bluuebunny/crossref_metadata_2025 huggingface dataset:
Efficacy of sodium hypochlorite and peracetic acid in reducing cross‐contamination during washing of baby spinach at different water quality levels
Authors: Gao Zhujun , Jha Aprajeeta , Hudson Claire L. , Hopper Adam L. , Critzer Faith J. , Micallef Shirley A. , Schaffner Donald W. , Tikekar Rohan V.
Published: January 2025

We evaluated the antimicrobial performance of sodium hypochlorite (NaOCl) and peracetic acid (PAA) during washing of baby spinach in water of varying levels of organic load, as measured by its chemical oxygen demand (COD). Escherichia coli TVS353 was spot inoculated onto one unwashed leaf. Sanitizers were added into water with preadjusted COD (300 or 2500 ppm) to achieve concentrations from 20 to 80 ppm. One inoculated leaf was washed with nine uninoculated leaves in 500 mL water (n = 6). Bacterial load on inoculated leaves was lowered by sanitizers in a dose‐dependent manner (p < 0.05) and the lowest bacterial survivor levels were observed at 80 ppm with 2.7 ± 1.2 and 5.1 ± 0.5 Log MPN/leaf for PAA and NaOCl, respectively, at low CODs. PAA was more effective in reducing bacterial load from the inoculated leaf than NaOCl at high CODs (p < 0.05), with 2.9 ± 2.8 and 5.3 ± 0.8 Log MPN/leaf survivors for PAA and NaOCl, respectively. At 80 ppm sanitizer levels, the bacteria was not detected in wash water at any condition but was detected at 20 and 40 ppm at high CODs. The lowest levels of bacteria transferred to uninoculated leaves were observed at 80 ppm sanitizer, at 0.3 ± 0.2 and 0.2 ± 0.1 Log MPN/leaf for PAA and 1.1 ± 1.0 and 0.3 ± 0.3 Log MPN/leaf for NaOCl at low and high CODs, respectively. The log percentage of bacteria transferred ranged from −1.1 at 0 ppm to over −4.5 at 80 ppm, highlighting a reduction in cross‐contamination by the sanitizers.

Practical Application

This study provides effective data on sanitizer usage to fresh produce industry for ensuring food safety during washing of produce. It evaluated the sanitizer effect in a broad range of scenarios including various sanitizer concentrations, and wash water with low and high organic load that is common when recirculating wash water. The results also revealed the differences in two common sanitizers (PAA and NaOCl) in terms of their effectiveness.
Role of protein and lipid oxidation in hardening of high‐protein bars during storage
Authors: Dietrich Rachel B. , Lincoln Lily , Momen Shima , Minkoff Benjamin B. , Sussman Michael R. , Girard Audrey L.
Published: January 2025

Protein bar hardening negatively impacts shelf life, quality, and consumer acceptance. Although oxidation is known to negatively affect the flavor and texture of foods, the specific roles of lipid and protein oxidation in bar hardening have not been thoroughly investigated. Furthermore, most research has concentrated on dairy proteins, with a notable lack of studies addressing the hardening of plant‐based protein bars. We investigated the role of protein and lipid oxidation, Maillard reactions, moisture loss, protein aggregation, and microstructural changes in the hardening of pea, whey, and rice protein bars over a storage period of 6 weeks (hardness increased 7.2×, 5.4×, and 4.4×, respectively). Changes in tryptophan fluorescence, free sulfhydryl content (e.g., loss of 57% for pea and 44% for whey), and carbonyl content demonstrated that pea and whey bars underwent protein oxidation. Lipid oxidation also occurred, demonstrated by increased peroxide and thiobarbituric acid‐reactive substance values. Rice bars, however, did not undergo oxidation. Mass spectrometry indicated greater Maillard‐reaction‐related protein glycations formed in pea and whey bars (6.9% and 7.7%, respectively) than in rice bars (2.1%). SDS‐PAGE revealed that pea and whey, but not rice, proteins aggregated during storage. Overall, this study found that moisture loss, protein and lipid oxidation, Maillard reactions, and protein aggregation correlated with bar hardening. Chemical changes may cause protein aggregation, resulting in hardening. Likely because of rice proteins’ innate insolubility and disulfide linkages, rice protein bars were less susceptible to chemical changes and aggregation and hardened more slowly than whey and pea protein bars.

Practical Application

This study shows that lipid and protein oxidation are correlated with protein bar hardening in both pea and whey protein bars. Additionally, this work suggests that rice protein bars may harden more slowly than pea and whey bars. These findings suggest that potential strategies to prevent bar hardening and extend shelf life include (1) adding antioxidants to prevent oxidation and (2) using rice proteins to partially or fully substitute other protein isolates.
Oral barriers to food‐derived active peptides and nano‐delivery strategies
Authors: Wang Xinyu , Cao Zhaoxin , Su Jingyi , Ge Xuemei , Zhou Zhiyong
Published: January 2025

Food‐derived bioactive peptides are a class of peptides from natural protein. It may have biological effects on the human body and play a significant role in protecting human physiological health and regulating physiological metabolism, such as lowering blood pressure, lowering cholesterol, antioxidant, antibacterial, regulating immune activity, and so on. However, most of the natural food‐derived functional peptides need to overcome a variety of barriers in the body to enter the blood circulation system and target to specific tissues to generate physiological activity. During this process, the bioavailability of the functional peptides will be reduced. The nano‐delivery system can offer the feasibility to overcome these obstacles and improve the stability and bioavailability of food‐derived active peptides by nanoencapsulation. This work summarizes the application of food‐derived bioactive peptides and the obstacles during the delivery pathway in vivo. Moreover, the different nano‐delivery systems used for bioactive peptides and their application were summarized, which could provide ideas for oral delivery of food‐derived bioactive peptides.
Bacteria as Precision Tools for Cancer Therapy
Authors: Michán Carmen , Prados José , Ramos Juan‐Luis
Published: January 2025

The discovery at the end of the 20th century of genes that induce cell death revolutionised the biocontaintment of genetically manipulated bacteria for environmental or agricultural applications. These bacterial ‘killer’ genes were then assayed for their potential to target and control malignant cells in human cancers. The identification of the bacteriomes in different human organs and tissues, coupled with the observation that bacteria tend to accumulate near tumours, has opened new avenues for anti‐cancer strategies. This progress, along with recent insights into how cancer cells evade immune response, has prompted innovative therapeutic approaches. Tumour microenvironments are typically nutrient‐rich, characterised by low oxygen tensions and very resistant to immune responses. Two recent studies in MBT highlight the promise of using Salmonella typhimurium and Escherichia coli as vectors in novel cancer treatments. Engineered S. typhimurium strains can generate adjuvant flagellin‐antigen complexes that function as in situ vaccines, hence increasing the immunogenic responses within tumour environment. Similarly, gut E. coli can be used as vectors to targert tumour cells in colon cancer, enabling both diagnostic applications and localised treatments. Both approaches hold significant potential to improve patient survival outcomes.
Streamflow and Groundwater Response to Stream Restoration Using Beaver Dam Analogues in a Semi‐Arid Perennial Stream
Authors: Bosworth Lily , Smith Rose M. , Culbertson Adam , Jamison Logan R. , Burnett Paul , Brooks Paul D.
Published: January 2025

Degradation of stream and riparian environments across the western United States has damaged critical ecosystems, water quality, and increased sedimentation of reservoirs. Low tech, process‐based restoration methods such as beaver dam analogues (BDAs) have been adopted by practitioners because they restore ecosystem function, improve habitat, and reduce downstream sediment delivery at a low cost. However, few studies have examined the potential impacts of beaver dam analogues on the quantity and timing of streamflow, which is of primary concern for water stakeholders. We address this gap with 2 years of streamflow and riparian groundwater observations before and after installing a series of 17 BDAs on Fish Creek, a first‐order stream in semi‐arid northern Utah. Within 8 weeks of BDA installation, and 1 year later, we found no significant change in streamflow compared with control and Regional Reference sites. Shallow groundwater table elevations within 7 m of the stream edge increased significantly, up to 14 cm, relative to the control reach. Our results suggest that the small, local hydrological changes from BDA installation are superimposed on much larger‐scale snowmelt and alluvial groundwater controls on streamflow, suggesting BDA installation can restore ecosystem function without deleterious impacts on streamflow. These findings are especially relevant to restoration and water stakeholders with concerns about impacts of BDAs on downstream flows.
Advancements and Challenges in Immune Protection Strategies for Islet Transplantation
Authors: Wang Xue , Zeng Ziyuan , Li Dayan , Wang Kai , Zhang Wei , Yu Yang , Wang Xi
Published: January 2025

Pancreatic islet transplantation is a crucial treatment for managing type 1 diabetes (T1D) in clinical settings. However, the limited availability of human cadaveric islet donors and the need for ongoing administration of immunosuppressive agents post‐transplantation hinder the widespread use of this treatment. Stem cell‐derived islet organoids have emerged as an effective alternative to primary human islets. Nevertheless, implementing this cell replacement therapy still requires chronic immune suppression, which may result in life‐long side effects. To address these challenges, innovations such as encapsulation devices, universal stem cells, and immunomodulatory strategies are being developed to mitigate immune rejection and prolong the function of the transplant. This review outlines the contemporary challenges in pancreatic β cell therapy, particularly immune rejection, and recent progress in immune‐isolation devices, hypoimmunogenic stem cells, and immune regulation of transplants. A comprehensive evaluation of the advantages and limitations of these approaches will contribute to improved future clinical investigations. With these promising advancements, the application of pancreatic β cell therapy holds the potential to effectively treat T1D and benefit a larger population of T1D patients.
From Clinical to Benchside: Lacticaseibacillus and Faecalibacterium Are Positively Associated With Muscle Health and Alleviate Age‐Related Muscle Disorder
Authors: Liu Chaoran , Wong Pui Yan , Barua Nilakshi , Li Baoqi , Wong Hei Yuet , Zhang Ning , Chow Simon Kwoon Ho , Wong Sunny Hei , Yu Jun , Ip Margaret , Cheung Wing Hoi , Duque Gustavo , Brochhausen Christoph , Sung Joseph Jao Yiu , Wong Ronald Man Yeung
Published: January 2025

Sarcopenia is an age‐related muscle disorder that increases risks of adverse clinical outcomes, but its treatments are still limited. Gut microbiota is potentially associated with sarcopenia, and its role is still unclear. To investigate the role of gut microbiota in sarcopenia, we first compared gut microbiota and metabolites composition in old participants with or without sarcopenia. Fecal microbiota transplantation (FMT) from human donors to antibiotic‐treated recipient mice was then performed. Specific probiotics and their mechanisms to treat aged mice were identified. Old people with sarcopenia had different microbial composition and metabolites, including Paraprevotella, Lachnospira, short‐chain fatty acids, and purine. After FMT, mice receiving microbes from people with sarcopenia displayed lower muscle mass and strength compared with those receiving microbes from non‐sarcopenic donors. Lacticaseibacillus rhamnosus (LR) and Faecalibacterium prausnitzii (FP) were positively related to muscle health of old people, and enhanced muscle mass and function of aged mice. Transcriptomics showed that genes related to tricarboxylic acid cycle (TCA) were enriched after treatments. Metabolic analysis showed increased substrates of TCA cycle in both LR and FP supernatants. Muscle mitochondria density, ATP content, NAD+/NADH, mitochondrial dynamics and biogenesis proteins, as well as colon tight junction proteins of aged mice were improved by both probiotics. LR and the combination of two probiotics also benefit intestinal immune health by reducing CD8+ IFNγ+ T cells. Gut microbiota dysbiosis is a pathogenesis of sarcopenia, and muscle‐related probiotics could alleviate age‐related muscle disorders mainly through mitochondria improvement. Further clinical translation is warranted.
Does bank digitalisation reduce corporate agency costs? Evidence from China
Authors: He Shuxian , Sheng Xin
Published: January 2025

This paper explores the impact of bank digitalisation on corporate agency costs. The findings show that: (i) bank digitalisation significantly reduces agency costs by improving banks' monitoring capabilities; (ii) financial regulation further lowers agency costs for firms engaging with more digitised banks; and (iii) the effect between bank digitalisation and corporate agency costs is weaker in state‐owned enterprises and those with short‐sighted management, but stronger in firms with greater financing constraints and shorter listing durations. These results provide new evidence of bank digitalisation's positive impact on the real economy and highlight the benefits of financial regulation in China.
Aniridia associated keratopathy – molecular and clinical mechanisms leading to keratopathy development
Authors: Szentmary Nora
Published: January 2025

Congenital aniridia is a rare, panocular disease with PAX6 haploinsufficiency in 90% of the cases. In addition to PAX6, other factors appear to play a role in aniridia associated keratopathy (AAK) development. FOXC1, DSG1, FOSL2 and FABP5 are genes that are crucial for eye development, differentiation of limbal stem cells and for the regulation of lipid metabolism in epithelial cells. Recent data also indicate the importance of microRNA 204‐5p and microRNA 138‐5p in AAK. The Lagali AAK classification well describes the progressive nature of AAK, from a mild peripheral to a central corneal vascularization with corneal pannus formation. Ocular surface inflammation, as much as previous ocular surgery seem to have a role in AAK progression. Nevertheless, the corneal endothelium might possess slightly better quantitative reserves in congenital aniridia, than in healthy subjects and corneal endothelial deposition seems to be independent from the developmental abnormalities but may be related to up to date undescribed endothelial inflammatory or metabolic changes.
New insights in the regulation of ocular angiogenesis by vegf and other mediators: therapeutic implications
Authors: Ferrara Napoleone
Published: January 2025

Angiogenesis plays a key role in the pathogenesis of numerous disorders. In 1989, we reported the isolation and cloning of vascular endothelial growth factor (VEGF) as an endothelial cell‐specific mitogen and angiogenic factor. VEGF is now recognized as a central player in the regulation of angiogenesis. VEGF inhibitors result in significant clinical benefits in intraocular vascular disorders such as age‐related macular degeneration (AMD) and today such drugs represent the most effective therapy for intraocular neovascular disorders. Since the introduction of anti‐VEGF therapy, neovascular AMD is no longer the leading cause of severe vision loss secondary to AMD, which at present is geographic atrophy (GA). Novel approaches to prevent choriocapillaris dropout and GA will be discussed.
Keratoconus staging, monitoring & corvis biomechanical analysis
Authors: Flockerzi Elias , Seitz Berthold
Published: January 2025

Keratoconus remains one of the leading indications for penetrating keratoplasty worldwide. Since it can be effectively stabilized today, its early diagnosis and progression assessment is of particular importance. Detection and progression assessment are no longer limited to slit lamp examination and corneal topography, but comprise nowadays increasingly corneal tomography and biomechanics. Because keratoconus has a decreased resistance to deformation when compared to healthy corneas, measuring and analyzing the corneal deformation behavior has been reported to enable distinction between healthy and keratoconus corneas. The main Corneal Visualization Scheimpflug Technology (Corvis ST®, Oculus, Wetzlar, Germany) indices Corvis Biomechanical Index (CBI) and Tomographic Biomechanical Index (TBI) aim to distinguish between healthy and keratoconus corneas in a quasi‐binary manner, whereas the Biomechanical E‐Staging is based on the linear term of the CBI and provides information about keratoconus severity, thus enabling a biomechanical severity staging and progression assessment.
Identification of biomarkers reflecting opa1‐related dominant optic atrophy severity to infer patient cohorts eligible to clinical trials
Authors: Bouzidi Aymane , Eckmann‐Hansen Christina , Larsen Mickael , Zanlonghi Xavier , Bocquet Béatrice , Meunier Isabelle , Lenaers Guy
Published: January 2025

Dominant Optic Atrophy is a blinding disease related to optic nerve degeneration, which is caused in more than 50% of cases, by a heterozygous mutation in the OPA1 gene. Since its molecular identification in 2000, one striking observation of DOA patients is the high variability of the visual defect, ranging from almost unaffected individuals to legally blind patients, even among relatives with the same mutation. This variability is further connected to a disease progression which is also highly variable, some patients evolving quickly to blindness, whereas others display a very slow progression. Together, this situation hampers the recruitment of OPA1‐related patients for clinical trials.

To tackle this problem, we have recruited 100 OPA1 patients and 40 controls to identify biomarkers segregating with DOA severity. All patients and controls had a full ophthalmological examination and a mean follow‐up of 5 years, and all answered a visual quality of life and a nutritional questionnaire. Blood and stools were collected to perform transcriptomic, metabolomic and microbiota analyses.

My talk will present the results of this multi‐OMIC analyses, the correlation between clinical and biological data and the identification of the most relevant biomarkers segregating with the evolution of DOA disease.
Challenging case III
Authors: Salas Ester Carreno
Published: January 2025

The aim of this SIS id to explore intricate clinical scenarios to bridge the gap between bedside observations and bench research in the field of uveitis. This will consist in a case of a patient suffering from macular telangiectasias type 3 (MacTel Type 3). This presentation focuses on the detailed analysis of macrophage‐like cells present on the surface of the retina in the context of MacTel Type 3 progression. Macrophages, known for their pivotal role in immune response and tissue homeostasis, are increasingly recognized as key players in retinal diseases. Through a multidimensional approach, we will scrutinize the dynamic behavior of these cells as the disease advances, aiming to elucidate their involvement in the pathophysiology of MacTel Type 3. This comprehensive analysis not only enhances our understanding of the disease mechanisms but also paves the way for potential therapeutic interventions targeting macrophage‐mediated pathways.
Can tear analysis drive a clinical decision? – A real‐world experience
Authors: Versura Piera , Ciavarella Carmen
Published: January 2025

Ophthalmologists take advantage from imaging in their daily practice. Limiting the field to Dry Eye Disease (DED), they take their decision on clinical assessment parameters obtained primarily from observation at the slit lamp, and this makes them de facto microscopists and laboratorysts on their own. However, since it is well recognized that the biological failure precedes morphological changes observable with the current instruments, at any level, the search for markers in fluids and cells is an urgent need so far.

The role of tear analysis in the research setting is well established, and data emerging from the literature highlight the role of analysis performed on tears in the search for biomarkers for ocular and systemic/neurodegenerative diseases.

Tear analysis is an emerging technique in the clinical scenario, but with great potential expansion as an easily accessible body fluid acquired in a totally non‐invasive procedure, which contains several molecular constituents changing in levels in pathological conditions. Tears are a reliable clinical sample to be probed by highly sensitive techniques now available, even in the sensor‐based Point of Care equipments, to diagnose different conditions, with special emphasis on eye diseases.

A subjective symptom‐based algorithm for variable an heterogeneous symptoms of DED has been attempted to guide clinical decision tailored to individual patients. However, subjective evaluation might be related to a great variety of confounding factors, including personal feelings, mood, and pain threshold. An objective tool to stratify patients in DED aetiology and severity represents an unmet medical need, and tears certainly represent a biological matrix to be explored. This presentation is about the experience conducted at the Laboratory for Cornea and Ocular Surface Analysis, active at the University of Bologna and IRCCS AOU di Bologna since 1990. Records of patients stored in the lab archive are now around 12K, with DED patient proportion around 80% out of the total. Analyses on tear quantity, evaporation rate, ocular protection index, meibomian gland impairment, protein content (lysozyme C, lactoferrin, serum albumin, lipocalin 1, Zinc alpha 2‐glycoprotein, transferrin, and total protein level), scraping and imprint cytology, and inflammatory indexes (including tear cytokines) are delivered in a report. Clinicians from other disciplines beyond Ophthalmologists (Rheumatologists, Haematologists, Dermatologists, Allergologists, among others) are guided to diagnosis and therapeutic programme, including personalized management.
Retinal microglia and intraocular inflammation in human
Authors: Salas Ester Carreno
Published: January 2025

The presentation will offer a comprehensive exploration of retinal microglia dynamics with a particular focus on intraocular inflammation. Applying cutting‐edge techniques for visualizing macrophage‐like cells in the retina in vivo in humans, we can provide unprecedented insights into their behavior and distribution in various ocular pathologies. Central to the discussion is the dynamic nature of retinal microglia and their response to inflammatory stimuli. By employing advanced imaging modalities, we will elucidate how these immune cells undergo changes in density, morphology, and spatial distribution in uveitis.
Transfer of mitochondria in rpe cells – proof of concept of mitotherapy
Authors: Hytti Maria
Published: January 2025

It is a long‐held belief that mitochondria are constrained within cells. However, in recent years research has shown that cells can not only transfer mitochondria between each other using tunneling nanotubes but can even take up mitochondria directly. The transfer of isolated, functional mitochondria to dysfunctional or stressed cells has been termed ‘mitophagy’ and has been found to be beneficial in multiple models of disease. In fact, injection of isolated mitochondria into affected tissue was found to prevent the progression of experimental Parkinson's disease, to maintain tissue bioenergetics after spinal cord injury and to increase electrophysiological activity, ganglion cell survival and axon outgrowth in a mouse model of optical nerve injury.

Age‐related macular degeneration (AMD) has been extensively linked to mitochondrial damage: the number and size of mitochondria, as well as mitochondrial matrix density are reduced in the retinal pigment epithelium (RPE) of AMD patients [3‐5]. Furthermore, mitochondrial DNA damage is increased [6,7] while protein expression levels of ATP synthase and cytochrome c oxidase are reduced. This might make AMD a promising target for mitotherapy.

In vivo, RPE cells were found to take up isolated, functional mitochondria supplemented to the medium, but the benefit of this transfer has not been studied in depth. This proof of concept explores the benefits of mitotherapy on RPE cells suffering from induced mitochondrial dysfunction, looking at mitochondrial uptake and fate, cell survival, bioenergetics, and inflammation.
Species‐specific mechanisms of vertebrate eye formation
Authors: Bovolenta Paola
Published: January 2025

Neurodevelopmental visual disorders (NDVD) are complex conditions that frequently arise from failure of proper embryonic eye formation, thereby directly impacting on the functional organization of the visual areas of the brain and, indirectly, on that of other brain regions. As a consequence, NDVD are heterogeneous in nature with clinical features that often include defects other than the visual ones. Understanding how the eye forms and what does interfere with the acquisition of its characteristic cup shape is thus a prerequisite to understand how NDVD arise. I will discuss our recent studies analyzing the mechanisms that different vertebrate species uses to shape the eye primordium focusing on the role of the retina pigment epithelium (RPE). The vertebrate eye‐primordium consists of a pseudostratified neuroepithelium, the optic vesicle, in which cells acquire neural retina or RPE fates. As these fates arise, the optic vesicle assumes a cup‐shape, influenced by mechanical forces generated within the neural retina. Whether the RPE passively adapts to retinal changes or actively contributes to optic vesicle morphogenesis remained unexplored. We generated a zebrafish Tg(E1‐bhlhe40:GFP) line to track RPE morphogenesis and interrogate its participation in optic vesicle folding. We have shown that, in virtual absence of proliferation, RPE cells stretch and flatten, thereby matching the retinal curvature and promoting optic vesicle folding. Localized interference with the RPE cytoskeleton disrupts tissue stretching and optic vesicle folding. This mechanism differs from that present in amniotes, in which proliferation drives RPE expansion with a much‐reduced need of cell flattening. Thus, extreme RPE flattening and accelerated differentiation are efficient solutions adopted by fast‐developing species to enable timely optic cup formation. Analysis of transcriptomic dynamics (RNA‐seq) and chromatin accessibility (ATAC‐seq) in segregating neural retina and RPE populations highlights an early recruitment of desmosomal genes in the flattening RPE, revealing Tead factors as upstream regulators. Investigation of GRNs dynamics uncovers an unexpected sequence of TF recruitment during RPE specification, which is conserved in human‐derived organoids, despite mechanistic differences in their RPE expansion. Taking this information together we are now developing hiPSC‐derived eye organoids carrying mutations in genes responsible for congenital eye malformations, such as microphthalmia or anophthalmia, to identify which are the morphogenetic events that fail, thereby originating NDVD.
Involvement of the neurokinin 1 receptor pathway in aniridia associated keratopathy: Evidence from pre‐clinical models
Authors: Palombella Silvia , Fonteyne Philippe , Javidjam Seyedehdina , Suanno Giuseppe , Bartolomeo Nicolò , Tilaro Gianluca , Lagali Neil , Ferrari Giulio
Published: January 2025

Aniridia is a rare ocular disorder characterized by the hypoplasia or absence of the iris associated with severe visual loss. Two‐thirds of cases are generated by mutations in the PAX6 gene and are frequently associated with severe aniridia‐associated keratopathy (AAK), which is characterized by pain, persistent epitheliopathy, corneal neovascularization and progressive loss of limbal stem cell function. The pathophysiology of AAK is still unclear; however, leukocyte infiltration and nerve degeneration have been reported. Substance P (SP) is a neuropeptide secreted by nerve fibers, immune and epithelial cells in the cornea and exerts its pro‐inflammatory functions by binding to neurokinin‐1 receptor (NK1R), promoting nerve activation, leukocyte chemotaxis and extravasation, thus contributing to the inflammatory response. In this context, the modulation of NK1R activation could be used as an innovative therapy to control inflammation and pain associated with AAK.

Our main aim was to quantify the expression of NK1R and SP in the cornea of a PAX 6+/− mouse model of AAK compared to unaffected animals at different ages. This model express PAX6 gene in heterozygosis and is characterized by iris hypoplasia, abnormal lens morphology, cataracts, corneal opacification, and incomplete separation of lens from the cornea, typical features associated with AAK patients. Corneal samples were collected from mice at different ages, that is 1 month old, 2 months old, 4 months old, and 7 months old, and immunohistochemistry was performed on cross‐sections to quantify the expression of SP and NK1R. In the control samples, NK1R decreased over time albeit not significant. The expression level in the PAX6 +/− samples was comparable to controls at 1, 4, and 7 months old. However, the expression level of NK1R obtained from 2 months old mice was significantly higher compared not only to 1, 4 and 7 months old PAX6 +/− mice but also to all control groups (p < 0.001). The same trend was observed also for SP. Indeed, SP was significantly more expressed in the corneas of 2 months old mice compared to other time points and also to control groups (p < 0.01). These data confirm that the expression of SP goes together with the expression of NK1R during the development of healthy animals. Our data suggest that there may be a window of opportunity for the modulation of SP activity at 2 months of age. Future experiments will be performed in order to investigate the actual efficacy of NK1R blockade for the treatment of AAK.
Dalk – when and how
Authors: Goweida Mohamed
Published: January 2025

Deep anterior lamellar keratoplasty (DALK) is a surgical procedure that involves replacing the anterior portion of the cornea with donor tissue while preserving the patient's endothelium. This technique has gained popularity in recent years as an alternative to traditional penetrating keratoplasty (PK) due to its potential advantages in terms of visual outcomes, graft survival and reduced risk of endothelial rejection.

The decision to perform DALK in keratoconus is based on various factors, including the keratometric readings, corneal thickness, corneal clarity and RGP tolerability.

Surgical technique plays a critical role in the success of DALK. Several approaches have been described to perform the DALKs; the big bubble technique is considered the most convenient for all surgeons. Other techniques were used according to the associated pathology.

This presentation is a quick journey into DALK indications and various techniques used in eyes with keratoconus.
Choroidal neovascularization in best disease: A pediatric ophthalmology challenge
Authors: Bacci Giacomo
Published: January 2025

Choroidal neovascularisation (CNV) is a rare but serious complication of Best disease. However, visualisation of the neovascular membrane is difficult on fluorescein angiography (FA) and indocyanine green angiography (ICGA) due to dye leakage from material accumulation. The authors' study reports a case series of paediatric Best's disease where optical coherence tomography angiography (OCT‐A) contributed to the diagnosis of CNV and prompt treatment.

Three eyes of three patients with Best's disease were included. The mean age at diagnosis was 5.6 years (range 4‐7 years). OCT‐A showed the typical sea fan‐shaped neovascular membrane in three eyes, whereas in most cases conventional FA and ICGA imaging did not clearly show neovascularisation due to the masking effect of vitelliform material. OCT‐A appears to be a good alternative for diagnosing CNV in Best's disease, particularly in children, as it is a non‐invasive and rapid imaging technique that does not require the administration of dyes.

The aim was to evaluate the effect of intravitreal injections on choroidal neovascularisation in patients with Best's disease using OCT and OCT‐A and to assess the increase in visual acuity after rehabilitation. During the talk, a literature data review on pathogenesis, evolution and future perspectives will be presented.
Genetic complexity of inherited retinal diseases in a large italian cohort
Authors: Karali Marianthi , Testa Francesco , Di Iorio Valentina , Torella Annalaura , Zeuli Roberta , Scarpato Margherita , Romano Francesca , Onore Maria Elena , Pizzo Mariateresa , Melillo Paolo , Brunetti‐Pierri Raffaella , Passerini Ilaria , Pelo Elisabetta , Cremers Frans P. M. , Esposito Gabriella , Nigro Vincenzo , Simonelli Francesca , Banfi Sandro
Published: January 2025

Introduction: Inherited retinal diseases (IRDs) are the leading cause of vision loss in the working‐age population and represent one of the most genetically heterogeneous, rare Mendelian diseases. This heterogeneity mirrors a spectrum of clinical phenotypes which vary in terms of cell/tissue involvement, disease onset, severity, and progression.

Aim: To determine the genetic basis of IRD pathogenesis in a large Italian cohort (n = 2790) followed at a single referral center.

Methods: We performed a retrospective epidemiological study by reviewing the clinical records and genetic data of the IRD patients followed at the Eye Clinic of the University of Campania ‘Luigi Vanvitelli’ (Naples, Italy). Almost 70% of cases were analysed by Next Generation Sequencing approaches using panels of known retinopathy genes, clinical exome or whole exome sequencing. A third of the molecular diagnoses were performed by candidate gene sequencing, APEX‐based genotyping microarrays and single molecule Molecular Inversion Probes.

Results: We provided conclusive molecular diagnosis for 2036 patients (from 1683 unrelated families). The overall diagnostic success rate was 73%. We identified a total of 1327 causative sequence variations in 127 genes, including 348 novel variants, and 866 potentially actionable genotypes for therapeutic approaches. ABCA4 was the most frequently mutated gene (n = 535; 26.3% of solved cases), followed by USH2A (n = 228; 11.2%) and RPGR (n = 102; 5%). About 60% of RPGR variants were detected in ORF15, a mutational hotspot refractory to NGS analyses. The other 124 genes had a lower contribution to IRD pathogenesis (e.g. CHM 3.5%, RHO 3.5%; MYO7A 3.4%; CRB1 2.7%; RPE65 2%, RP1 1.8%; GUCY2D 1.7%). Seventy‐eight genes were mutated in five patients or less. Mitochondrial DNA variants were responsible for 2.1% of cases. The combined genetic and clinical assessment uncovered novel genotype‐phenotype associations and phenotypic expansions.

Conclusions: We report on the largest cohort of molecularly characterised IRD patients from Italy to date. Our findings confirm the complex genetic etiology of IRDs and reveal the high prevalence of ABCA4 and USH2A mutations. The results suggest that mutations in RPGR‐ORF15 likely account for a significant fraction of male IRD patients who remain undiagnosed after a first tier NGS analysis. This study also uncovers novel genetic associations with a spectrum of clinical phenotypes and identifies numerous cases potentially eligible for clinical trials and, ultimately, for molecular therapies.

Keywords: Inherited retinal diseases, genetic epidemiology, retinitis pigmentosa, Stargardt disease, Leber Congenital Amaurosis
Panretinal photocoagulation for the treatment of proliferative diabetic retinopathy
Authors: Karakosta Christina , Chatziralli Irini , Kabanarou Stamatina A.
Published: January 2025

After the completion of DRS study in 1979, Panretinal Photocoagulation (PRP) became the gold standard for the treatment of proliferative diabetic retinopathy.

Light from the laser is absorbed by the retinal pigment epithelium (RPE), and the underlying choroid and it is converted to thermal energy. Those thermal burns denature tissue protein, causing local retinal cell death, coagulative necrosis and eventually scarring. In that respect, laser photocoagulation leads to diminished oxygen demands of the areas of capillary non perfusion, and reduced expression of vasoactive factors such as vascular endothelial growth factor (VEGF) and protein kinase C (PKC) in the retina. As a result, adequate oxygen and nutrients are diverted towards the non‐ischemic retinal areas.

As documented by the DRS study, after a 5‐year follow‐up period, the rates of severe visual loss were diminished by at least 50% in all eyes treated with PRP in comparison with untreated control eyes. The greatest beneficial effect was shown for treated eyes with high risk PDR characteristics (any NVD with extent greater than or equal to one‐fourth disc area without vitreous or preretinal hemorrhage, any NVD with vitreous or preretinal hemorrhage,any NVE with extent greater than or equal to one‐half disc area, with vitreous or preretinal hemorrhage). Harmful effects of PRP includes peripheral visual field loss, decrease in visual acuity, nyctalopia etc. Treatment of non‐high risk PDR cases, could also be proposed under certain circumstances such as in patients with difficulty to attend regular appointments, advanced diabetic disease in the fellow eye etc.

Although anti‐VEGF agents have been approved for the treatment of PDR, PRP remains an effective therapeutic option and the choice of treatment could be made on an individualized basis. However, in high risk PDR patients with co‐existing maculopathy the addition of anti‐VEGF injections to PRP treatment is advisable.
Ophthalmic trauma
Authors: Blatsios Georgios
Published: January 2025

Ophthalmic trauma presents a medical challenge for every ophthalmic surgeon. Every trauma is unique, thus it is difficult to establish invariable treatment standards. Meticulous primary globe repair is a prerequisite for successful further surgeries. During secondary repair the surgeon has to achieve best possible visualisation, to create space and drain the choroid if necessary, for maximum tamponade effect, to deal with the haemorrhage the possible retinal detachment, to remove an intraocular foreign body if present and to apply the proper tamponade to stabilize the globe. Additional scleral buckle or more extreme manoeuvres, such as choroidal suturing or intraocular amniotic membrane transplantation to cover bare sclera are other options, yet to be validated. The greatest risk for failure is PVR and subsequent hypotony. At the moment there is insufficient data about the role of intravitreal Methotrexate to prevent PVR. Advanced anterior segment rehabilitation, such as scleral fixated IOLs, artificial iris etc. should follow long term successful posterior segment treatment in cases of severe trauma.
Visual evoked potential
Authors: Neveu Magella
Published: January 2025

The visual evoked potential (VEP) is a measure of time locked visual cortical activity, measured at the scalp, to a controlled stimulus that enables objective assessment of the integrity of the retino‐cortical visual pathway. VEP waveform characteristics and morphology are dependant upon signal generators, in response to pattern or flash, achromatic or chromatic stimuli, and positioning of the active recording electrodes over the visual area of interest (e.g. V1 or MT).

In clinical electrophysiology, the VEP is recorded according to the ISCEV (International Society of Clinical Electrophysiology of Vision) standards using pattern reversal, pattern onset‐offset or flash stimulation. The clinical uses of the VEP include assessment of optic nerve dysfunction consequent upon demyelination, ischaemia, or nutritional deficiency; genetically determined optic neuropathy such as Autosomal Dominant Optic Atrophy (DOA), Leber Hereditary Optic Neuropathy, and intracranial misrouting of the retino‐cortical projection, as associated with albinism.

Although the VEP is used clinically primarily to assess optic nerve, chiasmal and cortical function, any disturbance in the visual pathway may result in an abnormal VEP, such as eye closure, optical degradation and retinal dysfunction. In addition, the clinical presentation in optic nerve disease and macular/retinal is often similar. Clinical features common in both include visual acuity loss, a pale disc, colour vision defect and less commonly, a RAPD. These factors must be taken into consideration when interpreting the VEP, therefore recording of the pattern ERG, multifocal ERG (mfERG) and/or ERG in conjunction with the VEP enables more accurate assessment of visual pathway function and the distinction between optic nerve disease and retinal disease.
Visual acuity outcomes by sex in a historical population of patients with leber hereditary optic neuropathy from case record survey‐2 (CRS‐2)
Authors: Votruba Marcela , Leroy Bart , Yu‐Wai‐Man Patrick , Van Everdingen Judith , Krawczynski Maciej , Lamperti Costanza , Carelli Valerio , Llòria Xavier , Klopstock Thomas
Published: January 2025

Aims/Purpose: Leber hereditary optic neuropathy (LHON) is a rare mitochondrial disorder resulting in severe bilateral vision loss. Age at symptom onset and estrogen levels are thought to influence disease progression. The Case Record Survey‐2 (CRS‐2; NCT02796274) was conducted to establish the natural history of LHON in idebenone‐naïve patients. Here, we report 12‐month clinical outcomes by sex and age at symptom onset.

Methods: Retrospective clinical data were extracted from case records of LHON patients. Eligibility: aged ≥12y, m.11778G>A, m.3460G>A, or m.14484T>C mitochondrial DNA mutation, symptom onset after 1999, ≥2 visual acuity (VA) assessments within 5y of symptom onset. Patients were subset by sex and age at symptom onset. Spontaneous clinically relevant benefit (sCRB), recovery (sCRR), stabilisation (sCRS), and worsening (sCRW) at 12 ± 3 months from baseline (first VA assessment after symptom onset) were calculated in subacute/dynamic eyes (≤1y since symptom onset) as previously described1.

Results: Overall, male and female eyes had similar rates of sCRB (male: 21.0% [17/81]; female: 20.0% [3/15]) and sCRW (male: 65.1% [41/63]; female: 69.2% [9/13]); however male eyes had a lower rate of sCRR (male: 11.1% [9/81]; female: 20.0% [3/15]) and a higher rate of sCRS (male: 32.4% [11/34]; female: 0.0% [0/8]). In females, there were no eyes aged <15y at symptom onset. Eyes aged 15–50y at onset had a higher rate of sCRR (15–50y: 33.3% [3/9] vs >50y: 0.0% [0/6]) and a lower rate of sCRW (15–50y: 50.0% [4/8] vs >50y: 100.0% [5/5]) compared to eyes aged >50y. In males, eyes aged 15–50y at onset had the lowest rate of sCRR (<15y: 19.0% [4/21]; 15–50y: 6.0% [3/50]; >50y: 20.0% [2/10]) and the highest rate of sCRW (<15y: 47.4% [9/19]; 15–50y: 77.1% [27/35]; >50y: 55.6% [5/9]) compared to eyes aged <15y and >50y.

Conclusions: The clinical course of LHON may differ based on sex and age at symptom onset.

References: 1. Yu‐Wai‐Man P, et al. Cell Reports Medicine 2024;5:101437.
Comparative efficacy of receptor tyrosine kinase inhibitors in a mouse model of alkali burn‐induced corneal neovascularization
Authors: Gladkauskas Titas , Rundgren Ida , Cristea Ileana , Rødahl Eyvind , Bredrup Cecilie
Published: January 2025

Aims/Purpose: This study evaluates the efficacy of topical dasatinib and axitinib for treating experimentally induced corneal neovascularization (CNV) in a mouse alkali burn model, comparing these treatments to established therapies like dexamethasone and bevacizumab. The study aims to determine which treatment provides superior efficacy in inhibiting CNV in this model.

Methods: Thirty‐six C57BL/6;129Sv mice underwent a standardized alkali burn to induce CNV in both eyes using a paper disc soaked in 1M NaOH. The mice were randomly assigned to one of six treatment groups: saline (0.9% sodium chloride), DMSO (5%), dexamethasone (0.1%), bevacizumab (0.5%), dasatinib (0.5%), or axitinib (0.5%). Treatments were applied topically three times daily. CNV assessments, including corneal neovascularization area (CNA), vessel length index (VLI), and limbus vasculature thickness, were conducted post‐mortem using corneal flat‐mounts stained with CD31 antibody for immunohistochemistry.

Results: Dexamethasone proved the most effective in inhibiting alkali burn‐induced CNA (p < 0.0001), with bevacizumab showing comparable efficacy (p < 0.001). Axitinib also effectively reduced CNA (p < 0.001) and VLI (p < 0.01). In contrast, dasatinib did not significantly reduce CNA (p = 0.74) or VLI (p = 0.98).

Conclusions: While dexamethasone was most effective, both bevacizumab and axitinib successfully reduced CNA, with axitinib additionally decreasing VLI. Axitinib has a broad‐spectrum inhibition of vascular endothelial growth factor receptors 1, 2, and 3 (VEGFR1, VEGFR2, VEGFR3), and platelet‐derived growth factor receptor β (PDGFRβ). This may offer superior benefits compared to narrower‐spectrum TKIs like dasatinib, which primarily target PDGFRβ and fibroblast growth factor receptor (FGFR). The results highlight the effectiveness of targeting the VEGF‐VEGFR pathway for managing corneal neovascularization, presenting a viable option for patients who are sensitive to steroids.
Efficacy and tolerability of a new riboflavin‐based dye compared to double staining with fluorescein and lissamine green in the evaluation of corneal‐conjunctival epithelial defects: preliminary data
Authors: Troisi Mario , Caruso Ciro , Troisi Salvatore , Bellucci Carlo , Costagliola Ciro
Published: January 2025

Aims/Purpose: To evaluate the diagnostic sensitivity and tolerability of a new 0.5% riboflavin vital dye (Remark®, Servimed Ind. SpA, Rome, IT), on corneal and conjunctival epithelial defects, compared to standard stains (fluorescein and lissamine green).

Methods: 56 eyes of 28 patients (15M, 13F; mean age 51.3 ± 9) with bilateral ocular surface epithelial defects were evaluated. In one eye of each patient selected randomly, after the application in the conjunctival fornix of Lissamine Green Ophthalmic strip (GreenGloTM, Stag Medical, Raleigh, NC) moistened with an eye drop of fluorescein 0.5% solution (Fluorofta®, Fidia Farmaceutici S.p.A., Abano Terme, IT), SLE was carried out using cobalt blue filter to evaluate corneal staining and white filter for the conjunctiva. Grading was assessed according to NEI score. After 30 minutes and washing with 5 drops of saline instilled every 10 minutes, a similar evaluation was carried out with cobalt blue light and yellow filter in the eyepiece after instillation of Remark® (Group A). The same evaluation was performed in the fellow eyes by reversing the sequence of dyes (Group B). A symptom score questionnaire was obtained for each group.

Results: The average corneal NEI score was 3.32 (Gr. A) and 3.12 (Gr. B) for double staining, 3.38 (Gr. A) and 3.22 (Gr. B) for Remark®; average conjunctival score was 5.12 (Gr. A) / 4.88 (Gr. B) for double staining and 5.04 (Gr. A) / 4.76 (Gr.B) for Remark®. The differences between the two staining methods were not statistically significant in both groups (p>0.05). In no case an intolerance to the products used has been recorded.

Conclusions: The new riboflavin‐based dye was effective in detecting both corneal and conjunctival defects, without statistically significant differences compared to double staining, and with a good tolerability. The study highlights the potential of this new dye overcoming the limit of the need of 2 different dyes, reducing costs and evaluation time.