Transcriptome Analysis of Pterygium and Pinguecula Reveals Evidence of Genomic Instability Associated with Chronic Inflammation.

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.

Intestinal microbiota regulates tryptophan metabolism following oral infection with Toxoplasma gondii.

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.

Phospholipidomic Studies in Human Cornea From Climatic Droplet Keratopathy.

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. J. Cell. Biochem. 118: 3920-3931, 2017. © 2017 Wiley Periodicals, Inc.

Myopia as a risk factor for subsequent retinal tears in the course of a symptomatic posterior vitreous detachment.

BACKGROUND: Retinal tears complicating the course of a posterior vitreous detachment (PVD) may be unique or multiple, and when multiple they may occur simultaneously or subsequently at different moments in the evolution of a PVD. The purpose of our study was to analyze the prevalence of subsequent retinal tears (SRT) in patients with a PVD, and to identify possible risk factors for SRT. METHODS: One hundred and seventy six eyes in 165 consecutive patients that presented one or more retinal tears in the evolution of a symptomatic PVD, with a minimum follow-up of 12 months, were retrospectively evaluated. The primary outcome measure was to characterize the clinical features associated with SRT formation against those eyes with non-subsequent retinal tear (NSRT-retinal tear/s diagnosed at initial examination) formation. For that purpose, this cohort of patients was divided into two different groups: group 1 included eyes presenting one or multiple retinal tears only at initial examination (NSRT), and group 2 eyes that progressed to a further retinal tear/s (SRT) during follow-up. RESULTS: Group 1 comprised 154 eyes from 145 patients, 48.7% males and 51.3% females with a mean age of 56.9 ± 14.0 years (range = 15-89); 17.2% of patients had a previous retinal tear or retinal detachment in the fellow eye; mean number of retinal tears per eye 1.42 ± 0.8 (range = 1-5); 20.8% presented bilateral retinal tears; 59.1% were myopic eyes (p < 0.05). Group 2 comprised 22 eyes from 20 patients; mean age was 53.3 ± 13.6 years (range = 30-69); 63.6% were male (p = 0.13), and 7 patients (31.8%) had a history of SRT or retinal detachment in the fellow eye (p = 0.13). The mean number of retinal tears per eye was 1.36 ± 0.5 (range = 1-2); bilateral retinal tears were noted in 18.2% of eyes; 86.4% were myopic eyes (p = 0.01); 81.8% occurred within a 120 days-period following diagnosis of the first retinal tear. CONCLUSIONS: Multiple retinal tears may be diagnosed in the evolution of a PVD. SRT are most frequently observed in myopic patients, and are usually symptomatic. Follow-up must extend for at least 4 months after the initial symptoms.

Deimination level and peptidyl arginine deiminase 2 expression are elevated in astrocytes with increased incubation temperature.

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, S100ß, 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.

Systemic amyloidosis with bilateral conjunctival involvement: a case report.

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 Is a Misnomer for This Corneal Degeneration.

PURPOSE: To describe a new insight into the Climate Droplet Keratopathy (CDK) pathophysiology and its major predisposing factors. METHOD: A literature search was undertaken on PubMed to compile papers published on CDK. The following is a focused opinion tempered by synthesis of current evidence, and research of the authors. RESULTS: CDK is a multifactorial rural disease common in regions with high incidence of pterygium, but not related to the type of climate or ozone concentrations. Although it has been thought that climate is the cause of this disease, recent investigations deny that and reveal that other environmental factors such as dietary intake, eye protection, oxidative stress, and ocular inflammatory pathways play an important role in the pathogenesis of CDK. CONCLUSION: Considering the negligible effect of climate, the present name " CDK" for this illness can be confusing for young ophthalmologists. Based on these remarks, it is imperative to start using an accurate name like "Environmental Corneal Degeneration (ECD)" that fits the most recent evidence related to its etiology.

The effect of vitamin C deficiency and chronic ultraviolet-B exposure on corneal ultrastructure: a preliminary investigation.

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<0.001), but this was not accompanied by a widespread expansion of the collagen fibrillar array, and in the case of ascorbate-deficient animals, stromal thickening was associated with the compaction of collagen fibrils (p<0.01). Neither UVB exposure nor ascorbic acid deficiency caused any change in the average diameter or D-periodicity of the stromal collagen fibrils. CONCLUSIONS: UVB-induced changes in the corneal ultrastructure were most pronounced in animals fed an ascorbic acid-deficient diet. This suggests that ascorbic acid may play a vital role in protecting the corneal stroma from the harmful effects of UVB.

Peptidylarginine deiminase type 2 is over expressed in the glaucomatous optic nerve.

PURPOSE: To determine levels of Peptidyl arginine deiminase 2 (PAD2) and its product protein-bound citrulline in cadaver eyes that suffered from normal tension glaucoma (NTG) compared to primary open angle glaucoma (POAG), and controls. METHODS: Western analysis, ELISA, and immunohistochemical analysis were performed with human tissues. RESULTS: We report over expression of PAD2 and higher levels of its product protein-bound citrulline in the optic nerve of normal tension glaucoma patients (NTG). CONCLUSIONS: This is the first report demonstrating that like in POAG, NTG also possesses elevated levels of both PAD2 and protein-bound citrulline.

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.

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.

The eye and the skin in endocrine metabolic diseases.

The eye and skin may offer critical clues to the diagnosis of a varied spectrum of metabolic diseases from endocrine origin and their different stages of severity, such as diabetes mellitus and Graves disease. On the other hand, such entities may compromise the eye and visual function severely, and awareness of these possible associations is an important step in their diagnosis and management. A large number of less common endocrine diseases may also have significant ocular/visual or skin involvement. Often the etiologic relationship between the endocrine metabolic disease and the ocular compromise is unknown, but diverse pathogenetic mechanisms may act through a common pathologic pathway producing ocular damage, as occur in diabetic retinopathy. This review emphasizes the ocular and skin manifestations of different metabolic diseases of endocrine origin.

The eye and the skin in nonendocrine metabolic disorders.

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.

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.

Climatic droplet keratopathy: an old disease in new clothes.

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.

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.

Alternaria keratitis and hypopyon after clear-cornea phacoemulsification.

UNLABELLED: 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.

Genetic background and climatic droplet keratopathy incidence in a Mapuche population from Argentina.

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.