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To compare two different secondary IOL fixation techniques, either flanged or hooked, regarding the least required force to dislocate the haptic in human corneoscleral donor tissue (CST). Experimental laboratory investigation. The least required dislocation force (LRDF) of two different fixation techniques, namely the flanged haptics (FH, as described by Yamane) and the harpoon haptic technique (HH, as described by Carlevale) were investigated using 20 three-piece IOLs (KOWA PU6AS) and 20 single-piece IOLs (SOLEKO CARLEVALE) fixated to human scleral tissue. The main outcome, differences in LRDF of the investigated techniques, was measured with a tensiometer. The dislocation force needed to dislocate the flanged haptics was significantly higher (p < 0.001) in the flanged 3-piece IOL (0.93 ± 0.43 N) than the specialized, harpoon haptics single-piece IOL (0.45 ± 0.18 N). During externalization, breakage occurred in three harpoon haptics. However, no breakage was observed in either haptics during dislocation. The flanged haptic technique proved to be the stronger form of secondary IOL fixation regarding dislocation force in this in vitro study. The harpoon haptics fixation technique showed significantly less resistance to axial traction and a susceptibility to breakage during externalization., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Purpose: To translate, validate, and evaluate the psychometric properties of the German version of Keratoconus Outcomes Research Questionnaire (KORQ)., Methods: This prospective study included 3 phases. In phase 1, the KORQ was translated in a standardized way; in phase 2, Rasch validation was performed; in phase 3, the validated questionnaire was evaluated in a separate patient cohort. All patients answered the German KORQ. Scheimpflug imaging (Pentacam) and optical coherence tomography scans (MS-39) were conducted. Corneal imaging measurements were correlated with patient-reported outcome measures., Results: After standardized translation, 100 patients self-administered the German KORQ. Rasch analysis was conducted for psychometric testing of the KORQ. Based on the results of the Rasch analysis, the original German version of the KORQ was modified and any items showing a misfit were excluded. This resulted in a new shortened version of the KORQ, which was evaluated in 30 patients. The revised German KORQ met the criteria of Rasch analysis and showed excellent internal consistency reliability and convergent validity. The correlation between the KORQ score and various clinical measurements was examined using Spearman correlation. A significant moderate correlation was observed between the D (r = 0.237; P = 0.006) and K max (r = 0.162; P = 0.065) values of Pentacam. Inverse correlations were observed for thinnest pachymetry measured with MS-39 (r = -0.167; P = 0.058) and best corrected visual acuity (r = -0.210; P = 0.016)., Conclusions: The shortened German KORQ met the assumptions of the Rasch model and displayed satisfactory psychometric properties. The German KORQ could be adopted as a powerful patient-reported outcome measures tool for German-speaking patients with keratoconus in the future., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: To compare interindividual differences in visual performance of an advanced monofocal with a nondiffractive extended depth of focus intraocular lens (IOL) using a mini-monovision approach., Design: Single-center, randomized, controlled, double-masked study., Methods: In total, 48 patients (96 eyes) with bilateral age-related cataract were enrolled. One group received an advanced monofocal IOL RAO200E (RayOne EMV; Rayner) and the other group a non-diffractive extended depth of focus IOL DFT015 (Acrysof IQ Vivity; Alcon) in both eyes. Target refraction for both groups was mini-monovision. After 3 months, monocular and binocular distance corrected and uncorrected distance (CDVA/UDVA), intermediate (DCIVA/UIVA), and near visual acuity (DCNVA/UNVA); contrast sensitivity; binocular defocus curves; halometry; and Quality of Vision questionnaire scores were compared., Results: Binocular mean CDVA, DCIVA at 66 cm, and DCNVA at 40 cm were -0.10±0.10, 0.15±0.11, and 0.32±0.16 logMAR for the RAO200E and -0.10±0.10, 0.12±0.10, and 0.27±0.16 logMAR for the DFT015, respectively, with no significant difference. A significant difference could be shown for the dominant eye in monocular DCIVA and DCNVA and for the dominant and nondominant eye in monocular UNVA, with 0.28±0.14, 0.48±0.22, 0.46±0.21, and 0.41±0.20 logMAR for the RAO200E and 0.14±0.10 (P = .023), 0.35±0.16 (P = .008), 0.30±0.14 (P = .001), and 0.21±0.10 (P = .003) logMAR for the DFT015, respectively. Significantly smaller halo size in the RAO200E group and significantly better distance-corrected defocus curve at -2.5 (P = .031), -2.0 (P = .03), and -1 diopters (P = .03) of defocus in the DFT015 group could be shown., Conclusions: Distance corrected or uncorrected binocular visual acuity for far, intermediate, and near distance between the advanced monofocal IOL RAO200E and the non-diffractive extended depth of focus IOL DFT015, when compared in a mini-monovision setting, showed no significant differences., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose: To compare the two most used digital alignment systems regarding precision, repeatability and loss of track., Methods: 15 eyes of 15 patients older than 21 years with cataracts were included in this prospective study. The two systems were intraoperatively superimposed and recorded, and the alignment of the two displayed alignment axes was analysed regarding precision, repeatability and loss of track., Results: There was a significant difference in precision and repeatability between the two digital alignment systems regarding the projected alignment axis. The deviation from the actual target axis was significantly different, with a mean of 0.34°±0.75° for the Zeiss system and 1.60°±1.08° for the Alcon system (p=0.03, n=14). The within-subject SD was significantly lower with 0.21° for the Zeiss system and 0.34° for the Alcon system (p=0.03, n=14)., Conclusions: The Zeiss Callisto system showed a significantly lower deviation from the target axis, higher stability with eye movements and less need for microscope illumination than the Alcon system. Both systems showed high precision when compared with manual marking methods., Trial Registration Number: NCT05220683., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: To compare 4 different secondary intraocular lens (IOL) fixation techniques regarding the least required force to dislocate a scleral fixated 3-piece IOL in human corneoscleral donor tissue., Design: Experimental laboratory investigation., Methods: The least required dislocation force (LRDF) of 4 different secondary IOL fixation techniques, namely, the techniques using transscleral tunnels (TTs; as described by Scharioth), glued haptics (GHs; Agarwal), flanged haptics (FHs; Yamane), and bent haptic ends (BH; Behera/Bolz), were investigated using 40 three-piece IOLs (Sensar AR40) fixated to human scleral tissue. The main outcome of the study, dislocation force between different techniques, was measured with a tensiometer., Results: The force needed to dislocate the haptics was highest with the FH technique and was significantly higher than with all the other techniques (GH vs FH: -1.02±0.02 N, P < .001; TT vs FH: -1.08±0.21 N, P < .001; BH vs FH: -1.00±0.25 N, P = .044). There was no significant difference regarding the dislocation force between the other techniques: GH vs TT (-0.06±0.100 N, P = .988), GH vs BH (-0.02±0.03 N, P = .60), TT vs BH (-0.08±0.04 N, P > .99)., Conclusions: The FH technique as described by Yamane proved to be the strongest form of secondary IOL fixation regarding dislocation force in this in vitro study. The other fixation techniques showed significantly less resistance to axial traction., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: To compare the difference in rebubbling rates between patients undergoing Descemet membrane endothelial keratoplasty (DMEK) with endothelium-in using a standard IOL cartridge and those with endothelium-out DMEK utilizing a no-touch technique with borosilicate glass cartridge transplantation., Methods: This retrospective study included all eyes that underwent preloaded endothelium-in or endothelium-out DMEK transplantation from June 2019 to December 2023 at the Hanusch Hospital, Vienna, Austria. All DMEKs were harvested, prepared and preloaded at the European Eye Bank of Venice, Italy. DMEK surgeries were done by one experienced surgeon and the procedure was completed by air tamponade of the anterior chamber., Results: Overall, 32 eyes each of 31 endothelium-out patients and of 29 endothelium-in patients were included. 32 preloaded endothelium-in procedures were followed by 32 preloaded endothelium-out procedures. Rebubbling rate for endothelium-in was 15/32 (47%) and for endothelium-out was 7/25 (28%) (p = 0.035, Pearson's chi-squared test). Donor age was the most important variable for rebubbling in a random forest algorithm model (ROC: 0.69)., Conclusions: Rebubbling rate in endothelium-out DMEK was less than two-thirds compared to endothelium-in DMEK favoring no-touch endothelium-out DMEK as the preferred technique of DMEK transplantation., (© 2024. The Author(s).)

Purpose: To measure the dislocation forces in relation to haptic material, flange size and needle used., Setting: Hanusch Hospital, Vienna, Austria., Design: Laboratory Investigation., Methods, Main Outcome Measures: 30 G (gauge) thin wall and 27 G standard needles were used for a 2 mm tangential scleral tunnel in combination with different PVDF (polyvinylidene fluoride) and PMMA (polymethylmethacrylate haptics). Flanges were created by heating 1 mm of the haptic end, non-forceps assisted in PVDF and forceps assisted in PMMA haptics. The dislocation force was measured in non-preserved cadaver sclera using a tensiometer device., Results: PVDF flanges achieved were of a mushroom-like shape and PMMA flanges were of a conic shape. For 30 G needle tunnels the dislocation forces for PVDF and PMMA haptic flanges were 1.58 ± 0.68 N (n = 10) and 0.70 ± 0.14 N (n = 9) (p = 0.003) respectively. For 27 G needle tunnels the dislocation forces for PVDF and PMMA haptic flanges were 0.31 ± 0.35 N (n = 3) and 0.0 N (n = 4), respectively. The flange size correlated with the occurring dislocation force in experiments with 30 G needle tunnels (r = 0.92), when flanges were bigger than 384 micrometres., Conclusions: The highest dislocation forces were found for PVDF haptic flanges and their characteristic mushroom-like shape for 30 G thin wall needle scleral tunnels. Forceps assisted flange creation in PMMA haptics did not compensate the disadvantage of PMMA haptics with their characteristic conic shape flange., (© 2024. The Author(s).)

Purpose: To investigate the flange properties of different iris hooks., Setting: Vienna Institute for Research in Ocular Surgery (VIROS), Hanusch Hospital, Vienna, Austria., Design: Laboratory study., Methods: The flanging properties of 4 different iris hooks made from polypropylene (PP), elastic polymer (EP), and nylon were investigated with different heating distances and both with and without forceps gripping. The maximum diameter of the flanges was measured, and the shape of the flanges was evaluated., Results: Although both nylon and EP iris hooks had too small flange diameters for intrascleral fixation, PP iris hooks had a sufficient flange diameter (>330 μm) and mushroom-like shape. Furthermore, in PP hooks, heating distance was directly proportional to flange diameter., Conclusions: The findings of this study suggest that only PP iris hooks are suitable for flanged intrascleral fixation, which is off-label, to secure adequate fixation., (Copyright © 2023 Published by Wolters Kluwer on behalf of ASCRS and ESCRS.)

Purpose: To assess the diameter of different 30-gauge thin-wall needles and 3-piece intraocular lens (IOL) haptics readily used for the flanged-haptic intrascleral fixation technique., Setting: Hanusch Hospital, Vienna, Austria., Design: Laboratory investigation., Methods: 5 30-gauge thin-wall needles and 5 3-piece IOLs were assessed. An upright light microscopy was used for measurements. The inner and outer diameters of the needles and the end thickness of the haptics were analyzed and compared for haptic fitting into the needle., Results: Among the needles, the inner diameter of the T-lab needle was significantly wider compared with all the others (mean 209.3 ± 8.0 μm, P < .001), followed by TSK (194.8 ± 5.0 μm), MST (194.7 ± 5.8 μm), Sterimedix (187.5 ± 9.0 μm) and significantly narrower Meso-relle (mean 178.7 ± 7.0 μm, P < .05). The outer diameter of the T-lab needle was significantly larger of all (mean 316.0 ± 2.0 μm, P < .001). Concerning the IOLs, the AvanseePreset Kowa's haptic was significantly thinner (mean 127.2 ± 0.7 μm) than all the others, such as the TecnisZA900 Johnson & Johnson (143.5 ± 3.1 μm), the CTLucia202 Zeiss (143.8 ± 1.3 μm), and the AcrysofMA60AC Alcon (143.9 ± 1.4 μm). The only haptic that was thicker than all the others assessed was that of SensarAR40 Johnson & Johnson (170.7 ± 1.7 μm, P < .001)., Conclusions: Most of the analyzed haptics would fit into most of the measured needles, with the exception of the Sensar AR40 in combination with the Meso-relle or Sterimedix needles. The combination of a larger needle lumen and a thinner haptic could result in more ease of insertion during surgery. If the dimensions of the needle and IOL haptics used are unknown, we recommend trying insertion before beginning surgery., (Copyright © 2023 Published by Wolters Kluwer on behalf of ASCRS and ESCRS.)

A technique for achieving an optimal flange size with 5-0 polypropylene and 6-0 polypropylene used for flanged intrascleral intraocular lens fixation is described. Flange size in polypropylene sutures is dependent on heating length and independent of forceps grip during heating. It was identified that heating of 1 mm created the optimal flange size for a 5-0 polypropylene suture when used for a 27-gauge needle scleral tunnel and for a 6-0 polypropylene suture when used for a 30-gauge needle scleral tunnel. Alternatively, 2 mm heating of a 6-0 polypropylene suture fits well for a 27-gauge needle tunnel. Even gentle forceps grip caused flattening of the polypropylene sutures but did not influence shaping and sizing of the flange., (Copyright © 2022 Published by Wolters Kluwer on behalf of ASCRS and ESCRS.)

Cataract is the leading cause of blindness worldwide and surgery is the only option to treat the disease. Although the surgery is considered to be relatively safe, complications may occur in a subset of patients and access to ophthalmic care may be limited. Due to a growing and ageing population, an increase in cataract prevalence is expected and its management will become a socioeconomic challenge. Hence, there is a need for an alternative to cataract surgery. It is well known that oxidative stress is one of the main pathological processes leading to the generation of the disease. Antioxidant supplementation may, therefore, be a strategy to delay or to prevent the progression of cataract. Caffeine is a widely consumed high-potency antioxidant and may be of interest for the prevention of the disease. This review aims to give an overview of the anatomy and function of the lens, its antioxidant and reactive oxygen species (ROS) composition, and the role of oxidative stress in cataractogenesis. Also, the pharmacokinetics and -dynamics of caffeine will be described and the literature will be reviewed to give an overview of its anti-cataract potential and its possible role in the prevention of the disease., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: To train and validate a code-free deep learning system (CFDLS) on classifying high-resolution digital retroillumination images of posterior capsule opacification (PCO) and to discriminate between clinically significant and non-significant PCOs., Methods and Analysis: For this retrospective registry study, three expert observers graded two independent datasets of 279 images three separate times with no PCO to severe PCO, providing binary labels for clinical significance. The CFDLS was trained and internally validated using 179 images of a training dataset and externally validated with 100 images. Model development was through Google Cloud AutoML Vision. Intraobserver and interobserver variabilities were assessed using Fleiss kappa (κ) coefficients and model performance through sensitivity, specificity and area under the curve (AUC)., Results: Intraobserver variability κ values for observers 1, 2 and 3 were 0.90 (95% CI 0.86 to 0.95), 0.94 (95% CI 0.90 to 0.97) and 0.88 (95% CI 0.82 to 0.93). Interobserver agreement was high, ranging from 0.85 (95% CI 0.79 to 0.90) between observers 1 and 2 to 0.90 (95% CI 0.85 to 0.94) for observers 1 and 3. On internal validation, the AUC of the CFDLS was 0.99 (95% CI 0.92 to 1.0); sensitivity was 0.89 at a specificity of 1. On external validation, the AUC was 0.97 (95% CI 0.93 to 0.99); sensitivity was 0.84 and specificity was 0.92., Conclusion: This CFDLS provides highly accurate discrimination between clinically significant and non-significant PCO equivalent to human expert graders. The clinical value as a potential decision support tool in different models of care warrants further research., Competing Interests: Competing interests: JH received travel grants from Bayer, speaker fees from Carl Zeiss Meditec AG and Bayer, and served on advisory boards for Roche outside of this work. DS received speaker fees from Allergan, Bayer, Novartis and Haag Streit. PAK acted as a consultant for DeepMind, Roche, Novartis and Apellis; was an equity owner in Big Picture Medical; and received speaker fees from Heidelberg Engineering, Topcon, Allergan and Bayer. OF is a scientific advisor to Alcon, Carl Zeiss Meditec AG, Croma Pharma, Johnson & Johnson and Merck., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY. Published by BMJ.)

At the moment, cataract, which is the opacification of the eye's lens, can only be treated by surgery. In order to develop and test new pharmacological treatment strategies for the disease, there is a need for an appropriate in vitro model using ex vivo animal lenses. In this study, porcine lenses were incubated in either culture medium, glucose, triamcinolone acetonide, sodium chloride, hydrogen peroxide, sodium selenite, neutral buffered formalin, or were exposed to microwave heating to experimentally induce lens opacification. Changes in the lens morphology, weight, size, and elasticity were monitored 7 days after treatment. The fastest induction of dense opacification was seen in lenses exposed to sodium chloride, neutral buffered formalin, and microwave heating. No change in the size and weight of the lenses were detected, whereas loss in elasticity could be detected in lenses treated with formalin solution or microwave heating. Thus, neutral buffered formalin- and microwave-treated ex vivo porcine lenses seem to be a suitable model for mature cataracts, whereas hypertonic sodium chloride may be useful for studies on osmolarity-induced lens opacification., (© 2021. The Author(s).)

Purpose : To investigate the effect of NKR-1 antagonists in an established UVR-B-induced cataract mouse model. Furthermore, to examine the expression of pro-inflammatory cytokines/chemokines in mouse eyes following unilateral UVR-B exposure. Methods : Mice received intraperitoneally injections of Fosaprepitant and Spantide I, before and after unilateral exposure to UVR-B. After day 3 and 7 post-exposure, ocular tissues were extracted for the detection of NKR-1 protein level by ELISA. Results : Pretreatment with Fosaprepitant decreases NKR-1 expression in exposed ocular tissues as well as in the unexposed lens epithelium compared to the saline group. Spantide I treatment showed a tendency of NKR-1 overexpression in ocular tissues. Conclusion : The clinically approved NKR-1 receptor antagonist Fosaprepitant decreases NKR-1 protein expression effectively not only in the exposed but also in the unexposed partner eye in a UVR-B irradiation mouse model. No effect was seen on the protein concentration of pro-inflammatory cytokines/chemokines in either eye.

Purpose: The aim of the present study was to determine whether caffeine concentrations in human lens epithelial cells (LECs) achieved from acute peroral caffeine intake inhibit ultraviolet radiation-induced apoptosis in vitro., Methods: Patients were planned for cataract surgery of both eyes with a caffeine abstinence of 2 weeks in total, starting 1 week before surgery of the first eye. The second eye was scheduled 1 week after the first eye. At the day of the second eye surgery, patients were given coffee containing 180 mg caffeine shortly before surgery. Lens capsules including LEC, harvested after capsulorhexis, were transferred to a cell culture dish and immediately exposed to close to threshold ultraviolet radiation (UVR). At 24 hr after UVR exposure, apoptotic LECs were analysed by TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining., Results: TUNEL-positive cells were detected in UVR-exposed lens capsules both after caffeine intake and in controls. The mean difference in TUNEL-positive cells between caffeine intake and contralateral controls (no caffeine) resulted in a 95% CI 15.3 ± 10.4% (degrees of freedom: 16)., Conclusion: Peroral caffeine consumption significantly decreased UVR-induced apoptosis in LEC supporting epidemiological findings that caffeine delays the onset of cataract., (© 2020 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.)

Background: Remnant interface fluid following Descemet stripping automated endothelial keratoplasty (DSAEK) is associated with postoperative detachments. The aim of this study was to assess outcomes of intraoperative optical coherence tomography (iOCT) guided meticulous peripheral corneal sweeping for removal of interface fluid during ultra-thin (UT) DSAEK., Methods: This retrospective study included all eyes underwent iOCT guided UT-DSAEK from October 2016 to February 2018 at the Hanusch Hospital, Vienna, Austria. Peripheral meticulous corneal sweeping was performed to remove excess fluid. Central graft thickness (CGT) was measured prior to surgery, after graft bubbling and after corneal sweeping. Remnant interface fluid rates were compared between eyes that underwent rebubbling and those that did not., Results: Overall, 28 eyes of 28 patients with a mean age of 73.9 ± 10.0 years were included. An iOCT guided meticulous peripheral sweeping was performed in 89.3% (n = 25) of the cases. Following 84% (n = 21) of the peripheral sweeping performed, remnant fluid was no longer identified. Following peripheral sweeping the interface fluid height was reduced from 17.31 ± 15.96 μm to 3.46 ± 9.52 μm (p < 0.001) and CGT was reduced by 7% (p < 0.001). Rebubbling was performed in 17.9% (n = 5) of the cases. The rebubbling group had a greater proportion of patients that had remnant fluid identified with iOCT at the end of surgery despite meticulous peripheral sweeping (60.0% versus 4.4%, p = 0.01)., Conclusion: The iOCT identified subclinical remnant fluid in nearly 90% of UT-DSAEK cases. An iOCT guided peripheral corneal sweeping led to resolution of interface fluid in a majority of cases. Eyes with persistent remnant fluid despite peripheral corneal sweeping are more likely to require subsequent rebubbling.

Background: Epiretinal membranes are a disorder leading to metamorphopsia and loss in visual function. The gold standard in therapy is vitrectomy with membrane peeling, usually performed with chromovitrectomy. The aim of this study was to examine whether dyes containing lutein are capable of enhancing visualization of epiretinal tissue in intraoperative optical coherence tomography (iOCT)., Patients and Methods: This was a prospective study that included 20 eyes of 20 patients with idiopathic epiretinal membranes scheduled for surgery. 23 G pars plana vitrectomy with intraoperative assistance of iOCT was performed in all cases. Staining of epiretinal membranes was performed with dyes containing trypan blue, brilliant blue G and lutein (tripledyne and dualdyne, both Kemin Industries Inc., USA)., Results: In all patients (n = 20), staining of epiretinal tissue was good, and crystalline lutein particles could be well depicted in iOCT compared to soluble lutein that does not enhance visualisation of epiretinal tissue in iOCT., Conclusions: The addition of lutein to commonly used dye formulations offers good staining properties and, in case of crystalline lutein, also enhances epiretinal tissue in iOCT., Competing Interests: Prof. Findl ist ein wissenschaftlicher Berater der Carl Zeiss Meditec AG, hat aber keine Interessenkonflikte, die die vorgestellte Studie betreffen, die anderen Autoren haben ebenfalls keine Interessenkonflikte., (Thieme. All rights reserved.)

Introduction: Caffeine and its metabolites have antioxidant activity, scavenging reactive oxygen species. The aim of our study was to measure caffeine concentrations in vitreous samples after peroral caffeine intake., Methods: This prospective study included patients scheduled for 23-G pars plana vitrectomy with membrane peeling due to epiretinal membranes. The study was performed in two parts: in the first part, patients were recruited into three different groups: group A consisted of habitual coffee drinkers who agreed to drink coffee containing 180 mg caffeine 1 h before surgery (n = 10), group B consisted of habitual coffee drinkers who were not offered coffee before surgery (n = 5), and group C consisted of non-habitual coffee drinkers, forming the control group (n = 5). In the second part (group D) patients (habitual coffee drinkers) agreed to give additional blood serum samples for measurement of caffeine concentration. Harvested samples of vitreous (groups A-D), epiretinal membranes (groups A-C), and blood serum samples (group D) were examined for concentrations of caffeine with gas chromatography-mass spectrometry., Results: Samples of 40 eyes of 40 patients were harvested. The concentrations of caffeine in the vitreous samples were 1,998 ± 967 ng/mL in group A and 1,108 ± 874 ng/mL in group B. In group C, caffeine concentrations were below 176 ng/mL in all vitreous samples. Both groups A and B had significantly higher concentrations of caffeine in the vitreous samples than group C (p < 0.002, p < 0.01, Mann-Whitney U test). Caffeine concentrations in epiretinal membranes were below the limits of detection. Correlation of caffeine concentrations between blood serum samples and vitreous samples in group D was high, with significantly higher caffeine concentrations in the blood serum., Conclusion: Coffee consumption leads to significant caffeine levels in the vitreous compared to patients in the control group, and caffeine concentrations in the vitreous showed a high correlation to blood serum concentrations of caffeine after peroral coffee consumption., (© 2020 S. Karger AG, Basel.)

Purpose: To investigate the influence of unilateral ultraviolet radiation (UVR) exposure on the unexposed, partner eye in vivo. To characterize the immunological cross-talk between the eyes and verify a sympathizing reaction of the partner eye via a neurokinin-dependent signaling pathway of substance P and its neurokinin-1 receptor (NKR-1) and/or monocyte chemoattractant protein-1 (MCP-1)., Methods: C57BL/6 mice were unilaterally exposed in vivo to UVR-B to a 5-fold cataract threshold equivalent dose of 14.5 kJ/m2 with a UV irradiation Bio-Spectra system. The unexposed contralateral eye was completely shielded during irradiation. After 3 and 7 days post exposure, eyes were stained with fluorescence-coupled antibody for substance P NKR-1. The same was performed in control animals receiving only anesthesia but no UVR-B exposure. NKR-1 and MCP-1 levels in ocular tissue lysates were quantified by enzyme-linked immunosorbent assay., Results: UVR-B induces NKR-1 upregulation after 3 and 7 days in the exposed and in the unexposed, contralateral mouse eye. NKR-1 protein level was upregulated in the exposed and contralateral iris/ciliary body complex, choroidea and in the contralateral retina as well as in the exposed cornea. MCP-1 levels were elevated in the exposed cornea, iris/ciliary body complex, and aqueous humor but not in contralateral ocular tissues., Conclusions: UVR-B exposure triggers NKR-1 upregulation not only in the exposed but also in the unexposed, partner eye in various ocular tissues. Following UVR-B exposure, MCP-1 protein levels are upregulated in the exposed eye, but the contralateral side remains unaffected., (© 2019 S. Karger AG, Basel.)

Background: The present study aims to investigate an automated qualitative and quantitative assessment system (Automated Quantification of After-Cataract [AQUA II]) of posterior capsule opacification (PCO) in high-resolution digital retroillumination images and consequently reduce observer bias and increase accuracy of PCO grading., Methods: A data set of 100 eyes with no to severe PCO was analysed. Ten eyes were consecutively photographed twice and ten images were rotated to give a total of 120 images for PCO assessment. Validity was determined by including subjective grading and repeatability was determined by evaluating the 20 additional images. Evaluation of posterior capsular opacification (EPCO), posterior capsule opacity (POCO) and AQUA I methods were included for comparative analysis of the data., Results: The system developed proved to classify six types of PCO. Validity was confirmed by a Pearson correlation coefficient of r = 0.95 (EPCO r = 0.93; POCO r = 0.72 and AQUA I r = 0.94). Repeatability was better in AQUA II (95% confidence interval [CI] for mean difference: 0.5 ± 1.2) than in subjective grading (95% CI for mean difference: 0.6 ± 1.7), in EPCO grading (95% CI for mean difference: - 0.2 ± 1.5), in POCO grading (95% CI for mean difference: 1.6 ± 2.7) and in AQUA I (95% CI for mean difference: - 1.1 ± 1.9)., Conclusions: AQUA II is a system that for the first time not only objectively quantifies PCO, but also qualitatively assesses PCO in an automated manner with texture classification. AQUA II showed an excellent validity and repeatability.

We describe a technique for making an optimal flange in intraocular lenses (IOLs) used for flanged intrascleral IOL fixation. The flange shape varies in poly(methyl methacrylate) (PMMA) haptics of different IOLs of different manufacturers. We identified the distance between the forceps grip of the haptic and the end of the haptic during heating with a cauter as a critical factor for the optimal flange shape in 5 PMMA haptics but not in 2 polyvinylidene fluoride haptics., (Copyright © 2018 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.)

The purpose of this study was to investigate the neurokinin receptor-1 (NKR-1) protein expression in ocular tissues before and after supra-cataract threshold ultraviolet radiation (UVR-B peak at 312 nm) exposure in vivo in a mouse model. Six-week-old C57Bl/6 mice were unilaterally exposed to a single (2.9 kJ/m 2 ) and an above 3-fold UVR-B cataract threshold dose (9.4 kJ/m 2 ) of UVR. UVR-exposure (λpeak = 312 nm) was performed in mydriasis using a Bio-Spectra exposure system. After latency periods of 3 and 7 days, eyes were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned and stained with fluorescence coupled antibody for NKR-1 and DAPI for cell nuclei staining. Control animals received only anesthesia but no UVR-exposure. Cataract development was documented with a Leica dark-field microscope and quantified as integrated optical density (IOD). NKR-1 is ubiquitously present in ocular tissues. An above 3-fold cataract threshold dose of UV-radiation induced NKR-1 upregulation after days 3 and 7 in the epithelium and endothelium of the cornea, the endothelial cells of the iris vessels, the pigmented epithelium/stroma of the ciliary body, the lens epithelium, pronounced in the nuclear bow region and the inner plexiform layer of the retina. A significant upregulation of NKR-1 could not be provoked with a single cataract threshold dose (2.9 kJ/m 2 UVR-B) ultraviolet irradiation. All exposed eyes developed anterior subcapsular cataracts. Neurokinin-1 receptor is present ubiquitously in ocular tissues including the lens epithelium and the nuclear bow region of the lens. UV-radiation exposure to an above 3-fold UVR-B cataract threshold dose triggers NKR-1 upregulation in the eye in vivo. The involvement of inflammation in ultraviolet radiation induced cataract and the role of neuroinflammatory peptides such as substance P and its receptor, NKR-1, might have been underestimated to date., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Purpose: To determine the pharmacokinetics of perorally administered caffeine, a widely consumed and potent dietary antioxidant, in the anterior lens capsule and lens epithelial cells, a crucial cell monolayer for cataract development., Methods: Bilateral cataract patients were scheduled for cataract surgery with a caffeine abstinence of 1 week before surgery of each eye. At the day of surgery of the second eye patients were administered no drink (0-mg group) or coffee with 60-, 120-, or 180-mg caffeine. After capsulorhexis the lens capsule including lens epithelial cells was transferred to a test tube for analysis of caffeine concentration by gas chromatography-mass spectrometry (GC-MS/MS)., Results: Coffee consumption significantly (P < 0.05) increased caffeine levels of the lens capsule/epithelium in the 60-, 120-, and 180-mg group. Caffeine concentrations (caffeine ng/lens capsule/epithelium) measured as difference between 1st and 2nd eye were -0.52 ± 1.16 (0-mg group, n = 7), 1.88 ± 2.02 (60-mg group, n = 8), 2.09 ± 0.67 (120-mg group, n = 9), and 3.68 ± 1.86 (180-mg group, n = 9). The increase constant of caffeine in a linear regression model was estimated as a 95% CI 0.02 ± 0.0046 (degrees of freedom; 25; r = 0.85)., Conclusions: Peroral intake of coffee significantly increased caffeine concentrations in the lens capsule and lens epithelial cells in a dose-dependent manner. This information is important for further investigations on preventing cataract.

Visual quantification and classification of fluorescent signals is the gold standard in microscopy. The purpose of this study was to develop an automated method to delineate cells and to quantify expression of fluorescent signal of biomarkers in each nucleus and cytoplasm of lens epithelial cells in a histological section. A region of interest representing the lens epithelium was manually demarcated in each input image. Thereafter, individual cell nuclei within the region of interest were automatically delineated based on watershed segmentation and thresholding with an algorithm developed in Matlab™. Fluorescence signal was quantified within nuclei, cytoplasms and juxtaposed backgrounds. The classification of cells as labelled or not labelled was based on comparison of the fluorescence signal within cells with local background. The classification rule was thereafter optimized as compared with visual classification of a limited dataset. The performance of the automated classification was evaluated by asking 11 independent blinded observers to classify all cells (n = 395) in one lens image. Time consumed by the automatic algorithm and visual classification of cells was recorded. On an average, 77% of the cells were correctly classified as compared with the majority vote of the visual observers. The average agreement among visual observers was 83%. However, variation among visual observers was high, and agreement between two visual observers was as low as 71% in the worst case. Automated classification was on average 10 times faster than visual scoring. The presented method enables objective and fast detection of lens epithelial cells and quantification of expression of fluorescent signal with an accuracy comparable with the variability among visual observers. © 2017 International Society for Advancement of Cytometry., (© 2017 International Society for Advancement of Cytometry.)

The current study aims to experimentally estimate the temperature in the lens due to heat load indirectly from the measurement of increases in the rate of temperature-induced light scattering. The lens was extracted from Sprague–Dawley rats and put into a temperature-controlled cuvette filled with a balanced salt solution. Altogether, 80 lenses were equally divided into four temperature groups. Each lens was exposed for 5 min to temperature depending on the group to which it belonged while the intensity of forward light scattering was recorded. The inclination coefficients of light scattering increase at the temperature of 37°C, 40°C, 43°C, and 46°C were estimated as a CI(0.95), 3.1 ± 0.8 , 4.4 ± 0.8 , 5.5 ± 0.9 , and 7.0 ± 0.8 × 10 ? 4 ?? tEDC / s , respectively. The Arrhenius equation implies that the natural logarithm of the inclination coefficient is linearly dependent on the inverse of the temperature. The proportionality constant and the intercept were 9.6 ± 2.4 × 10 3 ?? K and 22.8 ± 7.7 , respectively. The activation energy was 8.0 ± 2.0 × 10 1 ?? kJ · mol ? 1 . The current experiment implies that if averaging 20 measurements of inclination coefficients in a new experiment at constant heat load, the confidence limits for predicted temperature correspond to ± 1.9°C. With the proportionality constant and the intercept estimated in the current experiment, the in vivo temperature in the lens can be determined retrospectively with sufficient resolution.

Purpose: Peak toxicity for in vivo ultraviolet radiation (UVR) exposure to the lens is in the 300-nm wavelength region. However, little is known about corneal cell damage at 300 nm. The purpose of the study was to determine the time evolution of apoptosis in the cornea after in vivo exposure to 300-nm UVR., Methods: Altogether, 16 Sprague Dawley rats were divided into 4 groups and unilaterally exposed to 5 kJ/m UVR (λmax: 300 nm; λ0.5: 10 nm) for 15 minutes. After a predetermined latency period of 1, 5, 24, and 120 hours, depending on the group, the animals were killed and eyes were enucleated. Eye globes were further cryosectioned in 10-μm thick midsagittal sections. For the detection of apoptosis, the TUNEL method was applied., Results: TUNEL-positive signals were observed in the superficial epithelial cells in the exposed and control eyes at all latency periods. At 5 hours, TUNEL staining was detected in the exposed corneas in epithelial cells, keratocytes, and endothelial cells with a maximum signal at 24 hours. At 120 hours, no TUNEL staining was found in endothelial cells and only occasionally in keratocytes in exposed corneas. Signs of ulceration and stromal thinning were observed at 120 hours., Conclusions: UVR in the 300-nm wavelength region induces TUNEL staining in all 3 corneal layers. TUNEL staining of all 3 corneal layers is an early postexposure event observed after a 5-hour latency period. Corneal sterile keratolysis occurs in the time window of 24 to 120 hours probably induced by neutrophils.

Purpose: To determine the distribution of active caspase-3 in rat eye lens epithelium., Methods: In total, 120 sagittal sections from forty rats were assessed for active caspase-3 labelling using immunohistochemistry. Lens epithelial cells were counted, and the fraction of active caspase-3 labelled cells and their relative positions were identified in each section., Results: Active caspase-3 is present in normal lens epithelium. The active caspase-3 expression was higher in the anterior pole of the lens. Probability of radial spatial distribution of labelling was fitted with a logistic model. The increase rate and the inflection point were estimated as CI (0.95) to 23 ± 3 cells and 114 ± 3 cells, respectively., Conclusion: The gradually decreasing active caspase-3 labelling from the anterior pole to the periphery suggests that active caspase-3 may be involved in normal protein turnover caused by, for example, incident light., (© 2014 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.)

Purpose: To investigate whether infrared radiation (IRR)-induced cataract is instant or is associated with a time delay between the exposure and the onset of lens light scattering after an exposure to just above threshold dose., Methods: Six-weeks-old albino Sprague-Dawley female rats were unilaterally exposed to 197 W/cm2 IRR at 1090 nm within the dilated pupil. In the first experiment, the animals were exposed with four exposure times of 5, 8, 13 and 20 second, respectively. At 24 hr after exposure, the light scattering in both exposed and contralateral not exposed lenses was measured. Based on the first experiment, four postexposure time groups were exposed unilaterally to 1090 nm IRR of 197 W/cm2 for 8 second. At 6, 18, 55 and 168 hr after exposure, the light scattering in both lenses was measured., Results: A 197 W/cm2 IRR-induced light scattering in the lens with exposures of at least 8 second. Further, after exposure to IRR of 197 W/cm2 for 8 second, the light-scattering increase in the lens was delayed approximately 16 hr after the exposure., Conclusion: There is a time delay between the exposure and the onset of cataract after exposure to close to threshold dose implicating that either near IRR cataract is photochemical or there is a time delay in the biological expression of thermally induced damage., (© 2014 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.)

The damage mechanism for near-infrared radiation (IRR) induced cataract is unclear. Both a photochemical and a thermal mechanism were suggested. The current paper aims to elucidate a photochemical effect based on investigation of irradiance-exposure time reciprocity. Groups of 20 rats were unilaterally exposed to 96-W/cm(2) IRR at 1090 nm within the dilated pupil accumulating 57, 103, 198, and 344 kJ/cm(2), respectively. Temperature was recorded at the limbus of the exposed eye. Seven days after exposure, the lenses were macroscopically imaged and light scattering was quantitatively measured. The average maximum temperature increases for exposure times of 10, 18, 33, and 60 min were expressed as 7.0 ± 1.1, 6.8 ± 1.1, 7.6 ± 1.3, and 7.4 ± 1.1 °C [CI (0.95)] at the limbus of the exposed eye. The difference of light scattering in the lenses between exposed and contralateral not-exposed eyes was 0.00 ± 0.02, 0.01 ± 0.03, -0.01 ± 0.02, and -0.01 ± 0.03 transformed equivalent diazepam concentration (tEDC), respectively, and no apparent morphological changes in the lens were observed. An exposure to 96-W/cm(2) 1090-nm IRR projected on the cornea within the dilated pupil accumulating radiant exposures up to 344 kJ/cm(2) does not induce cataract if the temperature rise at the limbus is <8 °C. This is consistent with a thermal damage mechanism for IRR-induced cataract.

Purpose: To determine the time evolution of active caspase-3 protein expression in albino rat lens after in vivo exposure to low-dose UVR-300 nm, as detected by immunofluorescence., Methods: Forty Sprague-Dawley rats were unilaterally exposed in vivo to 1 kJ/m(2) UVR-300 nm for 15 min. At 0.5, 8, 16 and 24 hr after the UVR exposure, the exposed and contralateral nonexposed lenses were removed and processed for immunohistochemistry. Three mid-sagittal sections from each lens were stained. The cells labelled for active caspase-3 in each section of both the exposed and nonexposed lenses were counted and recorded three times. The difference of the proportion of labelling between the exposed and contralateral nonexposed lenses within each animal was calculated. The differences of active caspase-3 labelling at four different time-points after exposure were used to determine the time evolution of active caspase-3 expression., Results: Caspase-3 expression was higher in the exposed than in contralateral nonexposed lenses. The mean difference between the exposed and contralateral nonexposed lenses, including all lenses from all time intervals, was 0.12 ± 0.01 (= CI 95%). The mean differences between the exposed and contralateral nonexposed lenses were 0.11 ± 0.02, 0.13 ± 0.02, 0.14 ± 0.01 and 0.09 ± 0.03 (= CI 95%) for the 0.5-, 8-, 16- and 24-hr time groups, respectively. The orthogonal comparison showed no difference in the expression of active caspase-3 between the 0.5- and the 24-hr groups (Test statistic 1.50, F1,36 = 4.11, p < 0.05) or between the 8- and the 16-hr groups (test statistic 0.05, F1,36 = 4.11, p < 0.05). There was a difference when comparing the 0.5- and 24-hr groups to the 8- and 16-hr groups (test statistic 7.01, F1,36 = 4.11, p < 0.05)., Conclusion: The expression of active caspase-3 in the lens epithelium increases after UVR exposure. There is a peak of expression approximately 16 hr after the exposure., (© 2014 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.)

Purpose: The aim of the study is to investigate and visualize the ultrastructure of cataract morphology and repair, after in vivo exposure to double threshold dose UVR-B in the C57BL/6 mouse lens., Methods: Twenty-six-week-old C57BL/6 mice received in vivo double threshold dose (6.4 kJ/m2) UVR-B for 15 min. The radiation output of the UVR-source had λMAX at 302.6 nm. After a latency period of 1, 2, 4 and 8 days following UVR-B exposure, the induced cataract was visualized with electron microscopy techniques. Induced, cataract was quantified as forward lens light scattering. Damage to the lens epithelium and the anterior cortex was investigated with light microscopy in toluidine blue-stained semi-thin sections, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and dark field illumination photography., Results: UVR-B-exposed lenses developed anterior subcapsular and/or cortical and nuclear cataract after 1 day. Lens light scattering peaked 2 days after exposure. Lens epithelial cell damage was seen in TEM as apoptotic cells, apoptotic bodies, nuclear chromatin condensation, and swollen and disrupted anterior cortex fibres throughout the sections of the whole anterior lens surface. These morphologic changes were also visualized with SEM. Within 8 days, anterior subcapsular cataract was repaired towards the anterior sutures., Conclusion: UVR-B exposure of double cataract threshold dose induces a subtotal loss of epithelial cells across the whole anterior surface of the lens. This damage to the epithelium is repaired by epithelial cell movement from the equator towards the lens sutures, thus in retrograde direction to regular epithelial cell differentiation., (© 2014 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.)

The aim of the present study was to examine if topically applied caffeine influences pupil size in ketamine/xylazine anesthetized animals. Two experiments were carried out. In the first experiment, caffeine was topically applied to one of the eyes of 10 ketamine/xylazine anesthetized animals, while vehicle only was topically applied to the contralateral eye. In the second experiment, caffeine was topically applied to both eyes in one group of 10 ketamine/xylazine anesthetized rats, while in another group both eyes vehicle only was topically applied to both eyes. In both experiments pupil diameter was measured at 0, 10, 20, 40 and 60 min after topical application. In three of the animals, the pupil was dilated with tropicamide 5 mg/ml at 60 min after the topical application of caffeine and the pupil diameter was measured. The first experiment showed a relative miosis in caffeine treated eyes as compared to the vehicle treated eye, that changed over time. The second experiment in line with the first experiment, also showed that topically applied caffeine causes a relative miosis as compared to vehicle only that changes over time. Eyes treated with caffeine reacted with quick dilatation after tropicamide application. Topical caffeine antagonizes ketamine/xylazine anesthesia induced mydriasis in a time dependent manner., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Purpose: To introduce a model for the time evolution of active caspase-3 protein expression in albino rat lens up to 24 hours after in vivo exposure to low dose UVR in the 300 nm wavelength region (UVR-300 nm)., Methods: Forty Sprague-Dawley rats were unilaterally exposed in vivo to 1 kJ/m2 UVR-300 nm for 15 minutes. At 0.5, 8, 16, and 24 hours after the UVR exposure, the exposed and contralateral not-exposed lenses were removed and processed for immunohistochemistry. The differences in the probability of active caspase-3 expression at four different time points after exposure were used to determine the time evolution of active caspase-3 expression. A logistic model was introduced for the expression of active caspase-3. The parameters for the exposed and the not exposed lenses were estimated for the observation time points., Results: The exposure to UVR-300 nm impacted on the parameters of the logistic model. Further, the parameters of the model varied with time after exposure to UVR-300 nm., Conclusion: The logistic model predicts the impact of exposure to UVR-300 nm on the spatial distribution of probability of active caspase-3 protein expression, depending on time.

Topically applied caffeine was recently identified as a promising candidate molecule for cataract prevention. Little is known about the pharmacokinetics for topically applied caffeine. Potential toxicity of 72 mM caffeine on the ocular surface and the lens was qualitatively monitored and no toxic effects were observed. The concentration of caffeine was measured in the lens and the blood after topical application of 72 mM caffeine to groups of 10 animals sacrificed at 30, 60, 90 and 120 min after topical application. The lens concentration decreased throughout the observation period while the blood concentration increased up to 120 min. Further, the concentration of caffeine in the lens and blood was measured 30 min after topical application of caffeine, the concentration of caffeine being 0.72, 3.34, 15.51 and 72 mM depending on group belonging, in groups of 10 animals. The caffeine concentration in lens and blood, respectively, increased proportionally to the caffeine concentration topically applied. The rat blood concentrations achieved were far below the equivalent threshold dose of FDA recommended daily dose for humans. This information is important for further development of caffeine eye drops for cataract prevention., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

An in vivo exposure to 197  W/cm 2 1090-nm infrared radiation (IRR) requires a minimum 8 s for cataract induction. The present study aims to determine the ocular temperature evolution and the associated heat flow at the same exposure conditions. Two groups of 12 rats were unilaterally exposed within the dilated pupil with a close to collimated beam between lens and retina. Temperature was recorded with thermocouples. Within 5 min after exposure, the lens light scattering was measured. In one group, the temperature rise in the exposed eye, expressed as a confidence interval (0.95), was 11±3°C at the limbus, 16±6°C in the vitreous behind lens, and 16±7°C on the sclera next to the optic nerve, respectively. In the other group, the temperature rise in the exposed eye was 9±1°C at the limbus and 26±11°C on the sclera next to the optic nerve, respectively. The difference of forward light scattering between exposed and contralateral not exposed eye was 0.01±0.09 tEDC. An exposure to 197  W/cm 2 1090-nm IRR for 8 s induces a temperature increase of 10°C at the limbus and 26°C close to the retina. IRR cataract is probably of thermal origin.

Purpose/aim: To quantitatively analyse the evolution of TUNEL-labeling, after in vivo exposure to UVB., Methods: Altogether, 16 Sprague Dawley rats were unilaterally exposed in vivo for 15 min to close to threshold dose, 5 kJ/m(2), of ultraviolet radiation in the 300 nm wavelength region. Animals were sacrificed in groups of 4 at 1, 5, 24 and 120 h after exposure. For each animal, both eye globes were removed and frozen. The frozen eye was cryo-sectioned in 10 µm thick midsagittal sections. From each globe, three midsagittal sections with at least five sections interval in between were mounted on a microscope slide. Sections were TUNEL-labeled and counter stained with DAPI. For quantification of apoptosis, a fluorescence microscope was used. In sections with a continuous epithelial cell surface, the number of lens epithelial cell nuclei and the number of TUNEL-positive epithelial cell nuclei was counted. The total number of TUNEL-positive epithelial cell nuclei for all three sections of one lens in relation to the total number of epithelial cell nuclei for all three sections of the same lens was compared between exposed and contralateral not exposed lens for each animal., Results: The relative difference of the fraction of TUNEL-positive nuclei between exposed and contralateral not exposed lens increased gradually, peaked in the time interval 5-120 h after exposure, and then declined., Conclusions: Close to threshold dose of UVB induces TUNEL-labeling that peaks in the time window 5-120 h after exposure to UVB.