Activation of Dendritic Cells in Dry Eye Mouse Model.

Purpose: The immune system plays a major role in the pathogenesis of dry eye diseases (DED), and dendritic cells (DCs) are known to be important initiators of acquired immunity. Thus, the purpose of this study was to determine the contribution of DCs to the development of DED. Methods: Mouse dry eye model was induced by subcutaneous injections of scopolamine and was euthanized at the baseline, and 2, 4, and 7 days postinjection. The activation of the DCs was determined by the mixed leukocyte reaction (MLR), and the number of activated CD86+ DCs in the lymph nodes was determined by flow cytometry. Upregulation of cytokines in the culture supernatant of MLR was determined by ELISA. Results: Significantly increased superficial corneal punctate lesions and decreased number of goblet cells in the conjunctiva were observed in scopolamine-injected mice. The number of activated CD86+ DCs was significantly increased in the cervical lymph nodes but not in the inguinal lymph nodes of the dry eye mice. The stimulatory activity of the DCs derived from the cervical lymph nodes of dry eye mice was significantly higher than that of control mice, and upregulations of IL-17, IL-2, and IL-4 were observed in the culture supernatant of MLR. These results indicate that the DCs of the cervical lymph nodes were activated by the scopolamine injections. Conclusions: Our results indicate that DCs in our dry eye model were sufficiently activated to stimulate the T cells that participate in the onset and progression of DED.

Case of progressive hyperopia due to flattening of cornea.

PURPOSE: To report a case that had a progressive decrease in the visual acuity caused by a progressive increase in the hyperopia due to a flattening of the cornea. OBSERVATIONS: A 50-year-old woman complained of decreased vision in both eyes. Her decimal best-corrected visual acuity was 1.2 in the right and 0.5 in the left eyes, and the refractive error (spherical equivalent) was +3.75 diopters (D) for the right eye and +6.5 D for the left eye. Slit-lamp examinations showed clear corneas but Descemet's folds and fine pigmentations and opacities were present beneath the corneal epithelium in both eyes. Analysis of the corneal shape by anterior segment optical coherence tomography showed that the corneas were flattened, and the corneal refractive power was decreased in both eyes. The large values of the higher-order aberration in the cornea and total eye displayed bilateral irregular astigmatism. She obtained good vision by wearing hard contact lenses but her refractive power continued to decrease for at least 1 year. CONCLUSIONS: and Importance: We report a rare case of progressive hyperopia and irregular astigmatism due to a flattening of the cornea. Folds were present in Descemet's membrane but the cornea was not edematous.

The effect of blue-blocking intraocular lenses on circadian biological rhythm: protocol for a randomised controlled trial (CLOCK-IOL colour study).

INTRODUCTION: Blue light information plays an important role in synchronising internal biological rhythm within the external environment. Circadian misalignment is associated with the increased risk of sleep disturbance, obesity, diabetes mellitus, depression, ischaemic heart disease, stroke and cancer. Meanwhile, blue light causes photochemical damage to the retina, and may be associated with age-related macular degeneration (AMD). At present, clear intraocular lenses (IOLs) and blue-blocking IOLs are both widely used for cataract surgery; there is currently a lack of randomised controlled trials to determine whether clear or blue-blocking IOLs should be used. METHODS AND ANALYSIS: This randomised controlled trial will recruit 1000 cataract patients and randomly allocate them to receive clear IOLs or blue-blocking IOLs in a ratio of 1:1. The primary outcomes are mortality and the incidence of cardiovascular disease, cancer and AMD. Secondary outcomes are fasting plasma glucose, triglycerides, cholesterol, glycated haemoglobin, sleep quality, daytime sleepiness depressive symptoms, light sensitivity, the circadian rhythm of physical activity, wrist skin temperature and urinary melatonin metabolite. Primary outcomes will be followed until 20 years after surgery, and secondary outcomes will be assessed at baseline and 1 year after surgery. ETHICS AND DISSEMINATION: Ethical approval has been obtained from the Institutional Review Board of Nara Medical University (No. 13-032). The findings of this study will be communicated to healthcare professionals, participants and the public through peer-reviewed publications, scientific conferences and the University Hospital Medical Information Network Clinical Trials Registry (UMIN-CTR) home page. TRIAL REGISTRATION NUMBER: UMIN000014680.

Short-term influence of cataract surgery on circadian biological rhythm and related health outcomes (CLOCK-IOL trial): study protocol for a randomized controlled trial.

BACKGROUND: Light information is the most important cue of circadian rhythm which synchronizes biological rhythm with external environment. Circadian misalignment of biological rhythm and external environment is associated with increased risk of depression, insomnia, obesity, diabetes, cardiovascular disease, and cancer. Increased light transmission by cataract surgery may improve circadian misalignment and related health outcomes. Although some observational studies have shown improvement of depression and insomnia after cataract surgery, randomized controlled trials are lacking. We will conduct a parallel-group, assessor-blinded, simple randomized controlled study comparing a cataract surgery group at three months after surgery with a control group to determine whether cataract surgery improves depressive symptoms, sleep quality, body mass regulation, and glucose and lipid metabolism. METHODS/DESIGN: We will recruit patients who are aged 60 years and over, scheduled to receive their first cataract surgery, and have grade 2 or higher nuclear opacification as defined by the lens opacities classification system III. Exclusion criteria will be patients with major depression, severe corneal opacity, severe glaucoma, vitreous haemorrhage, proliferative diabetic retinopathy, macular oedema, age-related macular degeneration, and patients needing immediate or combined cataract surgery. After baseline participants will be randomized to two groups. Outcomes will be measured at three months after surgery among the intervention group, and three months after baseline among the control group. We will assess depressive symptoms as a primary outcome, using the short version geriatric depression scale (GDS-15). Secondary outcomes will be subjective and actigraph-measured sleep quality, sleepiness, glycated haemoglobin, fasting plasma glucose and triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, body mass index, abdominal circumference, circadian rhythms of physical activity and wrist skin temperature, and urinary melatonin metabolite. Chronotype and visual function will be assessed using the 'morningness-eveningness' questionnaire, the Munich chronotype questionnaire, and the National Eye Institute Visual Function Questionnaire. DISCUSSION: Although there are potential limitations due to the difference in duration from baseline survey to outcome measurements between two groups, any seasonal effect on the outcome measurement will be balanced as a result of continuous inclusion of participants through the year, and outcomes will be adjusted for day length at outcome measurements at analysis. TRIAL REGISTRATION: UMIN000014559, UMIN Clinical Trials Registry, registered on 15 July 2014.

Synthesis of phosphorothioate oligonucleotide-peptide conjugates by solid phase fragment condensation.

Phosphorothioate oligonucleotide-peptide conjugates were synthesized by solid phase fragment condensation (SPFC). Arginine rich peptides could be successfully conjugated in 2.8-13.4% isolated yields. All the products were fully characterized by reversed phase HPLC and MALDI-TOF-MS to give satisfactory results.

Biocompatibility of polyvinylalcohol gel as a vitreous substitute.

Polyvinylalcohol (PVA) hydrogel cross-linked by gamma irradiation was assessed as a possible vitreous substitute. From a series of experiments, rise of intraocular pressure and inflammatory changes in the vitreous cavity after operation were observed in some cases. Crab-eating macaques were used for this experiment. PVA gels were injected into vitreous cavity after vitrectomy and followed clinically by opthalmoscopy, tonometry, fundus photography, electroretinogram (ERG), chemotaxis, and flare cell meter. Histopathologic examination by light and electron microscopy was performed after 3 months. As a result, there were no significant changes in ophthalmoscopic findings. No abnormal rising of intraocular pressure (IOP) was recognized. ERG did not show meaningful amplitude weakness. From the photon counting of flare cell meter, significant break of blood-aqueous barrier and blood-retinal barrier was not observed. Histopathologic examination revealed that all layers of the retina were intact and no loss of tissue was evident. However, in PVA gel-injected eyes, some vacuolations of the inner retina were found in some specimens. To conclude, PVA gel was well tolerated in these experiments. The gel with a network similar to the vitreous body showed the best biocompatibility, though it is necessary to investigate the biocompatibility for the long-term. PVA gel is a good candidate for a vitreous substitute.

[Objective evaluation of the visual field defects in a patient with cerebral infarction using multifocal VEPs].

PURPOSE: We investigated whether visual field defects can be objectively evaluated using multifocal visual evoked potentials (mVEP) in a patient with cerebral infarction in whom it is difficult to measure the visual field. METHODS: To determine normal waves in mVEP recording was performed using a VERIS Junior Science (Mayo, Aichi, Japan) in 20 healthy subjects (20 eyes), peak latency and amplitude were used for assessment. In a patient with cerebral infarction, mVEP were recorded, and compared with the lesion observed by computed tomography. RESULTS: In 20 healthy subjects, the waveforms in the nasal and temporal quadrants were very similar but the waveforms in the superior and inferior quadrants were mirror images. The mVEP in patient with cerebral infarction showed abnormal waves, corresponding to the visual field defects in the lesion observed by computed tomography. CONCLUSIONS: Objective evaluation of visual field defects using mVEP may be useful in patients with cerebral infarction in whom kinetic/static perimetry as a subjective examination is difficult.

Photoreceptor cells from mouse ES cells by co-culture with chick embryonic retina.

Degeneration of photoreceptors is a consistent and common endpoint in retinal diseases. Herein, we report the efficient induction of photoreceptor-like cells from mouse embryonic stem (ES) cells using chick embryonic retina tissue. Undifferentiated mouse ES cells were initially cultured in a preferential condition into a neural lineage, and ES cells were then co-cultured with chick embryonic day 6 (E6) retina tissues. After a 10-day co-culture, approximately 20% of the mouse ES derivatives became immuno-positive for rhodopsin. RT-PCR analysis demonstrated expression of the transcription factor crx and a distinct increase of rod photoreceptor-specific markers, IRBP and recoverin, after the start of the co-culture. These results indicate that co-culture of ES cells with chick embryonic retina tissue is a useful and efficient method for the induction of photoreceptor-like cells.

Corneal epithelial barrier function in diabetic patients.

PURPOSE: To evaluate the corneal epithelial barrier function in diabetic patients. METHODS: In 29 eyes of 29 diabetic patients and 55 eyes of 55 nondiabetic controls, corneal epithelial permeability to fluorescein was measured using an anterior fluorophotometer. The average fluorescein concentration in the central cornea was compared between diabetic patients and controls. Multiple regression analysis was used to assess the factors that affect corneal epithelial barrier function in diabetic patients. RESULTS: The average fluorescein concentrations in diabetic patients and nondiabetic controls were 44.1 +/- 25.3 ng/mL and 29.9 +/- 19.8 ng/mL (mean +/- SD), respectively (P = 0.0057, unpaired t test). An explanatory variable relevant to the impaired corneal epithelial barrier function was the serum hemoglobin A1c (HbA1c) concentration (standardized partial regression coefficient = 0.466, P = 0.0163). CONCLUSIONS: The corneal epithelial barrier function is impaired in diabetic patients. Diabetic patients with higher serum HbA1c levels are more predisposed to impaired barrier function in the corneal epithelium.

Efficacy and safety of the soft-shell technique in cases with a hard lens nucleus.

PURPOSE: To evaluate the efficacy and safety of the soft-shell technique in reducing corneal endothelial cell damage during cataract surgery in patients with a hard lens nucleus. SETTING: Miyata Eye Hospital, Miyakonojo, Miyazaki, Japan. METHODS: Sixty eyes of 57 cataract patients with a hard lens nucleus (Emery-Little classification grade 3 or higher) had phacoemulsification using the soft-shell technique with Healon((R)) (sodium hyaluronate 1%) and Viscoat (sodium hyaluronate 3.0%-chondroitin sulfate 4.0%) (soft-shell group) or with Healon alone (control group). The visual acuity, intraocular pressure (IOP), flare intensity in the anterior chamber, central corneal thickness, and corneal endothelial cell density were evaluated postoperatively. RESULTS: There were no significant IOP elevations in either group. The mean central corneal thickness in the control group was 539 microm +/- 26.0 (SD) preoperatively and 578 +/- 52.0 microm 1 day after surgery; the increase was significant (P =.0154). There was no significant change in the central corneal thickness in the soft-shell group. There were no statistically significant differences between the 2 groups in uncorrected visual acuity, best corrected visual acuity, IOP, flare intensity in the anterior chamber, and central corneal thickness throughout the follow-up. The rate of endothelial cell loss 3 months after surgery was 6.4% +/- 9.6% in the soft-shell group and 16.3% +/- 9.8% in the control group (P =.0003). CONCLUSION: The results suggest that the soft-shell technique is safe and effective in protecting corneal endothelial cells during cataract surgery in patients with a hard lens nucleus.

Corneal endothelial cell protection during phacoemulsification: low- versus high-molecular-weight sodium hyaluronate.

PURPOSE: To compare the efficacy of low- and high-molecular-weight sodium hyaluronate in protecting corneal endothelial cells during phacoemulsification. SETTING: Miyata Eye Hospital, Miyakonojo, Miyazaki, Japan. METHODS: One hundred forty-nine eyes of 136 cataract patients were randomly assigned to have cataract surgery using sodium hyaluronate 1% with a low molecular weight (0.6 to 1.2 million d, Opegan) or sodium hyaluronate 1% with a high molecular weight (4 million d, Healon) during phacoemulsification. Each group was divided into 2 subgroups depending on the amount of ultrasound (% min) used during phacoemulsification, which was defined as the mean phacoemulsification energy (%) multiplied by phacoemulsification time (minutes). Corneal endothelial cell density was examined preoperatively and 3 months after surgery. The rate of cell loss was compared between the subgroups in the Opegan group and the Healon group. RESULTS: In the subgroups with ultrasound of 50% min or less, the mean rate of endothelial cell loss 3 months after surgery was 3.2% +/- 4.1% (SD) in the Opegan group and 5.9% +/- 5.3% in the Healon group (P =.0214). In the subgroups with ultrasound over 50% min, the mean rate of endothelial cell loss 3 months after surgery was 7.5% +/- 10.6% in the Opegan group and 14.8 +/- 9.0% in the Healon group (P =.0029). CONCLUSIONS: The results suggest that Opegan is more effective than Healon in protecting corneal endothelial cells during phacoemulsification regardless of the amount of ultrasound energy used.