Additional file 1: of Coping with dry eyes: a qualitative approach

Supplementary materials. Script for Dry Eye Focus Group. Table S1. Examples of strategies used in the themes uncovered in the focus groups. Table S2. The number of times focus group themes were mentioned by participants. (DOCX 21 kb

Need for Animal Models of Meibomian Gland Dysfunction

<p><b>Article full text</b></p> <p><br></p> <p>The full text of this article can be found here<b>.</b> <a href="https://link.springer.com/article/10.1007/s40123-016-0070-6">https://link.springer.com/article/10.1007/s40123-016-0070-6</a></p><p></p> <p><br></p> <p><b>Provide enhanced content for this article</b></p> <p><br></p> <p>If you are an author of this publication and would like to provide additional enhanced content for your article then please contact <a href="http://www.medengine.com/Redeem/”mailto:[email protected]”"><b>[email protected]</b></a>.</p> <p><br></p> <p>The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.</p> <p><br></p> <p>Other enhanced features include, but are not limited to:</p> <p><br></p> <p>• Slide decks</p> <p>• Videos and animations</p> <p>• Audio abstracts</p> <p>• Audio slides</p

MicroRNA-215 Regulates Fibroblast Function: Insights from a Human Fibrotic Disease

<p>MicroRNAs are implicated in the regulation of gene expression via various mechanisms in health and disease, including fibrotic processes. Pterygium is an ocular surface condition characterized by abnormal fibroblast proliferation and matrix deposition. We aimed to investigate the role of microRNAs in pterygium and understand the relevant cellular and molecular mechanisms. To achieve this objective, a combination of approaches using surgically excised paired human pterygium and conjunctival tissues as well as cultured primary fibroblast cells from tissue explants were evaluated. Fibroblast dysfunction has been shown to play a central role in pterygium pathology. Here we show that miR-215, among a few others, was down-regulated (2-fold) in pterygium compared to control, and this was consistent in microarray, real-time PCR and fluorescent <i>in-situ</i> hybridization. The effects of increased miR-215 were investigated by adding exogenous miR-215 to fibroblasts, and this showed a decrease in cell proliferation but no significant apoptosis compared to control. Further cell cycle analysis showed that miR-215 depressed progression of cells at G1/S as well as G2/M. A few cell cycle related transcripts were downregulated (2.2–4.5-fold) on addition of miR-215: Mcm3, Dicer1, Cdc25A, Ick, Trip13 and Mcm10. Theoretic binding energies were used to predict miR-215 binding targets and luciferase reporter studies confirmed Mcm10 and Cdc25A as direct targets. In summary, mir-215 could play a role in inhibiting fibroblast proliferation in ocular surface conjunctiva. Dampening of this mir-215 could result in increased fibroblast cell cycling and proliferation, with possibly increased fibroblastic production of matrix, inducing pterygium formation.</p

Dry Eye-Related Visual Blurring and Irritative Symptoms and Their Association with Depression and Anxiety in Eye Clinic Patients

<p><i>Purpose</i>: Dry eye is a common condition known to have many systemic co-morbidities. We aim to report the frequency of depression and anxiety suspects in eye clinic patients and explore the association between dry eye signs and symptoms with depression and anxiety.</p> <p><i>Materials and methods</i>: Adult outpatients were recruited from dry a public tertiary ophthalmology specialist center. Participants underwent socio-demographic and dry eye symptoms questionnaires, fluorescein dye tear break-up time (TBUT), fluorescein dye corneal staining and Schirmer’s test. Rasch analysis was used to internally validate the Hospital Anxiety and Depression Scale (HADS) and the Center for Epidemiologic Studies Depression Scale (CES-D) and determine its psychometric properties.</p> <p><i>Results</i>: A total of 96 participants (mean age 54.5 ± 10.8; 64.1% women) were recruited. The mean score for the CES-D was 12.1 ± 9.8 with 28 participants (31.5%) having a score of above 16 (published threshold). The mean score for depression-associated questions in the HADS was 3.6 ± 3.6 with 13 participants (14.1%) having a score ≥8. The mean score for the anxiety-associated questions in the HADS was 5.3 ± 3.6 with 24 participants (26.1%) having score ≥8. There was a weak inverse correlation between Schirmer’s test and CES-D scores. Elevated depression scores from the CES-D were significantly associated with visual blurring but not irritation, in dry eye patients with low TBUT.</p> <p><i>Conclusions</i>: In conclusion, specific types of dry eye symptoms, particularly those with increased frequency of visual blurring were associated with symptoms of depression. The optical aberrations induced by dry eye, associated with tear instability, may predispose to depressive tendency.</p

Cdc42 and TCL are required for HCET cell polarization.

<p>A. SiRNA transfected HCET cells stained with Golgi body (GM130) (green). Nuclei were counterstained with DAPI (blue). B. Typical cells that were polarized (left) and not polarized (right) were shown. C. Percentage of polarized cells. Percentage of polarized cells among cells transfected with Cdc42 or TCL siRNA were significantly lower than control siRNA transfected cells in both scratch assay and non-traumatic assay. * P<0.05.</p

Reverse transcription-PCR results showing the detection of various members of Rho proteins in human corneal epithelial cells (A), HeLa cells (B) and primary limbal cells (C).

<p>Reverse transcription-PCR results showing the detection of various members of Rho proteins in human corneal epithelial cells (A), HeLa cells (B) and primary limbal cells (C).</p

Inhibition of Cdc42 or TCL by dominant-negative plasmids delayed HCET cell migration.

<p>A. Transfection efficiency of control plasmids and dominant negative plasmids were similar when evaluated by observing GFP under fluorescence microscope. B. <i>In </i><i>vitro</i> scratch wounding assay performed on human corneal epithelial cells transfected with Cdc42, TCL, TC10 or Chp dominant-negative plasmids. Control plasmid transfected cells and cells transfected with TC10, Chp dominant-negative plasmids closed wound faster than Cdc42 or TCL dominant-negative plasmids transfected cells. C. Non-traumatic cell migration assay performed on human corneal epithelial cells transfected with Cdc42, TCL, TC10 or Chp dominant-negative plasmids. Results were similar with scratch wounding assay. </p

Inhibition of Cdc42 by siRNA impaired PAK4 localization.

<p>HCET cells transfected with control siRNA or Cdc42 siRNA were wounded and allowed to migrate for 1hr. Immunostaining for PAK4 were performed. Green: PAK4; Blue: Nulcei. PAK4 junction localization was destroyed at both central region and wound edge. Arrows point to leading edge.</p

Inhibition of Cdc42 or TCL by siRNA delayed Non-traumatic cell migration.

<p>A. Non-traumatic cell migration assays performed on human corneal epithelial cells transfected with siRNA. Migration of cells transfected with siRNA targeting Cdc42 or TCL was delayed compared to control siRNA transfected cells. B. Western blot showed that Cdc42 or TCL were inhibited efficiently by individual siRNA 48hrs after transfection. The ratio of Cdc42 and TCL to loading control were shown. C. Chart of the averaged wound size change against time. Results shown are for siRNA transfected non-traumatic cell migration assay over 12 hours. Wound closure rate in cells transfected with siRNA targeting Cdc42 were significantly slower than control cells. *p<0.05.</p

Evaluation of Global Differential Gene and Protein Expression in Primary Pterygium: S100A8 and S100A9 as Possible Drivers of a Signaling Network

<div><p>Purpose</p><p>Pterygium is a wing shaped fibrovascular growth on the ocular surface, characterized by fibrosis, angiogenesis, extracellular matrix remodeling, and inflammatory infiltrates. Epidemiologic studies have linked pterygium formation to various chronic inflammatory conditions, such as ultraviolet radiation, sawdust exposure, and dry eye disease. The purpose of this study is to identify proteins that are differentially expressed in primary pterygium by using a combination of gene microarray and proteomic platforms.</p><p>Methods</p><p>Paired pterygium and uninvolved conjunctiva tissues of four patients were evaluated for differences in global gene transcript levels using a genechip microarray. Proteins extracted from another four pairs of tissues were quantified by iTRAQ approach. Western blot and immunofluorescent staining on additional patients were used to validate dysregulated protein expression obtained from microarray and proteomics data. In addition, primary conjunctival fibroblasts were treated with recombinant S100A8, S100A9 or both. Transcript level changes of a panel of potential target genes were evaluated by real time-PCR.</p><p>Results</p><p>The following were up-regulated at both protein and transcript levels S100 A8 and A9, aldehyde dehydrogenase 3 family, member1 (ALDH3A1) and vimentin (VIM). Conversely, serpin peptidase inhibitor clade A member 1 (SERPINA1) and transferrin (TF) were down-regulated. Upon adding S100A8, S100A9 or both, the inflammatory chemokine CXCL1, matrix proteins vimentin, biglycan, and gelsolin, as well as annexin-A2, thymosin-β4, chymase (CMA1), member of Ras oncogene family RAB10 and SERPINA1 were found to be up-regulated.</p><p>Conclusions</p><p>We identified 3 up-regulated and 2 down-regulated proteins by using a stringent approach comparing microarray and proteomic data. On stimulating cells with S100A8/9, a repertoire of key genes found to be up-regulated in pterygium tissue, were induced in these cells. S100A8/9 may be an upstream trigger for inflammation and other disease pathways in pterygium.</p></div