Veneto Eye Bank Foundation

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T he term biomicroscopy entered ophthalmology in the early 20th century with the revolutionary slit lamp, which brought microscopic examination to the living eye without tissue removal. For decades, biomicroscopy meant slit-lamp examination of the anterior segment. However, as imaging technologies such as ultrasound, confocal microscopy, and optical coherence tomography (OCT) advanced, the term expanded to encompass any microscopic observation of living tissues.

NHSBT have already exceeded the number of precut corneas supplied in 2023.The implementation of the OCT Analyser as a quality control step has improved the reliability of the quality of the Tissue being supplied and being able to provide an OCT image of the prepared Tissue has provided an Improved service for the surgeons. 5.5.

PurposeTo compare the difference of long-term clinical outcomes between eye bank prepared pre-loaded (pl)-DMEK (endo-in technique) and pre-cut (pc)-ultra-thin (UT)-DSAEK grafts.MethodIn this retrospective single center interventional case series, patients were treated for Fuchs endothelial corneal dystrophy (FECD) using pl-DMEK (n=47) and pc-UT-DSAEK grafts (n=42). The grafts were prepared, loaded and transported in an IOL cartridge (pl-DMEK) or in the storage media (pc-UT-DSAEK) by a single eye-bank. All the grafts were delivered by a single surgeon using bimanual pull-through technique. Post-operative outcomes were monitored up to 3 years. The main outcome measures were early post-operative complications, best corrected visual acuity (BCVA) and endothelial cell density (ECD) at years 1, 2 and 3.Resultpl-DMEK showed a significantly higher rebubbling rate (21.3%) compared to pc-UT-DSAEK (4.8%; p < 0.001). pl-DMEK group showed a significant improvement in visual acuity (logMAR) at years 1 (0.06 vs 0.13), 2 (0.05 vs 0.12) and 3 (0.05 vs 0.12) (p < 0.05) compared to the pc-UT-DSAEK group. Mean endothelial cell density (cells/mm2) of pl-DMEK grafts did not differ (p > 0.05) compared to pc-UT-DSAEK grafts at year 1 (1745 vs 1789), year 2 (1553 vs 1495) and year 3 (1422 vs 1280).ConclusionDespite higher immediate post-operative complications in pl-DMEK, long-term clinical outcomes remain comparable or better than pc-UT-DSAEK grafts. Eye bank prepared grafts are therefore safe and reliable for challenging EK procedures.

Sequencing technologies have long limited the comprehensive investigation of large transcripts associated with inherited retinal diseases (IRDs) like Usher syndrome, which involves 11 associated genes with transcripts up to 19.6 kb. To address this, we used PacBio long-read mRNA isoform sequencing (Iso-Seq) following standard library preparation and an optimized workflow to enrich for long transcripts in the human neural retina. While our workflow achieved sequencing of transcripts up to 15 kb, this was insufficient for Usher syndrome-associated genes USH2A and ADGRV1, with transcripts of 18.9 kb and 19.6 kb, respectively. To overcome this, we employed the Samplix Xdrop System for indirect target enrichment of cDNA, a technique typically used for genomic DNA capture. This method facilitated the successful capture and sequencing of ADGRV1 transcripts as well as full-length 18.9 kb USH2A transcripts. By combining algorithmic analysis with detailed manual curation of sequenced reads, we identified novel isoforms characterized by an alternative 5' transcription start site, the inclusion of previously unannotated exons, or alternative splicing events across the 11 Usher syndrome-associated genes. These findings have significant implications for genetic diagnostics and therapeutic development. The analysis applied here on Usher syndrome-associated transcripts exemplifies a valuable approach that can be extended to explore the transcriptomic complexity of other IRD-associated genes in the complete transcriptome data set generated within this study. Additionally, we demonstrate the adaptability of the Samplix Xdrop System for capturing cDNA, and the optimized methodologies described can be expanded to facilitate the enrichment of large transcripts from various tissues of interest.

This study aims to characterize the conjunctival flora of patients scheduled for cataract surgery and determine the susceptibility profile of isolated bacteria to several commonly used topical antibiotics. Conjunctival swabs were taken from 44 consecutive patients (25 males, 19 females; mean age of 75.0 ± 12.6 years) who were scheduled for senile cataract surgery at two Italian centers before starting any prophylactic preoperative treatment. Swabs were processed for the detection of the microbial growth and for species identification. Selective culture media were used, and bacteria were identified using the MicroScan Specialty ID Panels (Beckman Coulter®, Brea, CA, USA). Antimicrobial susceptibility for the following antibiotics (netilmicin, tobramycin, ofloxacin, oxacillin, levofloxacin, moxifloxacin, chloramphenicol, cefuroxime, and azithromycin) were assessed using the Kirby-Bauer disk diffusion method. Susceptibility for oxacillin was useful to identify methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE). Among the swabs analyzed, 61.4% showed only saprophytic flora, 30.7% showed only potential pathogenic flora, and 8.0% showed mixed flora. S. epidermidis (20.5%), S. intermedius (18.2%), and S. aureus (14.8%) were the most frequent isolates; MRSA and MRSE accounted for 8.0% and 6.8% of isolates. Less frequently (9%), Gram-negative bacteria such as Pseudomonas fluorescent, Serratia marcescens, Moraxella lacunata, Morganella morgani, and Stenotrophomonas maltophila were detected. All isolated organisms showed an excellent sensitivity to moxifloxacin and chloramphenicol (range 83-100%, range 67-100%, Gram-positive sensitivity for moxifloxacin and chloramphenicol, respectively; 100% Gram-negative sensitivity for both). A significant percentage of the eyes of candidates for surgery presented potential pathogenic flora alone or in association with saprophytic organisms. Staphylococci were the most frequently isolated bacteria. Tobramycin and Ofloxacin, widely used in the ophthalmic field, are confirmed to have a reduced sensitivity in vitro.

IntroductionSynthetic materials have traditionally been used to reconstruct the diaphragm during extensive surgery for pleural mesothelioma. However, new biomaterials have shown promising results in various surgical fields. This study describes our experience using homologous fascia lata for diaphragm reconstruction in patients undergoing surgery with radical intent for pleural mesothelioma. Material and methodsData from patients who underwent extensive surgery for pleural mesothelioma requiring diaphragm reconstruction from January 2021 to December 2023 were extracted and analyzed. Patients were divided in two groups based on the type of material used for diaphragm reconstruction: expanded polytetrafluoroethylene (ePTFE) and cadaveric homologous fascia lata. ResultsForty-three consecutive patients were included in the analysis. Of these, 22 patients (51 %) underwent diaphragm reconstruction with expanded ePTFE prostheses, while 21 patients (49 %) had reconstruction using homologous fascia lata. No significant differences were found between the two groups in terms of in-hospital stay (14.5 vs 17 days, p = 0.865) and early post-operative complications (19 % vs 18 %, p = 0.943). Concerning long-term complications, a lower rate was found in the group treated with fascia lata (5 % vs 23 %, p = 0.09). ConclusionsDiaphragm reconstruction using cadaveric homologous fascia lata after extended surgery for pleural mesothelioma is feasible and results in satisfactory post-operative outcomes. Although the trend suggests fewer long-term complications with fascia lata, further research is needed to confirm these findings and determine the ideal prosthetic material for diaphragm reconstruction.

The human neural retina is a complex tissue with abundant alternative splicing and more than 10% of genetic variants linked to inherited retinal diseases (IRDs) alter splicing. Traditional short-read RNA-sequencing methods have been used for understanding retina-specific splicing but have limitations in detailing transcript isoforms. To address this, we generated a proteogenomic atlas that combines PacBio long-read RNA-sequencing data with mass spectrometry and whole genome sequencing data of three healthy human neural retina samples. We identified nearly 60,000 transcript isoforms, of which approximately one-third are novel. Additionally, ten novel peptides confirmed novel transcript isoforms. For instance, we identified a novel IMPDH1 isoform with a novel combination of known exons that is supported by peptide evidence. Our research underscores the potential of in-depth tissue-specific transcriptomic analysis to enhance our grasp of tissue-specific alternative splicing. The data underlying the proteogenomic atlas are available via EGA with identifier EGAD50000000101, via ProteomeXchange with identifier PXD045187, and accessible through the UCSC genome browser.

PurposeThis study aims to evaluate a novel medium formulation for corneal culture (MCC, AL.CHI.MI.A. Srl) at 31°C for 25 days.MethodsEighteen (n=18) donor corneas recovered in standard hypothermic medium (FBOV) were transferred to the MCC medium (n=14) or in standard corneal culture medium (Storagix, FBOV, n=4) as controls, at 31°C for up to 25 days. Endothelial cell density (ECD), Central corneal thickness (CCT) and transparency, using light microscopy, optical coherence tomography and a luxmeter were measured at 0, 14, 21 and 25 days of storage. Additionally, ECD, hexagonality (HEX%) and coefficient of variation (CV%) were evaluated by specular microscopy at the same time intervals.ResultsSimilar ECD trend was observed in corneas stored in both media over time. The remaining critical corneal quality parameters in MCC group resulted also stable during the whole incubation with following final values at day 25: CV% 41 ± 8, HEX% 48 ± 6, with a final central corneal thickness (CCT) measurements corresponding to 568 ± 31 µm after 25 days at 31°C. CV% and HEX% could not be evaluated for controls due to the increase in corneal thickness, which showed an approximately doubled CCT value as compared to the MCC group corneas. The storage in MCC allowed better preservation of endothelial morphology and corneal transparency during 25-days period preservation.ConclusionsMCC medium formulation shows is a safe and effective option for corneal culture allowing the maintenance of a corneal quality and physiological corneal thickness for 25 days of storage at 31°C