What's the situation with ocular inflammation? A cross-seasonal investigation of proteomic changes in ocular allergy sufferers' tears in Victoria, Australia.

Background: Ocular allergy (OA) is a localized subset of allergy characterized by ocular surface itchiness, redness and inflammation. Inflammation and eye-rubbing, due to allergy-associated itch, are common in OA sufferers and may trigger changes to the ocular surface biochemistry. The primary aim of this study is to assess the differences in the human tear proteome between OA sufferers and Healthy Controls (HCs) across peak allergy season and off-peak season in Victoria, Australia. Methods: 19 participants (14 OA sufferers, 5 HCs) aged 18-45 were recruited for this study. Participants were grouped based on allergy symptom assessment questionnaire scoring. Proteins were extracted from human tear samples and were run on an Orbitrap Mass Spectrometer. Peaks were matched to a DIA library. Data was analyzed using the software MaxQuant, Perseus and IBM SPSS. Results: 1267 proteins were identified in tear samples of OA sufferers and HCs. 23 proteins were differentially expressed between peak allergy season OA suffers vs HCs, and 21 were differentially expressed in off-peak season. Decreased proteins in OA sufferers related to cell structure regulation, inflammatory regulation and antimicrobial regulation. In both seasons, OA sufferers were shown to have increased expression of proteins relating to inflammation, immune responses and cellular development. Conclusion: Tear protein identification showed dysregulation of proteins involved in inflammation, immunity and cellular structures. Proteins relating to cellular structure may suggest a possible link between OA-associated itch and the subsequent ocular surface damage via eye-rubbing, while inflammatory and immune protein changes highlight potential diagnostic and therapeutic biomarkers of OA.

The Plight of the Metabolite: Oxidative Stress and Tear Film Destabilisation Evident in Ocular Allergy Sufferers across Seasons in Victoria, Australia.

Ocular allergy (OA) is characterised by ocular surface itchiness, redness, and inflammation in response to allergen exposure. The primary aim of this study was to assess differences in the human tear metabolome and lipidome between OA and healthy controls (HCs) across peak allergy (spring-summer) and off-peak (autumn-winter) seasons in Victoria, Australia. A total of 19 participants (14 OA, 5 HCs) aged 18-45 were recruited and grouped by allergy questionnaire score. Metabolites and lipids from tear samples were analysed using mass spectrometry. Data were analysed using TraceFinder and Metaboanalyst. Metabolomics analysis showed 12 differentially expressed (DE) metabolites between those with OA and the HCs during the peak allergy season, and 24 DE metabolites were found in the off-peak season. The expression of niacinamide was upregulated in OA sufferers vs. HCs across both seasons (p ≤ 0.05). A total of 6 DE lipids were DE between those with OA and the HCs during the peak season, and 24 were DE in the off-peak season. Dysregulated metabolites affected oxidative stress, inflammation, and homeostasis across seasons, suggesting a link between OA-associated itch and ocular surface damage via eye rubbing. Tear lipidome changes were minimal between but suggested tear film destabilisation and thinning. Such metabolipodome findings may pave new and exciting ways for effective diagnostics and therapeutics for OA sufferers in the future.

Development and validation of a health practitioner survey on ocular allergy.

Survey studies have played a significant role in understanding the gaps in the knowledge and practices of health practitioners. However, there have been no such survey studies on Ocular Allergy (OA). Thus, the purpose of this study was to develop and validate a survey on OA to better understand the gaps in the diagnostic, treatment, and collaborative care approaches of health practitioners in OA. The survey is titled "Survey on Ocular Allergy for Health Practitioners (SOAHP)". SOAHP was developed in a five-stage process. First, item extraction via the use of a literature review, second, face and content validity, third, a pilot study, fourth, test-retest reliability, and fifth, finalisation of the survey. 65 items under 6 domains were initially generated in the item extraction phase. Content validity was conducted on 15 experts in the field. This was conducted twice to reach consensus whereby items and domains were added, edited, kept, or removed, resulting in 50 items under 7 domains. The pilot study was conducted on 15 participants from the five relevant health practitioner fields (Allergists/Immunologists, General Practitioners (GPs), Ophthalmologists, Optometrists and Pharmacists). This altered the survey further to 40 items under 7 domains. Test-retest reliability was conducted on 25 participants from the five health practitioner fields. Reliability was moderate to almost perfect for most (97%) investigated items. The finalised survey was 40 items under 7 domains. SOAHP is the first survey created to assess diagnostic, treatment and collaborative care approaches of Allergists/Immunologists, GPs, Ophthalmologists, Optometrists and Pharmacists on OA. SOAHP will be a useful tool in clinical research on OA.

Preserved Ophthalmic Anti-Allergy Medication in Cumulatively Increasing Risk Factors of Corneal Ectasia.

The prevalence of allergies is rising every year. For those who suffer from it, ocular inflammation and irritation can be inconvenient and unpleasant. Anti-allergy eyedrops are a readily available treatment for symptoms of ocular allergy (OA) and can help allergy sufferers regain normal function. However, the eye is a delicate organ, and multiuse eyedrops often utilise preservatives to deter microbial growth. Preservatives such as benzalkonium chloride (BAK) have been shown to induce decreased cell viability. Therefore, during a period of high localised inflammation and eye rubbing, it is important that the preservatives used in topical medicines do not contribute to the weakening of the corneal structure. This review explores ocular allergy and the thinning and protrusion of the cornea that is characteristic of the disease keratoconus (KC) and how it relates to a weakened corneal structure. It also describes the use of BAK and its documented effects on the integrity of the cornea. It was found that atopy and eye rubbing are significant risk factors for KC, and BAK can severely decrease the integrity of the corneal structure when compared to other preservatives and preservative-free alternatives.

A Review of Emerging Tear Proteomics Research on the Ocular Surface in Ocular Allergy.

Ocular allergy is an immunoglobulin E-mediated Type I hypersensitivity reaction localised to the ocular surface and surrounding tissues. Primary signs and symptoms of ocular allergy include itching, redness, irritation and inflammation. Eye-rubbing caused by itching has been shown to alter ocular surface protein concentrations in conditions linked to ocular allergy such as keratoconus. In keratoconus, the cornea begins to thin and sag over time, leading to progressive vision loss and blindness in severe conditions. Due to the high incidence of ocular allergy sufferers rubbing their eyes in response to symptoms of itching, the protein landscape of the ocular surface may be significantly altered. Differential protein expression caused by long-term inflammation and eye-rubbing may lead to subsequent changes in ocular surface structure and function over time. This review aims to summarise and explore the findings of current ocular allergy proteome research conducted using techniques such as gel electrophoresis, mass spectrometry and lab-on-a-chip proteomics. Proteins of interest for this review include differentially expressed immunoglobulins, mucins, functional proteins, enzymes and proteins with previously uncharacterised roles in ocular allergy. Additionally, potential applications of this research are addressed in terms of diagnostics, drug development and future research prospects.

Questionnaires Assessing the Quality of Life of Ocular Allergy Patients.

Over the last 25 years, health-related quality of life has received increasing recognition, as it aids health practitioners in understanding the way a patient may be impacted by their health condition. Specifically, ocular allergy has been found to affect an individual emotionally, physically, socially, and economically. Hence, scientists have developed multiple questionnaires, based on the different etiologies of ocular allergy, to assess the quality of life of individuals affected by the condition. One highly regarded questionnaire is the Rhinoconjunctivitis Quality of Life Questionnaire and its variations, namely the Standardised Rhinoconjunctivitis Quality of Life Questionnaire, Mini Rhinoconjunctivitis Quality of Life Questionnaire, Nocturnal Rhinoconjunctivitis Quality of Life Questionnaire, Adolescent Rhinoconjunctivitis Quality of Life Questionnaire, and Paediatric Rhinoconjunctivitis Quality of Life Questionnaire. Other questionnaires include the Eye Allergy Patient Impact Questionnaire and the Quality of Life of Children with Allergic Keratoconjunctivitis questionnaire, among others that are suitable for different countries. The purpose of this commentary was to identify the advantages and disadvantages of each questionnaire by critically analyzing psychometric properties, identifying which ocular allergy domains are present, and evaluating additional features that are important to a questionnaire.

To See or Not to See: A Systematic Review of the Importance of Human Ocular Surface Cytokine Biosignatures in Ocular Allergy.

Cytokines are key cell signalling proteins in a number of immune and homeostatic pathways of the human body. In particular, they mediate intracellular mechanisms of allergy on the ocular surface by triggering cellular responses that result in typical physiological ocular allergy symptoms, such as itchiness, watery eyes, irritation, and swelling. Given the recent research focus in optometry on the aetiology of corneal ectasia subtypes like keratoconus, there is an increasing need for the development of new clinical diagnostic methods. An increasing trend is evident among recent publications in cytokine studies, whereby the concentrations of cytokines in healthy and disease states are compared to derive a specific cytokine profile for that disease referred to as 'biosignatures'. Biosignatures have diagnostic applications in ocular allergy as a cheap, non-invasive alternative to current techniques like IgE antibody testing and skin prick tests. Cytokine detection from tear samples collected via microcapillary flow can be analysed either by enzyme-linked immunosorbent assays (ELISA), multiplex magnetic bead assays, or immunoblot assays. Characterising patient hypersensitivities through diagnostic tests is the first step to managing exposure to triggers. Investigating cytokine biosignatures in ocular allergy and their links to physiology are imperative and will be the focus of this systematic review article.

Non-invasive objective and contemporary methods for measuring ocular surface inflammation in soft contact lens wearers - A review.

Contact lens wear is one of the primary risk factors for the development of ocular surface inflammatory events. The purpose of this review is to examine and summarize existing knowledge on the mechanisms of contact lens related ocular surface inflammation and the evidence for the effectiveness of current objective methods to measure ocular surface inflammation. Contact lens wear is postulated to trigger an inflammatory response on the ocular surface due to mechanical, chemical, hypoxic stress, or by the introduction of microbes and their toxins. Apart from the traditional signs of inflammation, such as swelling, oedema, redness and heat, on the ocular surface, other methods to measure ocular surface inflammation in sub-clinical levels include tear inflammatory mediator concentrations, conjunctival cell morphology, and corneal epithelial dendritic cell density and morphology. Tear inflammatory mediator concentrations are up- or down-regulated during contact lens wear, with or without the presence of associated inflammatory events. There is higher conjunctival cell metaplasia observed with contact lens wear, but changes in goblet cell density are inconclusive. Dendritic cell density is seen to increase soon after initiating soft contact lens wear. The long term effects of contact lens wear on dendritic cell migration in the cornea and conjunctiva, including the lid wiper area, require further investigation. Currently patient factors, such as age, smoking, systemic diseases and genetic profile are being studied. A better understanding of these mechanisms may facilitate the development of new management options and strategies to minimize ocular surface inflammation related to contact lens wear.

Substance P in Flush Tears and Schirmer Strips of Healthy Participants.

PURPOSE: To determine the repeatability of the flush tear collection technique and the Schirmer strip for Substance P tear analysis. METHODS: The tears of 10 healthy non-contact-lens wearers were collected via Schirmer strip and microcapillary following instillation of either 20 µL (F-20) or 60 µL (F-60) of saline. Each technique was conducted on two occasions and in a randomized order. Total protein content (TPC) and Substance P concentrations were determined. The overall protein separation profile of each type of tears was examined using one-dimensional gel electrophoresis (1DGE). RESULTS: Collection rates were significantly faster for the F-60 compared to F-20 (17.3 ± 6.9 µL/min and 11.9 ± 5.3 µL/min, respectively, P < .001), with an average Schirmer strip length of 1.5 ± 2.1 mm/min. The coefficient of repeatability between days and eyes was greatest for the Schirmer strip, with eyes and days being significantly different (P = .03 and P = .03, respectively) for Schirmer strip Substance P. TPC was 3.8 ± 2.6 mg/mL, 3.3 ± 1.8 mg/mL, and 3.6 ± 3.0 mg/mL for F-20, F-60, and Schirmer strip techniques, respectively, with no significant difference between techniques (P = .85). Substance P concentration was 13.1 ± 14.8 ng/mL, 9.1 ± 6.1 ng/mL, and 14.9 ± 10.6 ng/mL for F-20, F-60, and Schirmer strip tears, respectively, with no significant difference between techniques (P = .57). 1DGE profile showed similar electrophoresis patterns among F-20, F-60, and basal tears. CONCLUSIONS: The F-60 method allows faster collection than F-20, but the latter results in better repeatability than both the F-60 and Schirmer sampling techniques. All three techniques return the same concentrations of TPC and Substance P. This indicates that tear collection using the F-20 may be more appropriate when conducting comparative analysis, whereas the F-60 may be more appropriate when more volume is required.

In situ osmometry: validation and effect of sample collection technique.

PURPOSE: To compare tear film osmolarity measurements between in situ and vapor pressure osmometers. Repeatability of in situ measurements and the effect of sample collection techniques on tear film osmolarity were also evaluated. METHODS: Osmolarity was measured in one randomly determined eye of 52 healthy participants using the in situ (TearLab Corporation, San Diego, CA) and the vapor pressure (Vapro 5520; Wescor, Inc., Logan, UT) osmometers. In a subset of 20 participants, tear osmolarity was measured twice on-eye with the in situ osmometer and was additionally determined on a sample of nonstimulated collected tears (3 µL) with both instruments. RESULTS: Mean (SD) tear film osmolarity with the in situ osmometer was 299.2 (10.3) mOsmol/L compared with 298.4 (10) mmol/kg with the vapor pressure osmometer, which correlated moderately (r = 0.5, P < 0.05). Limits of agreement between the two instruments were -19.7 to +20.5 mOsmol/L. Using collected tears, measurements with the vapor pressure osmometer were marginally higher (mean [SD], 303.0 [11.0] vs 299.3 [8.0] mOsmol/L; P > 0.05) but correlated well with those using the in situ osmometer (r = 0.9, P < 0.05). The mean (SD) osmolarity of on-eye tears was 5.0 (6.6) mOsmol/L higher than that of collected tears, when both measurements were conducted with the in situ osmometer. This was a consistent effect because the measurements correlated well (r = 0.65, P < 0.05).The in situ osmometer showed good repeatability with a coefficient of repeatability of 9.4 mOsmol/L (r = 0.8, P < 0.05). CONCLUSIONS: Correlation between the two instruments was better when compared on collected tear samples. Tear film osmolarity measurement is influenced by the sample collection technique with the osmolarity of on-eye tears being higher than that of collected tears. This highlights the importance of measuring tear film osmolarity directly on-eye. The in situ osmometer has good repeatability for conducting this measurement.