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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.

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

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

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

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.

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 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

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

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

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.

To identify the proteins in the corneal droplets of climatic droplet keratopathy (CDK), a disease that results in the formation of droplets on the cornea. Progressive accumulation of droplets in CDK leads to visual loss.Proteomic mass spectrometry of the CDK specimens was performed after fractionation of proteins in 4% to 20% SDS-polyacrylamide gels. Droplets were derived from two human donors. Immunohistochemistry with antibodies was performed to confirm the presence of identified proteins on donor tissues from patients with CDK and control subjects.Proteomic analyses revealed identification of 105 proteins in CDK specimens. Immunohistochemical analyses confirmed localization of annexin A2 and glyceraldehyde 3-dehydrogenase (GAPDH), proteins identified by proteomic analyses in CDK specimens. The proteins were subjected to analyses with the Kyoto Encyclopedia of Genes and Genomes (KEGG) Database which showed that a few biochemical pathways were more frequent for the identified proteins.Approximately 105 proteins were identified in CDK specimens, and a subset of them was confirmed by immunohistochemistry. Several of these may play a role in fibril or deposit formation.

Intraocular inflammatory diseases are a common cause of severe visual impairment and blindness. In this study, we investigated the immunoregulatory role of galectin-1 (Gal-1), an endogenous lectin found at sites of T cell activation and immune privilege, in experimental autoimmune uveitis (EAU), a Th1-mediated model of retinal disease. Treatment with rGal-1 either early or late during the course of interphotoreceptor retinoid-binding protein-induced EAU was sufficient to suppress ocular pathology, inhibit leukocyte infiltration, and counteract pathogenic Th1 cells. Administration of rGal-1 at the early or late phases of EAU ameliorated disease by skewing the uveitogenic response toward nonpathogenic Th2 or T regulatory-mediated anti-inflammatory responses. Consistently, adoptive transfer of CD4(+) regulatory T cells obtained from rGal-1-treated mice prevented the development of active EAU in syngeneic recipients. In addition, increased levels of apoptosis were detected in lymph nodes from mice treated with rGal-1 during the efferent phase of the disease. Our results underscore the ability of Gal-1 to counteract Th1-mediated responses through different, but potentially overlapping anti-inflammatory mechanisms and suggest a possible therapeutic use of this protein for the treatment of human uveitic diseases of autoimmune etiology. Fil: Toscano, Marta Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina Fil: Commodaro, Alessandra G.. Universidade de Sao Paulo; Brasil Fil: Ilarregui, Juan Martin. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Bianco, German Ariel. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Liberman, Ana Clara. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Serra, Horacio M.. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina Fil: Jun Hirabayashi. National Institute of Advanced Industrial Science and Technology; Japón Fil: Rizzo, Luiz V.. Universidade de Sao Paulo; Brasil Fil: Rabinovich, Gabriel Adrián. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina

Background The efficacy and safety of ketotifen eye drop treatment in allergic conjunctivitis (AC) management is perfectly known by several studies, but the mechanism of action at the biochemical levels is poorly understood so we decided to perform an open, uncontrolled study in order to investigate the effect of the topical administration of ketotifen fumarate 0.05% on biochemical markers of inflammation on conjunctival cells in patients with AC. Methods Nineteen patients with symptoms and signs of AC (itching, discharge, burning, redness, increase in the watery discharge, swelling and follicles) and with a history of allergy were prescribed with two daily instillation of one drop of eyewash ketotifen fumarate 0,05% in both eyes during thirty days. They were studied by measuring clinical and immunologic parameters. Results Ketotifen fumarate treatment significantly reduced the total symptoms and signs score for each patient as well as each symptoms and signs at all time points compared with day 0 (p < 0.0001 and p < 0.016, respectively). Although the percentage of HLA-DR+ epithelial cells diminished only in 58% of patients, the numbers of CD29+ and eotaxin+ epithelial cells dropped significantly in 68% and 73 % of them (p < 0.0062 and Conclusion Ketotifen besides the well-known effect in reducing signs and symptoms of AC significantly diminished production of eotaxin and expression of CD29 by epithelial cells in patients with seasonal AC.

Background Allergic Contact Dermatitis (ACD) is regarded as a T-cell-mediated delayed-type hypersensitivity reaction. We studied the kinetics of the expression of CS-1 fibronectin, thymus and activation-regulated chemokine (CCL17/ TARC) and different chemokine receptors (CR) in skin biopsies from individuals suffering from back problems, with the antigen responsible of their contact dermatitis and an irrelevant antigen. Methods Samples were taken at 2, 10, and 48 hours for histological and immunohistochemical studies using monoclonal antibodies against human CS-1 fibronectin, CCL17, CD3, CD68, CD49d, CXCR3, CCR5, and CCR3. Results At positive antigen stimulated sites there was an early expression of CS-1 fibronectin (2 hours), followed by CCL17 and a later accumulation of alplha4/beta1+ (CD49d), CD3+, CD68+, CXCR3+ and CCR5+ mononuclear cells. At 48 hours, approximately 59 % of infiltrating cells were CXCR3+, 42% CCR5+, and only 14 % CCR3+. Conclusions These results showed for the first time a very early expression of CS-1 fibronectin which preceded production of CCL17 in blood endothelial cells (BCEs) from patients' skin with ACD. The role of these molecules in recruitment of monocytes and effector T cells in ACD is discussed.

Expression of murine CCL21 by dermal lymphatic endothelial cells (LEC) has been demonstrated to be one of the most important steps in Langerhans cell emigration from skin. Previously, our group and others have found that this chemokine is up-regulated in different human inflammatory skin diseases mediated by diverse specific immune responses. This study was carried out to investigate the involvement of CCL21 in human skin after challenge with irritant agents responsible for inducing Irritant Contact Dermatitis (ICD). Eleven normal individuals were challenged with different chemical or physical irritants. Two patients with Allergic Contact Dermatitis (ACD) were also challenged with the relevant antigen in order to have a positive control for CCL21 expression. Macroscopic as well as microscopic responses were evaluated. We observed typical ICD responses with mostly mononuclear cells in perivascular areas, but a predominance of polymorphonuclear cells away from the inflamed blood vessels and in the epidermis at 24 hours. Immunohistochemical studies showed up-regulation of CCL21 by lymphatic endothelial cells in all the biopsies taken from ICD and ACD lesions compared to normal skin. Kinetic study at 10, 48, 96 and 168 hours after contact with a classical irritant (sodium lauryl sulphate) showed that the expression of CCL21 was increased in lymphatic vessels at 10 hours, peaked at 48 hours, and then gradually declined. There was a strong correlation between CCL21 expression and the macroscopic response (r = 0.69; p = 0.0008), but not between CCL21 and the number of infiltrating cells in the lesions. These results provide new evidence for the role of CCL21 in inflammatory processes. Since the up-regulation of this chemokine was observed in ICD and ACD, it is tempting to speculate that this mechanism operates independently of the type of dermal insult, facilitating the emigration of CCR7+ cells.

Cytotoxic T lymphocytes (CTLs) recognize peptide antigens associated with cell surface major histocompatibility complex (MHC) molecules. The identification of tumor cell-derived peptides capable of eliciting anti-tumor CTL responses would enable the design of antigen-specific immunotherapies. Our strategy to identify such potentially therapeutic peptides relies on selecting high-affinity MHC binders from known tumor-associated antigens. These peptides are subsequently tested for their ability to induce CTLs capable of killing tumor cells. With this strategy, we have identified a nine-residue epitope, derived from the product of the tumor-associated gene MAGE-3, which has the capacity to induce in vitro CTLs that kill melanoma and other tumor cell lines. These results show the primary in vitro induction of tumor-specific human CTLs and illustrate the feasibility of ex vivo antigen-specific approaches to the immunological therapy of cancer.