Experimental interventions attenuate a conjunctival epidermal metaplasia model.

The conjunctiva is a non-keratinized, stratified columnar epithelium with characteristics different from the cornea and eyelid epidermis. From development to adulthood, a distinguishing feature of ocular versus epidermal epithelia is the expression of the master regulator PAX6. A conditionally immortalized conjunctival epithelial cell line (iHCjEC) devoid of stromal or immune cells established in our laboratory spontaneously manifested epidermal metaplasia and upregulated expression of the keratinization-related genes SPRR1A/B and the epidermal cytokeratins KRT1 and KRT10 at the expense of the conjunctival trait. In addition, iHCjEC indicated a significant decrease in PAX6 expression. Dry eye syndrome (DES) and severe ocular surface diseases, such as Sjögren's syndrome and Stevens-Johnson syndrome, cause the keratinization of the entire ocular surface epithelia. We used iHCjECs as a conjunctiva epidermal metaplasia model to test PAX6, serum, and glucocorticoid interventions. Reintroducing PAX6 to iHCjECs resulted in upregulating genes related to cell adhesion and tight junctions, including MIR200CHG and CLDN1. The administration of glucocorticoids or serum resulted in the downregulation of epidermal genes (DSG1, SPRR1A/B, and KRT1) and partially corrected epidermal metaplasia. Our results using an isolated conjunctival epidermal metaplasia model point toward the possibility of rationally "repurposing" clinical interventions, such as glucocorticoid, serum, or PAX6 administration, for treating epidermal metaplasia of the conjunctiva.

Clinical characteristics of lacrimal drainage pathway disease-associated keratopathy.

PURPOSE: To report the clinical characteristics of 13 cases of noninfectious corneal ulceration related to lacrimal drainage pathway disease. METHODS: Medical records of 13 patients with lacrimal drainage pathway disease-associated keratopathy who were examined at Ehime University Hospital between April 2007 and December 2021 were analyzed. RESULTS: The predisposing lacrimal drainage pathway diseases for corneal ulceration were chronic dacryocystitis in seven patients and lacrimal canaliculitis in six patients. The corneal ulcers were located at the peripheral cornea in 10 patients and the paracentral cornea in three patients. All patients indicated few cellular infiltrations of the ulcerated area at the slit-lamp examination. Corneal perforation was found in seven patients. The primary identified organisms were Streptococcus spp. in chronic dacryocystitis and Actinomycetes spp. in lacrimal canaliculitis. All patients showed rapid healing of the epithelial defects after treatment of the lacrimal drainage pathway disease. The mean time elapsed between treatment of the lacrimal drainage pathway disease and re-epithelialization of corneal ulcer was 14.5 ± 4.8 days. CONCLUSION: Lacrimal drainage pathway disease-associated keratopathy may be characterized by peripheral corneal ulcer with few cellular infiltrations, occasionally leading to corneal perforation. Treatment of the lacrimal drainage pathway disease could be the most effective treatment for lacrimal drainage pathway disease-associated keratopathy.

Improvement of visual function and ocular and systemic symptoms following blepharoptosis surgery.

Purpose: To study the changes of the visual function and ocular and systemic symptoms following blepharoptosis surgery.Methods: Seventy-eight involutional blepharoptosis patients (72.1 ± 6.4 years) underwent levator advancement procedure. Before and at 2 months after the surgery, OPD-Scan III (Nidek) was used to measure corneal astigmatism, total higher order aberrations (HOAs), and area ratio (AR), an index of the objective contrast sensitivity. FVA-100 (Nidek) was used to determine the functional visual acuity (FVA) and visual maintenance ratio (VMR). The ocular and systemic symptoms were also determined by a questionnaire using visual analogue scale (VAS) scores.Results: Before surgery, the corneal astigmatism, HOAs, AR, FVA, and VMR were 1.56 ± 0.52 diopters (D), 0.23 ± 0.24 µm, 14.8 ± 4.2%, 0.68 ± 0.32 logMAR units and 0.76 ± 0.06, respectively. After surgery, these values were 1.29 ± 0.41 D, 0.19 ± 0.21 µm, 18.6 ± 3.4%, 0.31 ± 0.18 logMAR units and 0.88 ± 0.03, respectively. Corneal astigmatism and HOAs were significantly reduced after surgery (P=0.007 and P=0.023, paired t test, respectively), and AR, FVA and VMR were significantly improved after surgery (P=0.033, P=0.012 and P=0.016, respectively). The VAS scores significantly improved after surgery, and this improvement was positively and significantly correlated with the increase of the AR (r=0.421, P=0.003) and the VMR (r=0.497, P =0.005).Conclusions: Blepharoptosis surgery is visual functionally beneficial and can help to reduce the ocular and systemic symptoms.

Evaluation of a new method of irrigation and aspiration for removal of ophthalmic viscoelastic device during cataract surgery in a porcine model.

BACKGROUND: To determine if a method for irrigation and aspiration (I/A) during cataract surgery provides effective removal of ophthalmic viscoelastic device (OVD). METHODS: Japanese porcine eyes were used to evaluate I/A performance with Technique 1 (the I/A tip placed on the center of the anterior surface of the IOL), Technique 2 (the I/A tip alternately pressed near the edge of the IOL optic anterior surface on one side and then the other to tilt the IOL back and forth), and Technique 3 (the I/A tip inserted behind the IOL optic, between it and the posterior capsule). Techniques 1 and 2 were compared using the Miyake-Apple posterior view video technique to visualize the flow of irrigation fluid containing triamcinolone acetonide particles behind the IOL. To check the efficacy of OVD removal from behind the IOL for of all three I/A techniques, OVD with fluorescein beads were inserted inside the lens capsule before implantation of the IOL. After each I/A technique, eyes were prepared for Miyake-Apple viewing and pictures of the lens capsule were taken using fluorescent microscopy. Residual fluorescein beads in the capsular bag were analyzed. RESULTS: Technique 1 resulted in a straight flow of fluid behind the IOL, while Technique 2 resulted in a vortex flow. The average amount of OVD retained inside the capsule after using Technique 2 or 3 was significantly lower than after using Technique 1 (p <0.0001). CONCLUSIONS: Technique 2 proved to remove more effectively fluorescein bead-labelled OVD under the IOL than Technique 1.

Accuracy of the Lacrimal Syringing Test in Relation to Dacryocystography and Dacryoendoscopy.

Purpose: This study aimed to evaluate the accuracy of the lacrimal syringing test (syringing) in relation to dacryocystography (DCG) and dacryoendoscopy (DE) in lacrimal pathway (LP) obstruction. Methods: We retrospectively reviewed 101 patients (188 LPs) who underwent syringing, cone-beam computed tomography (CBCT)-DCG, and DE to diagnose the degree and site of LP obstruction. The degree of obstruction was classified into patent, partial obstruction, and complete obstruction. The degree of LP obstruction was determined by combining the findings of DE and CBCT-DCG and was designated as DCG+DE. When differentiating complete or partial obstruction on DE was uncertain, complete or partial obstruction was assigned if the DCG showed evidence of contrast media obstruction or passage, respectively. The consistency between syringing and DCG+DE was evaluated by calculating the weighted kappa coefficient. Results: The sensitivity and specificity of syringing were 94% and 89%, respectively. The weighted Cohen's kappa value of agreement between syringing and DCG+DE was κ = 0.73 (SE = 0.04, 95% confidence interval [CI]: 0.66-0.80, p < 0.01). When divided into presaccal and postsaccal obstruction categories, the kappa values were κ = 0.40 (SE = 0.11, 95% CI: 0.19-0.62, p < 0.01) and 0.55 (SE = 0.09, 95% CI: 0.37-0.73, p < 0.01), respectively. Furthermore, when limited to common canaliculus obstruction (stenoses), which accounted for 78% (46 LPs) of the total presaccal obstructions, the kappa value was κ = 0.29 (SE = 0.12, 95% CI: 0.05-0.54, p = 0.021), which demonstrated the lowest consistency between syringing and DCG+DE. Conclusion: Syringing was "substantially" in agreement with DCG+DE in detecting the degree of LP obstruction. Moreover, syringing was a sufficiently sensitive and specific test when performed by an experienced clinician. However, distinguishing between complete and partial obstructions by syringing was sometimes difficult, particularly in the common canaliculus obstruction (stenosis).

Comparison of the efficacies of 1.0 and 1.5 mm silicone tubes for the treatment of nasolacrimal duct obstruction.

This retrospective observational study analyzed the postoperative outcomes of bicanalicular intubation using different diameters of tube stents for treating postsaccal nasolacrimal duct obstruction. A total of 130 patients diagnosed with postsaccal obstruction who underwent endoscopic-assisted silicone tube intubation were included in the study. Patients intubated with a 1.5-mm large-diameter tube were designated as the LD group, and those with a 1.0-mm normal-diameter tube were designated as the ND group. The patency rates of the two groups at 1 year after tube removal were compared using the Kaplan-Meier curve and restricted mean survival time (RMST) method with τ = 365 days. Results demonstrated that the recurrence rate after tube removal was significantly lower in the LD group as compared with the ND group (p = 0.001). The patency rates at 1 year after removal in the LD and ND group were 85.7% (95% confidence interval [CI]: 75.4, 91.9) and 73.9% (95% CI: 61.7, 82.8), respectively. When comparing the patency rates by the RMST method at τ = 365 days, the RMST difference, RMST ratio, and RMTL ratio were higher in the LD group at p = 0.045, 0.052, and 0.046, respectively.

Analysis of Lacrimal Duct Morphology from Cone-Beam Computed Tomography Dacryocystography in a Japanese Population.

Purpose: The dacryoendoscope is a practical instrument for the examination and the treatment of lacrimal duct obstruction. Nevertheless, as it is a rigid fiberscope, manipulation of the endoscope is somewhat affected by the patient's lacrimal duct alignment and the skeletal structure of the face. The morphology and inclination of the lacrimal duct vary among individuals and ethnic groups. We aimed to evaluate the alignment of the lacrimal duct from the perspective of endoscopic maneuverability in a Japanese population. Methods: This retrospective study analyzed the cone-beam computed tomography dacryocystography (CBCT-DCG) images of 102 patients diagnosed with unilateral primary acquired nasolacrimal duct obstruction (PANDO) at Ehime University Hospital from December 2015 to May 2021. The following parameters of the lacrimal duct on the contralateral side of unilateral PANDO were investigated: (1) angle formed by the superior orbital rim-internal common punctum-nasolacrimal duct opening, (2) angle formed by the lacrimal sac and the nasolacrimal duct, (3) length of the lacrimal sac, and (4) length of the nasolacrimal duct. Results: Measurements of the above parameters were (1) 10.2° ± 7.8° (range, -11° to +27°), (2) -6.3° ± 14.1° (range, -43° to +40°), (3) 8.9 ± 2.3 mm (range, 4.3-17.1), and (4) 13.2 ± 2.7 mm (range, 5.7-20.7), respectively. The Shapiro-Wilk test demonstrated that the values of all parameters, except (3), followed a normal distribution (p = 0.55, 0.30, 0.0002, and 0.39, respectively). No significant difference was found between the female and male groups (p > 0.05). Conclusion: This study reported anthropometric analysis data of the morphology of the lacrimal ducts using CBCT-DCG in a Japanese population. In our cohort, the line from the superior orbital rim through the internal common punctum to the nasolacrimal duct opening inclined anteriorly in 92% of the patients.

Characterization of doxycycline-dependent inducible Simian Virus 40 large T antigen immortalized human conjunctival epithelial cell line.

PURPOSE: To present the properties of a newly developed immortalized human conjunctival epithelial cell (iHCjEC) line. METHODS: iHCjECs were developed to induce Simian Virus 40 large T-antigen (SV40LT) by incorporating lentivirus in a tetracycline (Tet)-regulated gene-expression system into primary cultures of human conjunctival epithelial cells. The population doubling time and morphology of the iHCjECs were analyzed. The expressions of CK13, CK19, CK12, and MUC1, MUC4, MUC16, and MUC5AC were determined by real time PCR and immunohistochemically under different culture conditions. The organotypic culture model in which iHCjECs were cultured on rabbit conjunctival fibroblast-embedded collagen gel was used to characterize the iHCjECs. RESULTS: The iHCjECs cultured with doxycycline (Dox) continued to proliferate for at least 20 passages and had a cobblestone-like appearance. The expressions of CK13 and CK19 but not CK12 were detected in the iHCjECs, and the expression of CK13 increased in culture media lacking Dox (Dox-). The expressions of MUC1, MUC4, MUC16, and MUC5AC were detected in iHCjECs, and a relatively strong immunostaining of MUC5AC was detected with Dox(-) added 5% FBS. Stratified iHCjECs were observed in organotypic culture at 5 days. CONCLUSION: The iHCjECs had high proliferation rates and abilities to control the differentiation potency to control the expression of SV40 LT-antigen with Tet-regulated gene-expression system. They are able to express the mucin gene repertoire of their native epithelia. The iHCjECs can be a useful experimental cell line to study conjunctival epithelial cell characteristics and for pathophysiological and toxicological studies.

Ocular penetration and efficacy of levofloxacin using different drug-delivery techniques for the prevention of endophthalmitis in rabbit eyes with posterior capsule rupture.

PURPOSE: To evaluate the effects of different drug-delivery techniques for levofloxacin (LVFX) in ocular penetration and the prevention of endophthalmitis using an aphakic rabbit model with posterior capsule rupture (PCR). METHODS: LVFX was administered to aphakic rabbit eyes with or without PCR using eye drops (EDs), subconjunctival injection (SCI), or intracameral (IC) injection. The concentration of the drug in the vitreous and aqueous humors was estimated at 2 h after injection. In another study, aphakic rabbit eyes with PCR were inoculated with Enterococcus faecalis, immediately followed by 0.5% LVFX ED, 0.5% moxifloxacin (MFLX) ED, LVFX IC (500 µg/0.1 mL), or IC saline. EDs were administered 0, 3, and 6 h after surgery. Changes on electroretinography (ERG) and intraocular bacterial growth were determined sequentially until 48 h after inoculation. RESULTS: The concentrations of LVFX at 2 h after IC were higher in the aqueous humor and the vitreous cavity of eyes with or without PCR, compared with EDs or SCI. Eyes treated with LVFX ED, MFLX ED, or IC saline showed a significantly greater reduction in b-wave amplitude on ERG at 48 h compared with eyes treated with LVFX IC. The number of bacteria recovered from the vitreous humor in eyes treated with IC LVFX at 48 h was significantly less than from eyes that received other treatments. CONCLUSION: The LVFX IC was effective at suppressing endophthalmitis caused by E. faecalis in eyes with a PCR.

Fungal keratitis caused by Beauveria bassiana: drug and temperature sensitivity profiles: a case report.

BACKGROUND: Beauveria bassiana is an entomopathogenic fungus and is a rare cause of keratitis. We present a case of fungal keratitis caused by B. bassiana that was diagnosed by in vivo confocal microscopy and in vitro corneal cultures. In addition, we determined the temperature- and drug-sensitivities of the isolated strain of B. bassiana. CASE PRESENTATION: A 59-year-old Japanese man with a 2-month history of keratitis was examined by slit-lamp biomicroscopy, in vivo confocal microscopy, and histology and cultures of corneal scrapings. The corneal scrapings were used to determine the minimal inhibitory concentrations of different antifungal drugs and also to determine the temperature-sensitivity. In vivo confocal microscopy and histological examinations showed filamentous fungal keratitis. The characteristics of the fungal growth indicated that the keratitis was caused by B. bassiana. The keratitis responded poorly to systemic and topical voriconazole and to natamycin ointment. However, it was resolved after changing the natamycin to micafungin combined with surgical debridement. The isolated strain was sensitive to itraconazole, miconazole, micafungin, voriconazole, and resistant to flucytosine and fluconazole. It was moderately sensitive to amphotericin B, and natamycin. After 7 days in culture, the isolate grew small white colonies at 25 °C, very small colonies at 35 °C and 37 °C. CONCLUSION: The drug-sensitivity and temperature-sensitivity profiles of B. bassiana should be helpful in the treatment of B. bassiana keratitis. Therapeutic surgery may be helpful for mycotic keratitis poorly responsive to medical therapy alone.

Clinical characteristics of keratitis due to Colletotrichum gloeosporioides.

PURPOSE: To determine the characteristics of the keratitis due to Colletotrichum gloeosporioides. METHODS: The medical records of 3 cases of fungal keratitis caused by C. gloeosporioides were reviewed to determine the clinical characteristics. The minimal inhibitory concentrations of different antifungal drugs for all 3 isolates were determined. All 3 isolates were grown on Sabouraud dextrose agar at 25°C, 35°C, and 37°C to determine the temperature-sensitive growth. RESULTS: All 3 patients lived in the southwestern part of Japan and had an ocular trauma involving organic materials. The infectious foci were localized in the anterior stroma, and they did not extend deep into the stroma in all cases. The keratitis was treated with antifungal medications including topical voriconazole and natamycin eye ointment, and was resolved in 2-3 weeks. All of the isolated strains grew well at 25°C but poorly at 35°C and 37°C. All isolated strains had similar drug-sensitivity profiles; they were sensitive to amphotericin B, itraconazole, miconazole, micafungin, and voriconazole, and relatively resistant to flucytosine, fluconazole, and natamycin. CONCLUSIONS: All 3 cases of C. gloeosporioides keratitis had similar clinical features. The similarities in the drug-sensitivity profiles should be helpful in treating C. gloeosporioides keratitis.

In vivo and in vitro investigations of fungal keratitis caused by Colletotrichum gloeosporioides.

PURPOSE: To report a case of fungal keratitis caused by Colletotrichum gloeosporioides, which is a rare pathogen in humans. METHODS: An 80-year-old woman developed fungal keratitis after having sustained a traumatic injury during field work. The patient was initially examined by slit-lamp biomicroscopy and the Heidelberg Retina Tomograph II-Rostock Cornea Module (HRT II-RCM). Corneal scrapings were collected and submitted for laboratory investigations. RESULTS: Many septate, hyphae-like interlocking and branching white lines were observed in the area of the infiltrate by HRT II-RCM. A tentative diagnosis of fungal keratitis was made, and the patient was treated with systemic and topical voriconazole and pimaricin ophthalmic ointment. The infectious focus resolved within 2 weeks, and there were no signs of a recurrence after 3 months of treatment with the antifungal agents. The culture of the corneal scraping grew C. gloeosporioides. CONCLUSIONS: HRT II-RCM was useful in detecting filamentous fungi in the cornea. The treatment with voriconazole and pimaricin was effective in the treatment of C. gloeosporioides keratitis.