The Use of Complementary Luminescent and Fluorescent Techniques for Imaging Ca2+ Signaling Events During the Early Development of Zebrafish (Danio rerio).

We have visualized many of the Ca2+ signaling events that occur during the early stages of zebrafish development using complementary luminescent and fluorescent imaging techniques. We initially microinject embryos with the luminescent Ca2+ reporter, f-holo-aequorin, and using a custom-designed luminescent imaging system, we can obtain pan-embryonic visual information continually for up to the first ~24 h postfertilization (hpf). Once we know approximately when and where to look for these Ca2+ signaling events within a complex developing embryo, we then repeat the experiment using a fluorescent Ca2+ reporter such as calcium green-1 dextran and use confocal laser scanning microscopy to provide time-lapse series of higher-resolution images. These protocols allow us to identify the specific cell types and even the particular subcellular domain (e.g., nucleus or cytoplasm) generating the Ca2+ signal. Here, we outline the techniques we use to precisely microinject f-holo-aequorin or calcium green-1 dextran into embryos without affecting their viability or development. We also describe how to inject specific regions of early embryos in order to load localized embryonic domains with a particular Ca2+ reporter. These same techniques can also be used to introduce other membrane-impermeable reagents into embryos, including Ca2+ channel antagonists, Ca2+ chelators, fluorescent dyes, RNA, and DNA.

Bilateral Effect of the Unilateral Corneal Nerve Cut on Both Ocular Surface and Lacrimal Gland.

Purpose: This study investigated the effect of a unilateral cut of the corneal nerve on the bilateral ocular surface and tear secretory function. Methods: Seven-week-old female BALB/c mice were divided into control and nerve-cutting (NC) groups (n = 60). The left cornea was partially incised with a 2-mm circular trephine through the upper half of the stromal layer. Lissamine green corneal staining and tear volume measurements were conducted, and corneal whole-mount staining using class III ß-tubulin antibody was performed to assess corneal nerves. Flow cytometric analyses for dendritic cells (DCs), CD4+/CD8+ and regulatory T cells and ELISA for neuropeptides were performed. Results: The grading of corneal staining increased in the NC group, while the tear volume decreased over the 4 weeks. The nerve density decreased in bilateral corneas over 2 weeks. At day 14, CD11b+ or CD11c+ DCs and the mature DCs expressing CD86 or MHCII increased in bilateral cornea/conjunctiva. At day 28, CD11c+CD86hi, CD11c+MHCIIhi, Th17 and IFN-γ-secreting CD8+ T cells highly increased in bilateral draining lymph nodes. CD4+CD25hiFoxp3hi and CD8+CD25hiFoxp3hi regulatory T cells notably increased in the spleen. In ELISA, neuropeptide Y, calcitonin gene-related peptide, and vasoactive intestinal peptide were generally suppressed in the extraorbital lacrimal glands at day14. Conclusions: The unilateral corneal nerve severing resulted in activation of the immune cells on the ocular surface and dysregulated lacrimal secretion bilaterally through the bidirectional neuronal signals. It suggests that the unilateral corneal nerve damage may alter immune homeostasis and mechanistically participate in the development of bilateral inflammatory disorders such as dry eye.

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.

Ocular Surface Alterations in the Context of Corneal In Vivo Confocal Microscopic Characteristics in Patients With Fibromyalgia.

PURPOSE: To quantify the morphology of corneal basal epithelium and subbasal nerves and to evaluate the ocular surface alterations in patients with fibromyalgia (FM). METHODS: Patients with FM (n = 34) and healthy controls (n = 42) were enrolled. All participants underwent ocular surface tests in the following order: corneal sensitivity, tear film breakup time, lissamine green staining, Schirmer test, and the Ocular Surface Disease Index questionnaire. Basal epithelial cells and subbasal nerves were evaluated using in vivo confocal microscopy (IVCM). Demographic characteristics, Visual Analog Scale for Pain (VAS), American College of Rheumatology 1990, the Widespread Pain Index (WPI), and the Symptom Impact Questionnaire (SIQR) scores of patients with FM were obtained. RESULTS: Corneal sensitivity was 0.4 g/mm (fiber length: 6.0 cm) in all eyes. Patients with FM had a higher Ocular Surface Disease Index (42.2 ± 18.9 vs. 1.2 ± 1.7, P < 0.001), higher lissamine green staining scores (0.5 ± 0.5 vs. 0.05 ± 0.2, P < 0.001), and lower tear breakup time scores (9.0 ± 3.6 vs. 10.3 ± 1.5, P = 0.003) than the controls. Basal epithelial cell density (2709 ± 494 vs. 4491 ± 724), total nerve density (1563 ± 620 vs. 2545 ± 973), long nerve fibers (3.4 ± 1.3 vs. 4.5 ± 1.0), and the number of nerves (5.0 ± 1.8 vs. 10.3 ± 2.1) were all lower in patients with FM compared with those of the controls (P < 0.001 for all). There was a statistically significant negative correlation between the WPI score and Schirmer test results (rho = -0.374, P = 0.03) and between WPI and total nerve density (rho = -0.334, P = 0.054). CONCLUSIONS: To the best of our knowledge, this is the first study that evaluated ocular surface alterations in the context of corneal IVCM characteristics. Patients with FM should be evaluated in terms of ocular surface diseases. IVCM may be used in FM to assess small fiber neuropathy.

Antiplasmodial activities of dyes against Plasmodium falciparum asexual and sexual stages: Contrasted uptakes of triarylmethanes Brilliant green, Green S (E142), and Patent Blue V (E131) by erythrocytes.

The search for safe antimalarial compounds acting against asexual symptom-responsible stages and sexual transmission-responsible forms of Plasmodium species is one of the major challenges in malaria elimination programs. So far, among current drugs approved for human use, only primaquine has transmission-blocking activity. The discovery of small molecules targeting different Plasmodium falciparum life stages remains a priority in antimalarial drug research. In this context, several independent studies have recently reported antiplasmodial and transmission-blocking activities of commonly used stains, dyes and fluorescent probes against P. falciparum including chloroquine-resistant isolates. Herein we have studied the antimalarial activities of dyes with different scaffold and we report that the triarylmethane dye (TRAM) Brilliant green inhibits the growth of asexual stages (IC50 ≤ 2 µM) and has exflagellation-blocking activity (IC50 ≤ 800 nM) against P. falciparum reference strains (3D7, 7G8) and chloroquine-resistant clinical isolate (Q206). In a second step we have investigated the antiplasmodial activities of two polysulfonated triarylmethane food dyes. Green S (E142) is weakly active against P. falciparum asexual stage (IC50 ≃ 17 µM) whereas Patent Blue V (E131) is inactive in both antimalarial assays. By applying liquid chromatography techniques for the culture supernatant analysis after cell washings and lysis, we report the detection of Brilliant green in erythrocytes, the selective uptake of Green S (E142) by infected erythrocytes, whereas Patent Blue V (E131) could not be detected within non-infected and 3D7-infected erythrocytes. Overall, our results suggest that two polysulfonated food dyes might display different affinity with transporters or channels on infected RBC membrane.

Biosorption of Food Green 3 by a novel green generation composite biosorbent from aqueous environment.

A green type composite biosorbent composed of pine, oak, hornbeam, and fir sawdust biomasses modified with cetyltrimethylammonium bromide (CTAB) was first used for biosorption of an unsafe synthetic food dye, Food Green 3 from liquid medium in this study. Batch studies were carried by observing the effects of pH, dye concentration, biosorbent amount, and contact time. The equilibrium data were analyzed using Freundlich, Langmuir, and Dubinin-Radushkevich equations. Freundlich model gave a better conformity than other equations. The maximum dye removal potential of biosorbent was found to be 36.6 mg/g based on Langmuir isotherm. The pseudo-first-order, pseudo-second-order, Elovich, and intra-particle diffusion models were applied to clarify the process kinetics of biosorption. The mechanism studies suggested the biosorption process obeying Elovich kinetics and involving pore diffusion. The estimated values of biosorption free energy from Dubinin-Radushkevich isotherm (E value <8 kJ/mol) and thermodynamic studies (0 < ΔG° < -20 kJ/mol) implied a spontaneous, feasible, and physical process. Hence, this investigation suggested that the CTAB modified mix sawdust biomass could be a promising biosorbent for biosorption of such problematic dyes from impacted media.

A Clinical Study of Meibomian Gland Dysfunction in Patients with Diabetes.

AIMS OF THE STUDY: To study the frequency of meibomian gland dysfunction (MGD) in patients of diabetes mellitus with dry eye. MATERIALS AND METHODS: A case-control cross sectional study. SAMPLING: Purposive random sampling. Totally, 200 eyes of 100 patients of diabetes mellitus and an equal number of eyes of normal subjects as control, who were gender and age matched and all of whom were symptomatic for dry eye were assessed for MGD by noting the symptoms and determining the meibomian gland expression scale for volume and viscosity, and ocular surface staining with Lissamine green, and Fluorescein sodium. All the subjects were graded for the severity of MGD. The results were compared in both the groups to ascertain whether the frequency of MGD in diabetics is significantly more as compared to nondiabetics. STATISTICAL ANALYSIS: The data were analyzed by Chi-square test for significance. P < 0.05 was considered as statistically significant. RESULTS: There was a significant increase in the frequency of MGD in diabetics as compared to the nondiabetics. CONCLUSION: Diabetes mellitus is associated with MGD.

Lid Margin Sensitivity and Staining in Contact Lens Wear Versus No Lens Wear.

PURPOSE: To map the sensitivity and staining of the marginal conjunctiva in soft contact lens (SCL) wearers and nonwearers and to investigate diurnal changes. METHODS: Thirty-five nonwearers and 35 SCL wearers were recruited. Mechanical sensitivity was measured at 8 locations on the ocular surface and adnexa (cornea, marginal, bulbar, and tarsal conjunctivae) using the Cochet-Bonnet aesthesiometer. Lid margin staining (lissamine green) was assessed through image analysis. Measurements were repeated at 12 hours in 11 nonwearers and 10 symptomatic SCL wearers. RESULTS: The marginal conjunctiva was the most sensitive conjunctival region (all P < 0.001). There was no significant difference in sensitivity at 12 hours (all P > 0.05). There was a significant difference in sensitivity between the 2 groups in the morning for the lower lid tarsal conjunctiva (P = 0.003). After 12 hours, the sensitivity of the lower lid marginal conjunctiva was found to be reduced in SCL wearers compared with nonwearers (all P < 0.05). Lower lid margin staining was greater than upper lid margin staining in both groups (all P < 0.05). In the SCL wearers, a significant increase in lower lid margin staining was found at 12 hours compared with the morning (P = 0.04). After 12 hours, lower lid margin staining was significantly greater in SCL wearers compared with nonwearers (P = 0.03). CONCLUSIONS: This work has demonstrated that the marginal conjunctiva is the most sensitive conjunctival region. After 12 hours, symptomatic lens wearers showed decreased sensitivity of the lower marginal conjunctiva and increased lower lid margin lissamine green staining compared with those of the nonwearers.

Topical glaucoma therapy and ocular surface disease: a prospective, controlled cohort study.

OBJECTIVE: To evaluate the association between the intensity and duration of glaucoma topical therapy and severity of signs and symptoms of ocular surface disease (OSD). DESIGN: Single-site, prospective, controlled, cross-sectional study. PARTICIPANTS: Sixty-one patients with no diagnosis of or previous treatment for OSD were identified. METHODS: Patients were assigned to 2 groups: the glaucoma group with 31 patients diagnosed with primary open-angle glaucoma and using at least 1 topical intraocular pressure (IOP)-lowering medication and the control group including 30 patients with no diagnosis of glaucoma or history of topical therapy usage. The right eye of each patient was arbitrarily chosen. Each patient completed an Ocular Surface Disease Index (OSDI) questionnaire and underwent evaluation of the ocular surface by conjunctival and corneal lissamine green (LG) staining and tear breakup time (TBUT). The intensity index (drops/wk × therapy duration in years) was calculated to quantify the topical therapy. RESULTS: OSDI scores of the glaucoma group correlated to the intensity index (r = 0.46, CI 0.13-0.69). The glaucoma group had a higher mean OSDI score than the control group (18.97 ± 9.5 versus 6.25 ± 5.7, p = 5.85E-08). Abnormal TBUT and LG staining scores were prevalent in the glaucoma group compared with the control group (68% vs 17%, p = 0.000078; 65% vs 3%, p = 2.9E-07). CONCLUSIONS: Patients on glaucoma therapy have a greater prevalence of OSD symptoms, and their intensity index correlates to the OSDI score. The intensity index reflects quantitatively the amount of treatment and can be further validated in future studies as a predicting tool for OSD development.

In vivo confocal microscopic characterisation of the cornea in chronic graft-versus-host disease related severe dry eye disease.

AIMS: To first describe in vivo confocal microscopic (IVCM) corneal findings in severe dry eye syndrome due to ocular chronic graft versus host disease (cGvHD) after allogeneic stem cell transplantation. METHODS: IVCM of the central cornea was performed in 12 prospectively recruited patients with severe ocular cGvHD associated dry eye syndrome and in six control patients with haematological malignancies without cGvHD. Within each examined corneal layer, at least three non-overlapping areas were selected for representative analysis. RESULTS: The number of sub basal nerve branches was markedly reduced in patients with cGvHD. Sub basal nerve morphology was characterised by increased tortuosity and reduced reflectivity. Accumulation of hyper-reflective extracellular matrix, significantly increased haze and increased keratocyte density were found in the anterior stroma of the study group. CONCLUSIONS: IVCM findings of the cornea in patients with severe ocular cGvHD include a rarefaction of the sub basal corneal nerve plexus and dense accumulation of hyper-reflective extracellular matrix in the anterior stroma.

Tear film osmolarity in ocular mucous membrane pemphigoid.

PURPOSE: The aim of this study was to evaluate tear film osmolarity in patients with ocular mucous membrane pemphigoid (MMP). METHODS: This observational cross-sectional study included 40 patients with biopsy-proven ocular MMP at Foster stage III referred to the tertiary-care Ocular Immunology and Uveitis Service at the San Raffaele Scientific Institute in Milan from June 2010 to August 2013. We evaluated the following clinical parameters: tear film osmolarity, ocular surface disease symptoms (OSDI) questionnaire, Schirmer test, tear film break-up time (TFBUT), and corneal and conjunctival staining. RESULTS: Forty patients (27 women and 13 men) were enrolled. All patients were undergoing systemic immunosuppressive therapy: 19 patients (47.5%) were on methotrexate, 9 (22.5%) were on mycophenolate mofetil, 9 (22.5%) were on low-dose corticosteroids, and 3 (7.5%) were on azathioprine. The mean osmolarity was 322.90 ± 33.39 mOsm/L, the mean OSDI score was 73.2 ± 17.9, the mean TFBUT was 6.60 ± 3.13 seconds, and the mean Schirmer test value was 4.07 ± 3.58 seconds. Tear film osmolarity significantly correlated with the TFBUT (r = 0.80; P < 0.0001), whereas no clinical correlation was found with the Schirmer test value (r = 0.01; P = 0.40) or with the OSDI score (r = 0.02; P = 0.29). Osmolarity did not turn out to be statistically different in the subgroups according to the Oxford corneal staining scale (P = 0.71) and to the Van Bijsterveld conjunctival staining score (P = 0.31). CONCLUSIONS: Tear osmolarity increased in patients with ocular MMP and correlated with the TFBUT. This result emphasizes the role of evaporative dry-eye condition in patients with ocular MMP. Tear osmolarity may be considered as a useful test in the diagnostic assessment of dry eye associated with MMP and for targeting therapeutic decisions.

Epigallocatechin gallate eye drops protect against ultraviolet B-induced corneal oxidative damage in mice.

PURPOSE: Ultraviolet B (UVB) radiation from sunlight is a known risk factor for human corneal injury. The aim of the present study was to investigate the protective effects of green tea polyphenol epigallocatechin gallate (EGCG) on UVB radiation-induced corneal oxidative damage in male imprinting control region (ICR) mice. METHODS: Corneal oxidative damage was induced by exposure to UVB radiation at 560 µW/cm(2). The animals received 0%, 0.1%, and 0.01% EGCG eye drops at a 5 mg/ml dose, twice daily for 8 days. Corneal surface damage was graded according to smoothness and the extent of lissamine green staining. Corneal glutathione (GSH), thiobarbituric acid-reactive substances (TBARS), and protein carbonyl levels, as well as superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px), and glutathione reductase (GSH-Rd) activity in the cornea, were measured to monitor corneal injury. RESULTS: UVB radiation caused significant damage to the corneas, including apparent corneal ulceration and severe epithelial exfoliation, leading to a decrease in SOD, catalase, GSH-Px, GSH-Rd, and GSH activity in the cornea. However, the corneal TBARS and protein carbonyls increased compared with the control group. Treatment with EGCG eye drops significantly (p<0.05) ameliorated corneal damage, increased SOD, catalase, GSH-Px, GSH-Rd, and GSH activity, and decreased the TBARS and protein carbonyls in the corneas compared with the UVB-treated group. CONCLUSIONS: EGCG eye drops exhibit potent protective effects on UVB radiation-induced corneal oxidative damage in mice, likely due to the increase in antioxidant defense system activity and the inhibition of lipid peroxidation and protein oxidation.

Lifitegrast ophthalmic solution 5.0% for treatment of dry eye disease: results of the OPUS-1 phase 3 study.

PURPOSE: To assess the efficacy and safety of lifitegrast ophthalmic solution 5.0% compared with placebo in subjects with dry eye disease. DESIGN: Prospective, randomized, double-masked, placebo-controlled, parallel arm, multicenter clinical trial. PARTICIPANTS: A total of 588 adult subjects with dry eye disease. METHODS: Eligible subjects were randomized 1:1 to receive topically administered lifitegrast (5.0%) or placebo (vehicle) twice daily for 84 days after a 14-day open-label placebo run-in period. After enrollment (day 0), subjects were evaluated at days 14, 42, and 84. Key objective (fluorescein and lissamine staining scores [Ora scales]) and subjective (Ocular Surface Disease Index [OSDI], 7-item visual analog scale, and ocular discomfort score [Ora scale]) measures were assessed at all visits. MAIN OUTCOME MEASURES: The primary objective efficacy measure (sign) was mean change from baseline inferior corneal staining score (ICSS) at day 84. The co-primary subjective efficacy measure (symptom) was the mean change from baseline in the visual-related function subscale score of the Ocular Surface Disease Index (VR-OSDI). Supportive measures included corneal fluorescein scores (superior, central, total region) and conjunctival lissamine scores (nasal, temporal, total region) and symptom scores at day 84. RESULTS: The study met the primary objective efficacy ICSS end point in demonstrating superiority of lifitegrast compared with placebo (P = 0.0007). Lifitegrast significantly reduced corneal fluorescein staining (superior, P = 0.0392; total cornea, P = 0.0148) and conjunctival lissamine staining (nasal, P = 0.0039; total conjunctiva, P = 0.0086) at day 84 versus placebo. Significant (P < 0.05) improvements in nasal and total lissamine scores were observed at day 14 and maintained through day 84. The study did not meet the co-primary subjective VR-OSDI measure (P = 0.7894). However, significant improvements were observed at day 84 in ocular discomfort (P = 0.0273) and eye dryness (P = 0.0291), the most common and severe symptoms reported at baseline in both groups. There were no unanticipated or serious ocular adverse events (AEs). The most frequent reported ocular AEs were transient intermittent instillation site symptoms (irritation, discomfort) primarily on the initial lifitegrast dose at day 0. CONCLUSIONS: Lifitegrast ophthalmic solution 5.0% significantly reduced corneal fluorescein and conjunctival lissamine staining and improved symptoms of ocular discomfort and eye dryness compared with placebo when administered twice daily over 84 days.

Bacterial-fungal interactions enhance power generation in microbial fuel cells and drive dye decolourisation by an ex situ and in situ electro-Fenton process.

In this work, the potential for sustainable energy production from wastes has been exploited using a combination fungus-bacterium in microbial fuel cell (MFC) and electro-Fenton technology. The fungus Trametes versicolor was grown with Shewanella oneidensis so that the bacterium would use the networks of the fungus to transport the electrons to the anode. This system generated stable electricity that was enhanced when the electro-Fenton reactions occurred in the cathode chamber. This configuration reached a stable voltage of approximately 1000 mV. Thus, the dual benefits of the in situ-designed MFC electro-Fenton, the simultaneous dye decolourisation and the electricity generation, were demonstrated. Moreover, the generated power was effectively used to drive an ex situ electro-Fenton process in batch and continuous mode. This newly developed MFC fungus-bacterium with an in situ electro-Fenton system can ensure a high power output and a continuous degradation of organic pollutants.

Factors predicting the ocular surface response to desiccating environmental stress.

PURPOSE: To identify factors predicting the ocular surface response to experimental desiccating stress. METHODS: The ocular surfaces of both eyes of 15 normal and 10 dry eye subjects wearing goggles were exposed to a controlled desiccating environment (15%-25% relative humidity and 2-5 L/min airflow) for 90 minutes. Eye irritation symptoms, blink rate, tear meniscus dimensions, noninvasive (RBUT) and invasive tear break-up time, and corneal fluorescein and conjunctival lissamine green-dye staining were recorded before and after desiccating stress. Pre- and postexposure measurements were compared, and Pearson correlations between clinical parameters before and after desiccating stress were calculated. RESULTS: Corneal and conjunctival dye staining significantly increased in all subjects following 90-minute exposure to desiccating environment, and the magnitude of change was similar in normal and dry eye subjects; except superior cornea staining was greater in dry eye. Irritation severity in the desiccating environment was associated with baseline dye staining, baseline tear meniscus height, and blink rate after 45 minutes. Desiccation-induced change in corneal fluorescein staining was inversely correlated to baseline tear meniscus width, whereas change in total ocular surface dye staining was inversely correlated to baseline dye staining, RBUT, and tear meniscus height and width. Blink rate from 30 to 90 minutes in desiccating environment was higher in the dry eye than normal group. Blink rate significantly correlated to baseline corneal fluorescein staining and environmental-induced change in corneal fluorescein staining. CONCLUSIONS: Ocular surface dye staining increases in response to desiccating stress. Baseline ocular surface dye staining, tear meniscus height, and blink rate predict severity of ocular surface dye staining following exposure to a desiccating environment.

A randomized, multicenter phase 3 study comparing 2% rebamipide (OPC-12759) with 0.1% sodium hyaluronate in the treatment of dry eye.

OBJECTIVE: To investigate the efficacy of 2% rebamipide ophthalmic suspension compared with 0.1% sodium hyaluronate ophthalmic solution for the treatment of patients with dry eye. DESIGN: Randomized, multicenter, active-controlled parallel-group study. PARTICIPANTS: One hundred eighty-eight patients with dry eye. METHODS: Following a 2-week screening period, patients were allocated randomly to receive 2% rebamipide or 0.1% sodium hyaluronate, administered as 1 drop in each eye 4 or 6 times daily, respectively, for 4 weeks. MAIN OUTCOME MEASURES: There were 2 primary end points: changes in the fluorescein corneal staining (FCS) score to determine noninferiority of 2% rebamipide and changes in the lissamine green conjunctival staining (LGCS) score to determine superiority. Secondary objective end points were Schirmer's test results and tear film breakup time (TBUT). Secondary subjective end points were dry eye-related ocular symptoms (foreign body sensation, dryness, photophobia, eye pain, and blurred vision) score and the patients' overall treatment impression score. RESULTS: In the primary analysis, the mean change from baseline in FCS scores verified noninferiority, indicated significant improvement, and, in LGCS scores, verified the superiority of 2% rebamipide to 0.1% sodium hyaluronate. Values for the Schirmer's test and TBUT were comparable between the 2 groups. For 2 dry eye-related ocular symptoms--foreign body sensation and eye pain--2% rebamipide showed significant improvements over 0.1% sodium hyaluronate. Patients had a significantly more favorable impression of 2% rebamipide than of 0.1% sodium hyaluronate; 64.5% rated treatment as improved or markedly improved versus 34.7%, respectively. No serious adverse events were observed. CONCLUSIONS: Administration of 2% rebamipide was effective in improving both the objective signs and subjective symptoms of dry eye. Those findings, in addition to the well-tolerated profile of 2% rebamipide, clearly show that it is an effective therapeutic method for dry eye. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Development of the 4-3-2-1 meibum expressibility scale.

OBJECTIVES: With increased interest in the assessment of meibomian gland dysfunction and evaporative dry eye, there remains a deficit in simple, clinically applicable grading scales for gland expression. A new scale to assess meibum expressibility is described. METHODS: A meibum expressibility scale was developed using a new standardized meibomian gland expression device, which provides constant pressure along the inferior lid. For the scale development, 30 patients (53.0±8.49 years; 93.33% female) with mild-to-moderate dry eye were compared with 13 normal, non-dry eye subjects (25.6±4.3 years; 46.1% female) using the meibum expression device developed by Korb and Blackie. The device was placed 4 glands lateral to the inferior punctum and 1 mm below the lash line and was held stable for 15 sec. The glands expressing meibum were counted. The weighted κ statistic was used to evaluate the extent of agreement, and a receiver operating characteristic curve was created to test the proposed scale. RESULTS: The mean number of glands that expressed from the worse lid in the normal group was 3.54±1.61, whereas 1.53±1.28 glands expressed in the dry eye group. In the dry eye group, 1 subject showed 5 glands expressing, and 29 demonstrated scores of 4 or less. In the normal group, 3 or more glands were expressible in 11 of 13 subjects. CONCLUSIONS: A 4-3-2-1 scoring system is proposed, whereby 4 or greater=normal expressibility, 3=mildly reduced expressibility, 2=moderately reduced expressibility, and 1 or lesser=severely reduced expressibility. Further validation of the scale is warranted.

Rapid detection of ciprofloxacin effects on Fusarium graminearum and F. avenaceum cells in modulating environmental pH using a reactive, non-toxic food-dye indicator.

The objective of the study was to assess the effect of ciprofloxacin antibiotic on the physiological or phenotypic characteristics of food-borne toxigenic Fusarium graminearum and F. avenaceum molds under in vitro conditions. In the presence of ciprofloxacin, Fusarium mycelia growth and morphology were altered based on the antibiotic concentration range used. Results showed that ciprofloxacin in concentrations ≥40µg/mL induced chlamydospore formation in Fusaria and as such, this antibiotic should be considered as an important abiotic stress factor and growth inhibitor. A novel method was investigated to correlate chlamydospore formation with the colour changes observed in FD&C Green Number 3, a common water soluble food dye. The antibiotic-treated F. graminearum and F. avenaceum isolates produced chamydospores, which in turn altered environmental pH with concomitant changes in the colour and intensity of the dye. The colour changes observed as a function of environmental pH were supported by instrumental methods (pH meter and spectroscopy), and a commercial pH indicator (thymol blue) results. In conclusion, we propose that FD&C Green Number 3 can be used as an accurate indicator for the rapid assessment of Fusarium molds when grown on ciprofloxacin antibiotic-containing substrate. Special emphasis should be given to an indirect risk assessment of antibiotic effects on toxic molds.

Predicting dry eye using noninvasive techniques of tear film surface assessment.

PURPOSE: To measure tear film surface quality in healthy and dry eye subjects using three noninvasive techniques of tear film quality assessment and to establish the ability of these noninvasive techniques to predict dry eye. METHODS: Thirty-four subjects participated in the study and were classified as dry eye or normal, based on standard clinical assessments. Three noninvasive techniques were applied for measurement of tear film surface quality: dynamic-area high-speed videokeratoscopy (HSV), wavefront sensing (DWS), and lateral shearing interferometry (LSI). The measurements were performed in both natural (NBC) and suppressed (SBC) blinking conditions. RESULTS: To investigate the capability of each method to discriminate dry eye subjects from normal subjects, the receiver operating curve (ROC) was calculated and then the area under the curve (AUC) was extracted. The best result was obtained for the LSI technique (AUC = 0.80 in SBC and AUC = 0.73 in NBC), which was followed by HSV (AUC = 0.72 in SBC and AUC = 0.71 in NBC). The best result for DWS was an AUC of 0.64 obtained for changes in vertical coma in SBC, whereas for NBC, the results were poorer. CONCLUSIONS: Noninvasive techniques of tear film surface assessment can be used for predicting dry eye, and such an assay can be achieved in NBC as well as SBC. In this study, LSI showed the best detection performance, closely followed by the dynamic-area HSV. The DWS technique was less powerful, particularly in NBC.

A multicenter, double-blind, parallel group, placebo-controlled clinical study to examine the safety and efficacy of T-Clair SPHP700-3 in the management of mild to moderate dry eye in adults.

PURPOSE: To study the safety and efficacy of T-Clair SPHP700-3, a new over-the-counter preservative-free formulation, in the management of mild to moderate dry eye in adults. METHODS: Sixty adult patients with mild to moderate dry eye were consecutively recruited in 2 eye clinics and randomized into 2 groups: treatment and placebo. Signs and symptoms of dry eye were compared along 28 days of treatment. RESULTS: No adverse events were reported during the study. Symptoms and signs of dry eye showed significant differences between the 2 groups after 2 and 4 weeks of treatment. CONCLUSIONS: SPHP700-3 preservative-free formulation showed to be safe and effective in mild to moderate dry eye, improving tear film stability, ocular surface lubrification, and patients' symptomatology.