Inhibition of Rho-associated protein kinase activity enhances oxidative phosphorylation to support corneal endothelial cell migration.

Corneal endothelial cell (CEC) dysfunction causes corneal edema and severe visual impairment that require transplantation to restore vision. To address the unmet need of organ shortage, descemetorhexis without endothelial keratoplasty has been specifically employed to treat early stage Fuchs endothelial corneal dystrophy, which is pathophysiologically related to oxidative stress and exhibits centrally located corneal guttae. After stripping off central Descemet's membrane, rho-associated protein kinase (ROCK) inhibitor has been found to facilitate CEC migration, an energy-demanding task, thereby achieving wound closure. However, the correlation between ROCK inhibition and the change in bioenergetic status of CECs remained to be elucidated. Through transcriptomic profiling, we found that the inhibition of ROCK activity by the selective inhibitor, ripasudil or Y27632, promoted enrichment of oxidative phosphorylation (OXPHOS) gene set in bovine CECs (BCECs). Functional analysis revealed that ripasudil, a clinically approved anti-glaucoma agent, enhanced mitochondrial respiration, increased spare respiratory capacity, and induced overexpression of electron transport chain components through upregulation of AMP-activated protein kinase (AMPK) pathway. Accelerated BCEC migration and in vitro wound healing by ripasudil were diminished by OXPHOS and AMPK inhibition, but not by glycolysis inhibition. Correspondingly, lamellipodial protrusion and actin assembly that were augmented by ripasudil became reduced with additional OXPHOS or AMPK inhibition. These results indicate that ROCK inhibition induces metabolic reprogramming toward OXPHOS to support migration of CECs.

An Integrated Community-Based Blood Pressure Telemonitoring Program - A Population-Based Observational Study.

Background: Home blood pressure telemonitoring (BPT) has been shown to improve blood pressure control. A community-based BPT program (the Health+ program) was launched in 2015 in an urban area around a medical center. Objectives: To examine the impact of the BPT program on the use of medical resources. Methods: We conducted a retrospective propensity-score (PS)-matched observational cohort study using the National Health Insurance Research Database (NHIRD) 2013-2016 in Taiwan. A total of 9,546 adults with a high risk of cardiovascular disease participated in the integrated BPT program, and 19,082 PS-matched controls were identified from the NHIRD. The primary and secondary outcome measures were changes in 1-year emergency department visit rate, hospitalization rate, duration of hospital stay, and healthcare costs. Results: The number of emergency department visits in the Health+ group significantly reduced (0.8 to 0.6 per year vs. 0.8 to 0.9 per year, p < 0.0001) along with a significant decrease in hospitalization rate (43.7% to 21.3% vs. 42.7% to 35.3%, p < 0.001). The duration of hospital stay was also lower in the Health+ group (4.3 to 3.3 days vs. 5.3 to 6.5 days, p < 0.0001). The annual healthcare costs decreased more in the Health+ group (USD 1642 to 1169 vs. 1466 to 1393 per year, p < 0.001), compared with the controls. Subgroup analysis of the Health+ group revealed that the improvements in outcomes were significantly greater among those who were younger and had fewer comorbidities, especially without diabetes or hypertension. Conclusions: A community-based integrated BPT program may improve patients' health outcomes and reduce healthcare costs.

Refinement strategies in photoscreening for the detection of amblyopia risk factors in 12-to-24-month-old children.

PURPOSE: To establish an appropriate clinical protocol for early photoscreening in 12-to-24-month-old children at pediatric well-baby clinics. METHODS: This prospective study included a total of 277 children aged 12-24 months who visited a pediatric well-baby clinic. All participants underwent: 1) inquiry of medical history; 2) photoscreening with PlusoptiX A12; and 3) comprehensive ophthalmologic examinations. The optimal referral cut-off point for PlusoptiX was determined by receiver operating characteristic analyses. A high-risk subgroup was defined as having a birth weight <3000 g or a history of major systemic diseases, based on the results of multivariable risk factor analysis from children's medical history. A strategy of selective photoscreening focusing on the high-risk subgroup was evaluated. The main outcome measures included sensitivity, specificity, and positive and negative predictive values. RESULTS: The prevalence of amblyopia risk factors in our study population was 12.3%. A total of 172 children (62.1%) were assigned to the high-risk subgroup. Compared with the nonselective photoscreening of all 277 children, selective photoscreening of the high-risk children yielded a higher positive predictive value (59.6% vs. 46.7%, p = 0.001) while showing no difference in sensitivity (82.3% vs. 85.3%, p = 0.32), specificity (92.2% vs, 86.4%, p = 0.05), and negative predictive value (97.4% vs. 97.6%, p = 0.50). CONCLUSION: At pediatric well-baby clinics, selective photoscreening among 12-to-24-month-old children with a birth weight <3000 g or a history of major systemic diseases helped reducing the number of children need to be screened and conserving medical resources yet identifying children at risk for timely eye care.

Characteristics of tear film lipid layer in young dry eye patients.

BACKGROUND/PURPOSE: To investigate the characteristics of subjective symptoms and objective parameters among young patients with dry eye disease (DED) and compare them with those of older patients. METHODS: We retrospectively enrolled 675 patients with DED who were divided into three age groups (20-41 years [younger], 41-60 years [middle], and >60 years [older]) (n = 143, 304, and 228, respectively). Subjective symptoms were evaluated using Standardized Patient Evaluation of Eye Dryness (SPEED) and Ocular Surface Disease Index (OSDI) questionnaires. Aqueous tear secretion was evaluated with the Schirmer test II. The number of expressible meibomian glands was evaluated with a slit-lamp-aided standardized evaluator. Lipid layer thickness (LLT), blink/incomplete blink rates and meibography were measured with the LipiView® II interferometer. The extent of the meibomian gland dropout was graded using a meiboscale. RESULTS: The younger age group had higher subjective symptom severity, as reflected by higher SPEED (p < 0.001) and OSDI scores (p = 0.051). The SPEED scores negatively correlated with LLT in all patients (r = -0.136, p < 0.001). Younger patients also had thinner average LLT (p < 0.001), lower meiboscale (p < 0.001) and a higher number of expressible meibomian glands (p < 0.001). Additionally, they had significantly more total blinks (p < 0.001), incomplete blinks (p < 0.001), and incomplete blink rate (p = 0.006). CONCLUSION: Manifestations of DED vary with age. In our cohort, younger age patients had more symptoms and blinks, which may have resulted from thinner LLT as the structure and function of the meibomian glands were affected less than in middle and older age patients.

Regulation of Adipogenesis and Lipid Deposits by Collapsin Response Mediator Protein 2.

As emerging evidence suggesting neurodegenerative diseases and metabolic diseases have common pathogenesis, we hypothesized that the neurite outgrowth-controlling collapsin response mediator protein 2 (CRMP2) was involved in energy homeostasis. Therefore, putative roles of CRMP2 in adipocyte differentiation (adipogenesis) and lipid metabolism were explored and addressed in this study. CRMP2 expression profiles were in vitro and in vivo characterized during adipogenic process of 3T3-L1 pre-adipocytes and diet-induced obese (DIO) mice, respectively. Effects of CRMP2 on lipid metabolism and deposits were also analyzed. Our data revealed that CRMP2 expression pattern was coupled with adipogenic stages. CRMP2 overexpression inhibited cell proliferation at MCE phase, and significantly reduced lipid contents by down-regulating adipogenesis-driving transcription factors and lipid-synthesizing enzymes. Interestingly, GLUT4 translocation and the lipid droplets fusion were disturbed in CRMP2-silencing cells by affecting actin polymerization. Moreover, adipose CRMP2 was significantly increased in DIO mice, indicating CRMP2 is associated with obesity. Accordingly, CRMP2 exerts multiple functions in adipogenesis and lipid deposits through mediating cell proliferation, glucose/lipid metabolism and cytoskeleton dynamics. The present study identifies novel roles of CRMP2 in mediating adipogenesis and possible implication in metabolic disorders, as well as provides molecular evidence supporting the link of pathogenesis between neurodegenerative diseases and metabolic abnormalities.

MicroRNA-146a Alleviates Experimental Autoimmune Anterior Uveitis in the Eyes of Lewis Rats.

PURPOSE: This study aimed to determine the effect and roles of microRNA (miRNA, miR) treatment in experimental autoimmune anterior uveitis (EAAU). MATERIALS AND METHODS: Uveitis was induced in Lewis rats by simultaneous injections of bovine melanin-associated antigen into the hind footpad and the intraperitoneal cavity. The animals were injected intravitreally with low-dose (0.5 µg) or high-dose (1.5 µg) miR-146a. The clinical scores, leukocyte count in the aqueous humor, and histology were assessed. Cytokine changes were evaluated by relative mRNA expression and enzyme-linked immunosorbent assay (ELISA). The expression of nuclear factor kappa B (NF-κB) was assessed by immunofluorescence and Western blotting. Evaluation of the DNA-binding activity of NF-κB was performed by electrophoretic mobility shift assay (EMSA). RESULTS: Treatment with miR-146a significantly attenuated clinical scores and leukocyte infiltration in a dose-dependent manner, a result that was compatible with histological findings. Following miR-146a injections, downregulation of interleukin- (IL-) 1ß, IL-6, and IL-12 and interferon- (IFN-) γ and upregulation of IL-10 and IL-17 were noted. The decreased NF-κB expression on immunofluorescence and Western blotting and reduced DNA-binding activity on EMSA were demonstrated following miR-146a treatment. CONCLUSIONS: miR-146a effectively reduced intraocular inflammation in EAAU through the inhibition of NF-κB. miR-146a might be a new treatment choice for uveitis.

Infrared thermography in the evaluation of meibomian gland dysfunction.

BACKGROUND/PURPOSE: To evaluate meibomian gland dysfunction (MGD) by infrared thermography. METHODS: An observational study was conducted at the Department of Ophthalmology, Far Eastern Memorial Hospital, New Taipei City, Taiwan. Participants included 89 MGD patients (30 in Grade 1, 49 in Grade 2, and 10 in Grade 3) and 65 controls. The close-eye thermographic images of the eyelid were obtained noninvasively by infrared thermography. Temperatures at 8 regions of interest (ROIs) of the eyelid margin and a reference temperature at the center of the upper eyelid were measured. The temperature ratio was defined as the temperature of ROI divided by the reference temperature. RESULTS: Eyelid margin temperature measured by infrared thermography increased from temporal side (ROI 1) to the nasal side (ROI 8) of the eye in both MGD patients and control groups. The temperature ratios were significantly higher in MGD participants than in controls, especially at ROI 8. CONCLUSION: The eyelid margin temperature measured by infrared thermography was higher in MGD participants. Further development of this infrared thermography system may become a rapid and non-invasive tool for MGD screening.

Surgical approach affects intraocular lens decentration.

BACKGROUND/PURPOSE: This study aims to quantify and identify risk factors for intraocular lens (IOL) tilt and decentration early after surgery using Scheimpflug imaging. METHODS: We prospectively included 268 eyes of 253 patients who underwent uneventful cataract surgery and one-piece IOL implantation using a superior or temporal approach. Scheimpflug imaging was used to evaluate the tilt and decentration of IOLs at 1 week, 1 month, and 3 months postoperatively. Differences in IOL tilt and decentration between the approaches were examined. Correlations of age and axial length with the magnitudes of IOL decentration and tilt were also examined. RESULTS: In total, 139 right and 129 left eyes were included. IOL displacement averaged 150 µm upward and 150 µm to the nasal side of the pupil. Over 50% of the eyes were tilted upward and approximately 90% to the temporal side. The surgical approach was significantly associated with horizontal decentration in both eyes, but significantly associated with only vertical decentration in the right eye 1 week postoperatively. In the left eyes, IOLs were shifted to the nasal side in 57.1% and 36.8% of the eyes that received the temporal and the superior approach, respectively, compared with 75.8% and 50% in the right eyes. The differences were significant only at 1-week follow-up (p = 0.035 and p = 0.003, respectively). Age or axial length was not associated with IOL tilt or decentration in either eye. CONCLUSION: Scheimpflug imaging can be used as a quantitative tool to evaluate IOL position. The incision site affected the IOL position, this finding was significant at 1 week postoperatively only.

Refractive results of phacoemulsification in vitrectomized patients.

The aim of this study was to compare the refractive errors after uneventful phacoemulsification and intraocular lens implantation in the cases of vitrectomized eyes and non-vitrectomized eyes. Vitrectomized eyes were divided into 2 groups: vitrectomy alone as Group 1, vitrectomy with gas tamponade as Group 2. Normal-controlled eyes were categorized as Group 3. Intraocular lens (IOL) power calculation and the measurements of axial length (AL) and keratometric value (K) were performed using the IOLMaster and the Haigis and SRK/T formula as subgroup A and B. The measurements of AL, K, and IOL power calculation were also performed by the applanation ultrasound and the SRK/T formula as comparison subgroup C. The mean numeric error (MNE) and median absolute error (MedAE) were compared between Groups and subgroups. The MedAE was not significantly different in Group 1A, 2A, and 3A, respectively (p > 0.05). The MNE was significantly different in Group 1A, 2A, and 3A (p = 0.04). Post hoc test showed an insignificant difference between Group 1A and 3A (p = 0.36), but significantly a higher MNE in Group 2A than that in Group 3A (p = 0.03). A significantly longer AL was noted in Group 1B and 2B, comparing with Group 1C and 2C (p < 0.01). Refractive outcomes after phacoemulsification sequential to vitrectomy alone were comparable to those in normal control. Hyperopic shift was found following phacoemulsification sequential to vitrectomy with gas tamponade comparing to normal eyes. A significantly longer AL was found using optical biometry compared to applanation ultrasound in vitrectomized eyes, but not in normal eyes.

Inhibition of matrix metalloproteinase activity reverses corneal endothelial-mesenchymal transition.

Ex vivo culture or regeneration of corneal endothelial cells often is subjected to gradual endothelial-mesenchymal transition and loss of function. Here, we found that during ex vivo culture, bovine corneal endothelial cells underwent endothelial-mesenchymal transition and had an up-regulated expression and activity of matrix metalloproteinases. Inhibition of matrix metalloproteinase activity in confluent bovine corneal endothelial cells decreased the level of endothelial-mesenchymal transition regulators: snail and slug. The phosphorylation and degradation of the key Wnt signaling pathway modulator active ß-catenin also were accelerated with the broad-spectrum matrix metalloproteinase inhibitor Marimastat, which may result from decreased N-cadherin shedding and increased intact N-cadherin molecules on the cell membrane. Intracameral injection of Marimastat also suppressed basic fibroblast growth factor-induced endothelial-mesenchymal transition in a rat corneal endothelium cryo-injury model and significantly diminished the corneal edema. Our study indicated that inhibition of matrix metalloproteinase activity can reverse endothelial-mesenchymal transition and preserve the function of corneal endothelial cells both during ex vivo culture and in vivo. This may offer a potential therapeutic target in regenerative medicine for the treatment of corneal endothelial dysfunctions.

Molecular mechanism of fluoroquinolones modulation on corneal fibroblast motility.

Topical fluoroquinolones are widely used to prevent ocular infections after ophthalmic surgery. However, they have been shown to affect the corneal cell motility, whose mechanism remains indefinite. The purpose of this study was to investigate how fluoroquinolones affect corneal stromal cell motility. Human corneal fibroblasts (HCFs) were incubated in ciprofloxacin (CIP), levofloxacin (LEV), or moxifloxacin (MOX) at 0, 10, 50, and 100 µg/ml for up to 3 days. Effect of CIP, LEV, or MOX on HCF migration was monitored using migration assay. HCF viability was determined by WST-1 assay. Expression of focal adhesion kinase (FAK), paxillin (PXN), and their phosphorylated forms were analyzed by immunoblotting. Binding affinity between FAK and PXN was determined by co-immunoprecipitation. Our results revealed that CIP and MOX, but not LEV, noticeably retarded HCF migration. HCF proliferation was significantly reduced by CIP (38.2%), LEV (29.5%), and MOX (21.3%), respectively (p = 0.002). CIP and MOX suppressed the phosphorylation of PXN at tyrosines (10.2 ± 4.3%, p < 0.001; 11.7 ± 2.4%, p < 0.001, respectively), including tyrosine 118 (33.3 ± 5.2%, p < 0.001; 34.0 ± 4.4%, p < 0.001, respectively). CIP and MOX diminished the binding affinity between FAK and PXN (8.2 ± 1.8%, p < 0.001; 9.0 ± 4.5%, p < 0.001, respectively). Nevertheless, tyrosine dephosphorylation and FAK dissociation of PXN were not found in LEV-treated HCFs. None of these fluoroquinolones affect phosphorylation of FAK-Y397. We conclude that CIP and MOX, but not LEV, might delay corneal fibroblast migration via interfering with recruitment of PXN to focal adhesions and dephosphorylation of PXN at the tyrosines.

Anterior Segment Biometry Changes with Cycloplegia in Myopic Adults.

PURPOSE: To determine the effect of cycloplegia on corneal thickness, corneal curvature, anterior chamber depth (ACD), angle-to-angle (ATA) and white-to-white (WTW) distances, and axial length (AL). METHODS: Changes in corneal thickness, corneal curvature, ACD, ATA and WTW distances, and AL with and without cycloplegia were analyzed in 31 eyes of 31 young myopic adults, aged 26.4 ± 3.0 years. Pentacam was used to measure the corneal thickness, corneal volume, and corneal curvatures. Visante optical coherent tomography (OCT) measured corneal thickness, ATA distance, ACD, and pupil size. The AL and WTW distance were measured using IOLMaster. RESULTS: Cycloplegia induced significant flattening of corneal curvatures (p = 0.019, 0.001, and 0.003 for anterior sagittal, posterior tangential, and posterior sagittal curvatures, respectively). The difference in the posterior corneal curvature was greater in corneas with steeper posterior curvatures. Cycloplegia also induced significant deepening of ACD (0.08 ± 0.06, p < 0.001) and widening of both WTW (0.42 ± 0.43, p < 0.001) and ATA (0.08 ± 0.17, p = 0.015) distances. The cycloplegia-related increase in the ATA distance correlated negatively with AL (r = -0.361, p = 0.046), whereas the cycloplegia-related increase in WTW distance correlated weakly with the increase in ACD (r = 0.347, p = 0.056) but not with AL. The AL did not change with cycloplegia. Pentacam measured a slightly thicker cornea than OCT (p = 0.002). Both Pentacam and OCT detected a significant increase in corneal thickness of 4 µm, which could be attributed to reflex tearing, after cycloplegia. CONCLUSIONS: Cycloplegia resulted in deeper ACD, greater ATA distance, and flatter corneal curvatures. Surgeons should be aware of these cycloplegia-related alterations for more accurate phakic/functional intraocular lens selection and better refraction results.

Expression Profile of Cationic Amino Acid Transporters in Rats with Endotoxin-Induced Uveitis.

Purpose. The transcellular arginine transportation via cationic amino acid transporter (CAT) is the rate-limiting step in nitric oxide (NO) synthesis, which is crucial in intraocular inflammation. In this study, CAT isoforms and inducible nitric oxide synthase (iNOS) expression was investigated in endotoxin-induced uveitis (EIU). Methods. EIU was induced in Lewis rats by lipopolysaccharide (LPS) injection. In the treatment group, the rats were injected intraperitoneally with the proteasome inhibitor bortezomib before EIU induction. After 24 hours, leukocyte quantification, NO measurement of the aqueous humor, and histopathological examination were evaluated. The expression of CAT isoforms and iNOS was determined by reverse transcription-polymerase chain reaction, western blotting, and immunofluorescence staining. Nuclear factor-kappa B (NF-κB) binding activity was evaluated by electrophoretic mobility shift assay. The mouse macrophage cell line RAW 264.7 was used to validate the in vivo findings. Results. LPS significantly stimulated iNOS, CAT-2A, and CAT-2B mRNA and protein expression but did not affect CAT-1 in EIU rats and RAW 264.7 cells. Bortezomib attenuated inflammation and inhibited iNOS, CAT-2A, and CAT-2B expression through NF-κB inhibition. Conclusions. CAT-2 and iNOS, but not CAT-1, are specifically involved in EIU. NF-κB is essential in the induction of CAT-2 and iNOS in EIU.

Protective effects of systemic treatment with methylprednisolone in a rodent model of non-arteritic anterior ischemic optic neuropathy (rAION).

This study investigated the protective effects of the administration of steroids on optic nerves (ON) and retinal ganglion cells (RGCs) in a rodent model of non-arteritic anterior ischemic optic neuropathy (rAION). We induced rAION using rose bengal and argon laser irradiation in a photodynamic procedure on the optic discs of rats. The treated groups received methylprednisolone (MP) via peritoneal injection for 2 weeks. The control group received intraperitoneal injections of phosphate-buffered saline (PBS) post-rAION. At the 4th week post-infarct, MP treatments significantly rescued the RGCs (mm(2)) in the central retinas (1920 ± 210, p < 0.001) and mid-peripheral retinas (950 ± 240, respectively, p = 0.018) compared with those of the PBS-treated rats (central: 900 ± 210 and mid-peripheral: 440 ± 180). Functional assessment with flash visual-evoked potentials demonstrated that P1 latency (ms) was shortened in the MP group compared to the PBS group (108 ± 14 and 147 ± 9, respectively, p < 0.001). In addition, the P1 amplitude (uV) was enhanced in the MP group compared to the PBS group (55 ± 12 and 41 ± 13, respectively, p < 0.05). TUNEL assays showed a decrease in the number of apoptotic cells in the RGC layers of MP-treated retinas compared to the PBS-treated group (p < 0.05). ED1 positive cells (/HPF) were significantly decreased in the ONs of the MP group compared to the PBS group (p < 0.001). In conclusion, systemic administration of MP had neuroprotective effects on RGC survival and ON function in the rAION animal model.

Influence of ocular features and incision width on surgically induced astigmatism after cataract surgery.

PURPOSE: To identify factors associated with surgically induced astigmatism (SIA) following phacoemulsification. METHODS: Six hundred five eyes underwent phacoemulsification with a 2.2-mm (the 2.2-mm group, n = 248) or 2.75-mm (the 2.75-mm group, n = 357) superior limbal incision. Preoperative axial length, anterior chamber depth, corneal curvature, and intra-ocular pressure were measured. Corneal curvature and intraocular pressure were measured at 1 day, 1 week, and 1, 2, and 3 months postoperatively. SIA, corneal flattening, and torque were calculated using the Alpins method. The effect of preoperative corneal astigmatism meridian on SIA was also examined. Differences in SIA between the 2.2- and 2.75-mm groups were explored, and correlations between SIA and preoperative corneal astigmatism, anterior chamber depth, axial length, age, and intraocular pressure were analyzed. RESULTS: SIA, corneal flattening, and torque were smaller in the 2.2-mm group than in the 2.75-mm group at 1 week (P = .003, .006, and .014, respectively), but not statistically different thereafter. Higher preoperative corneal astigmatism, older age, and shallower anterior chamber depth were associated with greater SIA in both groups. The effect of astigmatism meridian on SIA was more noticeable in the 2.75-mm group. Shorter axial length and lower intraocular pressures were associated with greater SIA in the 2.75-mm group but not in the 2.2-mm group. CONCLUSIONS: Reducing limbal incision width and considering patient age, the meridian and magnitude of corneal astigmatism, anterior chamber depth, axial length, and intraocular pressure, and adjusting the flattening component of SIA input for toric intraocular lens power calculation could potentially improve the astigmatism control in refractive lens surgery.

Expression of MicroRNAs in the Eyes of Lewis Rats with Experimental Autoimmune Anterior Uveitis.

Purpose. This study aimed to determine the dynamic changes of NF-κB-related microRNAs (miRNAs) and cytokines over the course of experimental autoimmune anterior uveitis (EAAU) and elucidate the possible immunopathogenesis. Materials and Methods. Uveitis was induced in Lewis rats using bovine melanin-associated antigen. The inflammatory activity of the anterior chamber was clinically scored, and leukocytes in the aqueous humor were quantified. RNA was extracted from the iris/ciliary bodies and popliteal lymph nodes to reveal the dynamic changes of eight target miRNAs (miR-155-5p, miR-146a-5p, miR-182-5p, miR-183-5p, miR-147b, miR-21-5p, miR-9-3p, and miR-223-3p) and six cytokine mRNAs (IFN-γ, IL-17, IL-12A, IL-1ß, IL-6, and IL-10). In situ hybridization of miRNA and enzyme-linked immunosorbent assay quantification of cytokines were performed to confirm the results. Results. Disease activity and leukocyte quantification were maximum at day 15 after immunization. The profiling of miRNA revealed downregulation of miR-146a-5p, miR-155-5p, miR-223-3p, and miR-147b and upregulation of miR-182-5p, miR-183-5p, and miR-9-3p. Cytokine analysis revealed IFN-γ, IL-17, IL-12A, IL-1ß, and IL-6 overexpression, with IL-10 downregulation. Conclusions. Dynamic changes of miRNAs were observed over the course of EAAU. By initiating NF-κB signaling, the expressions of downstream cytokines and effector cells from the Th17 and Th1 lineages were sequentially activated, contributing to the disease.

Dexamethasone modifies mitomycin C-triggered interleukin-8 secretion in isolated human Tenon's capsule fibroblasts.

Intraoperative mitomycin C (MMC) is widely used to prevent pterygium recurrence and glaucoma filtering bleb failure, but it has been shown to induce corneal inflammation and cell death. Postoperative dexamethasone (DEX) is advocated to reduce MMC-related inflammation and cell death in corneal fibroblasts. Nevertheless, its long-term regulation mechanism in Tenon's capsule remains to be explored. The purpose of this study was to investigate how DEX modifies MMC's effects in human Tenon's capsule fibroblasts (HTFs). HTFs isolated from the pterygium surgical patients (n = 6) were treated with MMC at 0, 0.1, 0.2, and 0.4 mg/ml for 5 min and incubated in DEX at 10 µM for 0, 1, 2, and 3 days. Recombinant interleukin-8 (IL-8) was used to verify the effect of MMC-related IL-8 secretion. Cell proliferation of all the treated cells was analyzed by WST-1 assay. The amount of IL-8 secretion in HTFs was determined by enzyme-linked immunosorbent assay. Immunoblotting assay was used to analyze the expression of peroxisome-proliferator-activated receptor gamma (PPARγ) and B-cell lymphoma-extra large (Bcl-xL). Our results revealed that MMC significantly reduced the HTF cell proliferation rate. Additionally, MMC significantly upregulated IL-8 secretion in HTFs concentration-dependently. At 3 days post treatment (dpt), 5-min exposures to 0.1, 0.2, and 0.4 mg/ml MMC resulted in 1.4-fold (p = 0.012), 1.6-fold (p = 0.012), and 2.5-fold (p = 0.001) increases of IL-8 secretion. In contrast, DEX reversed the MMC-retarded cell proliferation rate (p = 0.036) and repressed MMC-related IL-8 secretion by 33.5% at 3 dpt (p = 0.003). Addition of recombinant IL-8 noticeably suppressed HTF cell proliferation in a concentration-dependent manner. DEX stimulated upregulation of both PPARγ and Bcl-xL at 1 dpt in normal HTFs and at 2 dpt in MMC-treated HTFs. PPARγ silencing reduced expression of PPARγ and Bcl-xL, but enhanced IL-8 secretion (p < 0.001). On the other hand, Bcl-xL silencing enhanced IL-8 secretion (p < 0.001), but did not affect PPARγ expression. These revealed that IL-8 secretion in HTFs is modulated by PPARγ-dependent Bcl-xL signaling. We conclude that DEX reversed the MMC-inhibited HTF cell proliferation via diminishing the MMC-induced IL-8 secretion, which resulted from a late-phase upregulation of the PPARγ and Bcl-xL. These long-term effects suggest a beneficial postoperative DEX treatment following intraoperative MMC application.

Non-lethal exposure to short-wavelength light-emitting diodes modulates tight-junction structure in human corneal epithelial cells via cAMP-dependent signaling.

Light-emitting diodes (LED)-derived lights have been widely used as a medical treatment in photobiomodulation (PBM). However, the PBM effects in ophthalmology are less well investigated. Herein, we explored the effect of LED-generated light on the tight-junction (TJ) formation in human corneal epithelial cells (HCEs). The HCEs were separately exposed to monochromatic LEDs at wavelengths of 365 nm (UVA), 420 nm (violet), 470 nm (blue), 530 nm (green), 590 nm (amber), 660 nm (deep red), and 740 nm (far red) at 10 J/cm2/day for 1 and 2 days. Long-term cultivation of HCEs without LED exposure for up to 14 days was established as a control. The effects of both LED wavelength and culture duration on cell morphology, cAMP-regulated proteins, TJ-associated proteins, and cell growth-associated proteins were also analyzed. Together with the increase in cell number during prolonged cultivation, cAMP, ZO-1, ZO-2, CLDN1, and CLDN4 all increased significantly during long-term cultivation without LED exposure. There was no difference in HCE viability after exposure to all monochromatic LEDs at an accumulated dose of 20 J/cm2. As determined by immunoblotting, UVA, violet, and blue light increased intracellular cAMP, ZO-1, ZO-2, CLDN1, and CLDN4 expression, respectively. UVA and violet, but not blue, light increased PKAreg-pS77 expression. However, none of the other treatments changed the expression of PKAcat-pT197, VASP-pS157, Bax, Bcl-2, or Bcl-xL. Immunofluorescence staining confirmed the formation of TJ structures. The expressions of ZO-1, ZO-2, CLDN1, and CLDN4 as well as TJ structures 2 days following UVA, violet, and blue exposure were similar to those of control cells after 9 days of cultivation. We conclude that short-wavelength LEDs at non-lethal exposure intensities accelerated the formation of TJ structure in HCEs via a cAMP-dependent regulatory cascade.

The extraction and application of antisymmetric characteristics of the cornea during air-puff perturbations.

BACKGROUND: A non-contact tonometer is used to measure intraocular pressure, and studies have primarily relied on apex displacements to assess corneal properties. However, previous studies have overlooked the asymmetric characteristics of lateral corneal perturbations, leading to a gap in understanding of the lateral mechanical properties and its application. METHOD: To investigate these lateral perturbations, we designed an experiment to sequentially record the corneal profiles when two consecutive air-puffs were applied at the center of the same cornea within a short period. Moreover, we used modal decomposition to decompose anterior surface profiles into symmetric and antisymmetric modes to comprehensively analyze the asymmetric characteristics. To extract mechanical properties, we utilized high-pass frequency analysis (>250 Hz) to filter out noise and errors. RESULTS: Symmetric modes between the two consecutive air-puffs exhibited major similarities during vibration; however, antisymmetric modes exhibited minor differences in lateral perturbations of asymmetric vibration. The antisymmetric modes might be related to air-puff misalignment and mechanical properties. Through applying frequency analysis, the mechanical properties could be proven at high frequencies and misalignment shown at low frequencies. Furthermore, we compared the corneal vibration profiles of 259 healthy participants and 50 patients with keratoconus. Their properties showed that the antisymmetric modes of the keratoconus group exhibited a completely opposite direction of deformation compared to that in the healthy group. CONCLUSIONS: Our proposed algorithm not only extracts antisymmetric characteristics but also offers valuable insights into decompose misalignment and mechanical properties of healthy and keratoconus corneas, presenting a new perspective for corneal biomechanics.

High-resolution full-field optical coherence tomography microscope for the evaluation of freshly excised skin specimens during Mohs surgery: A feasibility study.

Histopathology for tumor margin assessment is time-consuming and expensive. High-resolution full-field optical coherence tomography (FF-OCT) images fresh tissues rapidly at cellular resolution and potentially facilitates evaluation. Here, we define FF-OCT features of normal and neoplastic skin lesions in fresh ex vivo tissues and assess its diagnostic accuracy for malignancies. For this, normal and neoplastic tissues were obtained from Mohs surgery, imaged using FF-OCT, and their features were described. Two expert OCT readers conducted a blinded analysis to evaluate their diagnostic accuracies, using histopathology as the ground truth. A convolutional neural network was built to distinguish and outline normal structures and tumors. Of the 113 tissues imaged, 95 (84%) had a tumor (75 basal cell carcinomas [BCCs] and 17 squamous cell carcinomas [SCCs]). The average reader diagnostic accuracy was 88.1%, with a sensitivity of 93.7%, and a specificity of 58.3%. The artificial intelligence (AI) model achieved a diagnostic accuracy of 87.6 ± 5.9%, sensitivity of 93.2 ± 2.1%, and specificity of 81.2 ± 9.2%. A mean intersection-over-union of 60.3 ± 10.1% was achieved when delineating the nodular BCC from normal structures. Limitation of the study was the small sample size for all tumors, especially SCCs. However, based on our preliminary results, we envision FF-OCT to rapidly image fresh tissues, facilitating surgical margin assessment. AI algorithms can aid in automated tumor detection, enabling widespread adoption of this technique.