R125H, W240S, C386R, and V507I SLC4A11 mutations associated with corneal endothelial dystrophy affect the transporter function but not trafficking in PS120 cells.

SLC4A11 mutations are associated with Fuchs' endothelial corneal dystrophy (FECD), congenital hereditary endothelial dystrophy (CHED) and Harboyan syndrome (endothelial dystrophy with auditory deficiency). Mice with genetically ablated Slc4a11 recapitulate CHED, exhibiting significant corneal edema and altered endothelial morphology. We recently demonstrated that SLC4A11 functions as an NH3 sensitive, electrogenic H+ transporter. Here, we investigated the properties of five clinically relevant SLC4A11 mutants: R125H, W240S, C386R, V507I and N693A, relative to wild type, expressed in a PS120 fibroblast cell line. The effect of these mutations on the NH4Cl-dependent transporter activity was investigated by intracellular pH and electrophysiology measurements. Relative to plasma membrane expression of NaK ATPase, there were no significant differences in plasma membrane SLC4A11 expression among each mutant and wild type. All mutants revealed a marked decrease in acidification in response to NH4Cl when compared to wild type, indicating a decreased H+ permeability in mutants. All mutants exhibited significantly reduced H+ currents at negative holding potentials as compared to wild type. Uniquely, the C386R and W240S mutants exhibited a different inward current profile upon NH4Cl challenges, suggesting an altered transport mode. Thus, our data suggest that these SLC4A11 mutants, rather than having impaired protein trafficking, show altered H+ flux properties.

Ten-year incidence and prevalence of clinically diagnosed blepharitis in South Korea: a nationwide population-based cohort study.

IMPORTANCE: Blepharitis is one of the most common conditions. However, no study has yet evaluated the epidemiology by evaluating a large population-based sample. BACKGROUND: To evaluate the incidence and prevalence of clinically diagnosed blepharitis in South Korea. DESIGN: Nationwide population-based study. PARTICIPANTS: We investigated the Korean National Health Insurance Service-National Sample Cohort, a representative one million-sample of the Korean population, for patients diagnosed with blepharitis according to the Korean Classification of Diseases. METHODS: Annual and overall incidence and prevalence of blepharitis during the study period (2004-2013) were estimated after excluding chronic blepharitis patients, diagnosed during 2002-2003. Sociodemographic factors and comorbidities associated with blepharitis were evaluated using Cox proportional hazard regression. MAIN OUTCOME MEASURES: The first occurrence of blepharitis. RESULTS: A total of 1 116 363 individuals over 9 698 118 person-years were evaluated (mean follow up: 8.7 years) from 2004 to 2013. The overall incidence was 1.1 (95% confidence interval, 1.1-1.1) per 100 person-years. The incidence increased with time (0.9 vs. 1.3 per 100 person-years, in 2004 and 2013, respectively) and was higher in female patients (1.3 vs. 0.9 per 100 person-years, respectively). The overall prevalence was 8.1% (95% confidence interval: 8.0-8.1) among subjects aged 40 years or older. Chalazion, gastritis, Sjögren's syndrome, pterygium, rosacea, prostatic hypertrophy, atopy, irritable bowel disease and peptic ulcer were associated with an increased incidence of blepharitis in the multivariable Cox model. CONCLUSIONS: We found that blepharitis was a relatively common disease and is associated with various ocular and systemic conditions.

Predictive model for recovery of visual field after surgery of pituitary adenoma.

Visual field defect is a major indication for surgery of pituitary adenoma, but visual outcome after surgery is difficult to predict. We developed a nomogram that predicts postoperative restoration of visual field defects in patients with pituitary adenoma. This study was a retrospective cohort investigation of patients who were treated for pituitary adenoma between January 2009 and December 2013. We enrolled 111 eyes of 57 patients who completed one ophthalmological evaluation preoperatively and at least two evaluations within 6 months after surgery. Serial changes in visual fields and retinal nerve fiber layer (RNFL) thickness were evaluated. Multiple logistic regression analysis was performed to select prognostic variables, and a nomogram to predict restoration of visual field defects was constructed. Visual field defects continuously improved until 3 months after surgery. However, average, superior, and inferior RNFL thickness continuously decreased until 6 months after surgery. Multiple logistic regression analysis revealed that worse preoperative visual field defect (p = 0.018), high MRI compression grade (p = 0.009), and inferior RNFL thinning (p = 0.011) were significantly associated with worse visual outcome. The nomogram that predicts the visual restoration showed an area under the receiver operating characteristic curve of 0.84. In conclusion, we developed a nomogram that predicted the restoration of visual field defects after removal of pituitary adenoma. This would allow tailored counseling of individual patients by precisely predicting visual recovery after surgery.

Fundus autofluorescence findings in central serous chorioretinopathy using two different confocal scanning laser ophthalmoscopes: correlation with functional and structural status.

PURPOSE: To compare autofluorescence (AF) findings using wide-field (Optomap) and conventional (HRA-AF) confocal scanning laser ophthalmoscopy (cSLO) systems in patients with central serous chorioretinopathy (CSC), and to investigate the correlations between AF findings and functional and anatomical status. METHODS: Optical coherence tomography (OCT) and AF images were compared in 73 eyes with serous retinal detachment (SRD) (group A) and 30 eyes without SRD (group B). We evaluated AF findings from the SRD region, atrophic area, and foveola. Correlations between AF findings and outer retinal abnormalities in OCT and visual acuity (VA) were analyzed. RESULTS: Optomap-AF was more effective than HRA-AF in identifying the margins of a detached area (P = 0.001) in group A, and for monitoring mild outer retinal damage (P = 0.041) in group B. The foveolar AF grades in both instruments were significantly correlated with VA and central foveal thickness (CFT) in both group A (Optomap, VA r s = 0.33, P = 0.012; CFT r s = -0.38, P = 0.002; HRA, VA r s = 0.62, P < 0.001; CFT r s = -0.70, P < 0.001) and group B (Optomap, VA r s = 0.71, P < 0.001, CFT r s = -0.78, P < 0.001; HRA, VA r s = 0.40, P = 0.026, CFT r s = -0.40, P = 0.030). CONCLUSIONS: Optomap-AF was found to be advantageous for monitoring subretinal status in eyes with SRD, and more accurately reflected mild outer retinal changes in eyes without SRD. Foveolar AF grades of both imaging modalities were significantly correlated with functional and anatomical status.

Anti-Vascular Endothelial Growth Factor with Gas for Submacular Hemorrhage.

PURPOSE: To investigate the treatment outcome of pneumatic displacement and intravitreal anti-vascular endothelial growth factor (VEGF) for submacular hemorrhage (SMH) from exudative age-related macular degeneration (AMD). METHODS: Best-corrected visual acuity (BCVA) and central foveal thickness (CFT) were measured at baseline and at 1, 3, and 6 months after initial treatment in 72 eyes of 72 patients treated with a combination of pneumatic displacement and anti-VEGF injection for SMH from exudative AMD. RESULTS: Best-corrected visual acuity and CFT showed significant improvement from baseline during the 6-month follow-up period (logarithm of the minimum angle of resolution BCVA from 1.80 to 1.00, CFT from 886 to 383 µm, p < 0.001, respectively). The decrease in subretinal hemorrhage was greater than that in subretinal pigment epithelial hemorrhage at 1 month after initial treatment (p < 0.001). In eyes with symptoms for less than 30 days, higher reflectivity of hemorrhage on optical coherence tomography and higher CFT were associated with lower BCVA after 6 months of treatment (reflectivity B = 0.335, p = 0.007; CFT B = 0.001, p = 0.003). CONCLUSIONS: The combination of pneumatic displacement and intravitreal anti-VEGF is a useful treatment option for SMH secondary to AMD. Higher baseline CFT and higher reflectivity of hemorrhage were associated with lower BCVA 6 months after initial treatment.

Pigment epithelial tears after ranibizumab injection in polypoidal choroidal vasculopathy and typical age-related macular degeneration.

PURPOSE: The purpose of the study was to compare the rates and characteristics of retinal pigment epithelial (RPE) tears between typical exudative age-related macular degeneration (tAMD) and polypoidal choroidal vasculopathy (PCV) after injection of intravitreal ranibizumab (IVR). METHODS: In total, 836 eyes from 784 patients with exudative AMD treated with IVR were analyzed. The presence, type, size, and height of pigment epithelial detachment (PED) in OCT before injection were evaluated, and the occurrence rate of RPE tears within three months of injection between tAMD and PCV was compared. RESULTS: In total, 515 eyes (61.6 %) had tAMD and 321 eyes (38.4 %) were diagnosed as PCV. RPE tears developed in 18 eyes (3.5 %) in the tAMD group, while only two eyes (0.62 %) were associated with PCV (p = 0.009, Chi-square test). Eleven of the eighteen eyes with RPE tears in tAMD had fibrovascular PED with contractile neovascular tissue under the surface of the RPE and a cleft at baseline. Two eyes with RPE tears in PCV showed large hemorrhagic PED before presenting with an RPE tear. CONCLUSIONS: RPE tears after IVR were significantly less common in PCV than in tAMD. The different characteristics of RPE tears between the two disease entities suggest differences in the pathogenesis underlying the event.

Comparison of three types of images for the detection of retinal nerve fiber layer defects.

PURPOSE: To compare the clinical effectiveness of three types of images for detecting retinal nerve fiber layer (RNFL) defects. METHODS: Three image sets of 100 subjects (9 normal control subjects, 16 glaucoma suspects, and 75 glaucoma patients) were produced using color fundus photography, typical red-free RNFL photography, and blue reflectance RNFL photography with confocal scanning laser ophthalmoscopy (CSLO). A total of 300 images were rated twice in random order by five independent evaluators who were masked to the patient characteristics; each image was rated as normal, having a diffuse RNFL defect, or showing a wedge RNFL defect. Intraobserver and interobserver agreement, sensitivity, specificity, accuracy, and area under the curve were assessed. An additional analysis was performed for identifying differences in two black-and-white RNFL photographs. RESULTS: The results showed high intraobserver agreement, with relatively low interobserver agreements among the five evaluators. Blue reflectance RNFL photography with CSLO demonstrated the best performance in sensitivity, specificity, accuracy, and area under the curve. Blue reflectance RNFL images showed better accuracy than red-free RNFL images especially in subjects with wedge defects and in advanced glaucomatous cases. CONCLUSIONS: The RNFL images produced using blue reflectance with CSLO showed the best performance for the detection of RNFL defects, especially in cases with wedge defects and advanced glaucoma stages.

SiO(2) /TiO(2) hollow nanoparticles decorated with Ag nanoparticles: enhanced visible light absorption and improved light scattering in dye-sensitized solar cells.

Hollow SiO2 /TiO2 nanoparticles decorated with Ag nanoparticles (NPs) of controlled size (Ag@HNPs) were fabricated in order to enhance visible-light absorption and improve light scattering in dye-sensitized solar cells (DSSCs). They exhibited localized surface plasmon resonance (LSPR) and the LSPR effects were significantly influenced by the size of the Ag NPs. The absorption peak of the LSPR band dramatically increased with increasing Ag NP size. The LSPR of the large Ag NPs mainly increased the light absorption at short wavelengths, whereas the scattering from the SiO2 /TiO2 HNPs improved the light absorption at long wavelengths. This enabled the working electrode to use the full solar spectrum. Furthermore, the SiO2 layer thickness was adjusted to maximize the LSPR from the Ag NPs and avoid corrosion of the Ag NPs by the electrolyte. Importantly, the power conversion efficiency (PCE) increased from 7.1 % with purely TiO2 -based DSSCs to 8.1 % with HNP-based DSSCs, which is an approximately 12 % enhancement and can be attributed to greater light scattering. Furthermore, the PCEs of Ag@HNP-based DSSCs were 11 % higher (8.1 vs. 9.0 %) than the bare-HNP-based DSSCs, which can be attributed to LSPR. Together, the PCE of Ag@HNP-based DSSCs improved by a total of 27 %, from 7.1 to 9.0 %, due to these two effects. This comparative research will offer guidance in the design of multifunctional nanomaterials and the optimization of solar-cell performance.

Arsenic(III) detection using electrochemical-chemical-chemical redox cycling at bare indium-tin oxide electrodes.

Sensitive As(III) detection in ground water is of great importance for evaluating the quality of drinking water. We report a sensitive electrochemical method for As(III) detection based on electrochemical-chemical-chemical (ECC) redox cycling involving Ru(IV) [an oxidized species of Ru(III)(NH3)5NH2(2+)], As(III), and tris(3-carboxyethyl)phosphine (TCEP). Electrochemical oxidation of Ru(III)(NH3)5NH2(2+) formed from Ru(III)(NH3)6(3+) generates Ru(IV), which quickly oxidizes As(III). This electro-mediated oxidation of As(III) produces As(V), which is reduced back to As(III) by TCEP. Electrochemically generated Ru(IV) then reoxidizes As(III), allowing ECC redox cycling to occur at a high rate on bare indium-tin oxide (ITO) electrodes without modifying the surfaces with electrocatalytic materials. Because most interfering metal ions precipitate in a carbonate buffer, water samples are mixed with carbonate buffers prior to electrochemical measurements, rendering the effects of Cu(+), Cu(2+), Fe(2+), Fe(3+), and Pb(2+) insignificant. The detection limit calculated by ECC redox cycling using a chronocoulogram is 1.2 µM, much lower than that obtained using only the electro-mediated oxidation of As(III) (90 µM).

Regional Conjunctival Differences in Glycocalyx Mucin Expression in Dry Eye and Normal Subjects.

Purpose: To compare regional conjunctival expression of membrane-associated mucins (MAMs) MUC1, MUC4, and MUC16 in normal and dry eye (DE) subjects. Methods: Adults with and without signs and symptoms of DE were recruited. Impression cytology was performed to collect MAMs from four bulbar and upper eyelid palpebral conjunctival regions of both eyes. After protein extraction, samples from both eyes of a single subject were pooled by region, and expression was analyzed using a capillary electrophoresis nano-immunoassay system. The chemiluminescence intensity of each antigen binding signal was calculated after normalization to the total protein amount. Statistical analyses were conducted using GraphPad Prime 9. Results: Samples from thirteen to sixteen DE and seven to eleven normal subjects were analyzed. In normal samples, MUC1 expression from the nasal bulbar conjunctiva was significantly greater than superior (P = 0.004) and inferior (P = 0.005). In DE samples, MUC1 expression was highest superiorly. Significant differences in MUC4 and MUC16 expression were not seen in normal samples. MUC4 and MUC16 expression was upregulated superiorly (P < 0.0001) and inferiorly (P < 0.0001) in DE compared with those regions in normal samples. Conclusions: Although MAMs form a hydrophilic barrier called the glycocalyx, each mucin may have unique functions that are currently unexplored. All MAMs were expressed in the upper palpebral conjunctiva. Increased MUC1 expression nasally in healthy subjects suggests a functional need for increased protection. When comparing DE with normal eyes, upregulation of MUC1 superiorly, and in both MUC4 and MUC16 both superiorly and inferiorly, may indicate a need to decrease eyelid friction during blinking, especially in DE.

Comparative Evaluation of Matrix Metalloproteinase-9 Immunoassay and Tear Osmolarity Measurement for Diagnosing Severity of Dry Eye Disease.

PURPOSE: To evaluate and compare the clinical efficacy of matrix metalloproteinase-9 (MMP-9) immunoassay and tear osmolarity measurement in diagnosing dry eye severity. METHODS: Dry eye disease (DED) patients underwent diagnostic tests including MMP-9 assay, tear osmolarity measurement, fluorescein tear breakup time, ocular surface staining, anesthetized Schirmer test, Ocular Surface Disease Index questionnaire, and slit-lamp examination. The dry eye parameters were compared according to positive MMP-9 status and increased tear osmolarity. The correlation between dry eye profiles and MMP-9 positivity and high tear osmolarity was also analyzed. RESULTS: Those who tested positive in MMP-9 immunoassay had significantly higher corneal fluorescein staining score and worse DED severity than those who tested negative. The intensity of MMP-9 positivity showed positive correlation with the corneal staining score and DED severity. However, DED patients with high tear osmolarity above 308 mOsm/L did not show significantly different dry eye signs and symptoms compared to those with lower tear osmolarity values. Tear osmolarity was associated with ocular surface staining score in severe DED patients. CONCLUSIONS: MMP-9 positivity was associated with ocular surface staining and worse dry eye severity. Therefore, it may be used as a useful indicator of disease severity in conjunction to other diagnostic tests.

Efficient intracellular delivery of camptothecin by silica/titania hollow nanoparticles.

Silica/titania hollow nanoparticles (HNPs) with 50 nm were fabricated by using the dissolution and redeposition method and modified with anti-[human epidermal growth factor receptor 2] monoclonal antibody (herceptin), and their application as camptothecin (CPT) delivery agents to human breast cancer SK-BR-3 cells was investigated. Although the diameter of herceptin-modified HNPs (HER-HNP) is smaller than that of other reported mesoporous silica nanoparticles, the extensive hollow cavity of HNPs (ca. 30 nm) allowed the loading of a large amount of CPT. CPT-loaded HER-HNP (HER-HNP-CPT) did not release CPT in phosphate-buffered saline over a period of 24 h, however, HER-HNP-CPT in a hydrophobic solvent released its entire load of CPT. In addition, HER-HNPs were efficiently internalized owing to their herceptin conjugation and optimized size. To evaluate in vitro antitumor efficacy, apoptosis/necrosis and viability of HER-HNP-CPT-treated cells were investigated. When the cells were treated with HER-HNP-CPT for 30 min, a few apoptotic cells were observed. After 24 h, the viability of HER-HNP-CPT-treated SK-BR-3 decreased to 60 %, which revealed highly efficient chemotherapy. However, CPT loaded into HNP or HER-HNP had no significant effects on the viability of macrophages. Judging from these data, HER-HNPs are very suitable for application in anticancer therapy. A HER-HNP-CPT drug delivery system offers a new direction for a hydrophobic anticancer drug carrier and can be expanded to practical applications with further studies.

The H+ Transporter SLC4A11: Roles in Metabolism, Oxidative Stress and Mitochondrial Uncoupling.

Solute-linked cotransporter, SLC4A11, a member of the bicarbonate transporter family, is an electrogenic H+ transporter activated by NH3 and alkaline pH. Although SLC4A11 does not transport bicarbonate, it shares many properties with other members of the SLC4 family. SLC4A11 mutations can lead to corneal endothelial dystrophy and hearing deficits that are recapitulated in SLC4A11 knock-out mice. SLC4A11, at the inner mitochondrial membrane, facilitates glutamine catabolism and suppresses the production of mitochondrial superoxide by providing ammonia-sensitive H+ uncoupling that reduces glutamine-driven mitochondrial membrane potential hyperpolarization. Mitochondrial oxidative stress in SLC4A11 KO also triggers dysfunctional autophagy and lysosomes, as well as ER stress. SLC4A11 expression is induced by oxidative stress through the transcription factor NRF2, the master regulator of antioxidant genes. Outside of the corneal endothelium, SLC4A11's function has been demonstrated in cochlear fibrocytes, salivary glands, and kidneys, but is largely unexplored overall. Increased SLC4A11 expression is a component of some "glutamine-addicted" cancers, and is possibly linked to cells and tissues that rely on glutamine catabolism.

Rescue of the Congenital Hereditary Endothelial Dystrophy Mouse Model by Adeno-Associated Viruse-Mediated Slc4a11 Replacement.

Purpose: Congenital hereditary endothelial dystrophy (CHED) is a rare condition that manifests at an early age showing corneal edema, increased oxidative stress, mitochondrial dysfunction, and eventually apoptosis of the endothelium due to loss of function of the membrane transport protein SLC4A11. This project tested whether replacing Slc4a11 into the Slc4a11 -/- CHED mouse model can reverse the disease-associated phenotypes. Design: Experimental study. Participants: Five-week-old or 11-week-old Slc4a11 -/- mice. Age- and gender-matched Slc4a11 +/+ animals were used as controls. A total of 124 animals (62 female, and 62 male) were used in this study. Fifty-three animals of the genotype Slc4a11 +/+ were used as age- and gender-matched noninjected controls. Seventy-one Slc4a11 -/- mice were administered anterior chamber injections of adeno-associated virus (AAV). Methods: Anterior chambers of young (5 weeks old) or older (11 weeks old) Slc4a11 -/- mice were injected once with adeno-associated virus serotype 9 (AAV9) mouse Slc4a11 or AAV9-Null vectors. Corneal thickness was measured using OCT. End point analysis included corneal endothelial cell density, mitochondrial oxidative stress, and corneal lactate concentration. Main Outcome Measures: Corneal thickness, endothelial cell loss, lactate levels, and mitochondrial oxidative stress. Results: In the young animals, AAV9-Slc4a11 reversed corneal edema, endothelial cell loss, mitochondrial oxidative stress, lactate transporter expression, and corneal lactate concentration to the levels observed in wild-type animals. In the older animals, gene replacement did not reverse the phenotype but prevented progression. Conclusions: Functional rescue of CHED phenotypes in the Slc4a11 -/- mouse is possible; however, early intervention is critical.

Evaluation of dry eye subtypes and characteristics using conventional assessments and dynamic tear interferometry.

BACKGROUND/AIMS: To evaluate subtypes and characteristics of dry eye (DE) using conventional tests and dynamic tear interferometry, and to investigate determinants of disease severity in each DE subtype. METHODS: 309 patients diagnosed with DE and 69 healthy controls were prospectively enrolled. All eyes were evaluated using Ocular Surface Disease Index (OSDI), Schirmer's test I (ST1) and Meibomian gland dysfunction (MGD) grade were analysed. The tear interferometric pattern and lipid layer thickness were determined using DR-1α and LipiView II, respectively. RESULTS: Dynamic interferometric analysis revealed 56.6% of patients with DE exhibited Jupiter patterns, indicative of aqueous-deficiency, while 43.4% exhibited crystal patterns, indicative of lipid deficiency. These findings were in accordance with classification based on ST1 scores and MGD grade. Conventional assessment indicated 286 patients exhibited evidence of evaporative DE (EDE) due to MGD, while only 11 exhibited signs of pure aqueous-deficient DE (pure ADDE, only ST1 ≤5 mm). Interestingly, of 286 patients with EDE, 144 were categorised into the mixed-ADDE/EDE group, in which ST1 was identified as a strong negative determinant of OSDI. In contrast, 72.2% of patients with mixed-ADDE/EDE exhibited Jupiter patterns (Jupiter mixed), while 27.8% exhibited crystal patterns (crystal mixed). OSDI values were significantly higher in the crystal-mixed group than in the Jupiter mixed, in which OSDI scores were independently associated with ST1 values only. CONCLUSIONS: Our findings indicate that majority of EDE patients also exhibit aqueous deficiency, which can aggravate symptoms even in patients with lipid-deficient mixed-ADDE/EDE. Conventional assessments should be combined with interferometric tear analysis to determine the most appropriate treatment for each DE patient.

Mitochondrial Targeting of the Ammonia-Sensitive Uncoupler SLC4A11 by the Chaperone-Mediated Carrier Pathway in Corneal Endothelium.

Purpose: SLC4A11, an electrogenic H+ transporter, is found in the plasma membrane and mitochondria of corneal endothelium. However, the underlying mechanism of SLC4A11 targeting to mitochondria is unknown. Methods: The presence of mitochondrial targeting sequences was examined using in silico mitochondrial proteomic analyses. Thiol crosslinked peptide binding to SLC4A11 was screened by untargeted liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis. Direct protein interactions between SLC4A11 and chaperones were examined using coimmunoprecipitation analysis and proximity ligation assay. Knockdown or pharmacologic inhibition of chaperones in human corneal endothelial cells (HCECs) or mouse corneal endothelial cells (MCECs), ex vivo kidney, or HA-SLC4A11-transfected fibroblasts was performed to investigate the functional consequences of interfering with mitochondrial SLC4A11 trafficking. Results: SLC4A11 does not contain canonical N-terminal mitochondrial targeting sequences. LC-MS/MS analysis showed that HSC70 and/or HSP90 are bound to HA-SLC4A11-transfected PS120 fibroblast whole-cell lysates or isolated mitochondria, suggesting trafficking through the chaperone-mediated carrier pathway. SLC4A11 and either HSP90 or HSC70 complexes are directly bound to the mitochondrial surface receptor, TOM70. Interference with this trafficking leads to dysfunctional mitochondrial glutamine catabolism and increased reactive oxygen species production. In addition, glutamine (Gln) use upregulated SLC4A11, HSP70, and HSP90 expression in whole-cell lysates or purified mitochondria of HCECs and HA-SLC4A11-transfected fibroblasts. Conclusions: HSP90 and HSC70 are critical in mediating mitochondrial SLC4A11 translocation in corneal endothelial cells and kidney. Gln promotes SLC4A11 import to the mitochondria, and the continuous oxidative stress derived from Gln catabolism induced HSP70 and HSP90, protecting cells against oxidative stress.

Mitochondrial ROS Induced Lysosomal Dysfunction and Autophagy Impairment in an Animal Model of Congenital Hereditary Endothelial Dystrophy.

Purpose: The Slc4a11 knock out (KO) mouse model recapitulates the human disease phenotype associated with congenital hereditary endothelial dystrophy (CHED). Increased mitochondrial reactive oxygen species (ROS) in the Slc4a11 KO mouse model is a major cause of edema and endothelial cell loss. Here, we asked if autophagy was activated by ROS in the KO mice. Methods: Immortalized cell lines and mouse corneal endothelia were used to measure autophagy and lysosome associated protein expressions using Protein Simple Wes immunoassay. Autophagy and lysosome functions were examined in wild type (WT) and KO cells as well as animals treated with the mitochondrial ROS quencher MitoQ. Results: Even though autophagy activation was evident, autophagy flux was aberrant in Slc4a11 KO cells and corneal endothelium. Expression of lysosomal proteins and lysosomal mass were decreased along with reduced nuclear translocation of lysosomal master regulator, transcription factor EB (TFEB). MitoQ reversed aberrant lysosomal functions and TFEB nuclear localization in KO cells. MitoQ injections in KO animals reduced corneal edema and decreased the rate of endothelial cell loss. Conclusions: Mitochondrial ROS disrupts TFEB signaling causing lysosomal dysfunction with impairment of autophagy in Slc4a11 KO corneal endothelium. Our study is the first to identify the presence as well as cause of lysosomal dysfunction in an animal model of CHED, and to identify a potential therapeutic approach.

Heat-mortality risk and the population concentration of metropolitan areas in Japan: a nationwide time-series study.

BACKGROUND: The complex role of urbanisation in heat-mortality risk has not been fully studied. Japan has experienced a rapid population increase and densification in metropolitan areas since the 2000s; we investigated the effects of population concentration in metropolitan areas on heat-mortality risk using nationwide data. METHODS: We collected time-series data for mortality and weather variables for all 47 prefectures in Japan (1980-2015). The prefectures were classified into three sub-areas based on population size: lowest (<1 500 000), intermediate (1 500 000 to 3 000 000), and highest (>3 000 000; i.e. metropolitan areas). Regional indicators associated with the population concentration of metropolitan areas were obtained. RESULTS: Since the 2000s, the population concentration intensified in the metropolitan areas, with the highest heat-mortality risk in prefectures with the highest population. Higher population density and apartment % as well as lower forest area and medical services were associated with higher heat-mortality risk; these associations have generally become stronger since the 2000s. CONCLUSIONS: Population concentration in metropolitan areas intensified interregional disparities in demography, living environments, and medical services in Japan; these disparities were associated with higher heat-mortality risk. Our results can contribute to policies to reduce vulnerability to high temperatures.

Visual Outcomes after Bilateral Implantation of an Extended Depth of Focus Intraocular Lens: A Multicenter Study.

PURPOSE: To evaluate visual performance after bilateral implantation of an extended depth of focus (EDOF) intraocular lens (IOL). METHODS: This multicenter, prospective, observational study included 100 patients who underwent bilateral cataract surgery with a toric or non-toric EDOF IOL (Tecnis Symfony), and 96 patients completed the final assessment at 4 to 6 months. Binocular corrected distance visual acuity and uncorrected distance visual acuity (UDVA), uncorrected intermediate visual acuity (UIVA), and uncorrected near visual acuity (UNVA), spectacle independence, visual symptoms, and patient satisfaction were evaluated. RESULTS: Mean decimal visual acuity results showed a binocular corrected distance visual acuity of 1.10 ± 0.18, UDVA of 1.04 ± 0.17, UIVA of 0.96 ± 0.16, and UNVA of 0.68 ± 0.18. Binocular UDVA and UIVA were 0.8 (decimal) or better in 98% and 94% of patients, respectively. Binocular UNVA was 0.63 (decimal) or better in 76% of patients. Overall, 76% of the patients achieved spectacle independence across all distances, and more than 85% reported no or mild dysphotoptic phenomena. On a scale of 0 to 10, the median patient satisfaction score was 9 for far, 9.5 for intermediate, and 8 for near vision. CONCLUSIONS: The Symfony EDOF IOL provided excellent distance, intermediate visual outcome, and functional near visual acuity. The visual results were associated with prominent levels of spectacle independence and patient satisfaction.

Comparing Dry Eye Disease After Small Incision Lenticule Extraction and Laser Subepithelial Keratomileusis.

PURPOSE: To compare dry eye disease parameters between small incision lenticule extraction (SMILE) and laser subepithelial keratomileusis (LASEK). METHODS: This prospective, nonrandomized study included 39 eyes of 39 patients who underwent SMILE (n = 19) and LASEK (n = 20). Dry eye disease parameters included Ocular Surface Disease Index (OSDI), tear breakup time, Schirmer I test (SIT) value, corneal esthesiometry result, tear meniscus area, meibomian gland evaluation result, lipid layer thickness, and blink rate. Evaluations were performed before surgery and at 1 and 6 months after surgery. RESULTS: Compared with the LASEK group, the SMILE group showed lower corneal esthesiometry values at 1 month and higher OSDI scores and lower SIT values at 6 months after surgery. The SMILE group showed significantly higher OSDI scores at 1 and 6 months after surgery than before, but significantly lower OSDI scores at 6 months than at 1 month after surgery. Both groups showed significantly decreased corneal sensitivity at 1 month after surgery, which increased to the preoperative level 6 months after surgery. The change in OSDI scores at 1 month after surgery positively correlated with changes in the total and partial blink rates, whereas the change in corneal sensitivity at 1 month after surgery negatively correlated with the total and partial blink rates. CONCLUSIONS: Both SMILE and LASEK groups showed temporary decreases in corneal sensitivity, which recovered to baseline levels. Although subjective symptoms worsened after SMILE, SIT values improved after LASEK. Compared with SMILE, LASEK yielded better results regarding subjective symptoms, tear production, and corneal sensitivity.