bluuebunny/crossref_metadata_2025 huggingface dataset:
The bioaccessibility of phenolic compounds, nutritional quality, and textural properties of gluten‐free muffins enriched with artichoke leaves and green lentil protein isolate
Authors: Gulsunoglu‐Konuskan Zehra , Yagdi Sevgi Deren , Ersoy Burcu
Published: January 2025

Although the gluten‐free market is expanding and offers a variety of products, there are still some deficiencies in the nutritional and sensory quality of these products. Therefore, this study explores the bioaccessibility of phenolic compounds, nutritional quality, and textural properties of gluten‐free muffins enriched with artichoke leaves and green lentil protein (GLP) isolate, two novel ingredients introduced together for the first time in this context. The incorporation of GLP isolate aims to enhance the protein content, while artichoke leaves are evaluated for its potential to improve phenolic content and antioxidant activity. The muffins were also subjected to in vitro digestion to assess the bioaccessibility of phenolic compounds. The results demonstrated that the addition of GLP isolate increased the protein content 1.4 times, while artichoke leaves contributed to enhance the total phenolic content (TPC), flavonoid content (TFC), antioxidant activity (TAA), and ash content by 2.1, 5.4, 3.2–3.5, and 1.3 times, respectively. Nevertheless, the addition of artichoke leaves increased the hardness, gumminess, and chewiness of gluten‐free muffins (p < 0.05). The bioaccessibility values of gluten‐free muffins varied between 138% and 220%, 142% and 206%, and 40% and 160% for TAA, TPC, and TFC, respectively. Chlorogenic acid, ferulic acid, and quercetin derivatives were found as main phenolic compounds in gluten‐free muffins enriched with artichoke leaves in undigested and all phase of digestion. This study provides valuable insights into the development of functional gluten‐free muffins, highlighting the potential of artichoke leaves and GLP isolate as innovative ingredients in gluten‐free food products.

Practical Application

This study shows how adding green lentil protein and artichoke leaves to gluten‐free muffins can make them healthier by boosting their protein, antioxidants, and other nutrients. However, while muffins became more nutritious, their texture changes. Food companies could use this information to improve gluten‐free products, but they may need to adjust the recipe to improve the sensory properties.
Chinese yam (Dioscorea opposita Thunb.) starch‐ferulic acid complexes: Preparation, characterization, and physicochemical properties
Authors: Tan Shuqiong , Chen Huiqing , Huang Yating , Liu Suqi , Zheng Ziyan , Guo Zebin , Xie Sandu
Published: January 2025

Polyphenols are known to interact with starch to form the V‐type inclusion complex or the noninclusive complex. It is hypothesized that the addition of polyphenols could improve the properties of Chinese yam (Dioscorea opposita Thunb.) starch, and the properties of the complexes could be regulated by controlling the additive amount of polyphenols. To test this hypothesis, the effect of varying ferulic acid (FA) concentrations (3%, 9%, and 15%) on the structural characterization and physicochemical properties of Chinese yam starch (CYS) was investigated. X‐ray diffraction analysis showed that CYS complexes with FA generated characteristic peaks comparable to those of native CYS, but FA caused the crystal morphology of CYS to change from a B‐type to V‐type. During the gelatinization process, FA increased the solubility, water‐holding capacity, and hardness of CYS complexes, which was possibly attributed to the CYS molecules reaggregating and the formation of ordered crystals. The transmittance of CYS complexes decreased from 12.21% to 7.46% when the FA concentration increased from 3% to 15%. In addition, FA increased the viscosity and elasticity of the CYS system but decreased retrogradation. Thus, FA improved the structure and properties of CYS complexes, which can provide new directions and ideas for the development of CYS‐based food.

Practical Application

This study showed that adding FA can affect the characterization and physicochemical properties of CYS. We expect that CYS‐FA complexes can be used for health and medicine products thanks to unique nutritional values.
Fiber and polyphenol enriched biscuits using date palm byproduct: Physiochemical characteristics, sensory properties, in vitro digestion, and storage stability
Authors: Ranasinghe Meththa , Alghaithi Mariam , Mugdil Priti , Sundarakani Balan , Stathopoulos Constantinos , Maqsood Sajid
Published: January 2025

Functional biscuit was formulated by fortifying them with polyphenolic extract and fiber‐rich residue of defatted date seed powder (DDSP) obtained through microwave‐assisted extraction. Effect of particle size (small, medium, and large) and substitution level (2.5%, 5%, and 7.5%) of fiber‐rich residue, along with the phenolic extract, on bioactive, physical, textural, and gastrointestinal digestion of fortified biscuits was studied followed by sensory and shelf‐life studies. The total phenolic content (TPC) and antioxidant properties of biscuits increased with increasing substitution levels and particle size. DDSP residue fortification increased the fiber content in biscuits. Large particles of the residue‐fortified biscuits showed significantly higher (p < 0.05) hardness compared to the control biscuits. Diameter of the biscuits decreased with increasing substitution level and particle size of fiber‐rich residue with the lowest value of 50.66 mm in 7.5% substitution of large particles. The 7.5% substitution level of small particles resulted in the lowest spread ratio of 8.97 and the highest thickness of 5.79 mm. Consumer perceptions were at an acceptable level after the fortification, with an average sensory score of 6.02 out of 9 for overall acceptability. After 24 weeks storage, TPC decreased in biscuits, but TPC retention increased with increasing substitution level and particle size of residue. Thiobarbituric acid reactive substances (TBARS) value of biscuits increased with storage. The highest phenolic recovery was observed in the intestinal phase of the gastrointestinal digestion with the highest recovery of 102.33 at 2.5% level of large particles. Thus, phenolic extract and fiber‐rich residue incorporation was effective to enhance the nutritional and functional properties of biscuits.

Practical Application

Date seeds are rich in bioactive components and fiber. This study demonstrated the feasibility of utilizing date seeds to improve nutritional and functional properties of bakery products. The incorporation of the microwave‐assisted polyphenolic extract and the fiber‐rich residue of defatted date seed powder into biscuit enhanced the bioactive, nutritional, and functional characteristics while maintaining the consumer acceptance. This research contributes to the valorization of byproducts in the agriculture and food industries, promoting sustainability and a bio‐circular economy.
Effectiveness of oregano essential oil vapor on shelf life extension of kai lan (Brassica oleracea var. alboglabra)
Authors: Shu Weichen , Deng Zhuoliang , Liu Lingdai , Zhang Jiaxuan , Li Dan
Published: January 2025

This study compared the antimicrobial activity of several essential oils (EOs) vapor against food spoilage microbiota and further investigated the potential of EO vapor in extending the shelf life of leafy green vegetables. Oregano EO vapor showed stronger antimicrobial activities than basil and clove EO vapors against common spoilage‐causing microorganisms in fresh produce, including Pantoea agglomerans, Pseudomonas cichorii, Pectobacterium carotovorum, Pantoea ananatis, Pseudomonas marginalis, Alterneria bassicicola, and Botrytis cinerea. When oregano EO vapor was applied to leafy greens, phytotoxic effects were observed on butter lettuce and iceberg lettuce but not on kai lan and kale. On the contrary, the green color and total chlorophylls of kai lan were preserved by oregano EO vapor. With the use of oregano EO vapor generated by 10 µL oregano EO in a 1.8‐L container, the shelf life of kai lan extended from 3 days to at least 7 days when stored at 25°C and from 5 days to at least 14 days when stored at 7°C. The antimicrobial effect was studied with the plating counting method to reflect the dose‐dependent amicrobial effect of oregano EO vapor on kai lan. High‐throughput 16S/18S DNA sequencing method was applied to evaluate the microbial ecological changes of kai lan under oregano EO vapor treatment. Taken together, our results suggest that oregano EO vapor could be used to extend the shelf life of kai lan. However, the dose of usage must be optimized with care, and one must realize the effectiveness might vary essentially when applying the EO vapor on other types of leafy green vegetables.
A review of enhancing noodle quality: Fortification with micronutrients and substitution with nutrient‐dense alternative
Authors: Ayustaningwarno Fitriyono , Suryaputra Aditya , Anjani Gemala
Published: January 2025

Noodles are usually rich in carbohydrates but lack essential nutrients such as protein, fiber, vitamins, and minerals, potentially causing metabolic problems if consumed in the long term. This review explores strategies to improve the quality of noodles through substitution and fortification. Substitution is replacing the main ingredient with a more nutrient‐dense alternative, such as sweet potato starch, which has been shown to improve the nutritional content of noodles, such as fiber and beta carotene. Meanwhile, fortification is the addition of micronutrients to food products to restore or enhance levels of micronutrients that are not present in the food, which is usually done during the production process. However, substitution and fortification have their challenges. The substitution method usually causes a decrease in the physical quality of the noodles due to changes in the raw materials, thus reducing consumer acceptance of the final product. Meanwhile, fortified nutrients usually undergo considerable degradation during production, reducing their effectiveness. Further research and development should focus on new methods to help maintain physical quality while minimizing the loss of fortified or substituted nutrients. Technological approaches such as microencapsulation have proven effective in maintaining the stability of micronutrients such as L‐5‐methyltetrahydrofolate added during production. The methods of cold plasma, γ‐irradiation, and multifrequency ultrasound, as well as the addition of xanthan gum and guar gum, are able to improve the moisture level, color, texture, elasticity, and chewiness, as well as consumer sensory acceptance. This technology enables manufacturers to meet customer demand for quality and nutritious noodle options.
Recovery trajectories after major abdominal surgery: A retrospective pooled cohort study
Authors: Ou‐Young Jared , Royse Colin , Clarke‐Errey Sandy , El‐Ansary Doa , Riedel Bernhard , Griffiths James , Bowyer Andrea
Published: January 2025

Background

Recovery from major surgery can be difficult to predict given the many factors involved in treating disease and restoring preoperative function. Postoperative recovery metrics such as length of stay, complications, and mortality are typically described. However, large data quantities for patient‐reported recovery are scarce. In this retrospective study, we aimed to describe the multidimensional recovery trajectory of patients undergoing major abdominal surgery 4–8 weeks after surgery and explore factors related to incomplete overall recovery.

Methods

We retrospectively analysed pooled cohort data of adults undergoing elective major abdominal surgery between 2018 and 2024 across three tertiary‐level hospitals. Recovery was measured at postoperative days 1, 3, 7, 14, weeks 4–8, and 3 months using the Postoperative Quality of Recovery Scale (PostopQRS). Physiological, nociceptive, emotive, activities of daily living (ADL), and cognitive domains were assessed, with recovery defined as a return to, or improvement of, preoperative baseline levels. Overall recovery was defined as recovery in all domains. Length of stay was assessed for patients who recovered overall, or did not recover, at postoperative weeks 4–8.

Results

Six hundred and fifty‐three patients were included, with mean (SD) age of 57.8 (14.4) years. Of these, 36% were aged ≥65 years and 58% were male. The incidence of overall recovery at postoperative week 4–8 was 42%. Domain‐level recovery at postoperative weeks 4–8 was 63% for nociception, 81% for emotion, 82% for ADLs, and 83% for cognition. Patients failing to achieve overall recovery at weeks 4–8 had longer mean (SD) lengths of stay compared to those who recovered (11.3 (10.3) vs. 7.3 (7.1) days, p < .001).

Conclusions

The incidence of overall recovery at postoperative week 4–8 was 42%. Patients with incomplete overall recovery had longer lengths of stay. Multidimensional strategies to improve the recovery trajectory warrant further investigation.

Editorial Comment

Major surgical procedures are often followed by a lengthy and difficult recovery period. Traditional measures such as mortality and complications are usually analysed, but this 653‐patient study investigated patient‐reported recovery scores after major abdominal surgery. Novel findings include that only 42% of patients have recovered fully in all recovery domains at weeks 4–8, and these patients also had longer hospital stays. Preoperative risk factors were analysed for associations with recovery trajectories.
Carbon Storage Variation of Plantation Forest and Their Management Practices in Amhara, Ethiopia
Authors: Kassie Yishak Adgo , Teshome Yirdaw Meride
Published: January 2025

The rise in atmospheric carbon dioxide (CO2) and its potential to alter the climate are currently major global concerns. Forest plants and soil may absorb and retain carbon from the atmosphere, forming significant terrestrial carbon storage. The aims of this study were to estimate the carbon storage variance of Eucalyptus globulus and Cupressus lusitanica species in community and small‐scale plantation forests under different management practices in the study area. A systematic sampling technique was used to collect field data. The transect line and the sample plot were positioned at 100 and 50 m, respectively, between them. The diameter at breast height (DBH) and total tree height were measured using a total of 60 plots (10 m × 10 m) from community plantation forests and 30 plots from small‐scale plantation forests. The total mean carbon storage of C. lusitanica was 312.2 ± 97.9 t C/ha, and that of E. globulus was 356.1 ± 117.6 t C/ha. This was equivalent to 1146.4 ± 359.2 t C/ha CO2 (g) of C. lusitanica and 1306.9 ± 431.8 t C/ha CO2 (g) of E. globulus. The total mean carbon storage of E. globulus was higher than that of C. lusitanica in all carbon pools, except for litterfall biomass. From the management practices in small‐scale plantations, the total mean carbon storage of C. lusitanica and E. globulus was 120.5 ± 37.5 t C/ha and 130.5 ± 40.8 t C/ha, respectively. This was equivalent to 442.2 ± 137.5 and 478.9 ± 149.7 t C/ha of CO2 (g), C. lusitanica and E. globulus, respectively. The results of this study will identify research gaps for future investigations and assist in addressing knowledge gaps for forest managers and organisations committed to providing enough attention to forest conservation.
Barriers to the Cross‐Border Diffusion of Climate Change Policies
Authors: Vu Trung V.
Published: January 2025

This paper establishes a statistically and economically significant relationship between national responses to climate change and genetic distance, which is a proxy for countries' dissimilarities in cultures, ancestry, and historical legacies associated with long‐term exposure to divergent historical trajectories. It finds that countries that are genetically distant to the world‐leading nation‐state of climate change mitigation tend to experience barriers to the cross‐border diffusion of climate change policies and hence exhibit worse responses to climate change. A potential explanation is that climate change polices are more likely to spread between closely related countries with more similar preferences for the provision of the public goods of environmental and climate protection. The findings imply that strengthening climate change mitigation requires overcoming obstacles to international policy diffusion.
The future of ocular imaging: functional OCT versus deep learning
Authors: Schmetterer Leopold
Published: January 2025

The future of ocular imaging lies at the intersection of two powerful technologies: Functional Optical Coherence Tomography (OCT) and Deep Learning. Functional OCT represents a leap forward in imaging technology by integrating traditional OCT with functional measurements like blood flow, tissue stiffness, tissue birefringence, oxygen metabolism and optoretinography. This comprehensive may offers deeper insights into ocular health and disease progression than conventional structural OCT. By capturing both structural and functional information, Functional OCT may our ability to detect and monitor conditions such as myopia, glaucoma, diabetic retinopathy, and age‐related macular degeneration. Deep Learning, on the other hand, is a subset of artificial intelligence, analyzing the vast amounts of data generated by ocular imaging modalities. Through sophisticated algorithms, Deep Learning can identify intricate patterns and features in images that may elude human interpretation. This capability not only enhances diagnostic accuracy but also aids in automating image analysis, improving efficiency, and reducing the burden on healthcare professionals. The synergy between Functional OCT and Deep Learning holds immense promise for the future of ocular imaging. By integrating these technologies, we can achieve a comprehensive understanding of ocular physiology and pathology while enhancing diagnostic accuracy and efficiency. This convergence may lead to the development of advanced systems capable of real‐time analysis and interpretation, facilitating point‐of‐care diagnostics and remote monitoring. However, realizing this potential requires addressing various challenges, including data privacy concerns, standardization of imaging protocols, and regulatory approval. Collaborative efforts between researchers, clinicians, and industry stakeholders are essential to overcoming these hurdles and harnessing the full benefits of Functional OCT and Deep Learning in ophthalmic practice.
Gene therapy – can we do better?
Authors: Yu‐Wai‐Man Patrick
Published: January 2025

Inherited optic neuropathies are genetically heterogeneous. The two main groups are Leber hereditary optic neuropathy (LHON) caused by primary mitochondrial DNA (mtDNA) point mutations and autosomal dominant optic atrophy (DOA) caused by mutations in the nuclear‐encoded OPA1 gene. Gene therapy is an obvious therapeutic approach aimed at correcting the underlying genetic defect to prevent the ongoing loss of retinal ganglion cells and stabilise or improve vision. Although promising results have been obtained from phase III clinical trials for LHON based on allotopic expression of the MT‐ND4 gene for the common m.11778G>A mtDNA mutation, several unexpected issues have emerged calling into question the optimal gene delivery system, the ideal therapeutic window for intervention, and whether interocular transfer of the gene therapy vector can occur. Resolving these controversies will be crucial given the fact that other gene therapy trials are in the pipeline for LHON and OPA1 DOA.
Mitochondrial rejuvenation as a multipurpose therapy in eye disease
Authors: Hombrebueno Jose R.
Published: January 2025

Restoration of mitochondrial quality control (MQC) has emerged as a promising therapeutic strategy in ocular disease, including diabetes, scarring and aging‐related disorders. However, the success of potential interventions depends upon our understanding of how MQC mechanisms, mainly mitochondrial dynamics (fusion/fission) and turnover (mitophagy and biogenesis) interplay in specific ocular cells during pathology. To date, this knowledge has been constrained by the lack of models to accurately assess such a complex interplay in vivo, and consequently, by the limited access to drug discovery platforms that closely recapitulate altered MQC in ocular diseases. Over the recent years, our lab has developed and integrated state‐of‐the‐art platforms to comprehensively understand how dysregulated MQC impacts ocular disease, offering unique opportunities for the strategic development of therapies aimed at rescuing mitochondrial fitness. I will discuss our main studies and recent advancements describing how altered MQC impacts ocular disease, particularly diabetes and ocular scarring, and how targeting its underlying molecular processes can help devising versatile mitochondrial‐targeted interventions.
Acanthamoeba keratitis – conservative and surgical treatment
Authors: Szentmary Nora
Published: January 2025

Clinical signs of acanthamoeba keratitis are in early stages grey‐dirty epithelium, pseudodendritiformic epitheliopathy, perineuritis, multifocal stromal infiltrates, ring infiltrate and in later stages scleritis, iris atrophy, anterior synechiae, secondary glaucoma, mature cataract, and chorioretinitis. Beside triple‐topical‐therapy, in therapy resistant cases surgical treatment options such as corneal cryotherapy, amniotic membrane transplantation, riboflavin‐UVA crosslinking, and penetrating keratoplasty are applied.
Potential therapeutic targets in human limbal epithelial cells in congenital aniridia
Authors: Stachon Tanja , Zimmermann Julia , Trusen Simon , Berger Maximilian , Amini Maryam , Suiwal Shweta , Fries Fabian N. , Seitz Berthold , Szentmáry Nóra
Published: January 2025

Subjects with congenital aniridia and PAX6 haploinsufficiency develop aniridia‐associated keratopathy (AAK) in most of the cases. Development of AAK is related to limbal stem cell insufficiency (LSCI), with lack of stem cells and destruction of their niche. Loss of limbal epithelial stem cell (LEC) function results in corneal neovascularization, inflammation and opacification. In AAK, alterations in corneal epithelial cell gene expression differ from other types of LSCI. Primary LECs of aniridia subjects lack KRT3 and KRT12 mRNA and protein expression and their DSG1, ADH7, FABP5, CRABP2, SPINK7 and ALDH1A1 mRNA expression is downregulated. However, miRNA 138‐5p expression is upregulated. In addition, there is an altered stress resistance of congenital aniridia stromal cells (AN‐LFCs) and an increased expression of inflammatory related cytokines such as Interleukin‐1β and Interleukin‐6 in PAX6‐knockdown LECs and AN‐LFCs. A better understanding of the sensitivity of AN‐LECs and AN‐LFCs to inflammation and oxidative stress, as well as the interaction of epithelial and stromal cells, offers new therapeutic targets.
Electrical synapses in the optic nerve head – A novel model of gap junctions' involvement in optic nerve function
Authors: Smedowski Adrian
Published: January 2025

Retinal neurons are part of the central nervous system, making them unable to regenerate spontaneously when damaged. Glaucoma, a progressive optic neuropathy, is believed to be the primary cause of severe vision impairment or permanent vision loss. Connexins, essential channel proteins that create gap junctions between neighboring cells, including neurons, play a significant role in maintaining cell stability. However, in neurons, they can also contribute to the spread of apoptotic insults, which can lead to secondary neurodegeneration.

The discovery of previously unknown electrical synapses known as gap junctions between ON axons, which directly connect axons within bundles in the ON head, potentially speeds up signal transmission along the ON and allows for modulation of signal passage from the retina to the brain. By enabling crosswise conduction within ON bundles, these synapses could potentially help bypass localized damage within axons. It is hypothesized that the density and conductivity of these synapses may play a crucial role in determining the susceptibility of the ON to developing various impairments that lead to symptomatic pathologies.

Research has shown that temporarily blocking ON electrical synapses chemically can slow down the conduction of visual signals. In cases of structural or functional impairments in axons, signals could potentially be transmitted crosswise through gap junctions to neighboring axons, thereby maintaining the continuity of the syncytial ON structure and preventing the blockage of visual information propagation. This discovery holds significant implications for understanding the development of different optic neuropathies and highlights a potential new target for therapeutic interventions.
An uncommon neuronal class conveys visual signals from rods and cones to retinal ganglion cells
Authors: Tian Ning , Du Rui , Prodzinski Quinn , Wang Ping
Published: January 2025

We recently described a previously not fully characterized interneuron in the mammalian retina, the Campana cells. These cells share some morphological, physiological, and molecular features with both retinal bipolar cells and amacrine cells. We further studied Campana cells by revealing their distribution in mammalian species, their synaptic connections, and their roles in visual recognition.

We show that all tested mammals, including rodents, non‐rodent mammals, primates, and humans except rabbits, express Campana cells in their retina. The density of Campana cells in the central retina is much higher than that of the peripheral retina. Also, the nasal and ventral retinas have more Campana cells than those of the temporal and dorsal retina. Further, the density of Campana cells increases significantly with age in adult mice.

We characterized presynaptic connections of Campana cells with photoreceptors using two‐photon calcium imaging of light‐evoked responses of Campana cells. We showed that Campana cells responded to both rod‐ and cone‐mediated synaptic signals, which is different from bipolar cells that selectively connect to either rods or cones. The synaptic connections between Campana cells and RGCs are determined by recording calcium transients of RGCs to optogenetic stimulation of single channel rhodopsin (ChR2)‐expressing Campana cell. We showed that Campana cells relay both excitatory and inhibitory synaptic signals to multiple functional RGC types, including rod‐, cone‐mediated, or mixed ON, OFF, and ON‐OFF RGCs, and around 50% RGCs of WT mice receive synaptic inputs from Campana cells.

To determine whether Campana cells could relay visual signals to RGCs without retinal bipolar cells, we recorded the light‐evoked calcium responses of Campana cells and RGCs of VSX2‐SEΔ/Δ mice, in which the bipolar cells are completely absent. In these mice, Campana cells are present and respond to both rod‐ and cone‐mediated synaptic signals. Around 25% of RGCs of VSX2‐SEΔ/Δ mice exhibited light‐evoked responses.

To further determine whether Campana cells could evoke visual perception without bipolar cells, we accessed the visual responses of VSX2‐SEΔ/Δ mice using a fear conditioning test, in which mice learn to associate conditional stimuli (CS, such as light or visual patterns) with aversive unconditional stimuli (US; a mild electrical foot shocks) first and subsequently tested the conditional response (CR/freezing of movement) to CS without US. Accordingly, we use uniform light and checkboard as conditional stimuli (CS) to determine the light perception and visual pattern perception of VSX2‐SEΔ/Δ mice. We showed that, in WT mice, the freezing time is increased by 12.5 folds to light stimuli and 2.2 folds to checkboard. In the VSX2‐SEΔ/Δ mice, the freezing time is increased by 8.2 folds to light and 2.4 folds to checkboard. Therefore, the Campana cells could relay visual signals to CNS through RGCs for visual perception without bipolar cells.
Technical innovations in liquid biopsies
Authors: Nell Rogier J.
Published: January 2025

Uveal melanoma is a rare and aggressive intraocular malignancy. The majority of diagnoses are made on the basis of clinical examination solely, without taking a tumour biopsy. As more patients are treated by local, eye‐preserving radiotherapy instead of an enucleation, tumour tissue becomes even less available for further analysis. Nevertheless, the molecular characterisation of a patient's primary uveal melanoma may yield clinically‐important information.

Over the past decades, prognostically‐relevant molecular tumour subtypes have been identified based on the genetic alterations present in a uveal melanoma. These subtypes present independently from established clinicopathological markers such as tumour diameter and prominence. Patients with an increased risk of metastatic dissemination may be offered a more intense follow‐up screening program, and could possibly benefit from earlier detection of metastasis. Additionally, knowledge about the molecular profile of the primary tumour has been positively associated with various aspects of psychosocial well‐being and might be of (future) clinical relevance with regard to novel (neo‐)adjuvant therapies.

Digital PCR is one of the most sensitive molecular technologies available to analyse solid and liquid biopsies from cancer patients. It is widely used in a variety of malignancies, and in different stages of disease. We developed novel assays and experimental setups to make this technology applicable in the context of uveal melanoma. These were validated and applied in primary and metastatic tumour specimens, and in vitreous fluid samples obtained from eyes with a uveal melanoma. Others used this technology for the analysis of blood from patients with primary or metastatic uveal melanoma.

In this talk, the challenges and opportunities of digital PCR in the context of uveal melanoma will be discussed, and compared to those of other technologies available in the field.
Integration of ipsc‐derived human photoreceptors into rodent models of retinal degeneration
Authors: Ader Marius
Published: January 2025

A main cause for vision impairment and blindness is the loss of light‐sensing photoreceptors in the retina. However, photoreceptors do not regenerate in mammals including humans, thus cell transplantation has emerged as a potential treatment approach for blinding diseases. Therefore, a pipeline for assessing the transplantation of human stem cell‐derived photoreceptors into mouse models of retinal generation was developed. Human photoreceptor‐specific reporter iPSC‐derived retinal organoids were used for isolation of donor cells via flow‐cytometry. After transplantation we observed extensive incorporation of human photoreceptors in mice with cone dysfunction/loss. This incorporative phenotype was validated in both cone‐only as well as pan‐photoreceptor transplantations. Rather than forming a glial barrier, Müller cells extend throughout the graft, and additionally form a series of adherens junctions between mouse and human cells, reminiscent of an outer limiting membrane. Donor‐host interaction appears to promote transplant polarization as well as development of morphological features critical for light detection, namely formation of inner‐ and outer‐segments mainly oriented towards the host retinal pigment epithelium. Putative synapse formation and graft function was observed both at the structural level and by electrophysiological analysis using micro‐electrode recordings. Transcriptomic analysis reveals increased expression of cone photoreceptor maturation genes with time after transplantation, in comparison to cones of age‐matched retinal organoids in vitro. While highly integrative donor cell clusters were observed in partially degenerated recipients, transplantation into hosts with late‐stage degeneration, i.e. with complete photoreceptor loss, lead to more wide‐spread distribution of donor cells within the sub‐retinal space and reduced signs for integration and maturation. Thus, interaction with the host retina appears to be important for graft integration, maturation, polarization and function. Overall, these results show that human photoreceptors interact readily with a partially degenerated mouse retina.

Funding:

BMBF – ReSight (01EK1613A)

DFG – AD375/7‐1 (SPP2127), SEQ2146

FFB – TA‐RM‐0522‐0824‐TUD‐TRAP
Inherited optic neuropathies – what you need to know
Authors: La Morgia Chiara
Published: January 2025

Inherited optic neuropathies (ION) are in the large majority of cases due to mitochondrial defects and characterized by a preferential damage of the papillo‐macular bundle of the optic nerve, which is the most vulnerable to mitochondrial dysfunction.

The two most common inherited optic disorders are Leber's Hereditary Optic Neuropathy (LHON) and Dominant Optic Atrophy (DOA) due to mutations in the OPA1 gene.

LHON is due in 90‐95% of cases to the 3 most common LHON mtDNA mutations (11778/ND4, 3460/ND1 and 14484/ND6) affecting complex I of the mitochondrial respiratory chain, but can be also due to rarer mtDNA mutations and also to recessive mutations in the DNAJC30 gene (especially in patients from East European countries). Clinically, LHON is characterized by subacute painless visual loss bilateral at onset or sequentially bilateral leading to usually severe visual impairment and diffuse optic atrophy. Males are more frequently affected than females. The only therapy approved for LHON is idebenone. Gene therapy options are currently under investigation.

DOA due to OPA1 mutations is characterized by bilateral progressive visual loss starting usually in childhood with a variable degree of severity even in the same family. There are no approved therapies currently available for this disease. DOA, as LHON is characterized by complex I defect and idebenone has been used with some promising results also in this disorder. Gene therapy options are also under evaluation.

In the last years, the number of genes associated with ION is continuously growing and in a recent Italian reappraisal of the 5 most common nuclear genes responsible genes for ION, besides OPA1 which is the most common there are also WFS1, ACO2, SPG7 and AFG3L2 (Fiorini et al., Brain 2023). Interestingly, all of these genes may be causative for ION also in dominant fashion. Interestingly, SSBP1, implicated in mtDNA replication has been recently associated with autosomal dominant isolated optic atrophy but also to more complex phenotypes.

Pathogenic mechanisms associated with these new genes are discussed with particular focus on mitochondrial dynamics and mtDNA maintenance/replication.
Abnormal findings
Authors: Hommer Anton
Published: January 2025

The aim of this part is to show the difference between normal and abnormal findings in the angle. In addition grading systems and their therapeutic consequences will be discussed.

Congenital disorders and acquired diseases affecting the eye and angle will be shown. The clinical findings will be discussed as well its context of recent advances in glaucoma surgery with microsurgical techniques requiring intraoperative gonioscopy for implants.
Optical coherence tomography – basic optical principles
Authors: Irsch Kristina
Published: January 2025

Optical Coherence Tomography (OCT) is an optical analogue to ultrasound imaging. The much higher speed of light compared with sound allows for finer cross‐sectional views of the retina and anterior segment. Since it is extremely difficult to directly detect the shorter “echo” times it takes light to travel from different structures at axial distances within the eye, interferometry is used. Incident light is thus split into two beams, and the beam backscattered from the ocular tissue is then compared (“interfered”) with the beam that has traveled a known time from the reference mirror. Broadband (i.e., low “coherence”) light sources are used, because they produce a wider band of wavelengths, and thereby enable greater sensitivity in comparing the travel time differences of the two beams. In time‐domain OCT, the reference mirror position is altered, so that interference patterns are generated whenever the two beams have traveled almost the same amount of time. In spectral‐domain OCT, the reference mirror position is fixed and the mixed interference patterns are separated via spectral wavelength analysis. This lecture will explain underlying concepts of OCT and its technological variations.
How can we ensure quality simulation based education?
Authors: Labuschagne Mathys
Published: January 2025

There are certain recommendations to guide institutions planning to incorporate simulation‐based education (SBE) into their training programs or institutions that have an established program but want to benchmark it against best practices. Following these recommendations ensures quality SBE.

Create an effective simulation learning environment that is safe, trustful, supportive, and ethical. Well trained educators and facilitators complying with educational requirements such as pre‐briefing, small group learning techniques, skills transference aligned with the clinical environment and effective debriefing will ensure quality training.

Ethical principles and values must be applied to SBE not only to address patient safety, students’ psychological safety, protection of simulated patients and staff and educators, but also the simulators and skills task trainers. The ethical values for all stakeholders should adhere to resect for autonomy, beneficence, nonmaleficence, justice, and human dignity.

Embedding SBE in curricula can address the challenges on the clinical training platform, shewed burden of disease, patient safety and an environment that is safe and supportive for learning. Some jurisdictions, such as the Royal College of Ophthalmologists, have adopted this concept and implemented mandatory, demonstrable simulation‐based learning requirements as part of professional credentialing. Continued advancement of the quality of education and the establishment of clear universal essential comparable quality standard milestones are therefore advised.

Accreditation of simulation centres play a crucial role in the quality of SBE. Accreditation must promote patient safety, develop organisational structures, and maintain program management. Educational components must provide an adequate number and variety of well‐structured, high‐quality simulation learning opportunities. Review is an integral part of any training program to keep a simulation training program relevant and appropriately contextualized. Research development to inform the educational programs, clinical applications, and product development.

The creation of collaborations and partnerships under mutual agreements upon the goals and objectives of a program. Long term sustainability can be ensured by mentoring of local faculty and conceding to local curriculum plans.

Well‐structured and well‐planned contextualized training programs, where simulation could be employed as an educational methodology, are required to address global eye care needs. Trainee ophthalmologists who use simulation are more confident and perform safe eye surgeries. The training should be based on sound simulation educational principles. One must also keep in mind that simulation‐based ophthalmic surgery training goes far beyond the simulators or task trainers alone, as outlined in these recommendations.
Defocus incorporated multiple segments lenses and 0.025% atropine for myopia control in European children: 12‐month results of a randomized controlled trial
Authors: Villahoz Noemi Guemes , Talavero‐Gonzalez Paula , Bella‐Gala Rafael , Porras‐Angel Paloma , Hernandez‐Garcia Elena , Martin‐Garcia Beatriz , Ruiz‐Pomeda Alicia , Gomez‐de‐Liano C. Nunila , Shah Rakhee , Feijoo Julian Garcia , Gomez‐de‐Liano Rosario
Published: January 2025

Purpose: Defocus Incorporated Multiple Segments (DIMS) lenses and atropine have proven standalone efficacy in controlling myopia progression in children. However, there is a scarcity of evidence of their efficacy when used in combination. This randomized controlled trial (RCT) aims to evaluate the efficacy of combination treatment using 0.025% atropine and DIMS spectacle lenses compared to 0.025% atropine and single vision (SV) lenses in slowing myopia progression in European myopic children.

Methods: RCT conducted on children aged 4‐16 years with myopia between ‐1.00 and ‐6.00D and astigmatism ≤2.00D. Children were randomly assigned to the 0.025% atropine and SV lenses treatment group (group A) or 0.025% atropine and DIMS lenses treatment group (group B). Cycloplegic spherical equivalent refraction (SER) and axial length (AL) were measured at baseline, 6 and 12 months. Statistical analyses (Mann‐Whitney U‐test or Chi‐squared test) were performed to test for significance between the two groups.

Results: 79 patients completed the 12 month‐follow‐up: n = 38 (48.1%) in group A (47.4% female), mean age 9.00 ± 2.74 years; n = 41 (51. 9%) in group B (46.3% female), mean age 9.68 ± 2.65 years. There was no significant association between group and sex (p = 0.93) or age difference between the groups (p = 0.26). At 12 months, mean AL ± SD change was 0.16 ± 0.13mm in group A and 0.09 ± 0.16mm in group B, mean difference 0.07 ± 0.02 mm (95%CI 0.03‐0.12, p < 0.001). Mean SER ± SD progression was ‐0.16 ± 0.31D and ‐0.16 ± 0.36D in group A and B, respectively (p = 0.61). 39.5% of the children in Group B had no axial elongation over 12 months, compared to 14.5% of the children in group A (p < 0.001).

Conclusions: Combination treatment with 0.025% atropine and DIMS spectacle lenses is more effective in controlling axial elongation than 0.025% atropine with SV lenses in European children. The differences in SER between the groups were not significant. The AL increase in group B was less than that previously reported with DIMS lenses alone in Asian and European children over 12 months, suggesting that other factors, such as pupil size, may enhance the DIMS lens efficacy in controlling axial elongation.
Extracellular vesicles from retinal pigment epithelium: Key players in the outer blood‐retinal barrier disruption and choroidal neovascularization in age‐related macular degeneration
Authors: Martins Beatriz , Rodrigues Teresa , Boia Raquel , Ramalho José , Ambrósio António Francisco , Girão Henrique , Fernandes Rosa
Published: January 2025

Aims/Purpose: Age‐related macular degeneration (AMD), a degenerative disease causing irreversible central vision loss in the elderly, is characterized by dysregulation of the retinal pigment epithelium (RPE). During AMD, stressed RPE releases extracellular vesicles (EVs) carrying bioactive cargo, potentially disrupting the outer blood‐retinal barrier (oBRB) and accelerating AMD progression. Mechanisms behind EV‐induced oBRB disruption and their role in choroidal neovascularization (CNV) are unclear. Our study aims to assess how RPE‐derived EVs under inflammatory conditions impact oBRB integrity and CNV in AMD.

Methods: We used highly polarized primary cultures of porcine RPE (pRPE), porcine eyecups with the RPE exposed, and human umbilical vein endothelial cells (HUVEC). RPE and HUVEC cells were treated with TNF, LPS, or EVs derived from inflamed RPE cells. Additionally, Balb/c mice were intravitreally injected with RPE‐derived EVs.

Results: Treatment with TNF or LPS reduced the transepithelial resistance (TER) of pRPE cells monolayer and disrupted the tight junction complexes. Although inflammatory stimuli reduced EVs release from the apical membrane of RPE cells, increased CD63 levels and MMPs levels and activity were found in EVs. Moreover, these EVs led to the disruption of the RPE monolayer. In the Matrigel assay of HUVEC, angiogenesis was induced by both TNF and LPS, and by RPE‐derived EVs isolated after the LPS stimulus. At seven days following intravitreal injection, EVs induced outer retinal structural changes in the Balb/c mice.

Conclusions: Our study indicates that EVs released from RPE cells following an inflammatory insult contribute to the oBRB disruption and induce neovascularization, indicating that EVs may play a key role in the onset and progression of AMD.

Support: GOAP, Bayer; FCT, Portugal: 2020.04811.BD (to BM), PEst UIDB/04539/Base/2020 and UIDP/04539/Programatico/2020 through POCI‐01‐0145‐FEDER‐007440, CENTRO‐01‐0145‐FEDER‐000008: BRAINHEALTH2020
Correlation of biomarkers of oxidative stress and inflammation in tears and aqueous humor of primary open‐angle glaucoma patients
Authors: Sanchez Javier Benitez‐del‐Castillo , Duran Maria Dolores Pinazo , Sáez Sara Mora , Ruiz David Peña , Zanon‐Moreno Vicente , Ramos‐Lopez Juan Francisco , Galarreta‐Mira David
Published: January 2025

Aims/Purpose: Biomarkers improve our knowledge of the risk factors and pathogenic mechanisms of eye disorders. Validation of biomarkers is crucial for addressing an effective translation to the clinical practice. We aimed to find the correlation between oxidative stress (OS) and inflammation (INF) biomarkers obtained from two commonly sampled ocular fluids: tears and aqueous humor.

Methods: A longitudinal, prospective, case‐control study was conducted in 220 participants that were divided into: patients diagnosed of primary open‐angle glaucoma (POAG; n = 115), and patients diagnosed of cataracts, as the comparative group (CG; n = 105). All participants underwent an interview and standardized ophthalmological examination. Reflex tears were collected from the inferior meniscus by capillarity. Aqueous humor was collected at the onset of the programmed glaucoma or cataract surgery. Both samples were stored at ‐80°, until processing to enzyme‐linked immunosorbent assay (ELISA) technique, to determine OS by products (malondialdehyde ‐MDA‐/4 hydroxynonenal ‐4HNE‐/total antioxidant capacity ‐TAC‐) and INF mediators (IL1b, IL6, TNFa, INFg). The R‐core team statistical program was used for data processing.

Results: Mean age was 72 + 9 years in the POAG group and 65 + 10 years in the CG. Gender distribution was homogeneous in both groups of participants. Significantly higher MDA, 4HNE, IL1b, IL6, TNFa, and INFg and lower TAC, were found in tears from the POAG group respect to the CG (p < 0.001). Also, a statistically significant differential profile of OS (MDA, 4HNE, TAC) and INF (IL1b, IL6, TNFa, and INFg) molecules was detected in the aqueous humor of the POAG patients versus the CG (p < 0.001). Specific significant correlations were found between the above molecules and the imaging data regarding disease stages, as well as, importantly, between the tears and aqueous humor results.

Conclusions: We described a series of OS and INF biomarkers that can provide information of the disease severity. Furthermore, the OS and INF biomarker levels in tears have been shown to correlate with the aqueous humor in POAG patients, but not in the CG. These data enhance the value of non‐invasive tear collection for biomarkers analyses, that may be one useful driver of glaucoma molecular diagnosis and prognosis.