biopsies from negative control challenge sites, AD and ICD were also taken. Samples were fixed, embedded in paraffin wax and processed in order to perform histological and immunohistochemical studies. Results: All subjects with ACD showed a positive clinical response and a perivascular mononuclear cell infiltration at 48 h, which was not seen in the negative controls. The majority of skin-infiltrating cells were CD4+ and CD8+ and up to 54% or 40% of them expressed CXCR3 or CCR5, respectively. We also showed expression of CS-1 fibronectin in inflamed endothelial cells not only in ACD but also in AC and ICD. In contrast TARC was only expressed in ACD and AC. Conclusion: We showed for the first time that CS-1 fibronectin is expressed in dermal vessels from allergic patch tests positive reactions, as well as irritant and atopic skin lesions.

Astrocytes respond to environmental cues, including changes in temperatures. Increased deimination, observed in many progressive neurological diseases, is thought to be contributed by astrocytes. We determined the level of deimination and expression of peptidyl arginine deiminase 2 (PAD2) in isolated primary astrocytes in response to changes on either side (31°C and 41°C) of the optimal temperature (37°C). We investigated changes in the astrocytes by using a number of established markers and accounted for cell death with the CellTiter-Blue assay. We found increased expression of glial fibrillary acidic protein, ALDH1L1, and J1-31, resulting from increased incubation temperature and increased expression of TSP1, S100β, and AQP4, resulting from decreased incubation temperature vs. optimal temperature, suggesting activation of different biochemical pathways in astrocytes associated with different incubation temperatures. Mass spectrometric analyses support such trends. The PAD2 level was increased only as a result of increased incubation temperature with a commensurate increased level of deimination. Actin cytoskeleton and iso[4]LGE, a lipid peroxidase modification, also showed an increase with higher incubation temperature. Altogether, these results suggest that temperature, as an environmental cue, activates astrocytes in a different manner on either side of the optimal temperature and that increase in deimination is associated only with the higher temperature side of the spectrum.

Climatic droplet keratopathy (CDK) is an acquired and potentially handicapping cornea degenerative disease that is highly prevalent in certain rural communities around the world. It predominantly affects males over their forties. It has many other names such as Bietti's band-shaped nodular dystrophy, Labrador keratopathy, spheroidal degeneration, chronic actinic keratopathy, oil droplet degeneration, elastoid degeneration and keratinoid corneal degeneration. CDK is characterized by the haziness and opalescence of the cornea's most anterior layers which go through three stages with increasing severity. Globular deposits of different sizes may be histopathologically observed under the corneal epithelium by means of light and electron microscopy. The coalescence and increased volume of these spherules may cause the disruption of Bowman's membrane and the elevation and thinning of the corneal epithelium. The exact aetiology and pathogenesis of CDK are unknown, but they are possibly multifactorial. The only treatment in CDK advanced cases is a corneal transplantation, which in different impoverished regions of the world is not an available option. Many years ago, the clinical and histological aspects of this disease were described in several articles. This review highlights new scientific evidence of the expanding knowledge on CDK's pathogenesis which will open the prospect for new therapeutic interventions.

PURPOSE: To present the findings of climatic droplet keratopathy (CDK) that affects people of a rural area of the Argentine Patagonia. DESIGN: Observational case series. METHODS: Five hundred seventy-seven individuals who live in settlements and villages in an inland area of the northwest Patagonia region in Argentina received a complete eye examination. RESULTS: The mean age was 36.02 years (r = 1.5 months to 89 years); 55.63% of the patients were female. The mean age of the 7.62% of the patients who had typical CDK was 65.31 years (r = 42 to 89 years); 86.36% of the patients were male. Of 66 eyes in 35 patients with CDK, 35 eyes had peripheral haziness (grade 1); 23 eyes had a band-shaped haziness (grade 2), and 8 eyes had the aggregate of yellow subepithelial droplets (grade 3). Pinguecula, pterygium, cataract, and pseudoexfoliation were frequent findings among patients with CDK. CONCLUSION: CDK is not infrequent among male adults in this area of the Argentine Patagonia and may be severely handicapping.

The cornea plays important roles in the eye since it not only maintains the ocular pressure and protects against different aggressions and forces but constitutes the primary lens in many animals’ eyes as well. In order to fulfill these extremely crucial functions, the cornea requires sophisticated structures called the epithelium, Bowman’s membrane, the stroma, Descement’s layer, and the endothelium. The complexity of this tissue is such that despite many efforts, it has not been possible to obtain clinically viable corneas in the laboratory yet. Vitamin C or L-ascorbic acid is an essential vitamin and a water-soluble important nutrient many animals chemically synthesize from glucose by means of enzymatic reactions, with L-gulono-γ-lactone oxidase being the last step. Among the multiple roles played by this vitamin, its primary functions are to act as a cofactor for reactions requiring reduced iron or copper metalloenzyme and as a protective antioxidant that operates in the aqueous phases, both intra- and extracellularly. Ocular tissues, particularly the cornea, concentrate vitamin C, which might suggest that the cornea’s different structures need this vitamin to fulfill their roles. In this work, we present scientific evidence of the critical roles that ascorbic acid performs in maintaining the functions and ultrastructures of the corneal epithelium, the stroma, and the endothelium.

Aims: To assess the impact of different oxygenation policies on the rate and severity of retinopathy of prematurity (ROP). Methods: Between January 2003 and December 2006, infants of 1500 g birthweight (BW) or less and/or 32 weeks gestational age (GA) or less, and larger, more mature infants with risk factors for ROP were examined through three different time periods: period 1: high target oxygen saturation levels (88–96%) and treatment at threshold ROP; period 2: low target oxygen saturation levels (83–93%) and treatment at threshold ROP; period 3: low target oxygen saturation and treatment at type 1 ROP. Results: Type 1 ROP was detected more frequently in babies of 32 weeks GA or less (50/365, 13.7%) than in more mature babies (15/1167, 1.3%; p

Purpose To determine whether the incidence of and susceptibility to climatic droplet keratopathy (CDK), an acquired, often bilateral degenerative corneal disease, is influenced by the genetic background of the individuals who exhibit the disorder. Methods To determine whether the disease expression was influenced by the genetic ancestry of CDK cases in native Mapuche of the northwest area of Patagonia in Argentina, we examined mitochondrial DNA and Y-chromosome variation in 53 unrelated individuals. Twenty-nine of them were part of the CDK (patient) population, while 24 were part of the control group. The analysis revealed the maternal and paternal lineages that were present in the two study groups. Results This analysis demonstrated that nearly all persons had a Native American mtDNA background, whereas 50% of the CDK group and 37% of the control group had Native American paternal ancestry, respectively. There was no significant difference in the frequencies of mtDNA haplogroups between the CDK patient and control groups. Although the Y-chromosome data revealed differences in specific haplogroup frequencies between these two groups, there was no statistically significant relationship between individual paternal genetic backgrounds and the incidence or stage of disease. Conclusions These results indicate a lack of correlation between genetic ancestry as represented by haploid genetic systems and the incidence of CDK in Mapuche populations. In addition, the mtDNA appears to play less of a role in CDK expression than for other complex diseases linked to bioenergetic processes. However, further analysis of the mtDNA genome sequence and other genes involved in corneal function may reveal the more precise role that mitochondria play in the expression of CDK. Fil: Schurr, Theodore G.. University Of Pennsylvania. Penn Museum Of Archaeology And Anthropology; United State of America; Fil: Dulik, Matthew C.. University Of Pennsylvania. Penn Museum Of Archaeology And Anthropology; United State of America; Fil: Cafaro, Thamara Analia. Consejo Nacional de Investigaciones Científicas y Técnicas.Centro Científico Tecnológico - CONICET - Cordoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina; Fil: Suarez, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas.Centro Científico Tecnológico - CONICET - Cordoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina; Fil: Urrets-Zavalia, Julio A.. Universidad Catolica de Cordoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Fil: Serra, Horacio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas.Centro Científico Tecnológico - CONICET - Cordoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina

We report a case of Alternaria keratitis and hypopyon following clear-corneal cataract surgery. A 66-year-old woman presented with a painful red left eye several months after uneventful self-sealing clear-corneal phacoemulsification that was unresponsive to prolonged treatment with topical/oral quinolones and topical corticosteroids. A full-thickness stromal white dense infiltrate in the area of the intrastromal tunnel incision and a 2.0 mm hypopyon were observed. Culture from corneal scrapings revealed Alternaria species. Treatment included topical and subconjunctival injections of amphotericin-B (5 mg/mL) and 200 mg of oral ketoconazole. Complete resolution of the corneal infiltration and hypopyon was observed after 30 days of treatment, with no recurrence during 6 years of follow-up. To our knowledge, this is the first report of Alternaria species keratitis complicating self-sealing clear-corneal cataract surgery. Topical and subconjunctival injections of amphotericin-B and oral ketoconazole were effective in resolving the corneal abscess and anterior chamber inflammatory reaction. Financial Disclosure No author has a financial or proprietary interest in any material or method mentioned. Fil: Esposito, Evangelina. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Maccio, J. Pablo. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Monti, Rodolfo. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina Fil: Cervi, Laura Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Serra, Horacio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Urrets Zavalía, Julio Alberto. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola; Argentina

PURPOSE. Climatic droplet keratopathy (CDK) is a degenerative disease of the cornea with possible involvement from matrix metalloproteinases (MMPs). Therefore, the authors investigated histologic distribution, levels, and molecular forms of MMP- 2 and MMP-9, as well as tear fluid levels of MMPs and cytokines in CDK patients. They additionally examined UV-B-irradiation effect on production of gelatinases and cytokines by human corneal epithelial (HCE) cell culture model. METHODS. Tears were collected from 20 unrelated individuals (10 with CDK and 10 controls). CDK affected corneas were haematoxylin-eosin stained and the presence and distribution of MMP-2 and MMP-9 was examined using immunohistochemistry. Gelatinases and cytokine secretion was measured in tears and supernatants from UV-B-exposed HCEs by immunoblotting, gelatin zymography, and protein array, respectively. RESULTS. MMP-2 and MMP-9 values were significantly higher in tears collected from CDK patients than healthy controls and were accompanied by pro-inflammatory cytokine secretion. Immunohistochemistry showed that MMP-2 was expressed at the basement membrane zone in both control and affected corneas, but also marked the edges of the granular CDK deposits; MMP-9 expression was restrained to basal layers of the epithelium and was markedly induced in CDK corneas. In HCE cells, UV-B increased gelatinase secretion, with a striking effect on MMP-9, and was preceded by pro-inflammatory cytokine release. CONCLUSIONS. The authors demonstrate that the corneal epithelium could participate in CDK development as a source of cytokines and gelatinases. Additionally, in HCE cells, UV-B- modulated cytokine and subsequent MMP secretion. Local inhibition of cytokine secretion and gelatinases may prevent CDK progression. © 2012 The Association for Research in Vision and Ophthalmology, Inc. Fil: Holopainen, Juha M.. University Of Helsinski; Finlandia Fil: Robciuc, Alexandra. University Of Helsinski; Finlandia Fil: Cafaro, Thamara Analia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Suarez, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Konttinen, Yrjö T.. University Of Helsinski; Finlandia Fil: Alkatan, Hind M.. The Eye Center and The Eye Foundation for Research in Ophthalmology; Arabia Saudita Fil: Tabbara, Khalid F.. The Eye Center and The Eye Foundation for Research in Ophthalmology; Arabia Saudita Fil: Tervahartiala, Taina. University Of Helsinski; Finlandia Fil: Sorsa, Timo. University Of Helsinski; Finlandia Fil: Urrets Zavalía, Julio Alberto. Universidad Catolica de Córdoba. Facultad de Medicina. Clinica Universitaria Reina Fabiola. Dto de Oftalmologia; Argentina Fil: Serra, Horacio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina

Sir, Climatic droplet keratopathy (CDK) is a corneal degenerative disease characterized by its progressive opacity because of accumulation of globular deposits in Bowman's layer (BL) and anterior stroma (AS), as well as abnormal corneal sensitivity.1, 2 We report herein for the first time the study of three patients each with different grades of CDK using in vivo confocal microscopy (iVCM), a technique that has allowed the measurement of normal and pathological corneal components.3, 4, 5 Table 1, Figures 1 and ​and22 summarize the corneal abnormalities found in these patients. Cochet-Bonnet aesthesiometry (CBA) demonstrated a decrease of the mechanical sensitivity of the cornea in eyes with grade II and III CDK. Grade I CDK was characterized by incipient changes in the BL and AS but normal sub-basal and stromal nerve plexus. Grade II CDK showed hyper-reflectivity and globular non-reflective deposits in the BL and AS. In grade III, the AS showed fibrosis with increment of diffused hyper-reflective deposits and large non-reflective deposits. Concomitant to these changes, there was an increased number of dendritic cells (DC) at the peripheral cornea and limbus, but their role in the progression of CDK needs further studies. No abnormalities were found in the rest of the stroma and endothelium.​endothelium. Figure 1 In vivo confocal laser scanning microscopy images. BL: (a) Grade 1: dot-like hyper-reflective deposits in the central cornea. (b) Grade 2: increased density of the deposits. (c) Grade 3: confluent hyper-reflective deposits. Sub-basal nerves: (d) Grade ... Figure 2 In vivo confocal laser scanning microscopy images. Deposits at the peripheral cornea: (a) Grade 1: reflective punctiform or dot-like deposits. (b) Grade 2: hyper-reflective punctate and homogeneous round globular non-reflective deposits. (c) Grade 3: ... Table 1 Abnormalities found in corneas of CDK eyes according to biomicroscopic grade of the disease Although we were unable to study sub-epithelial nerves because they occupied the same area of the deposits in CDK, we found that the early changes in CDK did not affect the sub-basal and stromal nerves, but the progression of the disease lead to a significant density decrease of sub-basal nerves, and some structural changes in stromal nerves, such as uneven thickness and irregular configuration, that might be responsible for corneal hyposensitivity found in advanced stages of CDK. In a recent study, Patel et al5 showed that sub-basal nerve tortuosity but not mean total sub-basal nerve density varied with age in normal individuals. The loss of corneal sensation could not be attributed to the age of our patients as we have previously found in a case–control study that corneal sensation was related to CDK grades and not to the age of individuals.2

lmThe isotype distribution of the antibody response against one Trypanosoma cruzi antigenic fraction, FIV, and the putative association to heart disease were analyzed in patients of two apparently genetically distinct Amerindian populations, Mataco (M) and Toba (T), infected with this parasite. The isotypes profiles were analyzed by ELISA, and the antigen specificity of IgG immune response was determined by the immunoblot method. The percentages of infected individuals with abnormal electrocardiograms (GII) were 50% for population M and 10% for population T. Many individuals from both populations had measureable IgG2, IgG3 and IgG4 antibodies to FIV, but the level and frequency (%) of positive sera in population T was considerably higher than in population M (70% vs 15% for IgG2; 75% vs 40% for IgG3; 85% vs 20% for IgG4). The level and frequency of IgG1 reactivity against FIV were similar in the two populations. When the sera were titrated, the most remarkable difference in isotype levels between populations T and M was seen for IgG2 and IgG4, the T population showing the highest titer. No association between clinical state and a particular isotype profile was found by ELISA in any population. When the antigen specificity of antibody response was determined by immunoblot, the antigen patterns recognized by sera from the two clinical groups showed some differences only in population M. All sera assayed from GII of population M fixed more IgG than those with normal electrocardiograms (GI). Two bands of 36 and 43 kD were revealed only in GII of this population. Similar antigenic patterns between the two clinical groups from population T were observed, and they were comparable with those obtained with GI from population M.

Purpose In the visually debilitating condition of climatic droplet keratopathy, corneal transparency is progressively lost. Although the precise cause of the disease and the mechanism by which it progresses are not known, a lifetime exposure to high solar radiation and a vitamin C–deficient diet may be involved in its development. This study examines the effect of dietary ascorbate levels and ultraviolet (UV)-B exposure on corneal stromal structure. Methods Eight guinea pigs were divided into four treatment groups (A, B, C, and D). For 15 weeks, Groups A and C were fed an ascorbate-rich diet (2 mg/100 g bodyweight/day), while Groups B and D received an ascorbate-deficient diet (0.07 mg/100 g bodyweight/day). For the last 12 weeks of the study, Groups C and D also experienced chronic UVB exposure (0.12 J/cm2 for 40 min/day). Following euthanasia, the corneas were enucleated and their stromal ultrastructure examined using X-ray scattering and electron microscopy. Results UVB exposure resulted in an increased corneal thickness (p

The role of polymorphic residues of the beta chain of human histocompatibility leukocyte antigen-DQw5/w6 in antigen presentation to a hepatitis B surface antigen-specific T cell clone was studied. The results obtained demonstrate that the residue situated at position 57 of the beta chain (a valine) is critical for presentation of antigen by antigen-presenting cells to the DQ-restricted T cell clone. Experiments were also done to study the feasibility of peptide blocking of antigen recognition by DQ-restricted T cells. The results indicate that peptides known to associate with DQ molecules are capable of blocking the presentation of antigen to the DQ-restricted T cell clone, presumably by competing with antigen for binding to major histocompatibility complex (MHC) molecules. Moreover, truncations of the stimulatory antigenic peptide resulted in the production of T cell receptor antagonists, which inhibited the response of the T cells to antigen at 10-100-fold lower concentrations than conventional MHC blockers. The role of DQ-restricted T cell responses and peptide blocking approaches in autoimmunity are discussed.

Purpose: Climatic droplet keratopathy (CDK) is an acquired corneal disease characterized by progressive scarring of the cornea. In several corneal diseases, matrix metalloproteinases (MMPs) are upregulated during the degradation of epithelial and stromal tissues. We investigated the levels, degree of activation and molecular forms of MMP-2, MMP-9, MMP-8 and MMP-13 and their tissue inhibitors TIMP-1 and TIMP-2 in tear fluid of patients with CDK. Methods: Seventeen CDK patients and 10 controls living in Argentine Patagonia received a complete eye examination, and MMPs and TIMP-1 ⁄ 2 were determined by immunofluorometric assay (IFMA), gelatin zymography and quantitative Western immunoblot analysis in tear samples. Results: The MMPs were detected mostly in their latent forms. The levels of MMP-9 and MMP-2 were found to be significantly elevated in CDK patients, whereas latent and active MMP-8 levels were significantly enhanced in controls. There was no significant difference in the level of MMP-13. TIMPs were found as part of complexes, and the TIMP-1 levels were significantly lower in patients than controls. Conclusion: Elevated MMP-2 and MMP-9 levels have been implicated in the failure of corneal re-epithelialization, and enhanced MMP-2 and MMP-9 levels in CDK patients suggest that these MMPs may play a role in corneal scarring in CDK. Elevated levels of MMP-8 suggest a defensive role for this MMP in inflammatory reactions associated with recurring corneal traumas. Decreased expression of TIMP-1 in CDK patients suggest deficient antiproteolytic shield likely to render the corneas of CDK patients vulnerable to enhanced MMPs. Overall, these data suggest a mechanistic link between MMPs and TIMP-1 level in cornea and tears with corneal scarring in CDK.

Glycoproteins are potentially important biomarkers of disease and therapeutic targets. In particular, the N-linked glycoproteins are a focus of interest as they can be found in the extracellular environment and body fluids. In this study, we have sampled the tears, the extracellular fluid of the epithelial cells covering the surface of the eye, of patients with climatic droplet keratopathy (CDK) using tears of unaffected normal patients for comparison. Prefractionation of the tear sample used a hydrazide-resin capture method, and the previously N-glycosylated peptides were then subjected to two-dimensional nano-LC-nano-ESI-MS/MS analysis to obtain peptide fragmentation patterns for identification through protein database searches. We have identified a total of 43 unique N-glycoproteins, 19 of which have not previously been reported in tear fluid. In addition, we have quantitatively compared N-glycoprotein profiles in tear fluid of patients with CDK to tears of nondiseased controls using glycopeptide capture, iTRAQ labeling and 2D nano-LC-nano-ESI-MS/MS analysis. In tears of CDK patients, increased levels of four N-glycosylated proteins including haptoglobin (at sites N207, N211 and N241), polymeric immunoglobulin receptor (at sites N83, N90, N135, N186, N421, and N469), immunoglobulin J chain (at site N49) and an uncharacterized protein DKFZp686M08189 (at site N470), as well as a decrease in the N-glycosylation level of one N-glycosylated protein, lacritin (at site N119) were observed. However, the overall levels of these five proteins showed no appreciable changes between control and CDK samples. The findings could be clinically significant in terms of disease etiology and biomarkers.

Objective To describe the functional and structural characteristics of the cornea in healthy Guinea pigs. Animals studied Healthy male and female pigmented and albino Guinea pigs (Caviaporcellus) aged 3-5 months old were used. Procedures The animals' corneas underwent different in vivo studies including: slit-lamp biomicroscopy, fluorescein staining (FS), break-up time test (BUT), confocal microscopy and pachymetry. The corneas were also studied histopathologically with light microscopy, immunohistochemistry and transmission electron microscopy. Results No significant differences were found between pigmented and albino animals, male and female, OD and OS in any study performed. The differences on corneal thickness values were not significant among central (227.85 +/- 14.09 microm) and upper and temporal peripheral regions (226.60 +/- 12.50 and 225.70 +/- 14.40 microm, respectively). All histological studies performed permitted identification and precise description of the different corneal structures in Guinea pigs: the stratified epithelium (45.52 +/- 5.26 microm), Bowman's layer (2.23 +/- 0.38 microm), stroma (163.69 +/- 4.90 microm), Descemet's membrane (3.96 +/- 0.46 microm) and the endothelium (5.09 +/- 0.71 microm). Combining results from all eyes mean and SD from corneal BUT values was 4.98 +/- 1.67 s. Corneas often showed discrete superficial erosions being the FS positive in both eyes from all the animals. Conclusion This study provides a detailed in vivo and postfixed histological description of the Guinea pig's cornea and information about the physiological tests.