Review for special issue: Corneal lamellar surgery: Present outcomes and future perspectives.

Since the establishment of the first eye bank in the 1940s, their role has evolved to face new challenges. With the recent development of lamellar keratoplasties, eye banks play an even bigger role in the selection and preparation of donor tissues. The increasing number of keratoplasty techniques and the high demand for "ready-to-use" tissues are challenging eye banks to improve and develop new preparation techniques. Besides necessary examinations, new approaches of tissue analysis in eye banks allow a better/optimized selection of corneal tissues. These new challenges in tissue preservation, preparation, and selection are propelling eye banks into a new era of modern eye banking.

Evaluation of dynamic corneal response parameters and the biomechanical E-staging after Intacs® SK implantation in keratoconus.

PURPOSE: This retrospective longitudinal study evaluated the biomechanical E-staging in KC corneas before and after intracorneal ring segment (ICRS) implantation (Intacs® SK, Addition Technology, Illinois, United States). METHODS: Biomechanical E-staging for ectatic corneal diseases was applied retrospectively on 49 KC corneas of 41 patients who underwent ICRS implantation. The main outcome parameters included the Corvis Biomechanical Factor (CBiF, the linearized Corvis Biomechanical Index and the biomechanical parameters included), the resulting biomechanical E-staging, the stress-strain index, thinnest corneal thickness (TCT), maximal anterior keratometry (Kmax), and the anterior radius of curvature (ARC). They were evaluated at 1.9 ± 1.1 months preoperatively and postoperatively after 2.8 ± 0.7, 5.8 ± 1.0, and 10.6 ± 2.3 months. RESULTS: The CBiF decreased (4.9 ± 0.5 | 4.7 ± 0.5, P = 0.0013), and the E-staging increased significantly (2.8 ± 0.8 | 3.1 ± 0.9, P = 0.0012, paired t-test) from preoperatively to the first postoperative follow-up. The difference remained significant after 6 months; however, there was no more difference after 11 months. TCT was stable, whereas Kmax and ARC significantly decreased after ICRS implantation (TCT: 464 ± 49, 470 ± 51, 467 ± 38, 461 ± 48; Kmax: 56.3 ± 4.5, 54.7 ± 4.5, 54.2 ± 4.8, 54.1 ± 4.3; ARC: 51.5 ± 3.4, 48.3 ± 3.8, 48.6 ± 3.0, 48.6 ± 3.2 preoperatively and 3, 6, and 11 months postoperatively, respectively). Besides Kmax and ARC, Ambrósio's relational thickness to the horizontal profile (ARTh) was the only parameter that was significantly lower than preoperatively at any follow-up (P ≤ 0.0024, Wilcoxon matched-pairs test). CONCLUSION: Intacs® SK implantation results in an increasing biomechanical E-staging in the first postoperative months with stabilization near preoperative values after 1 year. Significantly lower ARTh values at any follow-up document the ICRS effect and contribute to a slightly higher postoperative biomechanical E-staging value.

Analysis of Graft Detachments and Re-Bubblings After 450 Descemet Membrane Endothelial Keratoplasty Procedures.

PURPOSE: To objectify the indication for re-bubbling by analyzing graft detachments (GDs) after Descemet membrane endothelial keratoplasty. METHODS: In this retrospective monocentric observational study, re-bubbling cases of 450 Descemet membrane endothelial keratoplasties and the percentage of the residual gas filling (RGF) in the anterior chamber on the first postoperative day were collected. The number/location/extent of GDs and the corneal thickness above GDs were analyzed using anterior segment optical coherence tomography. RESULTS: From a total of 450 grafts, 384 (85.3%) had at least a minimal degree GD. One hundred twenty-two of 450 grafts (27.1%) underwent at least 1 re-bubbling. The mean RGF was significantly lower in eyes with GD (67.7 ± 12.6%) than in eyes without GD (74.2 ± 11.3%). GDs occurred most frequently in the inferotemporal quadrant (46.0%). GDs were significantly more likely to require a re-bubbling when the central parts of the graft were affected (94.0% vs. 35.7%). The number of detachments per graft was directly proportional to the re-bubbling rate. The GDs which required a re-bubbling were on average 56 µm higher and 461 µm wider than the untreated ones. The cornea above the GDs that needed a re-bubbling was significantly thicker than above the untreated GDs (mean 988 ± 102 µm vs. 951 ± 99 µm). CONCLUSIONS: The RGF seems to be a major influencing factor for graft attachment. The most susceptible location of the GD is inferotemporal. The main factors that need to be investigated to decide if a re-bubbling is required are the number of detachments per graft, their dimensions, whether the central portions of the graft are involved, and the corneal thickness above GDs.

RNA and the RNA-binding protein FUS act in concert to prevent TDP-43 spatial segregation.

FUS and TDP-43 are two self-adhesive aggregation-prone mRNA-binding proteins whose pathological mutations have been linked to neurodegeneration. While TDP-43 and FUS form reversible mRNA-rich compartments in the nucleus, pathological mutations promote their respective cytoplasmic aggregation in neurons with no apparent link between the two proteins except their intertwined function in mRNA processing. By combining analyses in cellular context and at high resolution in vitro, we unraveled that TDP-43 is specifically recruited in FUS assemblies to form TDP-43-rich subcompartments but without reciprocity. The presence of mRNA provides an additional scaffold to promote the mixing between TDP-43 and FUS. Accordingly, we also found that the pathological truncated form of TDP-43, TDP-25, which has an impaired RNA-binding ability, no longer mixes with FUS. Together, these results suggest that the binding of FUS along nascent mRNAs enables TDP-43, which is highly aggregation-prone, to mix with FUS phase to form mRNA-rich subcompartments. A functional link between FUS and TDP-43 may explain their common implication in amyotrophic lateral sclerosis.

Leadership in ophthalmology training : Opportunities and risks of medical specialist education.

Medical specialist training requires constant improvement and adaptation of the contents to the current situation. Nowadays, young physicians have the opportunity to select among the most renowned institutions and can choose the one most qualified for their training. Hospitals on the other hand still have the desire to recruit highly qualified physicians for their resident programs, which requires a good, well-rounded and reliable offer by the department under good leadership. Thus, among other issues a modern and multilingual homepage is already an important instrument for successfully addressing applicants and winning them over for the department. In addition to a well-planned and structured training plan (e.g., the "Homburg Curriculum") and a so-called "resident guide", many other additional offers are nowadays part of a successful training, such as structured internal and external specialist training courses, well thought out research concepts available to all interested parties, wet labs for practical exercises on pig's eyes and as the latest most innovative addition, a virtual reality simulator. Due to a structured curriculum with regular continuous education during the daily early morning meetings and an exchange program with another university eye hospital, not only the residents can benefit but ultimately also the department itself. In addition, future specialists are involved in the respective organization (so-called "service teams") from the very beginning. This conveys a great deal of knowledge and expertise but also organizational skills and thus improves the quality of training. In any case, standardized residency training with a view beyond the horizon, which is transparently organized and reliably carried out, improves the quality of training in order to become a certified ophthalmologist and increases the satisfaction of the residents. A department which is committed and can offer a wide range of services will benefit from motivated and satisfied employees in a good interpersonal climate, which in the end benefits not only the team but also the patients.

Descemet Membrane Endothelial Keratoplasty after failed penetrating keratoplasty- case series and review of the literature.

BACKGROUND: This study aims to evaluate visual outcome, central corneal thickness, and re-bubbling rate in a cohort with undersized sequential Descemet Membrane Endothelial Keratoplasty (DMEK) due to endothelial graft decompensation following primary penetrating keratoplasty (PK). METHODS: All patients who received a sequential DMEK (n = 16) or triple DMEK (n = 2) after failed primary PK between November 2020 and June 2022 were retrospectively evaluated. Analyzed parameters were corrected distance visual acuity (CDVA), central corneal thickness (CCT), re-bubbling rate and graft survival. RESULTS: 18 eyes of 18 patients were included. All patients underwent a DMEK with undersized graft after failed PK(s). Mean time between the last PK and DMEK was 102 ± 82 weeks. Mean follow-up time was 8.9 ± 4.6 months. CDVA increased significantly from 1.12 ± 0.60 logMAR preoperatively to 0.64 ± 0.49 logMAR 6 weeks postoperatively (p = 0.013). Mean CCT decreased significantly from 807 ± 224 µm before to 573 ± 151 µm 6 weeks after DMEK (p = 0.003). Re-bubbling was necessary in eight eyes (44.4%) after a median time of 7 days. The 12-month Kaplan Meier survival was 66.7%. CONCLUSION: In case of endothelial graft decompensation without stromal scars after primary PK, a DMEK can be performed for selected patients who had satisfying CDVA before the endothelial decompensation. Prior to DMEK indication, an AS-OCT should routinely be performed to circularly search for posterior steps at the PK graft margin, as well as shortly after DMEK to exclude a detachment of the endothelial graft. All patients should be informed about a higher re-bubbling rate in comparison to primary DMEK.

Impact of cleanroom status on the reasons for discarding organ-cultured corneal transplants in a modern eye bank - More donor corneas thanks to astronaut suit?

PURPOSE: This study investigated the influence of cleanroom conditions on the discard rates of donor corneas in a German university eye bank. METHODS: Discard rates were analysed from 2017 to 2020 at the LIONS Cornea Bank at Saarland University Medical Center. 1941 corneas from 971 donors were included. 1262 corneas (65.1%) were stored in a class D cleanroom from 2017 to 2019 and processed in a cleanroom class A sterile bank (group 1). 679 corneas (34.9%) were continuously stored in a class B cleanroom and processed in a class A cleanroom safety cabinet in the same room from 2019 to 2020 (group 2). The target parameter of this work was the number of contamination-related discards. Although they cannot be influenced by the spatial conditions, the discards due to insufficient endothelial quality, serology, contraindications, scars and technical causes were also recorded. Statistical analysis was performed using SPSS and various testing procedures. RESULTS: In group 1, significantly more corneas were discarded due to positive serology (6.9%|3.8%, p = 0.020). There was no significant change between both groups for either contamination or the other reasons for discard. CONCLUSION: Optimization of hygiene standards from cleanroom class D to B did not reduce contamination. Serology, endothelial quality, medical contraindications and the presence of scars cannot be influenced by cleanroom conditions.

Anterior chamber intraocular lens removal combined with triple DMEK: Quadruple DMEK for bullous keratopathy.

PURPOSE: To present the outcomes of a patient with anterior chamber intraocular lens (ACIOL) related endothelial decompensation who underwent Descemet membrane endothelial keratoplasty (DMEK) and cataract surgery with intraocular lens (IOL) implantation in the capsular bag (so-called triple DMEK) combined with ACIOL removal (quadruple DMEK) in both eyes. METHODS: Case report. RESULTS: A 58-year-old female patient was referred due to decreased visual acuity within the last 18 months. She had a history of iris-claw ACIOL implantation 17 years before. The corrected distance visual acuity (CDVA) was 20/40 in both eyes. Due to low endothelial cell density and increased corneal thickness, ACIOL removal combined with triple DMEK (as quadruple DMEK) was performed for both eyes. Despite a graft detachment that was successfully managed with re-bubbling in the first eye, both eyes showed an increase in the CDVA (20/25 and 20/32, respectively) without any other significant complications in the follow-up of the patient. The corneas of both eyes were clear postoperatively. CONCLUSION: This case report demonstrated that quadruple DMEK may provide feasible management for chronic endothelial cell decompensation secondary to iris-claw ACIOL implantation.

Implantation of Intracorneal Ring Segments in Keratectasia: Effects on Corneal Biomechanics in 112 Eyes.

PURPOSE: The aim of this study was to analyze changes in corneal biomechanical properties after implantation of intracorneal ring segments (ICRSs) in keratectasia. METHODS: This retrospective single-center study included 112 patient eyes that underwent femtosecond laser-assisted ICRS implantation (Intacs SK; Addition Technology Inc, Des Plaines, IL) for keratectasia. Biomechanical analysis was performed using the Ocular Response Analyzer (ORA; Reichert Inc, Depew, NY), with determination of corneal resistance factor, corneal hysteresis, and Keratoconus Match Index, as well as by Corvis ST (OCULUS, Wetzlar, Germany), with determination of stiffness parameter A1, Ambrosio relational thickness to the horizontal profile (Arth), integrated radius, deformation amplitude ratio, and stress-strain index as well as Corvis Biomechanical Index and Tomographic Biomechanical Index. Data collection was performed preoperatively and 6 months postoperatively for ORA and Corvis ST and additionally after 1 and 2 years for ORA. RESULTS: The corneal resistance factor decreased significantly postoperatively (5.8 ± 1.7 mm Hg) compared with preoperatively (6.75 ± 3.7 mm Hg; P = 0.021) and increased again during follow-up (6.2 ± 1.9 mm Hg; P = 0.024), without regaining preoperative values. Corneal hysteresis and Keratoconus Match Index did not change significantly. Stiffness parameter A1 (P = 0.045) increased significantly after ICRS implantation and Arth decreased significantly from 181 ± 85 to 150 ± 92 (P = 0.016). However, there was no significant postoperative change for others Corvis parameters. CONCLUSIONS: Corneal biomechanical properties showed inconsistent changes after ICRS implantation. Classical corneal biomechanical parameters (using single central air-puff tonometers) do not seem to be suitable for follow-up after ICRS implantation.

FUS RRM regulates poly(ADP-ribose) levels after transcriptional arrest and PARP-1 activation on DNA damage.

PARP-1 activation at DNA damage sites leads to the synthesis of long poly(ADP-ribose) (PAR) chains, which serve as a signal for DNA repair. Here we show that FUS, an RNA-binding protein, is specifically directed to PAR through its RNA recognition motif (RRM) to increase PAR synthesis by PARP-1 in HeLa cells after genotoxic stress. Using a structural approach, we also identify specific residues located in the FUS RRM, which can be PARylated by PARP-1 to control the level of PAR synthesis. Based on the results of this work, we propose a model in which, following a transcriptional arrest that releases FUS from nascent mRNA, FUS can be recruited by PARP-1 activated by DNA damage to stimulate PAR synthesis. We anticipate that this model offers new perspectives to understand the role of FET proteins in cancers and in certain neurodegenerative diseases such as amyotrophic lateral sclerosis.

[Modern corneal diagnostics as the key for the correct classification of the disease and optimal treatment decisions]. / Moderne Hornhautdiagnostik als Schlüssel für die korrekte Einordnung der Erkrankung und optimale Therapieentscheidung.

Corneal diseases include a wide spectrum of different manifestations (inflammatory/noninflammatory) that need to be accurately classified for precise diagnosis and targeted treatment. In addition to the anamnesis and slit lamp biomicroscopy, further device-based examinations can be performed to narrow down the diagnosis. Nowadays, modern corneal imaging provides a variety of technologies, such as topography, tomography, in vivo confocal microscopy and analysis of biomechanics, which are able to reliably classify different pathologies. Knowledge of the available examination modalities helps to guide differential diagnostic considerations, facilitating the indication for stage-appropriate microsurgical intervention.

[Leadership in ophthalmology training : Opportunities and risks of medical specialist education]. / Führung in der augenärztlichen Ausbildung : Chancen und Risiken der Facharztweiterbildung.

Medical specialist training requires constant improvement and adaptation of the contents to the current situation. Nowadays, young physicians have the opportunity to select among the most renowned institutions and can choose the best qualified for their training. Hospitals on the other hand still have the desire to recruit highly qualified physicians for continuing education, which requires a good, well-rounded and reliable offer by the department under good leadership. Thus, among other issues a modern and multilingual homepage is already an important instrument for successfully addressing applicants and winning them over for the department. In addition to a well-planned and structured training plan (e.g., the Homburg curriculum) and a so-called resident guide, many other additional offers are nowadays part of a successful training, such as structured internal and external continuing education, well thought out research concepts available to all interested parties, wet labs for practical exercises on pig's eyes and as the latest most innovative addition, a virtual reality simulator. Due to a structured curriculum with regular continuous education during the daily early morning meetings and an exchange program with another university eye hospital, not only the residents can benefit but ultimately also the department itself. In addition, future specialists are involved in the respective organization (so-called service teams) from the very beginning. This conveys a great deal of knowledge and expertise but also organizational skills and thus improves the quality of training. In any case, standardized residency training with a view beyond the horizon, which is transparently organized and reliably carried out, improves the quality of training in order to become a certified ophthalmologist and increases the satisfaction of the residents. A department which is committed and can offer a wide range of services will benefit from motivated and satisfied employees in a good interpersonal climate, which in the end benefits not only the team but also the patients.

P23-A137†Cross-border cooperation for corneal donation between lions eye bank Saar-Lor-Lux, Trier/Westpfalz in Homburg/Saar and Luxembourg.

PURPOSE: With the increasing demand for corneas, eye banks must optimize and extend their sources of tissue donation. On the other hand, corneal transplantation is a specialized procedure performed in hospitals with high quality standards and ideally an integrated eye bank. In this report we would like to focus on an international win-win-win agreement between the Department of Ophthalmology at Saarland University Medical Center (Homburg/Saar, Germany), the LIONS Eye Bank Saar-Lor-Lux, Trier/Westpfalz and the four major non-university hospitals without corneal transplantation competence in Luxembourg. METHODS: In 2012, at the initiative of the Luxembourgish Ministry of Health and Department of Ophthalmology (Homburg/Saar, Germany), an international agreement was established with the Centre Hospitalier du Luxembourg (Luxembourg). Administrative and legislative rules were developed. Luxembourgish nursing personnel attended a practical training program for corneal excision at the Department of Ophthalmology in Homburg/Saar allowing them to harvest the two first corneal donors on site by themselves during the first year. In the following years two more hospitals, the Centre Hospitalier Emile Mayrisch (Esch-sur-Alzette, Luxembourg) and the Hôpitaux Robert Schuman (Kirchberg, Luxembourg), joined the cooperation. RESULTS: From 2012 until 2021, three hospitals in Luxembourg donated 779 corneas to the LIONS Eye Bank of the Saarland University Medical Center in Homburg/Saar (Germany). In return, 308 Luxembourgish patients have received a corneal transplantation at the Department of Ophthalmology in Homburg/Saar. In 2022, the extension continued and an agreement with a fourth hospital in Luxembourg at the Centre Hospitalier du Nord (Ettelbruck, Luxembourg) was signed providing even more donations. CONCLUSION: The cross-border collaboration for corneal donation and patient treatment has proven to be successful with both numbers of harvested donors and transplanted patients rising. However, international legislation for tissue donation needs to be accurately respected and a quality management system established to provide continuous quality of the donor tissue.

P42-A105†Sterile donor tomography for improvement of refractive results after keratoplasty.

PURPOSE: To evaluate the efficiency of using anterior segment optical coherence tomography (AS-OCT) as a non-invasive and sterile screening method in the eye bank to detect corneal grafts with curvature and/or thickness anomalies, thus improving the graft selection for corneal transplantation. METHODS: 1222 donor corneal tissues mounted in sterile organ culture flasks were imaged using an AS-OCT (CASIA 2 - Tomey, Nagoya, Japan) between January 2018 and September 2022. The corneal tissues were preserved at least 12 hours in organ culture medium 2 (containing 6% dextran T-500) before the measurement in order to allow deswelling prior to the examination. Depth scans were performed sterilely through the organ culture flask from the posterior surface of the corneal tissues within a 7 mm central zone to create 3D volume data. The volume data set was imported to MATLAB (MathWorks Inc., Natick, Massachusetts, USA) and, after preprocessing the data and defining the region of interest (ROI), the edge of the front and back surfaces of the corneal tissues was detected. Subsequently, the adaptation of a sphero-cylindrical surface model was carried out with raytracing. The radii of curvature for the front and back surfaces and the central corneal thickness were determined according to the method proposed by Mäurer, Eppig, Langenbucher et al at the Institute of Experimental Ophthalmology, Homburg/Saar, Germany. RESULTS: The mean steep/flat front surface radius was 7.46 ± 0.29 (6.07 - 9.29)/7.69 ± 0.24 (6.70 - 9.50) mm, the corresponding values for the back surface being 6.48 ± 0.32 (5.30 - 8.00)/6.80 ± 0.31 (5.81 - 8.00) mm and the mean central thickness was 611.5 ± 85.6 (378.5 - 1457.2) µm. Anomalies (beyond ± 2 or ± 3 standard deviations SD) were found in 111 or 41 corneas (9.1% or 3.4%) for anterior surface curvature, 135 or 38 for corneas (11.0% or 3.1%) for the posterior surface, and 53 or 15 corneas (4.3% or 1.2%) for central corneal thickness. CONCLUSION: The AS-OCT provides an objective, sterile and semi-automated screening method to identify corneal morphological and refractive alterations (e.g. keratoconus, status post keratorefractive surgery) to further optimize corneal donor selection in the eye bank. Corneal donors with curvature or thickness anomalies +/- 3 SD (eminence-based) do not have to be discarded but can be used for posterior lamellar keratoplasty, especially DMEK in Germany.

P08-A131†Comparison of sterile donor tomography in the eye bank and previous keratometric measurements during the donor's lifetime.

PURPOSE: Sterile donor tomography in the eye bank can be used to minimise refractive surprises after corneal transplantation.The aim of this study was to compare sterile tomography of donor corneas in the eye bank with keratometric measurements of the same donors performed prior their death. METHODS: Since 2018, 1246 donor sclerocorneal discs have been routinely measured using donor tomography, taken sterilely through their cell culture flask in medium II using the anterior segment optical coherence tomograph Casia 2 (Tomey Corp., Nagoya, Japan) and a custom-made Matlab software (The MathWorks Inc., Natick, Massachusetts, USA). Of all these donor corneas, 19 (1.5%) appeared to have been measured with Pentacam (Oculus Optikgeräte GmbH, Wetzlar, Germany) in the donors before death. Both measurements, taken at a mean interval of 35 ± 26 months, were compared using a Wilcoxon signed-rank test. RESULTS: The mean steepest/flattest front surface radius and anterior astigmatism of the corneas measured with Pentacam amounted 7.66±0.35/7.93±0.37 mm, and 0.27±0.43 mm. Corresponding values of sterile donor tomography were respectively 7.48±0.31 [p<0.01]/7.77±0.25 [p=0.01] mm, and 0.29±0.35 [p=0.78] mm.At the posterior corneal surface, the Pentacam measured a mean steepest/flattest surface radius and astigmatism of 6.27±0.33/6.72±0.48 mm and 0.45±0.47 mm, whereas values of sterile donor tomography amounted 6.55±0.30 [p<0.01]/6.94±0.33 [p=0.04] mm and 0.39±0.26 [p=0.63] mm, respectively.The central corneal thickness amounted 575±52 µm with Pentacam, and 597±80 µm [p=0.20] with sterile donor tomography. CONCLUSION: The front and back surface astigmatism as well as the central corneal thickness remained statistically unchanged after corneal excision and preservation in organ culture in comparison to measurement of the donor prior death. The statistically non-similar anterior and posterior radius of curvature between both methods must be seen in light of the known differing corneal topography between swept-source anterior segment optical coherence tomography and Scheimpflug imaging. These results suggest a merely minimal deformation caused by the storage and attachment of donor corneas to their holder in the cell culture flask for sterile donor tomography, causing a steeper anterior surface curvature but leaving the astigmatism still congruent with previous in situ conditions.

Baseline Characteristics of 1976 Patients With Ectatic Corneal Disorders at a Single Center From 2010 to 2021: A Cross-Sectional Study of the Homburg Keratoconus Center.

OBJECTIVES: The study objective was to analyze the baseline characteristics of keratoconus (KC) patients at the Homburg Keratoconus Center from 2010 to 2021. METHODS: This cross-sectional study included 3,674 eyes, with analysis of demographics, clinical findings, visual function, endothelial measurements, and topographic, tomographic, and corneal biomechanical data from the first visit. RESULTS: Mean patient age was 36.3±13.8 years. The mean uncorrected distance visual acuity in log of minimal angle of resolution was 0.60 (20/80, Snellen equivalent), and the corrected mean was 0.3 (20/40). Of 1976 patients, 48.9% reported eye rubbing. Mean values (ranges) were 49.4±6.3 (36.3-78.0) D for steep keratometry, 462.4±66.0 (48.0-659.0) µm for thinnest corneal thickness, 9.7±8.7 (-0.5 to 88.8) for Belin/Ambrósio enhanced ectasia total deviation, 0.8±0.4 (0.0-1.0) for the Corvis biomechanical index, 0.9±0.2 (0.0-1.0) for the tomographic biomechanical index, 0.1±0.5 (-0.9 to 2.0) for the KC match index, 8.3±1.8 (2.2-17.7) mm Hg for corneal hysteresis, 7.1±2.2 (0.0-17.0) mm Hg for corneal resistance factor, and 2,562.9±326.3 (1,011-3,937) cells/mm2 for endothelial cell density. The average ABCDE KC stage was A2B3C1D1E2. Distance-corrected visual acuity correlated strongly with topometric, tomographic, and biomechanical data ( P <0.001). CONCLUSIONS: This comprehensive description of baseline features of KC patients at a tertiary center provides a reference for further longitudinal and international multicentric studies.

FUS fibrillation occurs through a nucleation-based process below the critical concentration required for liquid-liquid phase separation.

FUS is an RNA-binding protein involved in familiar forms of ALS and FTLD that also assembles into fibrillar cytoplasmic aggregates in some neurodegenerative diseases without genetic causes. The self-adhesive prion-like domain in FUS generates reversible condensates via the liquid-liquid phase separation process (LLPS) whose maturation can lead to the formation of insoluble fibrillar aggregates in vitro, consistent with the appearance of cytoplasmic inclusions in ageing neurons. Using a single-molecule imaging approach, we reveal that FUS can assemble into nanofibrils at concentrations in the nanomolar range. These results suggest that the formation of fibrillar aggregates of FUS could occur in the cytoplasm at low concentrations of FUS, below the critical ones required to trigger the liquid-like condensate formation. Such nanofibrils may serve as seeds for the formation of pathological inclusions. Interestingly, the fibrillation of FUS at low concentrations is inhibited by its binding to mRNA or after the phosphorylation of its prion-like domain, in agreement with previous models.

In situ donor keratometry in deceased patients as a novel screening technique for eye banking.

PURPOSE: To investigate the potential role of keratometry on whole globes in situ of deceased patients by assessing its repeatability and comparing it with sterile donor tomography after excision and preservation in organ culture. METHODS: A sequence of 5 measurements was taken from 40 eyes in situ of deceased patients < 24 h after death using the portable Retinomax K-plus 3 (Bon, Tokyo, Japan). Keratometry of whole globes in situ, from which sclerocorneal discs were taken for organ culture, was compared to those obtained after measuring these sclerocorneal disks through their cell culture flask in medium I after 5 ± 4 days using the anterior segment optical coherence tomograph Casia 2 (Tomey Corp., Nagoya, Japan), and to 964 different donor corneas in medium II. RESULTS: Cronbach's alpha of the in situ keratometry was 0.891 and 0.942 for the steepest and flattest corneal power (P). The steepest (44.5D) and flattest (41.1D) P as well as the astigmatism (3.4D) of in situ corneas remained unchanged after preserving sclerocorneal discs in medium I (respectively 44.7D [p = 0.09]; 41.4D [p = 0.17]; 3.3D [p = 0.09]). The comparison of the in situ values with the 964 measured different donor corneas in medium II showed significantly (p < 0.001) higher P at the steep (45.4D) and flat (43.9D) meridian and smaller astigmatism (1.4D) for sterile donor tomography. CONCLUSIONS: Measuring deceased patients' eyes in situ with the portable Retinomax K-plus 3 represents a feasible and reliably repeatable screening method in the eye bank. In comparison to donor tomography in medium I, it measures a similar power and astigmatism.