Saturable and Reverse Saturable Absorption Features of Lissamine Green Dye in Different Solvent Media.

Organic dyes have shown a remarkable nonlinear optical (NLO) characteristics under low power laser regime because of chemical stability, large nonlinearity and high susceptibility. With this view in mind, herein we report the third-order NLO features of lissamine green dye using 5 mW power laser with operating wavelength of 650 nm. The lissamine green dye is dissolved into various polar solvents such as ethanol, methanol, acetone and DMSO. The closed aperture Z‒scan approach discloses the information about nonlinear refractive index, whereas the open aperture Z‒scan technique provides the information about nonlinear absorption coefficient. The closed aperture curve shows the peak-valley transmittance is the result of self-defocusing nonlinearity and the open aperture transmittance curve switchover from saturable absorption to reverse absorption in high polar solvent due to polarizability and dipole moment. The third-order NLO features such as nonlinear refractive index (n2), nonlinear absorption coefficient (ß), real and imaginary features of third-order NLO susceptibility of the dye is calculated to be the order of, 10-7 cm2/W 10-3 cm/W, and 10-7 esu respectively. The correlation between solvent polarizability and dipole moment on lissamine green dye is discussed. The results are revealed that the lissamine green dye is a good candidate for NLO applications.

Analysis of ocular surface damage and visual impact in patients with primary and secondary Sjögren syndrome.

The objective of this study is to review the ocular surface changes and complications of patients with Sjögren syndrome and assess their visual impact. A retrospective, cross-sectional, observational, and descriptive study of patients with Sjögren syndrome diagnosed according to the American-European Consensus Group criteria was designed. Data including age, gender, the reason for consultation, associated systemic disease, visual acuity, and ocular complications were recorded. Dry eye tests including tear meniscus thickness; tear film break-up time; ocular surface staining (fluorescein and lissamine green); and Schirmer I test were performed. A total of 249 patients, 233 women (93.6%) and 16 men (6.4%) were studied. Meibomian gland dysfunction was found in 46% (n = 229 eyes) patients; shortened tear film break-up time in 44% (n = 220 eyes); decreased tear meniscus in 49% (n = 243 eyes); significant superficial punctate keratopathy in 49% (n = 242 eyes); a mean ocular surface staining score of 5.92 points; and a low score for Schirmer I test (mean = 5.4 mm). Eyes with a 4 + corneal fluorescein score showed the worst BCVA (mean = 0.63 ± 0.66 LogMAR, ≤ 20/80 Snellen eq., 95% CI 0.29-0.97), compared to 1 + to 3 + scores (mean = 0.211 ± 0.37 LogMAR, 20/32 Snellen eq., 95% CI 0.53-1.15). Ten eyes (4.0%) presented central corneal ulceration with a mean visual acuity of 20/500 (96% visual loss). Ocular surface alterations related to severe dry eye and complications from Sjögren syndrome may have a significant impact on visual acuity. Secondary Sjögren syndrome to rheumatoid arthritis had the worse dry eye prognosis, visual outcome, and ocular complications.

Antimicrobial activity of topical dyes used in clinical veterinary ophthalmology.

OBJECTIVE: To evaluate in vitro the antibacterial effects of fluorescein, rose bengal, and lissamine green topical ophthalmic dyes against selected Gram-positive and Gram-negative bacteria, and to evaluate whether preserved or preservative-free fluorescein solutions are able to inhibit or potentiate bacterial growth. PROCEDURES: Susceptibility testing was performed using the Kirby-Bauer disk diffusion method plated with clinical ocular isolates of Staphylococcus aureus, Staphylococcus pseudintermedius, Streptococcus spp., Escherichia coli, and Pseudomonas aeruginosa. Bacterial growth inhibition was evaluated 24 hours following the addition of commercially available fluorescein, rose bengal, and lissamine green sterile strips. Antimicrobial effectiveness testing was performed by inoculation of compounded 1% dye solutions, both with and without preservatives (fluorescein and lissamine contained thiomersal, and rose bengal contained nipagin and nepazol), with the five previously mentioned bacteria. Growth was evaluated at days 7, 14, and 28. RESULTS: All dyes showed antibacterial activity against Gram-positive organisms. Preservative-free compounded 1% fluorescein solution inhibited growth of Gram-positive organisms but not of Gram-negative organisms. Preservative-free rose bengal and lissamine green inhibited growth of both types of organisms. CONCLUSIONS: Preferably, ocular surface samples for antimicrobial culture should be taken prior to the administration of topical dyes, due to their potential antibacterial activity, particularly if undiluted strips are applied directly or commercial fluorescein solutions are used and not immediately rinsed. Ophthalmic dye solutions containing preservative are safe from bacterial growth for up to 28 days if properly handled and stored. The use of preservative-free fluorescein solutions should be avoided and preservative-free rose bengal and lissamine green should be handled carefully.

Conjunctival staining with lissamine green as a predictor of tear film deficiency in dogs.

OBJECTIVE: To evaluate a grading scheme for conjunctival staining patterns with lissamine green ocular dye in the diagnosis of tear film deficiencies in dogs. PROCEDURES: Client-owned and research colony dogs were enrolled in a prospective study between February and October 2018 in which slit-lamp biomicroscopy, Schirmer tear test (STT), tear film breakup time (TFBUT), conjunctival lissamine green staining (LGS), and intraocular pressure (IOP) measurement were performed in both eyes of all dogs. Lissamine green staining of the temporal bulbar conjunctiva was graded from 0-3, with a higher grade corresponding to an increased stain intensity. RESULTS: Fifty-four dogs (107 eyes), comprising 31 males and 23 females with a mean age of 5.0 ± 3.9 years (range 0.5-14.3), were enrolled in the study. STT was <15 mm/min in 21 eyes and ≥15 mm/min in 86 eyes. Lissamine green staining grade for eyes with a STT of <15 mm/min (2.0 ± 0.9) was significantly higher than for eyes with a STT ≥15 mm/min (0.2 ± 0.7) (P < .001). TFBUT for eyes with a STT <15 mm/min (6.5 ± 4.4 seconds) was significantly shorter than for eyes with a STT ≥ 15 mm/min (16.1 ± 3.6 seconds) (P < .001). As LGS grade increased, both STT (P < .001) and TFBUT (P < .001) significantly decreased. CONCLUSIONS: A higher LGS grade was significantly associated with a lower STT and more rapid TFBUT in dogs. Lissamine green ocular dye can be considered as an adjunctive diagnostic test when evaluating tear film deficiency in dogs.

Association between objective signs and subjective symptoms of dry eye disease in patients with systemic sclerosis.

The aim of this study was to evaluate the association between clinical signs and symptoms of dry eye disease (DED) in patients with systemic sclerosis (SSc). This cross-sectional observational study included 19 SSc patients and 19 normal subjects with no ocular symptoms or ocular surface disorders. Clinical parameters included tear film break-up time (tBUT), Schirmer I, lissamine green (LG) dye, and tear film osmolarity tests, tear production, and tear secretion flow. For assessment of the dry eye symptoms, the Ocular Surface Disease Index (OSDI) questionnaire was administered to all patients. The following mean values were found in SSc patients: OSDI 33.6 ± 19.86; osmolarity of the tear fluid 310.8 mOsmol/l ± 14.47; tBUT time 5.158 ± 2.328 s; Schirmer I test 5.395 mm/5 min; LG grading score 2.026 ± 0.8893; collected tear fluid volume 6.397 ± 2.761 µl. The calculated average tear velocity was 4.654 ± 1.963 µl/min. A significant correlation was found between the OSDI as a subjective parameter and disease duration. Early recognition of dry eye symptoms, a possible extra-intestinal manifestation of SSc, should be included in the check up of the disease to reduce ocular complications. The objective tear functional tests were strongly influenced by individual factors like age and disease duration.

Non-invasive assessment of porcine oocyte quality by supravital staining of cumulus-oocyte complexes with lissamine green B.

We evaluated the usefulness of lissamine green B (LB) staining of cumulus-oocyte complexes (COC) as a non-invasive method of predicting maturational and developmental competence of slaughterhouse-derived porcine oocytes cultured in vitro. Cumulus cells of freshly aspirated COCs were evaluated either morphologically on the basis of thickness of cumulus cell layers, or stained with LB, which penetrates only non-viable cells. The extent of cumulus cell staining was taken as an inverse indicator of membrane integrity. The two methods of COC grading were then examined as predictors of nuclear maturation and development after parthenogenetic activation. In both cases LB staining proved a more reliable indicator than morphological assessment (P < 0.05). The relationship between LB staining and cumulus cell apoptosis was also examined. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay for DNA fragmentation revealed that oocytes within COCs graded as low quality by either LB staining or visual morphology showed significantly greater DNA fragmentation (P < 0.05) than higher grades, and that LB and visual grading were of similar predictive value. Expression of the stress response gene TP53 showed significantly higher expression in COCs graded as low quality by LB staining. However expression of the apoptosis-associated genes BAK and CASP3 was not significantly different between high or low grade COCs, suggesting that mRNA expression of BAK and CASP3 is not a reliable method of detecting apoptosis in porcine COCs. Evaluation of cumulus cell membrane integrity by lissamine green B staining thus provides a useful new tool to gain information about the maturational and developmental competence of porcine oocytes.

[Contemporary diagnostics of the primary Sjögren's syndrome].

Primary Sjögren's syndrome is one of the most common systemic autoimmune disorders, whose diagnosis is very often delayed. In most cases it is a mild disease with symptoms such as dryness and musculoskeletal pain and fatigue but 20-40% of patients suffer from severe systemic manifestations. Extraglandular manifestations can be the first sign of the disease, therefore it is very important to diagnose them as early as possible. Classification criteria established by the American-European Consensus Group (AECG) have been applied in primary Sjögren's syndrome diagnosis since 2002. They took both subjective - dryness of eyes and mouth - and objective tests - imaging and functional tests of salivary glands, ocular tests, histopathologic test of minor salivary glands and presence of typical autoantibodies - into account. New classification criteria proposed in 2012 by the American College of Rheumatology (ACR) include objective tests only. Most recent research data suggest that noninvasive ultrasound examination of salivary gland should be included in the diagnostics process of Sjögren's syndrome due to its high specificity values which are comparable to those obtained from minor salivary glands biopsy. Also, it is important to evaluate disease activity in patients with primary Sjögren's syndrome using a score index - ESSDAI (EULAR Sjögren's Syndrome Disease Activity Index).

Ocular surface changes following botulinum toxin injection for strabismus.

CONTEXT: Botulinum toxin injection is widely used for many purposes, including neuromuscular diseases, movement disorders and strabismus. OBJECTIVE: To evaluate and report the impact of botulinum toxin injection on ocular surface parameters in patients with strabismus. PARTICIPANTS: Twenty-six consecutive patients who underwent botulinum toxin A injection for ocular misalignment were recruited for this prospective study. MATERIALS AND METHODS: Testing of ocular surface parameters including tear break-up time (BUT), lissamine green (LG) staining and Schirmer test (under topical anesthesia) was performed, and Ocular Surface Disease Index (OSDI) questionnaire scores were recorded before and at certain time points after injection (one week, two weeks, one month, three months and six months after botulinum toxin A injection). Two-way analysis of variance (ANOVA) with repeated measures, Friedman's test and Wilcoxon test were used for statistical analysis. RESULTS: Twenty six patients (15 eso- and 11 exo-deviations) with a mean age of 32.46 ± 14.41 (17-65) years were recruited for the study. All injections were performed in one eye. The mean amount of deviation at near and distance reduced after injection. The change of BUT, LG staining and OSDI scores was found to be significant during follow-up in treated eyes (p = 0.001, p = 0.007 and p = 0.009, respectively) whereas the change was insignificant for the Schirmer test results (p = 0.266). CONCLUSIONS: The ocular surface parameters appear to be altered by botulinum toxin injection in strabismic patients. Even though these effects seemed to be temporary, the findings of the present study support the notion of botulinum toxin effects on ocular surface parameters.

Advantages of Lissamine Green Vital Staining as an Endpoint in Dry Eye Clinical Trials.

Purpose: The absence of a standardized diagnostic method for clinical signs of Dry Eye Disease (DED) complicates clinical trials for future treatments. This paper evaluated Lissamine Green (LG) conjunctival staining as a valid, stable and modifiable endpoint for both clinical practice and clinical trials. Methods: Screening and pre-randomization data from two identically designed clinical trials for DED resulted in a pooled dataset of 494 subjects. Inclusion was based on reported symptoms, lissamine green (LG) conjunctival staining, Fluorescein (Fl) corneal and conjunctival staining, and Schirmer's Test (ST). Outcome measures were assessed based on the modifiability of LG staining to exposure to a Controlled Adverse Environment (CAE®), correlation of LG to Fl staining, relative variation of LG staining scores and Schirmer test scores, and the correlation of LG staining with symptom scores. Results: The modifiability of LG conjunctival staining to environmental exposure was demonstrated, with nasal LG and FL staining displaying the most similar percent change. Nasal LG conjunctival staining scores for subjects with ST scores of less than 8mm were significantly higher than for subjects with ST greater than 8mm. LG staining scores were more consistent (25% change from baseline threshold) than ST scores. Finally, statistically significant correlations were found between LG staining and a number of symptom scores. Conclusion: This evaluation demonstrates the superiority of the utilization of a clinical endpoint focused on ocular surface damage. The reproducibility and modifiability of LG conjunctival staining to controlled adverse environment, coupled with its significant correlation with symptoms, positions it as an exemplary clinical sign endpoint for clinical management and in clinical trials. Our findings advocate for the adoption of LG conjunctival staining as a primary endpoint in both clinical research and drug development, offering a more effective means of identifying and addressing ocular surface damage in the realm of DED.

Development and validation of a new photographic scale to grade lid wiper epitheliopathy.

PURPOSE: Lid wiper epitheliopathy (LWE) is a clinical sign that has been associated with dry eye disease (DED) and contact lens discomfort (CLD). This study describes the development, validation and graders' preference of a new photographic scale for LWE, the Photographic Lid Wiper Epitheliopathy (PLWE) scale. METHODS: The PLWE grading scale was developed using LWE images selected from 57 screened patients (≥18 years of age) with confirmed LWE in both eyes. To validate the PLWE scale, a set including 20 images showing varying degrees of LWE from none to severe was chosen. To assess grading validity and grading reliability, observers were asked to grade the selected images using the PLWE and another commonly used subjective LWE grading protocol (Korb) on two separate sessions. RESULTS: The mean grade (±SD) of all images was not statistically significant different between the PLWE scale (1.55 ± 0.44) and the alternative grading scale (Korb, 1.47 ± 0.54) (ANOVA F1, p > 0.05). The average difference from the mean of all graders was 0.03 ± 0.53 using the PLWE scale and 0.06 ± 0.57 when using the Korb protocol (ANOVA F1, p > 0.05). The Coefficient of Repeatability was 1.04 and 1.12 for the PLWE and Korb scales (p > 0.05). Ninety-five percent of the graders found PLWE easier to use than Korb and the same percentage would consider using the PLWE scale in clinical practice. CONCLUSION: The format of the PLWE is similar to other anterior segment visual grading scales and this study revealed an ease of use preference for employing the PLWE by the graders. The presence of LWE has been associated with DED and CLD, and the addition of this new photographic scale could facilitate clinical judgement and record keeping of LWE in clinical practice.

The effect of time on grading corneal fluorescein and conjunctival lissamine green staining.

PURPOSE: To explore the effect of time on grading corneal fluorescein and conjunctival lissamine green staining in dry eye disease (DED). METHODS: Photographs of 68 subjects with non-Sjogren's DED (nSS DED) and 32 with Sjogren's DED (SS DED) were taken of corneal fluorescein staining, then conjunctival lissamine green staining every 30 s for at least 5 min. Photographs of one randomly selected eye were then randomly ordered and graded on a scale from 0 to 5 (severe staining) by two clinicians, masked to both site and subject. The average time required to reach the maximum grade of staining (Gmax) was calculated. RESULTS: The median time (upper and lower quartiles) to corneal fluorescein Gmax was 2.6 (1.3-5.3) minutes for nSS DED and 3.8 (2.6-5.4) minutes for SS DED, a statistically significant difference (Mann Whitney U test, p = 0.018). In contrast, the median time to the Gmax for lissamine green staining of the nasal and temporal conjunctiva was 0.5 (0.5-1.1 nasal, 0.5-0.8 temporal) minutes for nSS DED and 0.5 (0.5-0.8 nasal, 0.5-0.5 temporal) minutes for SS DED subjects, which was not statistically significant (p ≥ 0.383). CONCLUSIONS: The time required to reach the maximum grade of corneal fluorescein staining, but not conjunctival lissamine green staining, varied widely and was significantly longer in subjects with Sjögren's Syndrome. Early observation of corneal fluorescein staining can lead to under-grading, which may impact the diagnosis and assessment of treatment in DED. Further study of the best time to assess corneal fluorescein staining in various DED populations is warranted.

Natural course of lid wiper epitheliopathy (LWE) in symptomatic contact lens wearers.

PURPOSE: To establish the time course of lid wiper epitheliopathy (LWE) in established CL wearers after a single day (6-10 h) of daily disposable contact lens (CL) wear, the following day post-CL removal and 1-week after CL discontinuation. METHODS: Twenty-one symptomatic (CLDEQ-8 score ≥ 12) habitual wearers of MyDay® silicone hydrogel daily disposable were included. LWE staining was assessed prior to CL wear (Visit 1, V1) using semi-automated analysis after instillation of two drops of 1 % lissamine green (10 µL) that were applied to the superior bulbar conjunctiva. LWE measurements were repeated after 6-10 h of continuous CL wear (Visit 2, V2), post-CL removal the following day (Visit 3, V3) and after 1-week CL discontinuation (Visit 4, V4). At each visit, ocular symptoms were evaluated using the SPEED-8 questionnaire and set of 0-100 visual analogue scales (VAS). RESULTS: LWE showed no significant changes after 6-10 h of continuous CL wear (p = 0.536), post-CL removal the following day (p = 0.677) or following 1-week of CL discontinuation (p = 0.478). Analysis revealed a significant improvement in symptomatology between V1 and V2 (SPEED-8, p < 0.01) and also improvements in the 0-100 VAS scores between V2 and V4 for average daily dryness (p < 0.01), end-of-day dryness (p < 0.01) and frequency of end-of-day dryness (p < 0.05). CONCLUSION: The present data suggest that the etiology of LWE is multifactorial and the sole intervention of temporarily discontinuing CL wear does not lead to resolution of these clinical signs.

Effect of Autologous Platelet-Rich Plasma Drops in the Treatment of Ocular Surface Disease.

Purpose: Platelet rich plasma (PRP) is an autologous preparation that concentrates platelets in a small volume of plasma. The purpose of this study was to determine if PRP eye drops improved the symptoms and signs of ocular surface disease. Patients and Methods: A retrospective case series was conducted of patients who were prescribed PRP eye drops. Subjects were excluded if they did not have follow-up, underwent intraocular surgery prior to follow-up, received nerve growth factor treatments, or did not have a baseline examination with photography. Symptoms were assessed using the Ocular Surface Disease Index (OSDI). Patients also underwent a slit lamp exam, ocular surface staining with fluorescein and lissamine green, and Schirmer testing. Results: The charts of 47 patients treated with PRP drops for ocular surface disease were reviewed. Sixty-four eyes of 32 patients were included in the study who had photographs of lissamine green staining taken at baseline and at follow-up. Thirteen patients (28%) had ocular graft-versus-host disease, 16 patients (34%) had Sjögren's syndrome, and 4 patients (8.5%) had rheumatoid arthritis. There was a statistically significant decrease in OSDI score from baseline to follow-up (39.5 vs 30.8 points, p = 0.02). Among the 64 eyes included, 9 (14%) had an improvement in conjunctival lissamine green staining, while 6 (9%) had an increase in staining at follow-up. Among the 20 eyes with Schirmer testing, there was a borderline significant increase in score from baseline to follow-up (5.9 vs 9.7, p = 0.06). Among the 44 eyes that had corneal fluorescein staining (CFS) reported, 8 (18.2%) had decreased staining and 2 (4.5%) had increased staining at follow-up. Conclusion: Treatment with PRP drops was associated with a significant improvement in symptoms in patients with ocular surface disease. Future larger prospective studies are needed to further evaluate the efficacy of PRP drops for treating ocular surface disease.

Optimal methodology for lid wiper epitheliopathy identification.

PURPOSE: Lid wiper epitheliopathy (LWE) is a clinical sign that has been associated with dry eye disease. This study used a semi-automated method to identify the effect of drop instillation and post-dye viewing time on the absorption of lissamine green (LG) and sodium fluorescein (NaFl) on the upper eyelid in order to ascertain the optimal identification for LWE assessment. METHODS: In 37 participants with LWE, 1-drop of 1% LG (10 µL) was applied to the superior bulbar conjunctiva in the right eye, and photographs of the lid margin were taken 1, 3, and 5 min after instillation. Measurements were repeated in the same eye following instillations of 2-drops of 1% LG. The same procedures were followed for application of 2% NaFl (2 µL) to the left eye. Staining area was determined using software to detect and measure dye-stained images. Analysis used a linear mixed model with fixed effects of time, number of drops and their interaction. RESULTS: For LG, multivariate analysis showed that time of drop instillation was significant (p = 0.0091) as was the area of staining in the 2-drop versus 1-drop condition (p < 0.0001). For NaFl, there was a significant effect of time (p < 0.0001), drops (p < 0.0001), and a time/drops interaction (p < 0.0134), suggesting that both time and number of drops are important. CONCLUSION: A single drop of dye is insufficient to reveal the full extent of LWE staining. A 2-drop instillation is recommended and observation is recommended between 1 and 5 min (LG) and between 3 and 5 min (NaFl).

Are there Clinical Ways to Assess Inflammation in Dry Eye Disease?

In the diagnostic process of dry eye disease, the detection of inflammatory activity is critical in order to evaluate the risk of progression and immunologic shift of the disease, to predict patient response to treatment, and to design an efficient therapeutic strategy, including artificial tear replacement, punctal occlusion or anti-inflammatory therapy.Even if it is difficult to quantify, some indicators of the presence of inflammation are collectible during the examination of the ocular surface in a first-line clinical setting. This review presents and critically discusses the assessment of inflammation in dry eye disease in clinical practice.

Characterization of porcine oocytes stained with Lissamine Green B and their developmental potential in vitro.

Traditional methods for the evaluation of oocyte quality are based on morphological classification of the follicle, cumulus-oocyte complex, polar body and meiotic spindle. This study is focused on the differences between the morphological assessment of oocyte quality, the assessment based on Lissamine Green B (LB) staining and the analysis of oocytes using a proteomic approach. We evaluated the effectiveness of electrochemical and chemical parthenogenetic activation under our laboratory conditions and evaluated the applicability of Lissamine Green B staining of cumulus-oocyte complexes (COCs) as a non-invasive method for predicting the maturational and developmental competence of porcine oocytes cultured in vitro. We determined that chemical parthenogenetic activation using ionomycin and 6-dimethylaminopurine was slightly more effective than electrochemical activation. After oocyte selection according to LB staining, we found significant differences (P<0.05) between the LB- group and LB+ group and the control group in their maturation, cleavage rate and rate of blastocysts. Proteomic analyses identified a selection of proteins that were differentially expressed in each group of analysed oocytes. Oocytes of the LB- group exhibited an increased variability of proteins involved in transcription regulation, proteosynthesis and the protein folding crucial for oocyte maturation and further embryonic development. These results found a better competence of LB- oocytes in maturation, cleavage and ability to reach the blastocyst stage.

Impact of Improper Approach to Identify Lid Wiper Epitheliopathy (LWE).

PURPOSE: Variability in the use of ophthalmic dyes to diagnose lid wiper epitheliopathy (LWE) has led to division in the literature and clinical practice. The principal aim of this study was to evaluate whether the use of a non-optimal methodology to identify LWE had a potential for false negatives; in which LWE was overlooked. PATIENTS AND METHODS: A total of 20 participants were initially categorized to not have LWE and were enrolled in this study. The protocol examined whether or not LWE would later be revealed through the use of optimized methodology. Semi-automated analysis was performed of images taken after two different drop instillations with varying post-dye viewing times for both lissamine green (LG) and sodium fluorescein (NaFl). RESULTS: There was a significant increase in area of staining revealed when an optimal methodology for LWE identification was used. Comparisons for every non-optimal condition were statistically significantly different against the optimal condition (all p<0.01). The use of a non-optimal methodology resulted in a 70% false-negative rate when using LG and a 95% false-negative rate when using NaFl. CONCLUSION: The study demonstrated that using a double instillation of dye was statistically different from a single-dose, even with extended wait time for clinical observation. A single instillation did not offer adequate volume of dye for adequate lid margin uptake. A careful adherence to volume as well as a repeat administration is key to revealing the full area of LWE. A non-optimal approach to diagnose LWE can lead to false negatives.

Review and analysis of grading scales for ocular surface staining.

Vital dye staining has been used for over a century to assess the severity of ocular surface disease. However, despite common usage, a universally accepted "gold standard" grading scale does not exist for corneal and conjunctival staining, which can impact the ability to diagnose and monitor ocular surface conditions such as dry eye. The Food and Drug Administration (FDA) and other international regulatory agencies rely on ocular surface staining as a primary endpoint for new drug approvals, so that absence of a "gold standard" scale may affect approval of new drug treatments. To begin to address this problem, we review existing, published grading scales in an integrated fashion, highlighting their differences and similarities to emphasize common themes and the methods and elements that are important in creating a standardized scale. Our goal is to aid the field in moving towards an accepted standardized grading scale for ocular surface staining that can be applied in clinic and research settings for a variety of ocular conditions.

Lid wiper epitheliopathy: The influence of multiple lid eversions and exposure time.

PURPOSE: To investigate the effect of multiple lid eversions on lid wiper epitheliopathy (LWE), along with the effect of cumulative lid exposure time and the patterns of associated staining. METHODS: The increase in area of lid wiper staining with lissamine green was compared by everting both the upper eyelids of each subject (i.e. contralateral design), with one eye being everted once for 45 s and the fellow eyelid everted three times, each time for 15 s. This pattern of contralateral eversion was repeated with a total of three eversions in one eye and nine eversions in the fellow eye, with each eye totalling 135 s cumulative exposure to eversion over about 9 min. The LWE area of staining was objectively quantified from slit lamp photography images captured at every lid eversion by 2 masked observers. Two-way repeated measures ANOVAs were used to determine the effect of number of lid eversions and cumulative exposure time on the amount of staining caused. Each image was also categorized into its primary LWE staining pattern, by a masked observer. RESULTS: The multiple eversions condition caused significantly greater LWE than the single eversion condition (p < 0.001), while cumulative exposure time did not have a significant effect on LWE (p = 0.137). Classification of the primary staining patterns revealed that with more eyelid eversions there was a shift from mostly 'no staining' to minor patterns ('short horizontal bands' and 'vertical streaks') and then to more extensive patterns ('broad horizontal bands' and 'comb-shaped'). CONCLUSIONS: The number of eyelid eversions is a confounding factor that should be controlled when investigating LWE, in particular when considering the link with dry eye or contact lens discomfort. However the cumulative exposure time did not appear to influence the LWE magnitude.

Effect of upper eyelid blepharoplasty on the ocular surface, tear film, and corneal microstructure

ABSTRACT Purpose: This study aimed to investigate the effect of upper eyelid blepharoplasty with the removal of the skin and a strip of orbicularis oculi muscle on the ocular surface, tear film, and dry eye-related symptoms. Methods: Twenty-two eyes of 22 consecutive patients operated by a single surgeon (21 females; mean age, 61 years; age range, 41-75 years) were included. All subjects completed the Ocular Surface Disease Index questionnaire, underwent in vivo confocal microscopy, tear film breakup time measurements, the Schirmer test with anesthesia, and fluorescein and lissamine green staining measurements before, 1 month, and 6 months after upper blepharoplasty alone with preseptal orbicularis excision. Results: A significant increase in Ocular Surface Disease Index, and corneal fluorescein and lissamine green staining and a significant decrease in tear film breakup time were observed after 1 month (p=0.003, p=0.004, p=0.029, and p=0.024 respectively) and 6 months (p=0.001 for all findings). No significant difference in the Schirmer test score was observed during the follow-up. None of the in vivo confocal microscopy parameters showed significant changes during the study. Conclusions: An increase in dry eye symptoms and a decrease in tear film stability along with ocular surface staining were observed in patients undergoing upper eyelid blepharoplasty.