Altered expression of matrix metalloproteinases and their tissue inhibitors as possible contributors to corneal droplet formation in climatic droplet keratopathy.

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.

Scleral Graft and Muscle Transposition in the Treatment of Scleromalacia and Hypertropia Following Retinal Detachment Surgery.

Ocular motility disfunction and eye wall defects are possible complications of retinal detachment (RD) surgery that may interfere with an acceptable functional result. The case of a 41-year-old man that developed scleromalacia and hypertropia after an RD surgery is presented. A cadaveric scleral graft (CSG) was used for the treatment of scleromalacia. Because of absence of the anterior aspect of the inferior rectus muscle, an anterior transposition of the ipsilateral inferior oblique muscle (ATIIO) for the correction of hypertropia was performed in the same surgical procedure. After a follow-up of 22 months, a good evolution of CSG was observed, as well as orthotropia with the exception for extreme inferior gazes. CSG and ATIIO proved to be an effective initial treatment for scleromalacia and hypertropia secondary to RD surgery.

Quantitative analysis of N-linked glycoproteins in tear fluid of climatic droplet keratopathy by glycopeptide capture and iTRAQ.

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.

The cornea of Guinea pig: structural and functional studies.

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.

Macular and extramacular optical coherence tomography findings in X-linked retinoschisis.

Two brothers with X-linked retinoschisis were evaluated using optical coherence tomography. Macular cleavage was observed at the inner nuclear layer. Retinal nerve fiber layer loss or detachment was observed outside the macular area, even in areas of clinically normal retina. Macular thickening and cavitation with multiple septa was characteristic, decreasing rapidly from the center toward the macular periphery and being more pronounced in the younger patient. No abnormal vitreomacular relationship or traction over the macular and extramacular schisis areas was observed. Optical coherence tomography showed a characteristic pattern of macular schisis in X-linked retinoschisis and confirmed a widespread retinal splitting despite mild clinical findings. Middle and outer neurosensory retina was principally compromised in the macular area and the retinal nerve fiber layer in extramacular areas. No vitreous traction was apparently involved in the foveal and extramacular splitting and elevation.

Surface alterations, corneal hypoesthesia, and iris atrophy in patients with climatic droplet keratopathy.

PURPOSE: To present external eye findings and the observation of iris atrophy in patients with climatic droplet keratopathy (CDK). METHODS: 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 P < 0.05. RESULTS: 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. CONCLUSIONS: 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.

Climatic droplet keratopathy in the Argentine Patagonia.

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.

Parálisis unilateral de la elevación / Unilateral elevation palsy

Objetivos: analizar la incidencia y características clínicas de pacientes con parálisis unilateral de la elevación (PUE) en nuestra consulta privada. Pacientes y métodos: evaluamos las historias clínicas de 822 pacientes estrábicos estudiados consecutivamente. Estudiamos la presencia de estrabismo horizontal asociado, de inversión anómala, del fenómeno de Bell, tortícolis y ptosis o pseudoptosis. Estudiamos clínicamente los movimientos sacádicos en el plano vertical. Cuatro pacientes con doble parálisis de los elevadores (DPE) fueron operados mediante la técnica de Knapp modificada. Resultados: 7 pacientes presentaron una PUE. Seis casos correspondían a una DPE presentando hipotropia en el ojo afectado, pseudoptosis y ptosis y fénomeno de Bell negativo ipsilateral. El caso restante correspondía a una parálisis unilateral de la elevación de origen supranuclear presentando ortotropia en PPM y fenómeno de Bell positivo en el ojo afectado. Entre los casos de DPE dos estaban asociados al fenómeno de Marcus Gunn (FMG). Clínicamente no pudimos establecer diferencias en la velocidad sucádica entre ambos ejes en los 7 pacientes estudiados. Los cuatro pacientes con DPE operados mostraron ducciones pasivas normales y obtuvieron un buen alineamiento ocular en PPM. Conclusión: Las PUE son cuadros clínicos infrecuentes. La asociación de PUE y FMG confirmaría el diagnóstico de DPE.

Purpose: to analyze the incidence and clinical characteristics of patients with unilateral elevation palsy (UEP) in our private practice. Patients and methods: 822 clinical recordd of strabismus patients seen consecutively were evaluated. We studied the existence of horizontal strabismus, anomalous innervations, Bell's phenomenon, torticollis and ptosis or pseudoptosis. We studied vertical saccadic movements. Four patients with double elevator palsy (DEP) were operated with a Knapp modified procedure. Results: 7 patients had UEP. Six had double elevator paresis (DEP) presenting hypotropia of the involved eye, pseudoptosis, ptosis and ipsilateral negative Bell's phenomenon. The other case was a supranuclear elevation palsy with ortotropia in primary position and, positive Bell's phenomenon in the affected eye. Two DEP patients were associated with Marcus Gunn phenomenon (MGF). Clinically we couldn't find differences in saccadic velocity between both eyes of 7 patients. The four patients with DEP that were operated had normal passive ductions and attained good results. Conclusions: clinical causes of UEP are infrequent. The association of UEP and MGF would support DEP diagnosis.

Administration of a peptide inhibitor of alpha4-integrin inhibits the development of experimental autoimmune uveitis.

PURPOSE: Recruitment of lymphocytes into the retina and to the vitreous during the development of experimental autoimmune uveitis (EAU) is governed by factors such as the state of activation of inflammatory cells and the repertoire of adhesion molecules expressed by the local vascular endothelia. alpha4 Integrins and their receptors play an important role during homing of cells to the inflammatory site. In the present study, the effect of alpha4-integrin inhibitor on the development of EAU was investigated. METHODS: EAU was induced either by immunizing B10.RIII mice with the 161-180 peptide or by adoptive transfer of interphotoreceptor retinoid-binding protein (IRBP)-specific uveitogenic T cells. Animals were treated with an active peptide inhibitor (alpha4-api) or a peptide control at different time points after induction of disease. EAU was evaluated by histology 21 to 49 days after immunization. Antigen-specific cell proliferation was evaluated by thymidine incorporation. Cytokine synthesis in culture supernatants and anti-IRBP-specific serum IgG1 and IgG2a were evaluated by ELISA. Delayed-type hypersensitivity was evaluated by ear challenge 2 days before the termination of the experiment. RESULTS: Treatment with alpha4-api had a significant ameliorating effect on EAU. The anti-IRBP antibody response and cellular proliferation were not affected by the treatment, whereas delayed-type hypersensitivity was significantly diminished. Cytokine synthesis was not changed by treatment, except for a decrease in IL-10 levels. CONCLUSIONS: The results show that small-molecule inhibitors of alpha4-integrins can act therapeutically in EAU, possibly by interfering with cell adhesion events involved in the development of the disease.

The effect of ketotifen on inflammatory markers in allergic conjunctivitis: an open, uncontrolled study.

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 <0.0082, respectively) as a consequence of the treatment. In 9 out of 19 patients a simultaneous decrease in the percentage of epithelial cells positive for CD29 and eotaxin was observed. 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.