A New Method for Lateral Visualization of the Primary Cilia on the Surfaces of Cells Cultured on White Glass Rods.

Purpose: To investigate the motility of the primary cilia of corneal endothelial cells (CECs), which exist like a hair on the cell surface, using our new in vitro method.

Methods: A white glass rod was heated with a gas burner to produce a rod approximately 0.5 mm in diameter and 20 mm in length and then coated with collagen.

Membrane structures and functional correlates in the bi-segmented eye lens of the cephalopod.

The cephalopod eye lens is unique because it has evolved as a compound structure with two physiologically distinct segments. However, the detailed ultrastructure of this lens and precise optical role of each segment are far from clear. To help elucidate structure-function relationships in the cephalopod lens, we conducted multiple structural investigations on squid.

Identification and Analysis of Primary Cilia in the Corneal Endothelial Cells of Patients with Bullous Keratopathy.

Purpose: To identify primary cilia in human corneal endothelial cells (CECs) obtained from patients with bullous keratopathy (BK).

Methods: This study involved CEC specimens obtained from 10 eyes of 10 consecutive patients (three males and seven females; mean age: 74.5 years, range: 68-90 years) with BK who underwent Descemet's stripping automated endothelial keratoplasty at Baptist Eye Institute, Kyoto, Japan between August 2019 and September 2020.

Temperature effects on the disappearance and reappearance of corneal-endothelium primary cilia.

Introduction: To elucidate the specific functions of the primary cilia in corneal endothelial cells (CECs) by investigating the histological changes of corneal endothelium exposed at low temperature.

Study Design: Experimental study.

Methods: This study involved corneas freshly obtained from Japanese white rabbits preserved in Optisol™-GS (Bausch & Lomb) corneal storage medium at 4 °C for 0, 1, and 7 days.

Toxicity of Amphotericin B in Rabbit Corneal Epithelial Cells Stored in Optisol™-GS: Corneal Epithelial Cell Morphology and Migration.

Purpose: To evaluate the toxicity of Amphotericin B (AmB) in Optisol™-GS Corneal Storage Media (Bausch & Lomb) on corneal epithelial cell (CEC) morphology and migration ability.

Methods: Sclerocorneal strips were removed from male Japanese white rabbits, and then stored at 4 °C in Optisol™-GS containing 0 µg/ml of AmB (control group) and 2.5, 5, 25, and 50 µg/ml of AmB (AmB groups; four eyes per group).

Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina.

Despite efficient and specific in vitro knockdown, more reliable and convenient methods for in vivo knockdown of target genes remain to be developed particularly for retinal research. Using commercially available and chemically modified siRNA so-called Accell siRNA, we established a novel in vivo gene silencing approach in the rat retina. siRNA designed for knockdown of the house keeping gene Gapdh or four retinal cell type-specific genes (Nefl, Pvalb, Rho and Opn1sw) was injected into the vitreous body, and their retinal mRNA levels were quantified using real-time PCR.

The existence of dead cells in donor corneal endothelium preserved with storage media.

Aim: To investigate the viability of donor corneal endothelial cells (CECs) preserved in storage media by histological examination.

Methods: Twenty-eight donor corneas were obtained from SightLife Eye Bank (Seattle, Washington), and redundant peripheral portions of those corneas were used for histological examination after removal of the centre corneal graft for transplantation. To assess cell viability in the corneal endothelium, biostaining experiments were performed using propidium iodide, calcein-AM, Hoechst 33 342, annexin V, anti-vimentin antibody and toluidine blue.

Expression of the P2Y₂ receptor on the rat ocular surface during a 1-year rearing period.

Purpose: P2Y2 receptors are expressed on ocular surface tissues. Diquafosol ophthalmic solution (DIQUAS(®) ophthalmic solution 3 %; Santen Pharmaceutical Co., Ltd.

A comparison of glycosaminoglycan distributions, keratan sulphate sulphation patterns and collagen fibril architecture from central to peripheral regions of the bovine cornea.

This study investigated changes in collagen fibril architecture and the sulphation status of keratan sulphate (KS) glycosaminoglycan (GAG) epitopes from central to peripheral corneal regions. Freshly excised adult bovine corneal tissue was examined as a function of radial position from the centre of the cornea outwards. Corneal thickness, tissue hydration, hydroxyproline content, and the total amount of sulphated GAG were all measured.

Ocular toxicity of benzalkonium chloride homologs compared with their mixtures.

This study was performed to assess the in vivo ocular toxicity of benzalkonium chloride (BAK) homologs compared with commercially available BAK (BAK mixture) and to assess the ocular toxicity of BAK homolog after repeated ocular application. Rabbit eyes were examined by ophthalmology and scanning electron microscopy (SEM) after 10 applications of BAK homologs with C12 (C12-BAK) and C14 (C14-BAK) alkyl chain lengths and a BAK mixture at concentrations of 0.001% (w/v), 0.

Degeneration of retinal on bipolar cells induced by serum including autoantibody against TRPM1 in mouse model of paraneoplastic retinopathy.

The paraneoplastic retinopathies (PRs) are a group of eye diseases characterized by a sudden and progressive dysfunction of the retina caused by an antibody against a protein in a neoplasm. Evidence has been obtained that the transient receptor potential melastatin 1 (TRPM1) protein was one of the antigens for the autoantibody against the ON bipolar cells in PR patients. However, it has not been determined how the autoantibody causes the dysfunction of the ON bipolar cells.

P2X₇ receptor activation may be involved in neuronal loss in the retinal ganglion cell layer after acute elevation of intraocular pressure in rats.

Purpose: To investigate whether the P2X₇ receptor is involved in retinal ganglion cell (RGC) death after the intraocular pressure (IOP) is elevated in rats.

Methods: After the IOP was elevated to 90 mmHg for 1 h, the rats were subsequently administered oxidized adenosine triphosphate (OxATP) and brilliant blue G (BBG) as P2X₇ antagonists. The rats were euthanized 7 days after IOP elevation for histologic evaluation and at 1, 3, and 7 days after IOP elevation to immunostain for the P2X₇ receptor and neuron-specific class III β-tubulin in the retina.

A study of host corneal endothelial cells after non-Descemet stripping automated endothelial keratoplasty.

Purpose: To determine the short-term fate of the host endothelium and Descemet membrane after non-Descemet stripping automated endothelial keratoplasty (nDSAEK).

Methods: Eight unilateral DSAEK (n = 4) or nDSAEK (n = 4) surgeries were performed in the right eyes of 8 rabbits. Corneal transparency and thickness were followed-up by slit-lamp microscopy, and 2 weeks postoperatively, corneas were evaluated by immunohistochemistry and transmission electron microscopy.

Tumor-associated calcium signal transducer 2 is required for the proper subcellular localization of claudin 1 and 7: implications in the pathogenesis of gelatinous drop-like corneal dystrophy.

Gelatinous drop-like dystrophy (GDLD) is a rare autosomal recessive form of corneal dystrophy characterized by subepithelial amyloid depositions on the cornea. Previous clinical and laboratory observations have strongly suggested that epithelial barrier function is significantly decreased in GDLD. Despite the decade-old identification of the tumor-associated calcium signal transducer 2 (TACSTD2) gene as a causative gene for GDLD, the mechanism by which the loss of function of this causative gene leads to the pathological consequence of this disease remains unknown.

Structural collagen alterations in macular corneal dystrophy occur mainly in the posterior stroma.

Purpose: Collagen fibrils in the corneal stroma in macular corneal dystrophy, on average, are more closely spaced than in the normal cornea. This study was conducted to investigate if this occurs uniformly across the stroma or is more prevalent at certain stromal depths.

Methods: Microbeam synchrotron X-ray fiber diffraction patterns were obtained in 25 microm steps across the whole thickness of a thin strip of a macular corneal dystrophy cornea obtained at keratoplasty.

The relationship between preoperative clinical scores and immunohistological evaluation of surgically resected tissues in chronic severe ocular surface diseases.

Purpose: To clarify the relationship between clinical symptoms and histological status in patients with ocular cicatricial pemphigoid (OCP) and Stevens-Johnson syndrome (SJS).

Methods: Clinical symptoms of four OCP and eight SJS patients in the chronic phase were scored with our recently proposed grading system. The histological status of the pannus tissue removed from the corneal surface during surgery was investigated using immunohistological techniques.

[A case presenting peculiar staining with fluorescein on the unilateral cornea].

Background: Corneal staining with fluorescein resulting from a cause distinct from that of general epithelial barrier disturbance.

Case: A 66-year-old woman saw her doctor complaining of blurred vision and foreign-body sensation in her left eye. Superficial punctate keratopathy (SPK) was detected on her left cornea.

Functional role of thymic stromal lymphopoietin in chronic allergic keratoconjunctivitis.

Purpose: Previous reports have shown that thymic stromal lymphopoietin (TSLP) plays a role in atopic diseases. This study was undertaken to investigate the expression of TSLP in the giant papillae obtained from patients with vernal keratoconjunctivitis (VKC) or atopic keratoconjunctivitis (AKC), and its functional roles were analyzed.

Methods: TSLP mRNA expression was examined in resected conjunctival samples obtained from four patients with VKC/AKC and three control subjects by reverse transcription-polymerase chain reaction.

Cultivated human conjunctival epithelial transplantation for total limbal stem cell deficiency.

Purpose: To determine the feasibility of cultivated conjunctiva as a viable epithelial sheet for transplantation and corneal resurfacing in eyes with limbal stem cell deficiency (LSCD).

Methods: Human corneal epithelial (HCE) and human conjunctival epithelial (HCjE) cells were cultivated on human amniotic membrane (AM) to confluence and then air lifted to allow further stratification and differentiation. Denuded AM and cultivated HCE and cultivated HCjE cells were then transplanted into 18 eyes of rabbits with induced LSCD.

The role of interleukin-33 in chronic allergic conjunctivitis.

Purpose: The authors discovered a genetic association between the ST2L gene and atopy. The ST2L gene encodes a membrane-bound functional marker for Th2 cells. Recently, a novel Th2 cytokine, interleukin-33 (IL-33), was discovered to be a specific ligand for ST2L.

Investigation of the corneal filament in filamentary keratitis.

Purpose: To date, no studies have elucidated the composition of the corneal filament in detail. In this study, an immunohistochemical technique was used to clarify the exact composition of the corneal filament in filamentary keratitis. In addition, the mechanisms responsible for filament formation were identified.

Genomic aberrations and cellular heterogeneity in SV40-immortalized human corneal epithelial cells.

Purpose: Simian virus (SV)40-immortalized human corneal epithelial (HCE-T) cells have been widely used as an in vitro model of human corneal epithelial cells. The nature of this cell line was assessed for genomic aberrations and cellular heterogeneity.

Methods: For the quantitative measurement of genomic aberrations, array-based comparative genomic hybridization (CGH) analysis was performed.

Cultivated corneal endothelial transplantation in a primate: possible future clinical application in corneal endothelial regenerative medicine.

Purpose: To review our attempt to devise a method of cultivated corneal endothelial transplantation using primates in which corneal endothelium, like that of humans, has low proliferative ability.

Methods: Monkey corneal endothelial cells (MCECs) were cultivated, with subcultures grown on collagen type I carriers. The corneal endothelia of 6 eyes of 6 monkeys were scraped intensively, after which cultivated MCEC sheets were inserted into the anterior chamber of 4 eyes.

Diagnosis of multiple myeloma in a patient with atypical corneal findings.

Purpose: To report the case of a 67-year-old woman with a diagnosis of multiple myeloma (MM) and atypical corneal opacity as the first appearance.

Methods: The patient consulted an ophthalmologist regarding blurred vision that progressed gradually without any systemic symptoms. The corneal epithelium and anterior stroma of both eyes harbored diffuse gray-white deposits of unknown etiology.

A thermoreversible hydrogel as a biosynthetic bandage for corneal wound repair.

Ocular trauma and disorders that lead to corneal blindness account for over 2 million new cases of monocular blindness every year. A popular ocular surface reconstruction therapy, amniotic membrane transplantation, has been shown to aid corneal wound repair. However, the success rates of the procedure are variable.