Eye Washing Downregulated Angiotensin-Converting Enzyme 2 in Conjunctival Tissue Samples from Smokers.

This study aimed to (1) determine whether the expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 is increased in tobacco smokers, which potentially increases their susceptibility to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, and (2) assess whether eye rinsing can reduce susceptibility. This prospective study included 20 eyes of 10 smokers and 18 eyes of nine healthy non-smokers (control) for reverse-transcription polymerase chain reaction. This study also included 28 eyes of 14 smokers and 16 eyes of eight healthy non-smokers (control) for enzyme-linked immunosorbent assay. Tear and impression cytology samples were collected from the right eye of each patient. The left eye was then rinsed for 30 s, and after 5 min, the tear and impression cytology samples were collected in the same manner. The expression of the ACE2 gene was significantly higher in the conjunctiva of smokers (n = 17; median 3.07 copies/ng of total RNA) than in those of non-smokers (n = 17; median 1.92 copies/ng of total RNA, p = 0.003). Further, mRNA expression and protein levels of ACE2 were weakly correlated in smokers (r = 0.49). ACE2 protein levels in Schirmer's strip samples were significantly reduced from 5051 to 3202 pg/mL after eye washing (n = 10; p = 0.001). Ocular surface cells are susceptible to SARS-CoV-2 infection. Smoking may be a risk factor for SARS-CoV-2 infection, and eye rinsing may reduce the risk of infection.

Elevated levels of prostaglandin E2 in the tears of patients with severe allergic conjunctivitis and primary cultured conjunctival cells are suppressed by ketotifen and dexamethasone.

OBJECTIVE: We examined the production of prostaglandin E2 (PGE2), which is the key prostaglandin involved in inflammatory disorders of the ocular surface. Tears and conjunctival fibroblasts were evaluated in order to assess allergic inflammation and the effect of specific drugs. METHODS AND ANALYSIS: PGE2 was measured in tears from both patients and normal volunteers. Primary cultures of human conjunctival fibroblasts were incubated with interleukin (IL)-4 and tumour necrosis factor (TNF)-α with or without ketotifen fumarate or dexamethasone. The culture supernatants were removed 24 hours after exposure and the concentrations of PGE2 were quantified by ELISA. RESULTS: Significantly higher levels of PGE2 were observed in the tears of patients with severe allergic conjunctivitis than in those with post-surgical inflammation (p=0.02), and this production was reduced by eye drops. Stimulation with IL-4 and TNF-α induced the generation of PGE2 in supernatants of conjunctival fibroblasts, and this production was significantly downregulated by ketotifen fumarate or steroids. CONCLUSION: PGE2 may participate in the pathogenesis of severe ocular allergic disease, and both ketotifen fumarate and steroid reduce the production of PGE2.

Combination of Two Rapid Ophthalmic Test Kits for Improved Diagnosis in Cases of Severe Binocular Conjunctivitis.

INTRODUCTION: Diagnosis of conjunctivitis can be sometimes difficult, especially in cases of severe conjunctivitis and those involving both eyes. In this study, we performed commercial tests for adenovirus (Capilia Adeno Eye®) and total tear IgE (Allerwatch®) in a single visit in patients with bilateral conjunctivitis to examine if, and by how much, the combination of these two tests would improve the diagnostic accuracy of conjunctivitis. METHODS: The study included sixty patients with relatively severe conjunctivitis in both eyes within a week of consulting our clinic and who had no previous treatment. Capilia Adeno Eye® and Allerwatch® tests were performed. RESULTS: A significantly higher number of cases (55/60) were diagnosed when both tests were evaluated than with either test (Capilia Adeno Eye® (12/60; p < 0.001) or Allerwatch® (44/60; p < 0.005)) alone. The positivity rate of Allerwatch® was significantly higher than that of Capilia Adeno Eye® (p < 0.001). The diagnosis rate of atopic keratoconjunctivitis was 100% in patients with allergic conjunctivitis, but there was no significant difference in positivity compared with other types of allergic conjunctivitis. CONCLUSIONS: Testing patients with both Capilia Adeno Eye® and Allerwatch® improves the diagnostic accuracy for conjunctivitis and can diagnose more than 90% of cases. Detection of adenovirus antigen and IgE in tears, using these simple and rapid methods, will be useful for early diagnosis and prevention of adenoviral conjunctivitis.

Effects of Docosahexaenoic Acid on Chemokine Expression in Human Conjunctival Fibroblasts.

Purpose: We assessed the production of chemokines by human conjunctival fibroblasts in response to inflammation and the effects of omega (ω)-3 fatty acids on chemokine expression.Methods: Primary cultures of human conjunctival fibroblasts were incubated with interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-α). The expression of eotaxin-1 and RANTES in response to pretreatment with docosahexaenoic acid (DHA) was investigated. Moreover, western blotting was used to evaluate the effects of DHA on the activation of nuclear factor (NF)-κB and signal transducer and activator of transcription 6 (STAT6).Results: The expression of eotaxin-1 mRNA was significantly suppressed by pretreatment with DHA with IL-4 and TNF-α costimulation. RANTES expression was similarly suppressed, but the difference was not significant. The secretion of eotaxin-1 and RANTES was significantly lower in DHA-pretreated cells than in vehicle-treated cells. Western blotting for NF-κB and STAT6 showed that these proteins were downregulated in the DHA pretreatment group compared with those in the vehicle control group.Conclusion: The results of this study suggested that DHA could have applications in the management of allergic inflammation.

The Effect of Cytokine-Stimulation and Pharmacologic Intervention on PGE2 Production in Primary Human Conjunctival and Corneal Cells.

We studied the production of PGE2 by human conjunctival and corneal cells in response to inflammation, and reduction of inflammation with non-steroidal anti-inflammatory drugs. Primary cultures of human conjunctival epithelial cells, fibroblasts, corneal epithelial cells, and keratocytes were incubated with IL-4 and TNF-α. PGE2 and COX-2 levels were analyzed. Effects of anti-inflammatory and anti-immune drugs on PGE2 production were also investigated. IL-4 and TNF-α induced the generation of PGE2 and COX-2 in conjunctival and corneal cells. Epithelial PGE2 production was significantly lower than in keratocytes and fibroblasts, which was down-regulated by aspirin. IL-4 and TNF-α enhanced the inflammatory response via prostaglandin production which contributed to ocular surface inflammation. Prostaglandin production was higher in stromal cells than epithelial cells. These results suggest that the epithelial barrier disruption may contribute to ocular allergic inflammation by the PGE2 production from stromal cells. Moreover, NSAIDs were effective in suppressing PGE2 production in our experiment.

Advanced CUBIC tissue clearing for whole-organ cell profiling.

Tissue-clearing techniques are powerful tools for biological research and pathological diagnosis. Here, we describe advanced clear, unobstructed brain imaging cocktails and computational analysis (CUBIC) procedures that can be applied to biomedical research. This protocol enables preparation of high-transparency organs that retain fluorescent protein signals within 7-21 d by immersion in CUBIC reagents. A transparent mouse organ can then be imaged by a high-speed imaging system (>0.5 TB/h/color). In addition, to improve the understanding and simplify handling of the data, the positions of all detected cells in an organ (3-12 GB) can be extracted from a large image dataset (2.5-14 TB) within 3-12 h. As an example of how the protocol can be used, we counted the number of cells in an adult whole mouse brain and other distinct anatomical regions and determined the number of cells transduced with mCherry following whole-brain infection with adeno-associated virus (AAV)-PHP.eB. The improved throughput offered by this protocol allows analysis of numerous samples (e.g., >100 mouse brains per study), providing a platform for next-generation biomedical research.

Temperature-Sensitive Substrate and Product Binding Underlie Temperature-Compensated Phosphorylation in the Clock.

Temperature compensation is a striking feature of the circadian clock. Here we investigate biochemical mechanisms underlying temperature-compensated, CKIδ-dependent multi-site phosphorylation in mammals. We identify two mechanisms for temperature-insensitive phosphorylation at higher temperature: lower substrate affinity to CKIδ-ATP complex and higher product affinity to CKIδ-ADP complex. Inhibitor screening of ADP-dependent phosphatase activity of CKIδ identified aurintricarboxylic acid (ATA) as a temperature-sensitive kinase activator. Docking simulation of ATA and mutagenesis experiment revealed K224D/K224E mutations in CKIδ that impaired product binding and temperature-compensated primed phosphorylation. Importantly, K224D mutation shortens behavioral circadian rhythms and changes the temperature dependency of SCN's circadian period. Interestingly, temperature-compensated phosphorylation was evolutionary conserved in yeast. Molecular dynamics simulation and X-ray crystallography demonstrate that an evolutionally conserved CKI-specific domain around K224 can provide a structural basis for temperature-sensitive substrate and product binding. Surprisingly, this domain can confer temperature compensation on a temperature-sensitive TTBK1. These findings suggest the temperature-sensitive substrate- and product-binding mechanisms underlie temperature compensation.

Involvement of Ca(2+)-Dependent Hyperpolarization in Sleep Duration in Mammals.

The detailed molecular mechanisms underlying the regulation of sleep duration in mammals are still elusive. To address this challenge, we constructed a simple computational model, which recapitulates the electrophysiological characteristics of the slow-wave sleep and awake states. Comprehensive bifurcation analysis predicted that a Ca(2+)-dependent hyperpolarization pathway may play a role in slow-wave sleep and hence in the regulation of sleep duration. To experimentally validate the prediction, we generate and analyze 21 KO mice. Here we found that impaired Ca(2+)-dependent K(+) channels (Kcnn2 and Kcnn3), voltage-gated Ca(2+) channels (Cacna1g and Cacna1h), or Ca(2+)/calmodulin-dependent kinases (Camk2a and Camk2b) decrease sleep duration, while impaired plasma membrane Ca(2+) ATPase (Atp2b3) increases sleep duration. Pharmacological intervention and whole-brain imaging validated that impaired NMDA receptors reduce sleep duration and directly increase the excitability of cells. Based on these results, we propose a hypothesis that a Ca(2+)-dependent hyperpolarization pathway underlies the regulation of sleep duration in mammals.

The usefulness of measuring tear periostin for the diagnosis and management of ocular allergic diseases.

BACKGROUND: Chronic ocular allergic diseases such as vernal keratoconjunctivitis (VKC) and atopic keratoconjunctivitis (AKC) are accompanied by serious comorbidities; however, the underlying pathogenesis remains obscure. Furthermore, diagnosing conjunctival lesions in patients with atopic dermatitis and estimating the severity in AKC are important for the treatment of ocular allergic diseases. OBJECTIVE: We addressed whether periostin, a novel mediator and biomarker in allergic inflammation, is involved in the pathogenesis of ocular allergic diseases and whether periostin can be a biomarker for these diseases. METHODS: We investigated tear periostin in patients with seasonal allergic conjunctivitis (SAC), VKC, and AKC and allergic patients without conjunctivitis and compared it with tear IL-13 and serum periostin. Furthermore, in patients with AKC, we measured tear periostin before and after topical treatment with tacrolimus. RESULTS: Tears from patients with ocular allergic disease showed significantly high periostin levels than did tears from allergic patients without conjunctivitis and from patients with AKC, VKC, and SAC in descending order. Tear periostin was associated with serious comorbidities such as large papilla formation and corneal damage in AKC, although both tear IL-13 and serum periostin had little to no such abilities. Furthermore, after topical tacrolimus treatment, tear periostin tended to decrease in most patients with AKC along with their clinical improvement. CONCLUSIONS: Periostin produced in conjunctival tissues stimulated by IL-13 may contribute to the pathogenesis of ocular allergic diseases. Furthermore, tear periostin can be potentially applied as a biomarker to diagnose conjunctivitis in allergic patients and to evaluate disease severity as well as the efficacy of treatments in AKC.

Effects of diesel exhaust particles on primary cultured healthy human conjunctival epithelium.

BACKGROUND: Air pollution from road traffic is a serious public health problem. Epidemiologic studies have demonstrated adverse health effects associated with environmental pollution. Diesel exhaust is a major contributor to ambient particulate matter air pollution. We studied the effects of exposure to diesel exhaust particles on allergic conjunctivitis using cultured conjunctival epithelial cells obtained from healthy people. OBJECTIVE: To identify the factors involved in the human conjunctival epithelial response to diesel exhaust in vitro. METHODS: Healthy individuals underwent conjunctival biopsy, and the samples were incubated on conjunctival epithelial sheets. We investigated the effects of exposure to diesel exhaust using GeneChip arrays. The adhesion molecules and cytokines showing increased expression on GeneChip arrays were verified by real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: The GeneChip array showed increased expression of adhesion molecules, cytokines, chemokines, and growth factors after exposure to diesel exhaust. Real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay confirmed that the expression of intercellular adhesion molecule 1 and interleukin 6, in particular, were significantly upregulated. CONCLUSION: Our experimental data confirm that exposure to diesel exhaust particles increases inflammatory factor expression in human conjunctiva and thereby contributes to allergic conjunctival responses.

Naphthol derivatives as TRPV1 inhibitors for the treatment of urinary incontinence.

We have identified naphthol derivatives as inhibitors of the vanilloid receptor TRPV1 by high throughput screening. The initial lead showed high clearance in rats and has been optimized by enhancing the acidity of the phenol group. Compound 6b has reduced clearance, improved potency and is active in rat cystometry models of urinary incontinence after intravenous administration.

Evaluation of lipid oxidative stress status and inflammation in atopic ocular surface disease.

BACKGROUND: Although the oxidative stress status in atopic skin disease has been reported to be elevated, there are still no studies related to the status of oxidative stress in atopic ocular surface disease. The purpose of this study was to evaluate the ocular surface lipid oxidative stress status and inflammation in atopic keratoconjunctivitis (AKC) patients and normal subjects. METHODS: Twenty eight eyes of 14 patients (9 males, 5 females) with AKC and 18 eyes of 9 age and sex matched (4 males and 5 females) normal healthy controls were examined in this prospective study. The severity of atopic dermatitis (AD) was scored by the SCORing Atopic Dermatitis (SCORAD) index. All subjects underwent Schirmer test, tear film break up time (BUT), fluorescein/Rose Bengal stainings, tear collection, and brush cytology from the upper palpebral conjunctiva. The brush cytology samples were stained with Diff-Quik for differentiation of inflammatory cells and immunohistochemistry (IHC) staining with HEL (hexanoyl-lysine) and 4-HNE (4-hydroxy-2-nonenal) to study lipid oxidation. HEL and cytokine (interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ)) levels were measured by enzyme-linked immunosorbent assay (ELISA) from tear samples of AKC patients and control subjects. Toluidine Blue and IHC staining with HEL, 4-HNE and cluster of differentiation 45 (CD45) were performed on papillary samples of AKC patients. This study was conducted in compliance with the "Declaration of Helsinki." RESULTS: The tear stability and vital staining scores were significantly worse in eyes of AKC patients (p<0.05) compared to the controls. Inflammatory cells and positively stained conjunctival epithelial cells for HEL and 4-HNE showed a significant elevation in brush cytology samples of AKC patients. Significantly higher levels of HEL and cytokines were detected in tears of AKC patients compared to controls. Papillary specimens also revealed many CD45 inflammatory cells as well as many cells positively stained with HEL and 4-HNE in IHC. A strong significant linear positive correlation between conjunctival inflammation and epithelial lipid oxidative stress status was observed. Conjunctival lipid oxidative stress also correlated strongly with tear HEL levels and epithelial damage scores. CONCLUSIONS: The ocular surface disease in AKC was characterized by marked tear instability, ocular surface epithelial damage, increase in inflammatory infiltrates and presence of increased lipid oxidation.

Corneal and conjunctival fibroblasts are major sources of eosinophil-recruiting chemokines.

BACKGROUND: Differential expression of chemokine genes were investigated in various types of ocular surface cells. METHODS: Primary cultures of human corneal epithelial cells (n = 3), corneal fibroblasts (n = 2), conjunctival epithelial cells (n = 2) and conjunctival fibroblasts (n = 2) were established and incubated with or without interleukin (IL)-4 (30ng/ml) and tumor necrosis factor (TNF)-alpha(30ng/ml) for 24 hours. Gene transcription levels of 33 chemokines and production of 4 chemokines were analyzed. RESULTS: After stimulation, chemokine expression increased for 18 of 33 coded chemokine gene transcripts. In stimulated conjunctival and corneal cells, CC chemokine genes increased in fibroblasts (expression of 6 out of 8 genes), while CXC chemokine genes increased in both epithelial cells (expression of 4 out of 9 genes in conjunctival epithelial cells and 7 out of 9 genes in corneal epithelial cells) and in fibroblasts (expression of 8 out of 9 genes in conjunctival and corneal fibroblasts). Except for MCP-1, gene transcription levels for most CC chemokines were inducible and, except for IP-10 and I-TAC, most CXC chemokines were constitutively expressed. Corneal epithelial cell and fibroblast production patterns for eotaxin-1, MCP-1 and IP-10 were comparable to the mRNA expression pattern. CONCLUSIONS: Corneal and conjunctival fibroblasts exhibited marked increases in the expression of chemokines upon stimulation with TNF-alpha and IL-4, suggesting that fibroblasts may be one of the primary sources of chemokines in allergic conjunctival diseases. Therefore, regulation of chemokine production from these cells may be an effective strategy for treating such diseases.

Suppressive effects of tranilast on eotaxin-1 production from cultured conjunctival fibroblasts.

PURPOSE: Eotaxin, a CC-chemokine with selective chemotactic effects for eosinophils, has been reported to play an important role in allergic conjunctival diseases. We previously reported that eotaxin is produced by conjunctival fibroblasts and keratocytes on stimulation with Th2 cytokines. Tranilast is known to have anti-allergic properties. In this study, we examined the inhibitory effect of tranilast, an anti-allergic drug, on eotaxin-1 production from cultured conjunctival fibroblasts. METHODS: Conjunctival fibroblasts obtained from normal patients were cultured in DMEM/F12 medium. On the fifth passage, the cells were transferred to a 96-well plate and, after starvation for 24 hr, TNF-alpha, IL-4, and tranilast or dexamethasone were added. After 48 hr, the concentrations of eotaxin-1 in the supernatants were measured by ELISA, and the cells were tested for eotaxin-1 expression by real-time PCR. RESULTS: Eotaxin-1 production was observed on simultaneous stimulation with TNF-alpha and IL-4. This production was inhibited by both tranilast and dexamethasone. Inhibition of eotaxin-1 expression was also observed by real-time PCR. CONCLUSIONS: Eotaxin-1 production from conjunctival fibroblasts was inhibited by both tranilast and dexamethasone. These results suggest that the anti-allergic effect of tranilast may be partly due to the inhibition of eotaxin-1 production from conjunctival fibroblasts.

Mechanisms of giant papillary formation in vernal keratoconjunctivitis.

PURPOSE: Tissue remodeling, known to accompany exacerbation of bronchial asthma (BA) and characterized by thickening of reticular basement membrane, increased fibrosis, and angiogenesis, is believed to be an important mechanism in giant papillary formation in vernal keratoconjunctivitis (VKC). This study was conducted to confirm the difference of tissue remodeling between BA and VKC and to determine the most relevant factor for VKC exacerbation. METHODS: Histopathologic analysis of conjunctival specimens from 6 patients with VKC and 4 normal controls was performed. Immunohistochemistry tests for collagen types 1, 3, and 5, fibronectin, laminin (a marker of the basement membrane of the epithelium or vessels), fibroblast (prolyl 4-hydroxylase), and alpha-smooth muscle actin (alpha-SMA) were performed. Morphometric analysis evaluated immunoreactive areas for collagens and fibronectin and the number of cell nuclei in VKC papillae and normal control specimens. RESULTS: In histopathologic sections of VKC papillae, mononuclear cells were mainly observed in the central region and granulocytes preferentially in the peripheral stroma. Immunohistochemistry showed increased vascularity and existence of fibroblasts and alpha-SMA-positive cells in VKC papillae. Morphometric analysis showed that the number of cell nuclei in VKC papillae was 7.3 times higher versus in normal controls (P < 0.05). Furthermore, the immunoreactive area for fibronectin was 4.5 times larger versus in normal controls (P < 0.05), although collagens were not significantly different between the 2 groups. CONCLUSION: Giant papillary formation in VKC is thought to occur in relation to tissue remodeling. However, the most relevant factor for the exacerbation of VKC seems to be inflammation rather than fibrosis.

Quantitative evaluation of the early changes in ocular surface inflammation following MMC-aided papillary resection in severe allergic patients with corneal complications.

BACKGROUND: Atopic keratoconjunctivitis (AKC) and vernal keratoconjunctivitis (VKC) are chronic inflammatory allergic diseases that are associated with some common conjunctival and corneal complications.1 The clinical corneal manifestations of both entities may include superficial punctate keratitis, macroerosions, corneal ulceration, plaque formation, corneal neovascularization, and lipid infiltration. PURPOSE: To quantitatively evaluate the early ocular surface inflammation before and after mitomycin C (MMC)-aided papillary resection in severe allergy patients with corneal complications. METHODS: Three eyes with VKC and 5 eyes with AKC were included in this study. All eyes had cobblestone-like papillae on the upper tarsal conjunctiva and corneal complications such as corneal ulcers, defect, or erosions that were refractory to conventional treatment of more than 2 weeks. Papillary resection with intraoperative 0.05% MMC application for 5 minutes on the palpebral conjunctiva was carried out in all eyes. Corneal and conjunctival findings were evaluated before and 1 week and 2 weeks after surgery. Brush cytology (BC) and evaluation of tear eosinophilic cationic protein (ECP) levels by radioimmunoassay techniques were performed as well pre- and postoperatively. RESULTS: Corneal and conjunctival complications resolved in all patients within 7 days after resection. Postoperative tear ECP levels decreased significantly with papillary resection (P< 0.05). Concomitant brush cytology showed a significant decrease in the number of eosinophils and neutrophils following papillary resection (P < 0.05). CONCLUSION: MMC-aided papillary resection provided a dramatic decrease in ocular surface inflammation as evidenced by decrease in the number of inflammatory cells as well as tear ECP levels with a rapid improvement of clinical corneal and conjunctival findings.

The implications of the upregulation of ICAM-1/VCAM-1 expression of corneal fibroblasts on the pathogenesis of allergic keratopathy.

OBJECTIVE: The present study investigated the expression of ICAM-1 and VCAM-1 on fibroblasts with interleukin (IL)-4 and/or tumor necrosis factor (TNF)-alpha stimulation and assessed the effect of eosinophil adhesion on fibroblast viability. METHODS: Primary cultured human corneal fibroblasts were incubated with IL-4, TNF-alpha, or their combination for 24 hours. Expression of ICAM-1 and VCAM-1 was examined by real-time quantitative PCR and flow cytometric analysis. Purified eosinophils were cocultured with activated fibroblasts, and the number of eosinophils adhered to fibroblasts and the number of damaged fibroblasts were counted using microscopy. In a separate trial, conjunctival and corneal impression cytology was performed on patients with atopic keratoconjunctivitis and corneal ulcers (eight eyes) to assess the status of the ocular surface epithelium and the presence of inflammatory cell infiltrates. RESULTS: Real-time quantitative PCR and flow cytometric analysis revealed that both mRNA and protein of VCAM-1 and ICAM-1 were upregulated by IL-4 and TNF-alpha. IL-5-primed eosinophils adhered to the corneal fibroblasts treated with IL-4 and TNF-alpha, and the fibroblasts were damaged by eosinophil adherence. Anti-ICAM-1 antibody and anti-VCAM-1 antibody inhibited the eosinophil adherence to fibroblasts and the fibroblast damage. Impression cytology revealed extensive infiltration of neutrophil and eosinophils among isolated ocular surface epithelial cells with advanced squamous metaplasia. CONCLUSIONS: Corneal fibroblasts expressed ICAM-1 and VCAM-1 when activated with IL-4 and TNF-alpha. Eosinophils can adhere to the activated fibroblasts and can induce subsequent fibroblast damage through these adhesion molecules. Eosinophil adhesion to fibroblasts may possibly contribute to the pathogenesis of severe persistent allergic corneal ulcers.

Ocular surface and MUC5AC alterations in atopic patients with corneal shield ulcers.

PURPOSE: To describe MUC5AC alterations and the ocular surface disorder in atopic patients with or without corneal ulcers. METHODS: Atopic patients' eyes were divided into two groups according to the presence and absence of corneal ulceration. The subjects underwent corneal sensitivity measurements, Schirmer test, tear film break-up time (BUT), fluorescein and Rose Bengal staining of the ocular surface and conjunctival impression cytology and brush cytology. Impression cytology samples underwent PAS and immunohistochemical staining for MUC5AC. Brush cytology specimens underwent evaluation for inflammatory cell expression and quantitative real-time PCR for MUC5AC mRNA expression. The differences related to the tear function and ocular surface examination parameters between patients with and without corneal ulceration and healthy control subjects were studied. In addition, the differences of the study parameters related to ocular surface epithelial health and inflammatory status between patient eyes with atopic keratoconjunctivitis (AKC) and vernal keratoconjunctivitis (VKC) were investigated. RESULTS: The mean corneal sensitivity and BUT values were significantly lower in atopic patients with corneal ulcers, compared to patients without ulcers and controls (p < 0.001). Brush cytology specimens from patients with corneal ulcers revealed significantly higher expression of inflammatory cells compared to patients without ulcers and controls (p < 0.001). Impression cytology samples from eyes with corneal ulcers showed significant squamous metaplasia and reduction in goblet cell density compared to eyes without ulcers and eyes of control subjects. The mean squamous metaplasia grade was significantly higher in eyes with AKC compared to eyes with VKC (p < 0.02). The mean goblet cell density was significantly lower in eyes with AKC compared to eyes with VKC (p < 0.01). Specimens from eyes with corneal ulcers showed PAS positive mucin pickup and did not stain positive for MUC5AC. MUC5AC mRNA expression was significantly lower in eyes with corneal ulcers compared to eyes without ulcers and eyes of control subjects. MUC5AC mRNA expression was also significantly lower in eyes with AKC compared to eyes with VKC. CONCLUSIONS: Ocular surface inflammation, tear film instability, and decreased conjunctival MUC5AC mRNA expression were thought to be important in the pathogenesis of noninfectious corneal shield ulcers in atopic ocular surface disease.

Atopic ocular surface disease: implications on tear function and ocular surface mucins.

PURPOSE: To describe tear function, mucin alterations, and ocular surface disorder in patients with atopic diseases. METHODS: Subjects underwent corneal sensitivity measurements, Schirmer test, tear film break-up time (BUT) assay, and fluorescein and rose Bengal staining of the ocular surface. Conjunctival impression cytology and brush cytology were also conducted. Impression cytology samples underwent PAS and immunohistochemical staining for MUC5AC. Brush cytology specimens underwent evaluation for inflammatory cell expression and RT-PCR for MUC5AC mRNA expression. Differences related to tear function and ocular surface examination parameters among patients with and without corneal ulceration and healthy control subjects were studied. RESULTS: Mean corneal sensitivity and BUT values were significantly lower in atopic patients with corneal ulcers compared with patients without ulcers and controls (P<0.001). Brush cytology specimens from patients with corneal ulcers revealed significantly higher expression of inflammatory cells compared with patients without ulcers and controls (P<0.001). Impression cytology samples from eyes with corneal ulcers showed significant squamous metaplasia and reduction of goblet cell density compared with eyes without ulcers and control subjects. Specimens from eyes with corneal ulcers showed PAS (+) mucin pick up and did not stain positive for MUC5AC. MUC5AC mRNA expression was significantly lower in eyes with corneal ulcers compared with in eyes without ulcers and control subjects. CONCLUSIONS: Ocular surface inflammation, tear film instability, and decreased conjunctival MUC5AC mRNA expression are important in the pathogenesis of noninfectious corneal shield ulcers in atopic ocular surface disease.

Early visual results with the 1CU accommodating intraocular lens.

PURPOSE: To prospectively assess the clinical outcome after implantation of the 1CU accommodating intraocular lens (IOL) and a foldable acrylic IOL (AcrySof, Alcon). SETTING: Department of Ophthalmology, Tokyo Dental College, Ichikawa Hospital, Ichikawa, and Minami Aoyama Eye Clinics, Tokyo, Yokohama, Japan. METHODS: Twenty-two eyes of 16 patients with cataract had phacoemulsification implantation of 1CU accommodating IOL. Twenty eyes of 10 age-matched and sex-matched patients with cataract had the same surgery but with a foldable acrylic IOL. All patients had assessments of the amplitude of accommodation, refraction, uncorrected and best corrected distance and near visual acuity, and distance corrected near visual acuity before surgery up to 12 months after surgery. Contrast visual acuities were measured 1 year after surgery. Anterior segment photography, intraocular pressure measurements, specular microscopy, and computerized topography were also performed. RESULTS: The final best corrected distance visual acuity was above 20/25 in all eyes with the 1CU and the AcrySof IOLs. The mean distance corrected near visual acuity was significantly higher in the 1CU IOL group than in the acrylic IOL group after 3 months. None of the eyes with the AcrySof IOL implants displayed an accommodative response at any examination. The peak mean amplitude of accommodation with the 1CU IOLs was observed at 3 months and was 0.5 diopters +/- 0.44 (SD). Accommodation amplitude declined after 6 months. CONCLUSION: The 1CU IOL provided additional near acuity postoperatively, but the benefit disappeared at 12 months with a concomitant decrease in accommodation amplitude owing to an increase in anterior and posterior capsular opacities.