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Nitric oxide (NO) has a wide array of biological functions including the regulation of vascular tone, neurotransmission, immunomodulation, stimulation of proinflammatory cytokine expression and antimicrobial action. These functions may depend on the type of isoform that is responsible for the synthesis of NO. NO is found in various ocular tissues playing a pivotal role in physiological mechanisms, namely regulating vascular tone in the uvea, retinal blood circulation, aqueous humor dynamics, neurotransmission and phototransduction in retinal layers. Unregulated production of NO in ocular tissues may result in production of toxic superoxide free radicals that participate in ocular diseases such as endotoxin-induced uveitis, ischemic proliferative retinopathy and neurotoxicity of optic nerve head in glaucoma. However, the role of NO on the ocular surface in mediating physiology and pathophysiological processes is not fully understood. Moreover, methods used to measure levels of NO in the biological samples of the ocular surface are not well established due to its rapid oxidation. The purpose of this review is to highlight the role of NO in the physiology and pathophysiology of ocular surface and propose suitable techniques to measure NO levels in ocular surface tissues and tears. This will improve the understanding of NO's role in ocular surface biology and the development of new NO-based therapies to treat various ocular surface diseases. Further, this review summarizes the biochemistry underpinning NO's antimicrobial action.

SIGNIFICANCE A multipurpose disinfecting solution (MPDS), which contains povidone-iodine (PI) as a disinfectant, has high disinfecting efficacy not only on planktonic bacterium but also on the case biofilms. The addition of case hygiene practice removed more bacteria from cases than MPDS alone. PURPOSE This study compared the ability of two MPDSs, one containing PI and another containing polyaminopropyl biguanide and polyquaternium, to reduce bacterial numbers in solution or adhered to the cases following case hygiene procedures. METHODS Bacterial strains (Delftia acidovorans, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Serratia marcescens, Staphylococcus aureus, and Staphylococcus epidermidis) were exposed to the MPDSs for the recommended disinfection times, and the viable number evaluated according to ISO 14729. Cases were inoculated with bacterial strains and incubated for 24 hours to allow for biofilm formation. Cases were disinfected with both disinfecting solutions for 4 hours and rinsed, followed by recapping or air-drying, or tissue-wiping and air-drying for 18 hours. The number of survivors was counted using standard culture techniques. RESULTS Both products exceeded the recommended 3-log reduction against planktonic bacteria. Regarding biofilm, after rinsing and recapping wet, the numbers of D. acidovorans (mean difference [95% confidence interval] log10 colony-forming units per case, -2.9 [0.8 to -4.6], P < .01), P. aeruginosa (-2.0 [0.5 to -3.1], P < .01), S. marcescens (-1.7 [0.8 to -3.5], P < .05), and S. epidermidis (-2.1 [0.6 to -3.5], P < .05) in PI cases were significantly lower than in the dual-disinfectant MPDS storage cases. After air-drying, the PI storage cases had significantly lower numbers of S. maltophilia (-2.6 [0.6 to -4.0], P < .01), D. acidovorans (-1.6 [0.7 to -3.3], P < .05), and S. aureus (-1.6 [0.7 to -3.1], P < .05). The addition of tissue-wiping reduced the bacterial numbers in the MPDS storage cases to levels in the PI storage cases. CONCLUSIONS Contact lens users should be recommended to tissue-wipe and air-dry their lens storage cases after disinfection with regular MPDS.

To assess the six-month therapeutic profiles of lipid and non-lipid-based artificial tear supplements in managing dry eye disease (DED).Ninety-nine participants fulfilling the TFOS DEWS II diagnostic criteria for DED (64% females; mean ± SD age, 44 ± 16 years) were enrolled in a prospective, multicentre, double-masked, parallel group, randomised controlled trial. Participants instilled lipid-based nanoemulsion drops or non-lipid-based aqueous drops for six months, at least four times daily. Symptomology, tear film and ocular surface characteristics were assessed at Days 0, 30, 60, 90, 120, 150 and 180.Sustained reductions in OSDI, DEQ-5, and SANDE symptom scores from baseline were observed from Day 30 onwards in both groups (all p 0.05) and decreased superior lid wiper epitheliopathy grades from Day 60 onwards (all p ≤ 0.01). Improvements in non-invasive tear film breakup time, and sodium fluorescein and lissamine green staining scores followed from Day 120 onwards in both groups (all p 0.05). Tear lipid layer grades increased from Day 90 onwards only with the lipid-based drops, and with significantly greater improvement in those with suboptimal lipid layer thickness at baseline (grade ≤3; p = 0.02). By Day 180, 19% of participants no longer fulfilled the diagnostic criteria for DED.Over a six-month treatment period, improvements in dry eye symptomology preceded tear film and ocular surface changes with regular use of both lipid and non-lipid-based artificial tear supplements. Both formulations addressed most mild-to-moderate forms of aqueous deficient and evaporative DED, while evaporative cases benefitted preferentially from lipid-based supplementation. This represents a first step towards mapping DED therapeutic strategies according to disease subtype and severity.

Isolation of antimicrobial-resistant microbes from ocular infections may be becoming more frequent. Infections caused by these microbes can be difficult to treat and lead to poor outcomes. However, new therapies are being developed which may help improve clinical outcomes. This review examines recent reports on the isolation of antibiotic-resistant microbes from ocular infections. In addition, an overview of the development of some new antibiotic therapies is given. The recent literature regarding antibiotic use and resistance, isolation of antibiotic-resistant microbes from ocular infections and the development of potential new antibiotics that can be used to treat these infections was reviewed. Ocular microbial infections are a global public health issue as they can result in vision loss which compromises quality of life. Approximately 70 per cent of ocular infections are caused by bacteria including Chlamydia trachomatis, Staphylococcus aureus, and Pseudomonas aeruginosa and fungi such as Candida albicans, Aspergillus spp. and Fusarium spp. Resistance to first-line antibiotics such as fluoroquinolones and azoles has increased, with resistance of S. aureus isolates from the USA to fluoroquinolones reaching 32 per cent of isolates and 35 per cent being methicillin-resistant (MRSA). Lower levels of MRSA (seven per cent) were isolated by an Australian study. Antimicrobial peptides, which are broad-spectrum alternatives to antibiotics, have been tested as possible new drugs. Several have shown promise in animal models of keratitis, especially treating P. aeruginosa, S. aureus or C. albicans infections. Reports of increasing resistance of ocular isolates to mainstay antibiotics are a concern, and there is evidence that for ocular surface disease this resistance translates into worse clinical outcomes. New antibiotics are being developed, but not by large pharmaceutical companies and mostly in university research laboratories and smaller biotech companies. Antimicrobial peptides show promise in treating keratitis.

Purpose The ocular surface microbiota are recognised as one of causative microorganisms in post-procedural endophthalmitis but in many cases the vitreous tap is culture negative. This study investigated bacterial contamination of intravitreal (IVT) needles using multiple approaches covering culturing, 16S rRNA gene sequencing, fluorescent in situ hybridisation (FISH) and scanning electron microscopy (SEM). Methods IVT needles were obtained immediately after injection from patients undergoing treatment for predominantly age-related macular degeneration. Eighteen needles were analysed by culturing on chocolate blood agar. In addition, 40 needles were analysed by extracting DNA and paired-end sequencing of the 16S rRNA gene. Sequences were quality filtered (USEARCH), taxonomically classified (SILVA) and contaminant filtered (DECONTAM). Nine needles were analysed by either FISH using the bacterial probe EUB338 or SEM. Results Using culturing, three bacteria were identified from 5 of 18 needles (28%) - Kocuria kristinae, Staphylococcus hominis and Sphingomonas paucimobilis. The negative control needles showed no growth. Following rigorous data filtering, bacterial community analysis using 16S rRNA gene sequencing showed the presence of predominantly Corynebacterium but also Pseudomonas, Acinetobacter, Sphingomonas, Staphylococcus and Bacillus on the needles. Cocci-shaped cells in a tetrad formation were observed using FISH, while SEM images showed cocci-shaped bacteria in pairs and irregular tetrads. Conclusions The study showed evidence for a large diversity of bacteria on IVT needles and visually confirmed their adherence. The diversity was similar to that found on the ocular surface and in conjunctival tissue. This suggests the risk of exogenous endophthalmitis remains even with sterilization of the conjunctival surface.

PURPOSE Photoactivated chromophore for keratitis cross-linking (PACK-CXL) is used as an adjunct therapy to antibiotic medication in infectious keratitis. This experimental study aimed at quantifying the PACK-CXL efficacy as a function of UV fluence using several bacterial strains and irradiated volumes. METHODS Six distinct bacterial strains, including standardized strains and clinically isolated strains from patients with keratitis, were analyzed. Bacterial concentrations between 10 and 10 cells/mL were used (simulating small corneal ulcers). Volumes of either 11 μL (≈285 μm stromal thickness) or 40 μL (≈1000 μm stromal thickness) were irradiated within a microtiter plate at different fluences (5.4-27 J/cm) and irradiances (3, 9 and 18 mW/cm). The ratio of bacterial killing (B†) was determined to evaluate the antimicrobial efficacy of PACK-CXL. RESULTS B† was similar (51 ± 11%) in bacterial concentrations between 10 and 10 per ml. In 11 μL volume, Staphylococcus aureus (SA) 8325-4 ATCC 29213, Bacillus subtilis (BS) 212901, and Pseudomonas aeruginosa (PA) 2016-866624 were most sensitive to PACK-CXL at 5.4 J/cm (on average B† = 49 ± 8%), whereas Klebsiella oxytoca (KO) 2016-86624 (B† = 25%) was least sensitive. When irradiating a larger volume, B† was on average lower in 40 μL (19 ± 18%), compared with 11 μL (45 ± 17%, P < 0.001). By contrast, applying a higher UV fluence increased B† of SA ATCC 29213, from 50% at 5.4 J/cm to 92% at 10.8 J/cm, to 100% at 16.2 J/cm and above. CONCLUSIONS Applying higher UV fluences substantially increases the bacterial killing rates. Safety limits for clinical application require further investigation.

Objective During use, contact lens disinfecting solutions are exposed to tears and clinical microbial isolates. The current study was designed to test the performance of several disinfecting solution in the presence of organic soils or clinical isolates. Methods Standard and clinical isolates were exposed to the disinfecting solutions in the presence or absence of different organic soils. The number of microbial cells killed during disinfection was established by growing cells after disinfection on agar plates. Results The disinfecting activity of the povidone-iodine or hydrogen peroxide solutions was not affected by the organic soils or clinical isolates. The presence of yeast organic soil did not affect the performance of the disinfecting solutions when tested with standard microbial strains, but the addition of a model tear organic soil significantly reduced the disinfecting activity of the solutions containing various combinations of polyhexamethylene biguanide, polyquaternium-1, alexidine, and myristamindopropyl dimethylamine especially when tested against the standard fungal strains (reducing the effectiveness by between 0.5-4 log10) or the clinical bacterial isolates (reducing the effectiveness by between 0.5-3.5 log10). One disinfecting solution that contained polyquaternium-1 and myristamindopropyl dimethylamine had very poor activity against the clinical bacterial isolates in the absence or presence of either organic soil. Conclusions Povidone-iodine or hydrogen peroxide disinfecting solutions are not affected by organic soils and are very active against clinical bacterial isolates. Disinfecting solutions containing combinations of polyhexamethylene biguanide, polyquaternium-1, alexidine, and myristamindopropyl dimethylamine are affected by model tear organic soil and may have poor activity against clinical isolates.

Corneal confocal microscopy (CCM) has been identified as a non-invasive technique to assess corneal nerve fiber morphology. It is not known how corneal nerve changes relate to measures of peripheral nerve function in diabetic peripheral neuropathy (DPN). The present study investigates the relationship between nerve structure and function in DPN.Fifty participants with type 1 diabetes (T1DM) and 29 healthy controls underwent CCM to assess corneal nerve fiber density (CNFD), branch density (CNBD), fiber length (CNFL), total branch density (CTBD), nerve fractal dimension (CNFrD) and inferior whorl length (IWL). The severity of DPN was assessed as Total Neuropathy Score (TNS). Motor nerve axonal excitability tests were conducted to assess axonal function.Significant correlations were noted between CNFD (rho = -0.783; P 0.01) or superexcitability (rho = 0.435; P 0.01) and TNS. CNFrD was significantly correlated with peak response to stimulus (r = 0.414; P 0.01) and superexcitability (r = -0.467; P 0.01) measurements.Corneal nerve loss demonstrates a significant association with axonal ion channel dysfunction in T1DM.Detection of altered corneal nerve morphology may lead to the earlier diagnosis of DPN.

CLINICAL RELEVANCE: Optometrists have been advised to wear face masks during the COVID-19 pandemic. This study examined whether face masks were equally protective against transmission of microbes. BACKGROUND: The aim of the current study was to examine the ability of face masks to reduce transmission of microbes in aerosols and during speech. METHODS: Different face masks, surgical, medical 3-ply and cloth masks with different layers were used. The masks were tested under the ASTM standard F2101-1 to measure their ability to reduce the transmission of aerosolised Staphylococcus aureus. Bacterial cells in different sized aerosols were captured on agar plates. The ability of masks to reduce the transmission of bacteria during speech over 30 cm was measured. Bacteria were captured in masks or on agar plates at a distance of 30 cm during the speech. RESULTS: All masks reduced the transmission of aerosolised S. aureus (p ≤ 0.007). The medical 3-ply and cloth masks with three layers reduced the transmission of S. aureus aerosols (3.3 µm) by 98% and surgical and seven-layer masks reduced this by 100%. An antibacterial silver mask showed significantly greater transmission of S. aureus in aerosols of 4.7 µm (16 ± 6 cells) and 3.3 µm (122 ± 66 cells) compared to all other masks (0-3 cells and 0-15 cells, respectively; p ≤ 0.016). Surgical and three-layer masks had significantly worse filtration of 1.1 µm aerosols than for other aerosol sizes. Wearing a mask reduced the transmission of bacteria during speech, but the inner surface of these masks became contaminated with 528-3060 bacterial cells. CONCLUSIONS: Face masks effectively reduce the transmission of microbes in laboratory tests. Face masks made with seven layers were very effective at stopping transmission of S. aureus in all aerosol particle sizes. However, face masks become rapidly contaminated during wear. If masks are to be re-used, they should be regularly replaced or appropriately washed.

The widely used immortalised human meibomian gland epithelia cell (iHMGEC) line has made possible extensive studies of the biology and pathophysiology of meibomian glands (MG). Tissue culture protocols for iHMGEC have been revised and modified to optimise the growth conditions for cell differentiation and lipid accumulation. iHMGEC proliferate in serum-free medium but require serum or other appropriate exogenous factors to differentiate. Several supplements can enhance differentiation and neutral lipid accumulation in iHMGEC grown in serum-containing medium. In serum-free medium, rosiglitazone, a peroxisome proliferator activator receptor-γ (PPARγ) agonist, is reported to induce iHMGEC differentiation, neutral lipid accumulation and expression of key biomarkers of differentiation. iHMGEC cultured in serum-containing medium under hypoxia or with azithromycin increases DNAse 2 activity, a biomarker of terminal differentiation in sebocytes. The production of lipids with composition similar to meibum has not been observed in vitro and this remains a major challenge for iHMGEC culture. Innovative methodologies such as 3D ex vivo culture of MG and generation of MG organoids from stem cells are important for further developing a model that more closely mimics the in vivo biology of human MG and to facilitate the next generation of studies of MG disease and dry eye.

Purpose To evaluate the antiviral potential of five multipurpose disinfecting solutions against coronavirus (mouse hepatitis virus, a surrogate for SARS-CoV-2 human corona virus). Methods Test solutions (Biotrue, renu Advanced [Bausch and Lomb], ACUVUE RevitaLens [Johnson and Johnson Vision], cleadew [Ophtecs corp.] or AOSept Plus [Alcon]) were mixed with the coronavirus mouse hepatitis virus at 104 plaque forming units (PFU)/mL as the final concentration and incubated at room temperature for the specified disinfection time. Surviving virus from each sample was then quantified by standard plaque forming unit assay and the reduction of PFU for each disinfectant was compared to the phosphate buffer saline (PBS) treated negative control. A regimen test was also conducted using Biotrue. Results The three multipurpose disinfecting solutions Biotrue (containing PHMB and polyquaternium-1), renu Advanced (PHMB, polyquaternium-1 and alexidine) and ACUVUE RevitaLens (polyquaternium-1 and alexidine) did not kill the coronavirus at the manufacturers recommended disinfection time in the stand alone test. After treatment, the virus’s titer (3.8 ± 0.2 log10 for Biotrue, 3.7 ± 0.1 log10 for renu and 3.7 ± 0.2 log10 for RevitaLens) was similar to the negative control (3.7 ± 0.1 log10; p ≥ 0.996). AOSept Plus (hydrogen peroxide) and cleadew (povidone iodine) significantly (p

Purpose To determine if there is diurnal variation in gene expression in normal healthy conjunctival cells. Methods Bulbar conjunctival swab samples were collected from four healthy subjects in the morning and evening of the same day. The two swab samples were taken from one eye of each participant, with a minimum of five hours gap between the two samples. RNA was extracted and analysed using RNA sequencing (RNA-Seq). Results A total of 121 genes were differentially expressed between the morning and the evening conjunctival samples, of which 94 genes were upregulated in the morning, and 27 genes were upregulated in the evening. Many of the genes that were upregulated in the morning were involved in defence, cell turnover and regulation of gene expression, while the genes upregulated in the evening were involved in signalling and mucin production. Conclusions This study has identified several genes whose expression changes over the course of the day. Knowledge of diurnal variations of conjunctival gene expression provides an insight into the regulatory status of the healthy eye and provides a baseline for examining changes during ocular surface disease.

Two spectrophotometric microplate assays with dual staining for either fluorescent Nile red (NR) plus 4,6-diamidino-2-phenylindole (DAPI) or non-fluorescent Oil red O (ORO) plus Crystal violet (CV) were applied and optimised to evaluate the lipid producing capacity of immortalised human meibomian gland epithelial cells (iHMGEC). Cells were treated with rosiglitazone (Rosi, 10–50 μM), a known lipid producing inducer for iHMGEC, and were analysed for lipids using the NR-DAPI and ORO-CV microplate assays. The lipid producing capacity of iHMGEC after each treatment was determined by normalising lipid quantity (measured with NR or ORO) to cell number (measured with DAPI or CV). The dye concentrations of NR 1 μg/mL, DAPI 5 μg/mL, ORO 0.3% (v/v) and CV 0.5% (v/v), provided optimal linearity and coverage of signals over a range of cell densities (corresponding to 10–100% cell confluence). Both NR-DAPI and ORO-CV showed a dose-dependent effect of Rosi on lipid production in iHMGEC, consistent with the results reported previously using traditional microscopic imaging methods. The microplate assays offer a rapid, high throughput and objective measurement of the amount of lipids in iHMGEC (and potentially other lipid-producing cells) and can be used for screening the effects of biological agents or incubation changes on lipid production in cells in future studies.

Purpose: To evaluate the effect of age, gender and body mass index (BMI) on the levels of tear film neuromediators and corneal nerve parameters in healthy individuals.Methods: Twenty-six he...

This report aimed to explore whether certain inflammatory mediators were absorbed, extracted, or bound by various contact lens materials.Comfilcon A, balafilcon A, omafilcon A, and etafilcon A were soaked in 500 and 100 pg/mL of interleukin-8 (IL-8), matrix metalloproteinase-9 (MMP-9), or interleukin-1 receptor antagonist (IL-1Ra), and also in combined solutions of inflammatory mediators (500 pg/mL or 100 pg/mL) separately. Lenses were then extracted in 1:1 2% trifluoroacetic acid:acetonitrile. The extracted and residual concentrations of inflammatory mediators were determined using enzyme-linked immunosorbent assays. Absorbed (control-residual) and firmly bound (absorbed-extracted) concentrations were calculated for analysis.More MMP-9 was absorbed by omafilcon A (466±9 pg/mL) than balafilcon A (P=0.006; 437±11 pg/mL) or etafilcon A (P=0.001; 428±13 pg/mL) when soaked in 500 pg/mL, but no differences in 100 pg/mL. More MMP-9 remained firmly bound to omafilcon A (P=0.03; 174±3 pg/mL), comfilcon A (P=0.049; 168±34 pg/mL), and balafilcon A (P=0.01; 186±14 pg/mL) than etafilcon A (128±22 pg/mL). There were no differences in IL-8 absorption between lenses; however, more IL-8 remained firmly bound to omafilcon A (P=0.01; 336±25 pg/mL) than etafilcon A (106±133 pg/mL) when soaked in 500 pg/mL. No differences were found in concentrations of absorbed or firmly bound IL-1Ra between materials. When the mediators were combined, IL-8 was absorbed more in etafilcon A (P=0.03) than in other lens materials, but the absorbed IL-8 did not remain firmly bound.The uptake and extraction of inflammatory mediators from contact lenses was affected by competitive binding between the mediators.

Aim: The ocular surface is continually exposed to bacteria from the environment and traditional culture-based microbiological studies have isolated a low diversity of microorganisms from this region. The use of culture-independent methods to define the ocular microbiome, primarily involving 16S ribosomal RNA gene sequencing studies, have shown that the microbial communities present on the ocular surface have a greater diversity than previously reported. Method: A review of the literature on ocular microbiome research in health and disease. Results: Molecular techniques have been used to investigate the effect of contact lens wear and disease on the microbiota of the ocular surface and eyelids and the immunoregulatory role of the ocular surface microbiota. Studies have shown that compositional changes in the microbiota occur in ocular surface disorders such as blepharitis, trachoma and dry eye and also suggest a role of the ocular and non-ocular microbiome in retinal disease including age-related macular degeneration, glaucoma, uveitis and diabetic retinopathy. However, ocular microbiome studies need to recognise the potential for contamination to impact findings and carefully control each stage of the experimental procedure and to utilise statistical methods to identify contamination signals. Conclusion: The healthy ocular surface is characterised by a relatively stable, comparatively low diversity microbiome with recent findings that the bacteria of the ocular surface appear to have a role in maintaining homeostasis by modulating immune function.

The human eye is composed of numerous microhabitats. The aim of this study was to understand the communality and differences in the microbiomes of various regions of the eye.Four ocular sites from different subject groups were assessed including the eyelid margin tissue from patients with lid abnormalities (n = 20), fornix and limbus conjunctival tissue from patients with pterygia (n = 23), ocular (conjunctival) surface swabs (n = 45) and facial skin swabs (n = 16). Microbial communities were analysed by extracting total DNA from samples and sequencing the 16S ribosomal(r)RNA gene using the Illumina MiSeq platform. Sequences were quality filtered, clustered into unique sequences (zOTUs) using the UNOISE pipeline in USEARCH and taxonomically classified using SILVA.A difference in bacterial richness and diversity was found between sites (P 0.001) and for age (P 0.035) but not for sex (P 0.05). There was a difference in bacterial community structure and composition between sites (P 0.001). Bacterial distribution could be broadly classified into three groups - zOTUs resident on the skin and lid margin but with low abundances at other sites (Corynebacterium, Staphylococcus), zOTUs found mainly on the ocular surface (Acinetobacter, Aeribacillus) and zOTUs mostly present in the conjunctiva and lid margin (Pseudomonas).The microhabitats of the human eye (ocular surface, conjunctiva, lid margin and skin) have a distinct bacterial biogeography with some bacteria shared between multiple regions while other bacteria occupy a more confined niche.

Contact lens cases become contaminated with microbes during use. We wished to compare the adhesion of uncommon bacterial contaminants isolated from lens cases to contact lenses with and without organic soil.Strains of Delftia acidovorans (001), Stenotrophomonas maltophilia (002 and 006), and Achromobacter xylosoxidans (001) isolated from contact lens cases (test strains) and Pseudomonas aeruginosa (Paer1) isolated from eyes at the time of infiltrative response (control strain) were used. Bacteria were grown and resuspended in phosphate-buffered saline (PBS) or 10% organic soil (heat-killed Saccharomyces cerevisiae resuspended in complement inactivated bovine serum). Two silicone hydrogel (senofilcon A and comfilcon A) and one hydrogel lens (etafilcon A) lens materials were used. Bacteria (1.0×10 and 1.0×10 colony-forming units/mL; CFU/mL) adhered to lenses for 24 hr and the numbers of bacteria adherent to each lens type (with and without organic soil) were estimated by culture.All the four test strains adhered in significantly greater numbers to contact lenses after incubation in inoculum prepared with organic soil compared with PBS-D. acidovorans 001 (0.7 log10 CFU; P0.05), S. maltophilia 002 (1.7 log10 CFU; P0.05), S. maltophilia 006 (0.9 log10 CFU; P0.05), and A. xylosoxidans 001 (0.4 log10 CFU; P0.05). However, the presence of organic soil did not increase adhesion of P. aeruginosa Paer1 (-0.1 log10 CFU; P0.05). Achromobacter xylosoxidans 001 (P0.01), D. acidovorans 001 (P0.01), and S. maltophilia 002 (P0.01) significantly differed in their adhesion to the three contact lens materials.Bacteria that are commonly found in contact lens cases adhered to contact lenses in relatively high numbers in the presence of organic soil. This might indicate that a similar phenomenon occurs in the presence of tears. This may facilitate their transfer from the lens to the cornea and the production of corneal infiltrates.

Contact lenses in the future will likely have functions other than correction of refractive error. Lenses designed to control the development of myopia are already commercially available. Contact lenses as drug delivery devices and powered through advancements in nanotechnology will open up further opportunities for unique uses of contact lenses. This review examines the use, or potential use, of contact lenses aside from their role to correct refractive error. Contact lenses can be used to detect systemic and ocular surface diseases, treat and manage various ocular conditions and as devices that can correct presbyopia, control the development of myopia or be used for augmented vision. There is also discussion of new developments in contact lens packaging and storage cases. The use of contact lenses as devices to detect systemic disease has mostly focussed on detecting changes to glucose levels in tears for monitoring diabetic control. Glucose can be detected using changes in colour, fluorescence or generation of electric signals by embedded sensors such as boronic acid, concanavalin A or glucose oxidase. Contact lenses that have gained regulatory approval can measure changes in intraocular pressure to monitor glaucoma by measuring small changes in corneal shape. Challenges include integrating sensors into contact lenses and detecting the signals generated. Various techniques are used to optimise uptake and release of the drugs to the ocular surface to treat diseases such as dry eye, glaucoma, infection and allergy. Contact lenses that either mechanically or electronically change their shape are being investigated for the management of presbyopia. Contact lenses that slow the development of myopia are based upon incorporating concentric rings of plus power, peripheral optical zone(s) with add power or non-monotonic variations in power. Various forms of these lenses have shown a reduction in myopia in clinical trials and are available in various markets.

This paper outlines changes to the ocular surface caused by contact lenses and their degree of clinical significance. Substantial research and development to improve oxygen permeability of rigid and soft contact lenses has meant that in many countries the issues caused by hypoxia to the ocular surface have largely been negated. The ability of contact lenses to change the axial growth characteristics of the globe is being utilised to help reduce the myopia pandemic and several studies and meta-analyses have shown that wearing orthokeratology lenses or soft multifocal contact lenses can reduce axial length growth (and hence myopia).However, effects on blinking, ptosis, the function of Meibomian glands, fluorescein and lissamine green staining of the conjunctiva and cornea, production of lid-parallel conjunctival folds and lid wiper epitheliopathy have received less research attention. Contact lens wear produces a subclinical inflammatory response manifested by increases in the number of dendritiform cells in the conjunctiva, cornea and limbus. Papillary conjunctivitis is also a complication of all types of contact lenses. Changes to wear schedule (daily disposable from overnight wear) or lens materials (hydrogel from SiHy) can reduce papillary conjunctivitis, but the effect of such changes on dendritic cell migration needs further study. These changes may be associated with decreased comfort but confirmatory studies are needed. Contact lenses can affect the sensitivity of the ocular surface to mechanical stimulation, but whether these changes affect comfort requires further investigation.In conclusion, there have been changes to lens materials, design and wear schedules over the past 20+ years that have improved their safety and seen the development of lenses that can reduce the myopia development. However, several changes to the ocular surface still occur and warrant further research effort in order to optimise the lens wearing experience.

Significance Think Tank 2019 affirmed that the rate of infection associated with contact lenses has not changed in several decades. Also, there is a trend toward more serious infections associated with Acanthamoeba and fungi. The growing use of contact lenses in children demands our attention with surveillance and case-control studies. Purpose The American Academy of Optometry (AAO) gathered researchers and key opinion leaders from around the world to discuss contact lens-associated microbial keratitis at the 2019 AAO Annual Meeting. Methods Experts presented within four sessions. Session 1 covered the epidemiology of microbial keratitis, pathogenesis of Pseudomonas aeruginosa, and the role of lens care systems and storage cases in corneal disease. Session 2 covered nonbacterial forms of keratitis in contact lens wearers. Session 3 covered future needs, challenges, and research questions in relation to microbial keratitis in youth and myopia control, microbiome, antimicrobial surfaces, and genetic susceptibility. Session 4 covered compliance and communication imperatives. Results The absolute rate of microbial keratitis has remained very consistent for three decades despite new technologies, and extended wear significantly increases the risk. Improved oxygen delivery afforded by silicone hydrogel lenses has not impacted the rates, and although the introduction of daily disposable lenses has minimized the risk of severe disease, there is no consistent evidence that they have altered the overall rate of microbial keratitis. Overnight orthokeratology lenses may increase the risk of microbial keratitis, especially secondary to Acanthamoeba, in children. Compliance remains a concern and a significant risk factor for disease. New insights into host microbiome and genetic susceptibility may uncover new theories. More studies such as case-control designs suited for rare diseases and registries are needed. Conclusions The first annual AAO Think Tank acknowledged that the risk of microbial keratitis has not decreased over decades, despite innovation. Important questions and research directions remain.

Animal models are a critical element of ocular surface research for investigating therapeutic drops, surgical implants, and infection research. This study was a comparative analysis of the microbial communities on conjunctival tissue samples from humans compared to several commonly used laboratory animals (BALB/c mice, New Zealand white rabbits and IMVS colored stock guinea pigs). Microbial communities were analyzed by extracting total DNA from conjunctival tissue and sequencing the 16 S rRNA gene using the Illumina MiSeq platform. Sequences were quality filtered using the UNOISE pipeline in USEARCH and taxonomically classified using GTDB database. Sequences associated with blank extraction and sampling negative controls were removed with the decontam R software package prior to downstream analysis. There was a difference in the diversity measures of richness (P = 0.0124) and Shannon index (P = 0.0002) between humans and rabbits but not between human, mouse and guinea pigs. There was a difference between the human and any animal for bacterial community structure (P = 0.006). There was a higher degree of similarity between the bacterial composition of the human and mouse samples with each dominated by the phyla Proteobacteria and Firmicutes. The use of mouse models may be more appropriate for studies investigating changes to the ocular microbiome due to interventions such as application of antibiotics due to greater similarities in bacterial community structure and composition to humans.

Contact lens materials have undergone significant changes over the past 20 years, particularly with respect to the introduction of silicone hydrogel materials. Whilst this development addressed hypoxic issues, other important areas relating to contact lens success, notably comfort, require further research. Contact lens wettability remains a crucially important part of biocompatibility. Contact lenses can be made more wettable by incorporation of surfactants into blister packs, internal wetting agents, surface treatments or care solutions. However, there remains no clear association between contact lens wettability and comfort, making it challenging to determine the potential for these approaches to be of significant clinical benefit. Most contact lenses are used on a daily wear, reusable basis, which requires them to be disinfected when not worn. The ideal disinfecting solution would also improve comfort during wear. However, balancing these requirements with other factors, including biocompatibility, remains a challenge. Soft lens materials invariably take up and subsequently release certain components of disinfecting solutions onto the ocular surface. This may affect tear film stability and the normal ocular microbiome, and further research is needed in this area to determine whether this has any affect on comfort. Finally, contact lens materials sorb components of the tear film, and these interactions are complex and may change the biochemistry of the tear film, which in turn may affect their comfort. In conclusion, the interaction between lens materials, tear film and disinfection solution plays an important role in the biocompatibility of lenses. However, the exact role and whether this can be altered to improve biocompatibility and comfort during wear remains debatable. This report summarises the best available evidence to examine this complex relationship and the opportunities for practitioners to enhance in-eye comfort of contemporary lenses, along with providing suggestions for areas of study that may provide further information on this topic.

Soft contact lenses that have been prescribed by eye care practitioners are sometimes substituted for alternative lenses by unqualified, unregulated and sometimes even fully regulated lens suppliers, in the mistaken belief that there is essentially no difference between different soft lens types. This review considers the implications of inappropriately substituting soft contact lens types in terms of (a) lens properties: surface treatment, internal wetting agents, material, total diameter, back optic zone radius, thickness, edge profile, back surface design, optical design, power, colour (tint) and ultraviolet protection; and (b) lens usage: wearing modality (daily versus overnight wear) and replacement frequency. Potential aspects of patient dissatisfaction and adverse events when prescribed soft lenses are substituted for lenses with different properties or intended usage are considered. Substitution of 15 of the 16 lens properties considered (i.e. except for back surface design) was found to be related to at least one – and as many as six – potential sources of patient dissatisfaction and adverse ocular events. Contact lens are medical devices which are prescribed and fitted; they should never be substituted for another lens type in the absence of a new prescription further to a full finalised fitting, for the simple reason that all soft contact lenses are not created equal. A substituted lens may have properties that results in undesirable consequences in respect of vision, ocular health, comfort and cosmetic appearance, and may be incompatible with the lifestyle of the patient.

The purpose of this study was to investigate the effect of Mel4 antimicrobial peptide-coated contact lenses (MACL) on the microbiota of the conjunctiva and lenses during three months of extended wear.One hundred and seventy-six participants were recruited into a randomised, contralateral, double masked, biweekly extended wear MACL and uncoated control lens trial. At the one month and 3-month visit, the conjunctival microbiota was sampled using sterile cotton swabs and contact lenses were collected aseptically. Standard microbiological procedures were employed for culture of the swabs and contact lenses and identification of the isolated microorganisms.Gram-positive bacteria (predominantly coagulase-negative staphylococci) were the most frequently isolated microbes from both contact lenses and conjunctiva. There was no difference in the frequency of isolation of most bacteria or fungi from the conjunctival swabs of eyes wearing either MACL or control lenses. The only exception was a higher frequency of eyes harbouring Staphylococcus arlettae when wearing control lenses (5%) versus MACL (1%) (p = 0.002). There was no significant difference in the frequency of microbes isolated from MACL or control contact lenses. There were also no differences between lens types in the frequency of isolation of1 microbial type per sampling occasion for either conjunctiva swabs or contact lenses.MACL wear did not change the conjunctival microbiota during extended wear, and the types of microbes isolated from MACL were similar to those isolated from control lenses.

Corneal infection caused by a bacteria Pseudomonas aeruginosa is common cause of ocular morbidity. Increasing antibiotic resistance by ocular P. aeruginosa is an emerging concern. In this study the resistome of ocular isolates of Pseudomonas aeruginosa clone ST308 isolated in India in 1997 (PA31, PA32, PA33, PA35 and PA37) and 2018 (PA198 and PA219) were investigated. All the isolates of ST308 had >95% nucleotide similarity. The isolates from 2018 had larger genomes, coding sequences, accessory and pan genes compared to the older isolates from 1997. The 2018 isolate PA219 was resistant to all antibiotics except polymyxin B, while the 2018 isolate PA198 was resistant to ciprofloxacin, levofloxacin, gentamicin and tobramycin. Among the isolates from 1997, five were resistant to gentamicin, tobramycin and ciprofloxacin, four were resistant to levofloxacin while two were resistant to polymyxin B. Twenty-four acquired resistance genes were present in the 2018 isolates compared to 11 in the historical isolates. All isolates contained genes encoding for aminoglycoside (aph(6)-Id, aph(3')-lIb, aph(3″)-Ib), beta-lactam (blaPAO), tetracycline (tet(G)), fosfomycin (fosA), chloramphenicol (catB7), sulphonamide (sul1), quaternary ammonium (qacEdelta1) and fluoroquinolone (crpP) resistance. Isolate PA198 possessed aph(3')-VI, rmtD2, qnrVC1, blaOXA-488, blaPME-1, while PA219 possessed aadA1, rmtB, qnrVC1, aac(6')-Ib-cr, blaTEM-1B, blaVIM-2, blaPAO-1, mph(E), mph(A), msr(E). In both recent isolates qnrVC1 was present in Tn3 transposon. In 219 blaTEM-1 was carried on a transposon and blaOXA-10 on a class 1 integron. There were no notable differences in the number of single nucleotide polymorphisms, but recent isolates carried more insertions and deletions in their genes. These findings suggest that genomes of P. aeruginosa ocular clonal strains with >95% nucleotide identity isolated twenty years apart had changed over time with the acquisition of resistance genes. The pattern of gene mutations also varied with more insertions and deletions in their chromosomal genes which confer resistance to antibiotics.

Frequent replacement contact lenses made from the etafilcon A hydrogel lens material were introduced onto the market over 30 years ago, and etafilcon A remains the most widely used hydrogel lens material today. Although the prescribing of silicone hydrogel lenses is increasing, millions of lens wearers globally have been wearing hydrogel lenses for many years and exhibit a physiologically-stable ‘quiet eye’, with a low profile of adverse events. Hydrogel lenses are demonstrated to maintain a low inflammatory response and infection risk profile during daily wear, which in the case of etafilcon A, may be related to its low modulus, and the naturally-protective, anti-microbial, non-denatured lysozyme absorbed into the lens from the tear fluid. Although improved corneal physiology from decreased hypoxia with silicone hydrogel lenses is well accepted, equivalent levels of corneal oxygenation are maintained during daily wear of low to medium powered hydrogel lenses, which do not impede the daily corneal de-swelling process, and do not induce clinically significant changes in ocular health. Therefore, hydrogel lenses remain an important alternative for daily wear in modern contact lens practice.

A novel coronavirus (CoV), the Severe Acute Respiratory Syndrome Coronavirus - 2 (SARS-CoV-2), results in the coronavirus disease 2019 (COVID-19). As information concerning the COVID-19 disease continues to evolve, patients look to their eye care practitioners for accurate eye health guidance. There is currently no evidence to suggest an increased risk of contracting COVID-19 through contact lens (CL) wear compared to spectacle lens wear and no scientific evidence that wearing standard prescription spectacles provides protection against COVID-19 or other viral transmissions. During the pandemic there will potentially be significant changes in access to local eyecare. Thus, it is imperative CL wearers are reminded of the steps they should follow to minimise their risk of complications, to reduce their need to leave isolation and seek care. Management of adverse events should be retained within optometric systems if possible, to minimise the impact on the wider healthcare service, which will be stretched. Optimal CL care behaviours should be the same as those under normal circumstances, which include appropriate hand washing (thoroughly with soap and water) and drying (with paper towels) before both CL application and removal. Daily CL cleaning and correct case care for reusable CL should be followed according to appropriate guidelines, and CL exposure to water must be avoided. Where the availability of local clinical care is restricted, practitioners could consider advising patients to reduce or eliminate sleeping in their CL (where patients have the appropriate knowledge about correct daily care and access to suitable lens-care products) or consider the option of moving patients to daily disposable lenses (where patients have appropriate lens supplies available). Patients should also avoid touching their face, including their eyes, nose and mouth, with unwashed hands and avoid CL wear altogether if unwell (particularly with any cold or flu-like symptoms).

Purpose This study examined the rate and level of microbial contamination of silver and non-silver (control) barrel contact lens storage cases in vivo. Methods A prospective, single-centre, randomized, double blinded, crossover study was conducted to evaluate the efficacy of silver and control barrel cases used with an Oxipol™-based disinfecting solution over a two-month period. Lens cases were collected, and the frequency, type and level of microbial contamination evaluated using viable culture and standard identification methods. Questionnaires were used to assess the contact lens-related hygiene compliance of the participants. Results Forty eight daily contact lens wearers completed the study. Overall, 27 % of silver and 35 % control lens cases (P > 0.05) were contaminated, predominantly with Gram-positive bacteria. Only, 12 % of participants had good compliance, with 46 % having average compliance and 43 % poor compliance. Compliance score was not associated with level of microbial contamination. However, the use of hydrogel lenses in combination with silver cases compared to non-silver barrel cases resulted significantly greater numbers of bacteria colonising cases. Conclusion The present investigation demonstrated that the use of silver barrel cases does not reduce the overall rate and level of bacterial contamination, but that using hydrogel lenses in combination with silver cases resulted in higher numbers of bacteria isolated from cases.

Objective To investigate differences in SNPs in TLR genes between people who had keratitis and controls in an Indian population. Methods 145 cases of keratitis and 189 matched controls were recruited. DNA was extracted from peripheral blood. Single nucleotide polymorphisms (SNP) in TLR2 (n = 6), TLR4 (n = 15), TLR5 (n = 13) and TLR9 (n = 10) were analysed. The risk of developing keratitis was assessed based on allele, genotype and haplotype associations. Results For all cases of keratitis, the TLR4 SNP rs4986791 TC genotype frequency was significantly higher in cases (p = 0.006, OR = 1.96, 95 % CI 1.19–3.2). Including cases of only microbial keratitis (MK) revealed that genotypes in TLR2 SNP rs5743706 TA (p = 0.0001; OR = 8.61; 95 % CI 2.59–28.56)), TLR4 SNP s4986791 TC (p = 0.002; OR = 2.65; 95 % CI 1.39–5.07) were significantly more common for MK, whereas the TLR5 SNP rs2241096 A allele (p = 0.00316, OR = 0.42, 95 % CI 0.2−0.9286) and GA genotype (p = 0.016; OR = 0.45; 95 % CI 0.23−0.86) was significantly less common in MK cases. The TLR2 SNP rs5743706 genotype TA was significantly less common in the sterile keratitis (SK) group (p = 0.004, OR = 0.43, 95 %CI 0.24−0.77). Haplotype analysis of MK compared to controls showed that TLR2 AT was more common in controls (p = 0.003); TLR4 ACAC was more common in cases (p = 0.004); TLR5 TGGCA was more common in controls (p = 0.001). Conclusion The present study revealed multiple associations between variants across TLR genes, which may have implications for understanding the underlying host factors, risk of developing keratitis and molecular pathogenesis in keratitis.

Significance This study investigated the development of an antimicrobial coating on silicone hydrogel contact lenses that may have the capacity to reduce contact lens-related infection and inflammatory events. Purpose The purpose of this study was to develop an effective antimicrobial coating for silicone hydrogel contact lenses by attachment of Mel4 peptide. Methods Lotrafilcon A, comfilcon A, somofilcon A, senofilcon A, and lotrafilcon B silicone hydrogel contact lenses were plasma coated with acrylic acid followed by Mel4 antimicrobial peptide immobilization by covalent coupling. Peptide immobilization was quantified by x-ray electron spectroscopy. Contact lens diameter, base curve, center thickness, and lens surface wettability were measured by captive-bubble contact-angle technique. Antimicrobial activity of the lenses was determined against Pseudomonas aeruginosa and Staphylococcus aureus by viable plate count and also after soaking with artificial tears solution for 1 day. In vivo safety and biocompatibility were determined in an animal model for 1 week. Results Mel4 peptide-coated silicone hydrogel contact lenses were associated with high antimicrobial inhibition (>2 log), except for lotrafilcon B and senofilcon A. Lotrafilcon B did not exhibit any activity, whereas senofilcon A showed 1.4- and 0.7-log inhibition against P. aeruginosa and S. aureus, respectively. X-ray electron spectroscopy revealed significant increases in the lens surface-bound amide nitrogen in all contact lenses except for lotrafilcon B. All contact lens parameters remained unchanged except for the base curve and center thickness for senofilcon A. Mel4 immobilization was associated with a decrease in contact angle. Mel4-coated contact lens wear was not associated with any signs or symptoms of ocular irritation in a rabbit model study. Reduced antimicrobial activity was observed with all the lenses after soaking with artificial tears solution or rabbit wear. Conclusions Mel4 antimicrobial coating may be an effective option for development of antimicrobial silicone hydrogel contact lenses.

Objective To assess the incidence of adverse events during the use of a povidone-iodine disinfecting solution (cleadew) and the microbial contamination in contact lens cases. Methods A prospective, single-center, open-label, controlled study evaluating the use of cleadew cleaning and disinfecting system in existing daily wear soft contact lens wearers over a 3-month period was conducted. Ocular signs and symptoms during lens wear were assessed at baseline and after 1 and 3 months of using cleadew. Contact lens cases were assessed for the frequency of microbial contamination and the types of microbes, using traditional microbial culture, followed by identification of bacteria using 16S rRNA sequencing. Results Use of cleadew was well tolerated. There was reduction in corneal staining (0.5±0.5 at 3 months of use) compared with the participant's habitual multipurpose disinfecting system (1.1±1.0); all other clinical signs were not significantly different. There were no cases of solution-induced corneal staining. There was a low adverse event rate of 0.8% per 100 participant-months. Contact lens case contamination was low, with 30% of cases having no culturable microbes. Comparison with previously published data showed that use of cleadew resulted in low frequencies of Gram-positive (49%) and fungal (8%) contamination and a low, but higher than some other disinfecting solutions, level of Gram-negative bacteria. Conclusions Cleadew cleaning and disinfecting system was associated with low levels of adverse events during use. Contact lens cases were significantly less frequently contaminated than for some other types of disinfecting solutions.

To determine the utility of corneal confocal microscopy and tear neuromediator analysis in the diagnosis of diabetic peripheral neuropathy (DPN) as a result of type 1 and type 2 diabetes.Seventy individuals with either type 1 diabetes or type 2 diabetes (T1D/T2D) underwent corneal confocal microscopy to assess the corneal nerve morphology. The concentration of substance P and calcitonin gene-related peptide (CGRP) in tears was measured by enzyme-linked immunosorbent assay. Motor excitability studies were conducted on the median nerve to assess axonal ion channel function. Based on total neuropathy score (TNS), participants were stratified into DPN (DPN+ve; TNS ≥ 2; T1D, n = 19; T2D, n = 16) and without DPN (DPN-ve; TNS ≤ 1; T1D, n = 19; T2D, n = 16). Areas under the receiver operating characteristic curves (AUCs) were calculated to obtain specificity and sensitivity of the measures to diagnose DPN.In T1D, the concentration of substance P and confocal microscopy measures were significantly reduced (Corneal confocal microscopy parameters provide a better diagnostic ability to detect DPN in T1D and T2D than nerve excitability measures or concentrations of tear neuromediators. The concentration of substance P could also be useful in diagnosing DPN but for T1D only.

OBJECTIVES To evaluate the effect of microblepharon exfoliation on the number of eyelid bacteria and their lipase activity and the relationship of these to contact lens discomfort. METHODS Thirty experienced contact lens wearers had their eyelid margin physiology, tear properties, and comfort scores assessed. The number, type, and frequency of lower eyelid margin bacteria, and their lipase activity, were measured. Eyelids were treated with a foam cleanser or microblepharon exfoliation. Clinical and microbiological tests were repeated at each visit. Changes and correlations were examined. RESULTS Symptomatic lens wearers had a higher ratio for the number and frequency of gram-positive rods and cocci. Microblepharon exfoliation reduced the number and ratio of gram-positive rods to cocci from baseline for symptomatic wearers that lasted 7 to 10 days after treatment (P

Purpose To report the density and morphology of cells that are analogous to corneal Langerhans cells and their associations in keratoconus. Materials and methods This prospective cross-sectional study included a convenience sample of keratoconus subjects aged between 18-65 years. Corneal topography, assessment of ocular symptoms, tear variables, corneal sensitivity, in-vivo confocal microscopy were performed. The number of Langerhans cells were manually counted and averaged across three central corneal images. Cell morphology was graded on a 0-3 scale, where grade 3 indicates cells with long visible dendrites. Associations of Langerhans cells with other variables were evaluated using Spearman’s correlation. Results Twenty-one keratoconus subjects with a mean age of 43 ± 11 years were included. Eighty-one percent of them were males, 48% had mild keratoconus and 52% were contact lens wearers. Langerhans cells were present in the central cornea in 91% of subjects. Median cell density was 15 cells/mm2(IQR: 3-21). Cell morphology of grades 2 or 3 (with short or long dendrites) was seen in 71% of subjects. There was a significant association between Langerhans cell frequency and density with male gender (rho and p-values: -0.669, 0.001 and -0.441,0.045) and between Langerhans cell density and nerve fibre tortuosity (0.479,0.028). No significant association observed with age, contact lens wear or ocular symptoms. Conclusion Langerhans cells were present in a significant number of subjects suggesting the possibility of inflammation in keratoconus. Based on the association of Langerhans cells with nerve parameters, we propose inflammation as the underlying cause for corneal nerve changes in keratoconus.

Purpose To investigate characteristics of the eyelid margins, meibomian glands and the tear film of contact lens wearers, and to determine whether these characteristics were related to symptoms of contact lens discomfort. Methods A cross sectional study was performed on thirty existing daily wear soft contact lens wearers (6 male; 24 female) with median age of 23 years (range 18–41). Eyelid signs and tear film characteristics were evaluated during a single visit and subjects completed the contact lens and dry eye questionnaire (CLDEQ-8) to evaluate ocular discomfort. Results Based on the CLDEQ-8 responses, subjects were classified as symptomatic (n = 17) or asymptomatic (n = 13). Grades of foam at meibomian gland orifices (3 ± 1), expressibility (2 ± 1) and quality of secretions (2 ± 1), tear evaporation rate with (112 ± 54 g/m2/h) or without (88 ± 45 g/m2/h) contact lens wear, fluorescein tear breakup time (8 ± 2 seconds) and tear lipid layer thickness (45 ± 17 nm) were significantly associated with symptoms of discomfort in symptomatic lens wearers only (r2 > 0.45; p value Conclusions Morphological irregularities of the meibomian glands and alterations to tear film secretions that affect tear evaporative dynamics were associated with symptoms of discomfort amongst the symptomatic lens wearers.

Treatment of Pseudomonas aeruginosa eye infections often becomes a challenge due to the ability of this bacterium to be resistant to antibiotics via intrinsic and acquired mechanisms. Transfer of resistance due to interchangeable genetic elements is an important mechanism for the rapid transfer of antibiotic resistance in this pathogen. As a result, drug-resistant strains are becoming increasingly prevalent worldwide. This review systematically analyses data from recent publications to describe the global prevalence and antibiotic sensitivity of ocular P. aeruginosa. Thirty-seven studies were selected for review from PubMed-based searches using the criteria 'microbial keratitis OR eye infection AND Pseudomonas aeruginosa AND antibiotic resistance' and limiting to papers from 2011 onward, to demonstrate the antibiotic resistance from isolates from around the world. Subsequently, we reviewed the ways in which P. aeruginosa can become resistant to antibiotics. Both the rate of isolation of bacteria in general (79 per cent of cases), and prevalence of P. aeruginosa (68 per cent of all isolates) were highest in contact lens-related microbial keratitis. The average resistance rate to common ocular antibiotics such as ciprofloxacin (9 per cent), gentamicin (22 per cent) and ceftazidime (13 per cent) remained relatively low. However, there were large variations in resistance rates reported in studies from different countries, for example resistance to ciprofloxacin reached up to 33 per cent. We next reviewed the types of mobile genetic elements (MGEs) such as plasmids, integrons and transposons that are frequently associated with drug resistance in P. aeruginosa. MGEs are important for the transmission of resistance to beta-lactams and aminoglycosides and recently have been shown to be potential factors for the transmission of fluoroquinolone resistance. Studies on the molecular mechanisms of resistance transfer in ocular P. aeruginosa have begun to be reported and will provide valuable information on the emergence of new antibiotic resistance and potential to treat resistant strains.

Inflammasomes, key molecular regulators that play an important role in inflammation, consist of a central protein, an adaptor protein ASC (apoptosis speck-like protein) and a caspase-1 protein. Upon activation, caspase-1 induces maturation of cytokines such as interleukin-1β (IL-1β) and interleukin-18 (IL-18). The release of these cytokines can result in inflammation. Inflammasomes are activated by a variety of factors and their activation involves complex signalling leading to resolution of infection, but can also contribute to the pathology of inflammatory, autoimmune, and infectious diseases. The role of NLRP1, NLRP3, NLRC4 and AIM2 inflammasomes in the pathogenesis of ocular diseases such as glaucoma, age related macular degeneration (AMD), diabetic retinopathy, dry eye and infections of the eye has been established over the past decade. In experimental studies and models, inhibition of inflammasomes generally helps to reduce the inflammation associated with these eye diseases, but as yet the role of these inflammasomes in many human eye diseases is unknown. Therefore, a need exists to study and understand various aspects of inflammasomes and their contribution to the pathology of human eye diseases. The goal of this review is to discuss the role of inflammasomes in the pathology of eye diseases, scope for anti-inflammasome therapy, and current research gaps in inflammasome-related eye disease.

Purpose To determine whether Mel4-coated antimicrobial contact lenses (MACLs) can reduce the incidence of corneal infiltrative events (CIEs) during extended wear. Methods A prospective, randomized, double-masked, single-center, contralateral, extended contact lens wear clinical trial was conducted with 176 subjects. Each participant was randomly assigned to wear a MACL in one eye and an uncoated control contact lens in the contralateral eye or an extended-wear biweekly disposable modality for 3 months. The main outcome measures were the incidence of CIEs per 100 eye-months, identification of the microbial types colonizing the contact lenses or eyes at the time of the CIEs, and their susceptibility to Mel4. Results Nine participants (5.1%) experienced unilateral CIEs; six participants had contact lens acute red eye, and three participants had infiltrative keratitis. The incidence rate for CIEs (0.4 events per 100 participant months; 1.7%) in the Mel4-coated lenses (test) was 69% less than that of the control lenses (1.3 events per 100 participant months; 3.4%; P = 0.29). All Gram-negative bacteria isolated from lenses and lids of participants with CIEs (Citrobacter diversus, Acinetobacter haemolyticus, and Acinetobacter lwoffii) were susceptible to Mel4 peptide; minimum inhibitory concentrations ranged from 15.6 to 62.5 µg/mL. Reduction of adhesion of these bacteria by Mel4-coated lenses ranged from 2.1 to 2.2 log10 colony-forming units/lens. Conclusions MACLs had the capacity to reduce CIEs by at least 50% compared with uncoated control lenses during extended wear over 3 months; however, due to the relatively low rates of CIEs, the reduction was not statistically different compared with control lenses. Translational relevance This study provides evidence that antimicrobial contact lenses have the potential to reduce the incidence of corneal infiltrative events during extended wear.

Purpose This study explored whether the non-polar lipids in the human tear fluid lipidome show diurnal variation with and without contact lens wear. It also addressed the relationship between changes in ocular comfort during the day with the level of non-polar lipids. Methods Tear samples were collected in the morning and evening with and without contact lenses using fine glass capillary tubes and were analysed by chip-based nano-electrospray ionization tandem mass spectrometric techniques. Tear levels of cholesteryl esters (CE), wax esters (WE) and triacylglycerides (TAG) were quantified. Results TAG 48:0, 52:0 and WE 26:0/16:0, and 27:0/17:0 increased from morning to evening. TAG 52:2, WE 21:0/16:0, 21:0/18:1 and 28:0/18:1 decreased during the day when no lenses were worn. CE 21:0 was the only non-polar lipid that increased from morning to evening in contact lens wear. WE 21:0/16:0 and 27:0/17:0 were lower in the morning in contact lens wear compared to no lens wear (p ≤ 0.05). The level of non-polar lipids did not correlate with ocular comfort at the end of the day. Conclusion Even though the level of some of non-polar lipid species changed from morning to evening the total level of major tear non-polar lipids remained unchanged during the day with and without contact lens wear. The effect of change in the quantity and structure of lipid species on tear stability and ocular comfort warrants more investigation.

This article presents an Executive Summary of the conclusions and recommendations of the 10-chapter TFOS DEWS II report. The entire TFOS DEWS II report was published in the July 2017 issue of The Ocular Surface. A downloadable version of the document and additional material, including videos of diagnostic and management techniques, are available on the TFOS website: www.TearFilm.org.

Background and Objective: To investigate changes in ocular surface inflammatory markers after daily and overnight silicone hydrogel contact lens wear in healthy wearers. Material and Methods: Twenty-six experienced soft contact lens subjects were evaluated at baseline, after 1-day of silicone hydrogel lens wear, and after 1-night of wear. Basal tears were collected at each visit and tear cytokine concentrations were quantified using multiplex [interleukin (IL)-1β, IL-6, IL-10, IL-12(p70), IL-17A and tumor necrosis factor (TNF)-α] or ELISA (IL-8) kits. A historical control group of 27 non-contact lens wearers was used to compare absolute concentrations and diurnal variations in tear cytokine concentrations. Changes in cytokine concentrations were analyzed using linear mixed models. Linear regression with bootstrapping was used to assess whether changes in IL-1β concentrations were associated with changes in other cytokines. Results: IL-8 concentrations decreased after 1 day of silicone hydrogel contact lens wear and returned to baseline levels the next morning (p=0.04). This same diurnal fluctuation was seen in non-contact lens wearers (p=0.03). With daily contact lens wear, there was a significant positive correlation between the changes in IL-1β and IL-8, TNF-α, IL-10 and IL-12(p70) (all p

Objectives To describe the past 20 years' contemporary management modalities for keratoconus (KC) and their outcomes and failures and to propose recommendations in reporting their outcomes. Methods A systematic review of the literature on KC management options for the last 20 years was performed. Original articles that reported the outcome of any form of KC management other than full-thickness or lamellar corneal graft were reviewed to collect information on their outcomes and complications and the level of scientific evidence of the study. Results A total of 1,147 articles retrieved and of those, 241 satisfied the inclusion criteria, and 41.1% of them were prospective case series. The higher level of evidence studies, that is, randomized controlled trials (RCTs), were limited to only one intervention, that is, corneal collagen cross-linking (CXL). However, the quality of most RCTs was limited because of performance and detection bias. Contact lenses (CLs) remain the mainstream of KC management and were associated with reversible and non-sight-threatening complications. Surgical options such as intracorneal segment implantation and phakic intraocular lens are considered in the visual rehabilitation of CL intolerants, and CXL is the only available option to stop or delay the disease progression. Generally, these surgical procedures are associated with transient inflammatory events and permanent sequelae. Conclusions The published studies on KC management options varied significantly with respect to the study population, age of participants, severity of KC, clinical outcome measurements, and follow-up period. High-quality longer follow-up RCTs are required to evaluate the long-term effects of KC interventions.

PURPOSE To establish the effect of lipid supplements on the tear lipid layer and their influence on lens wear comfort in habitual lens wearers. METHODS Forty habitual soft contact lens wearers were recruited to a double-masked, randomized crossover trial. An emulsion drop containing phosphatidylglycerine (Systane Balance; Alcon) and a saline drop as a placebo or a liposomal spray containing phosphatidylcholine (Tears again; BioRevive) and a saline spray as a placebo were used three times a day for 2 weeks with 48 hours washout between each intervention. Ocular comfort, lipid layer grade, and stability of the tear film using a Tearscope and tear evaporation rate using a modified VapoMeter were assessed after 6 hours of lens wear with lenses in situ. RESULTS Neither of the lipid supplements improved lens wear comfort compared to baseline. The noninvasive surface drying time significantly reduced with the placebo spray at day 1 (P = .002) and day 14 (P = .01) whereas the lipid spray had no effect. With the lipid drop and placebo, noninvasive surface drying time was unchanged compared to baseline (P > .05) on day 1, but by day 14, noninvasive surface drying time was reduced with the lipid drop (P = .02) and placebo (P < .001). Symptomatic wearers showed shorter noninvasive surface drying time compared to asymptomatic wearers with the spray treatment on both days (P = .03) but not with the lipid drop (P = .64). The placebo drop significantly changed the lipid layer distribution (P = .03) with a higher percentage of thinner patterns compared to the baseline distribution at day 14. A weak but significant correlation was shown between ocular comfort and noninvasive surface drying time (r = -0.21, P = .003) and tear evaporation rate (r = 0.19, P = .008). Ocular comfort was not associated with lipid layer patterns (r = 0.13, P = .06). CONCLUSIONS Ocular comfort during contact lens wear improved with increased tear film stability and a reduced tear evaporation rate. However, the lipid supplements did not improve ocular comfort from baseline.

Purpose To evaluate corneal sensitivity and its association with other clinical parameters in keratoconus. Methods Twenty-four subjects with keratoconus aged between 18 and 65 years were recruited in this cross-sectional study. Ocular symptoms, corneal topography, tear variables such as tear osmolarity, volume and lower tear meniscus height, ocular surface staining, central sensitivity threshold (CST), and corneal subepithelial nerve mapping were obtained. Association between central CST and other clinical variables was examined using the Spearman correlation coefficient. Partial correlation was performed to control for effects of confounding factors. Results Data from the most severe eye of each subject were included in analyses. Based on the maximum simulated keratometry (Kmax) reading, subjects were graded as having mild (N = 11; K max ≤ 52 D) or severe (N = 13; K max > 52 D) keratoconus. Central corneal sensitivity was lower (ie, increased CST) in the severe keratoconus group compared with that in the mild keratoconus group (median, interquartile range: 1.09; 0.60-19.66 vs. 0.51; 0.39-1.51 g/mm, P = 0.035). In bivariate correlations, reduced corneal sensitivity in keratoconus was associated with age (ρ = 0.42, P = 0.040), disease duration (ρ = 0.49, P = 0.015) and severity (ρ = 0.44; P = 0.032), lower central nerve fiber density (ρ = -0.68, P = 0.014), contact lens wear (ρ = 0.44; P = 0.025), and contact lens tolerance (ρ = 0.46; P = 0.033). After adjusting for contact lens wear, reduced corneal sensitivity was negatively associated with ocular symptoms (ρ = -0.426, P = 0.048) and pain sensitivity (ρ = -0.423, P = 0.045) and positively associated with corneal staining (ρ = 0.52, P = 0.011). Conclusions Altered corneal sensitivity in keratoconus affected ocular symptoms and ocular surface health, which may have significant impact on the success of management options for keratoconus.

Contact lenses are associated with discomfort during wear. This may be the result of stimulation of the ocular surface and production of pro-inflammatory mediators which are then released into the tears. This study examined changes in the concentration in tears of arachidonic acid metabolites (AAM) prostaglandins, cysteinyl leukotrienes, and resolvin-DTears were collected twice a day (morning and evening) for up to 10 days on two different occasions (with and without contact lens wear) from 30 experienced contact lens wearers for analysis of AAM and a separate group (N = 33) for analysis of histamine. Ocular comfort was rated subjectively on an ordinal scale at each time of tear collection. Tears were analyzed using commercial immunoassay-based kits. Statistical analysis was performed using linear mixed model test.Ocular comfort decreased from morning to evening with and without contact lenses (p = 0.001), and the difference in comfort in the evening was greater with contact lens wear (80.9 ± 16.2 vs. 75.5 ± 16.8; p = 0.008). The total concentration of PGs (10.7 ± 10.8 ng/ml), cysteinyl leukotrienes (8.7 ± 0.38 ng/ml), resolvin-DProstaglandins, cysteinyl leukotrienes, resolvin-D1, or histamine concentrations did not alter in relation to changes in comfort of the eye during the day or during contact lens wear. These results suggest that release of these mediators is not responsible for contact lens discomfort.

Purpose The aim of this study was to examine the literature with respect to inflammation of the ocular surface and the presence of inflammatory mediators in the tear film during contact lens wear. Methods The literature on contact lens discomfort that relates to signs of inflammation was searched. Reference was paid to the cardinal signs of inflammation (pain, heat, redness, and swelling) as well as the appearance of inflammatory mediators in the tear film during contact lens wear. Results Contact lens wear does induce discomfort, which is a mild form of pain, and wearing of lenses can induce increases in limbal and conjunctival redness. However, there is little evidence for a direct relationship between limbal or conjunctival redness and contact lens discomfort. Contact lenses may increase the temperature of the ocular surface by a small amount (≤2.1°C). Corneal or conjunctival swelling has not been associated with contact lens discomfort. Complement or kinin breakdown products or histamine in tears are not associated with contact lens discomfort. Cytokines are generally not related to discomfort, although nerve growth factor levels were increased in tears of symptomatics in one report. The presence of degraded lipids, leukotriene B4 and peroxidation products, and the enzyme secretory phospholipase A2 in tears has been associated with contact lens discomfort, and this area is worthy of further research. Conclusions There have been only a few studies that have investigated a role for inflammation in contact lens-induced discomfort, and the strongest associations have been found with lipid degradation processes.