Ultraviolet Cross-linking Research Articles

Poly (lactic acid) electrospun nanofiber membranes: Advanced characterization for biomedical applications with drug loading performance studies

Traditional dressings have shortcomings such as poor moisture absorption and easy to adhere, making the development of new dressings crucial. In this work, a PLA/PVP crosslinked drug-loaded nanofiber membrane was prepared through electrospinning and ultraviolet crosslinking, with poly (lactic acid) (PLA), polyvinylpyrrolidone (PVP), and salicylic acid (SA) as starting materials. The results demonstrated that the inclusion of PVP notably boosted the viscosity and conductivity of the blend spinning solution. The roughness of the fabricated fiber was elevated, and the diameter of the fibers was more uniform. Additionally, the incorporation of PVP not only enhanced the porosity of the fiber membrane but also effectively decreased its contact angle. Notably, when the PVP content reached 40 %, the contact angle underwent a substantial reduction, decreasing significantly from 125.4° to 82.2°. The SA drug-loaded fiber membrane exhibited a notable bacteriostatic effect against Escherichia coli and Staphylococcus aureus, with its release behavior adhering to Fick's diffusion law. In the cell viability experiment, the cell proliferation rate increased from 94 % to 129 % after 3 days. This shows that the prepared membrane has good antibacterial effect and cell compatibility, which provides a theoretical basis for the construction of a new medical dressing.

IrCLIP-RNP and Re-CLIP reveal patterns of dynamic protein associations on RNA.

RNA binding proteins ( RBPs ) control varied processes, including RNA splicing, stability, transport, and translation 1-3 . Dysfunctional RNA-RBP interactions contribute to the pathogenesis of human disease 1,4,5 , however, characterizing the nature and dynamics of multiprotein assemblies on RNA has been challenging. To address this, non-isotopic ligation-based ultraviolet crosslinking immunoprecipitation 6 was combined with mass spectrometry ( irCLIP-RNP ) to identify RNA-dependent associated proteins ( RDAPs ) co-bound to RNA with any RBP of interest. irCLIP-RNP defined landscapes of multimeric protein assemblies on RNA, uncovering previously unknown patterns of RBP-RNA associations, including cell-type-selective combinatorial relationships between RDAPs and primary RBPs. irCLIP-RNP also defined dynamic RDAP remodeling in response to epidermal growth factor ( EGF ), uncovering EGF-induced recruitment of UPF1 adjacent to HNRNPC to effect splicing surveillance of cell proliferation mRNAs. To identify the RNAs simultaneously co-bound by multiple studied RBPs, a sequential immunoprecipitation irCLIP ( Re-CLIP ) method was also developed. Re-CLIP confirmed binding relationships seen in irCLIP-RNP and detected simultaneous HNRNPC and UPF1 co-binding on RND3 and DDX3X mRNAs. irCLIP-RNP and Re-CLIP provide a framework to identify and characterize dynamic RNA-protein assemblies in living cells.

Self-powered wearable remote control system based on self-adhesive, self-healing, and tough hydrogels

The advent of the fifth-generation mobile communication network era induces a great potential for the design and development of self-powered wearable electronic devices. Hydrogels, as a typical material for flexible electrodes, have been widely employed in self-powered wearable electronic devices. However, the hydrogel-based triboelectric nanogenerator (H-TENG) is usually compromised by the challenges in data collection, operational stability, and manufacturability due to the unstable combination between the friction layer and the electrode layer, the occurrence of hydrogel damage during prolonged operation, and also the difficulty of machining sticky hydrogel electrodes. In the present study, a composite hydrogel composed of acrylamide, crotonyl alcohol, and 4-carboxyphenylboronic acid was synthesized via ultraviolet crosslinking, which was specially developed for H-TENG and wearable wireless remote control interface, featuring with excellent stretchability and mechanical strength as well as self-healing and self-adhesive properties. The open-circuit voltage of the H-TENG prepared with the hydrogel reached up to 165 V, and the output voltage remained almost unchanged following 10,000 cycles of compression test. Additionally, a wearable wireless remote control system was designed that could accurately control the multiple appliances and adjust different working modes using the H-TENG. Consequently, the development of the H-TENG and wearable wireless remote control system has the potential to provide a new methodology for the application of next-generation wearable devices in various areas, such as smart homes, as a representative example.

Chemical crosslinking extends and complements UV crosslinking in analysis of RNA/DNA nucleic acid-protein interaction sites by mass spectrometry.

UV (ultra-violet) crosslinking with mass spectrometry (XL-MS) has been established for identifying RNA-and DNA-binding proteins along with their domains and amino acids involved. Here, we explore chemical XL-MS for RNA-protein, DNA-protein, and nucleotide-protein complexes in vitro and in vivo . We introduce a specialized nucleotide-protein-crosslink search engine, NuXL, for robust and fast identification of such crosslinks at amino acid resolution. Chemical XL-MS complements UV XL-MS by generating different crosslink species, increasing crosslinked protein yields in vivo almost four-fold and thus it expands the structural information accessible via XL-MS. Our workflow facilitates integrative structural modelling of nucleic acid-protein complexes and adds spatial information to the described RNA-binding properties of enzymes, for which crosslinking sites are often observed close to their cofactor-binding domains. In vivo UV and chemical XL-MS data from E. coli cells analysed by NuXL establish a comprehensive nucleic acid-protein crosslink inventory with crosslink sites at amino acid level for more than 1500 proteins. Our new workflow combined with the dedicated NuXL search engine identified RNA crosslinks that cover most RNA-binding proteins, with DNA and RNA crosslinks detected in transcriptional repressors and activators.

Solution-processed high-k photopatternable polymers for low-voltage electronics.

High dielectric constant (k) polymers have been widely explored for flexible, low-power-consumption electronic devices. In this work, solution-processable high-k polymers were designed and synthesized by ultraviolet (UV) triggered crosslinking at a low temperature (60 °C). The highly crosslinked network allows for high resistance to organic solvents and high breakdown strength over 2 MV cm-1. The UV-crosslinking capability of the polymers enables them to achieve a high-resolution pattern with a feature size down to 1 μm. Further investigation suggests that the polar cyano pendants in side chains are responsible for increasing the dielectric constant up to 10 in a large-area device array, thereby contributing to a low driving voltage of 5 V and high field-effect mobility exceeding 20 cm2 V-1 s-1 in indium gallium zinc oxide (IGZO) thin-film transistors (TFTs). In addition, the solution-processable high-k dielectric polymers were utilized to fabricate flexible low-voltage organic TFTs, which show highly reliable and reproducible mechanical stability at a bending radius of 5 mm after 1000 cycles. And also, the high radiation stability of the dielectric polymers was observed in a UV-sensitive TFT device, thereby achieving highly reproducible pattern recognition, which is promising for artificial optic nerve circuits.

RBP-Tar – a searchable database for experimental RBP binding sites

Background: RNA-binding proteins (RBPs) play a critical role in regulating gene expression by binding to specific sites on RNA molecules. Identifying these binding sites is crucial for understanding the many functions of RBPs in cellular function, development and disease. Current experimental methods for identifying RBP binding sites, such as ultra-violet (UV) crosslinking and immunoprecipitation (CLIP), and especially the enhanced CLIP (eCLIP) protocol, were developed to identify authentic RBP binding sites experimentally. Methods: To make this data more accessible to the scientific community, we have developed RBP-Tar (https://ncbr.muni.cz/RBP-Tar), a web server and database that utilises eCLIP data for 167 RBPs mapped on the human genome. The web server allows researchers to easily search and retrieve binding site information by genomic location and RBP name. Use case: Researchers can produce lists of all known RBP binding sites on a gene of interest, or produce lists of binding sites for one RBP on different genomic loci. Conclusions: Our future goal is to continue to populate the web server with additional experimental datasets from CLIP experiments as they become available and processed, making it an increasingly valuable resource for the scientific community.

Rapid and Efficient Isolation of Total RNA-Bound Proteomes by Liquid Emulsion-Assisted Purification of RNA-Bound Protein (LEAP-RBP).

The critical roles of RNA-binding proteins (RBPs) in all aspects of RNA biology fostered the development of methods utilizing ultraviolet (UV) crosslinking and method-specific RNA enrichment steps for proteome-wide identification and assessment of RBP function. Despite the substantial contributions of these UV-based RNA-centric methods to our understanding of RNA-protein interaction networks, their utility is constrained by biases in RBP recovery and significant noise contributions, which can confound meaningful interpretation. To overcome these issues, we recently developed a method termed Liquid Emulsion-Assisted Purification of RNA-Bound Protein (LEAP-RBP) and introduced quantitative signal-to-noise (S:N)-based metrics for the proteome-wide identification of RNA interactomes and accurate assessment of global RBP occupancy dynamics. Compared to existing methodologies, LEAP-RBP provides significant advantages in speed, cost, efficiency, and selectivity for RNA-bound proteins. In this work, we provide a step-by-step guide for the successful application of the LEAP-RBP method for both small- and large-scale investigations of RNA-bound proteomes. Key features • Unbiased and efficient isolation of total RNA-bound protein, RNA, and protein from biological samples. • Cost-effective identification of proteome-wide RNA interactomes and validation of direct RNA-binding protein functionality. • Robust and accurate assessment of context- and/or condition-dependent RBP occupancy state dynamics.

Decoding protein-RNA interactions using CLIP-based methodologies.

Protein-RNA interactions are central to all RNA processing events, with pivotal roles in the regulation of gene expression and cellular functions. Dysregulation of these interactions has been increasingly linked to the pathogenesis of human diseases. High-throughput approaches to identify RNA-binding proteins and their binding sites on RNA - in particular, ultraviolet crosslinking followed by immunoprecipitation (CLIP) - have helped to map the RNA interactome, yielding transcriptome-wide protein-RNA atlases that have contributed to key mechanistic insights into gene expression and gene-regulatory networks. Here, we review these recent advances, explore the effects of cellular context on RNA binding, and discuss how these insights are shaping our understanding of cellular biology. We also review the potential therapeutic applications arising from new knowledge of protein-RNA interactions.

IGF2BP2 Maintains Retinal Pigment Epithelium Homeostasis by Stabilizing PAX6 and OTX2.

N6-methyladenosine (m6A) methylation is a chemical modification that occurs on RNA molecules, where the hydrogen atom of adenine (A) nucleotides is replaced by a methyl group, forming N6-methyladenosine. This modification is a dynamic and reversible process that plays a crucial role in regulating various biological processes, including RNA stability, transport, translation, and degradation. Currently, there is a lack of research on the role of m6A modifications in maintaining the characteristics of RPE cells. m6A readers play a crucial role in executing the functions of m6A modifications, which prompted our investigation into their regulatory roles in the RPE. Phagocytosis assays, immunofluorescence staining, flow cytometry experiments, β-galactosidase staining, and RNA sequencing (RNA-seq) were conducted to assess the functional and cellular characteristics changes in retinal pigment epithelium (RPE) cells following short-hairpin RNA-mediated knockdown of insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). RNA-seq and ultraviolet crosslinking immunoprecipitation with high-throughput sequencing (HITS-CLIP) were employed to identify the target genes regulated by IGF2BP2. adeno-associated virus (AAV) subretinal injection was performed in 6- to 8-week-old C57 mice to reduce IGF2BP2 expression in the RPE, and the impact of IGF2BP2 knockdown on mouse visual function was assessed using immunofluorescence, quantitative real-time PCR, optical coherence tomography, and electroretinography. IGF2BP2 was found to have a pronounced effect on RPE phagocytosis. Subsequent in-depth exploration revealed that IGF2BP2 modulates the mRNA stability of PAX6 and OTX2, and the loss of IGF2BP2 induces inflammatory and aging phenotypes in RPE cells. IGF2BP2 knockdown impaired RPE function, leading to retinal dysfunction in vivo. Our data suggest a crucial role of IGF2BP2 as an m6A reader in maintaining RPE homeostasis by regulating the stability of PAX6 and OTX2, making it a potential target for preventing the occurrence of retinal diseases related to RPE malfunction.

Preparation and photocuring performance of itaconic acid-derived hyperbranched macromers with internal and terminal C[dbnd]C bonds catalyzed by tetramethylguanidine

A series of hyperbranched polymers derived from biomass itaconic acid was prepared via aliphatic nucleophilic substitution polycondensation reaction under mild conditions by using bis(2-bromoethyl) itaconate and trimeric acid as A2 and B3 monomers, respectively, and tetramethylguanidine as catalyst. On this basis, the hyperbranched macromers that contained C = C double bonds in the internal skeleton and the periphery were obtained by further adding A1 monomer (2-bromoethyl acrylate) directly into the reaction system. The successful synthesis of these macromers were confirmed via proton nuclear magnetic resonance, Fourier transform infrared and gel permeation chromatography. The results of photo-differential scanning calorimetry and rheological experiments showed that these macromers exerted a positive effect on the ultraviolet (UV) curing of commercial resin. The properties of cured splines and coatings, such as mechanical properties, pencil hardness, and thermal stability, were considerably improved with the addition of hyperbranched macromers. They were significantly better than those of linear macromers and macromers that contained polymerizable groups only in the inner framework or their terminal units. The results show that both the inner and terminal C = C double bonds of hyperbranched macromers can participate in UV cross-linking. Thus, such macromers may be used as potential additives for high-performance UV curable coatings.

Thermal aging behavior and heat resistance mechanism of ultraviolet crosslinked ultra‐high molecular weight polyethylene fiber

AbstractAs advances in heat resistant ultrahigh molecular weight polyethylene (UHMWPE) fiber via ultraviolet (UV) crosslinking, it is worth paying attention to its thermal aging behavior and heat resistance mechanism. UHMWPE fibers in different forms, the fully drawn yarn (FDY) and the UV crosslinked FDY (UVFDY), are subjected to isothermal aging at 135°C. The effects of thermal aging on their surface morphology and mechanical properties are studied and compared initially. The results demonstrate that UVFDY is a kind of a heat‐resistant fiber. Subsequently, to reveal the heat resistance mechanism of UVFDY, its crystalline and orientation structure evolution during thermal aging are further studied. It is indicated that the formation of dense‐packing crystals and disorientation behavior lead to fiber shrink, so that a number of grooves appear on the fiber surface. Since the crosslinked stucture of UVFDY hinders its molecules movement, the crystallization and disorientation behavior of UVFDY are weaker than those of FDY, resulting in UVFDY possesses slighter thermal shrinkage (Ts), fewer grooves and better mechanical properties than those of FDY. Therefore, the heat resistance of the fiber improves effectively after UV crosslinking. This work provides insights into the structure–property relationship of UVFDY during thermal aging, and offers basis data for its application in specific conditions.

Sticky Multicolor Mechanochromic Labels.

Sticky-colored labels are an efficient way to communicate visual information. However, most labels are static. Here, we propose a new category of dynamic sticky labels that change structural colors when stretched. The sticky mechanochromic labels can be pasted on flexible surfaces such as fabric and rubber or even on brittle materials. To enhance their applicability, we demonstrate a simple method for imprinting structural color patterns that are either always visible or reversibly revealed or concealed upon mechanical deformation. The mechanochromic patterns are imprinted with a photomask during the ultraviolet (UV) cross-linking of acrylate-terminated cholesteric liquid crystal oligomers in a single step at room temperature. The photomask locally controls the cross-linking degree and volumetric response of the cholesteric liquid crystal elastomers (CLCEs). A nonuniform thickness change induced by the Poisson's ratio contrast between the pattern and the surrounding background might lead to a color-separation effect. Our sticky multicolor mechanochromic labels may be utilized in stress-strain sensing, building environments, smart clothing, security labels, and decoration.

3D Foaming Printing Biomimetic Hierarchically Macro-Micronanoporous Hydrogels for Enhancing Cell Growth and Proliferation.

Hydrogel, recognized as a promising biomaterial for tissue engineering, possesses notable characteristics, including high water uptake, an interconnected porous structure, and excellent permeability. However, the intricate task of fabricating a hierarchically macro-micronanoporous structure, essential for providing adequate space for nutrient diffusion and cell growth within hydrogels, remains a formidable challenge. In response to these challenges, this study introduces a sustainable and straightforward three-dimensional (3D) foaming printing strategy to produce hierarchically macro-micronanoporous hydrogels (HPHs) without the utilization of porogens and post-etching process. This method entails the controlled generation of air bubbles within the hydrogels through the application of optimal mechanical stirring rates. Subsequent ultraviolet (UV) cross-linking serves to effectively stabilize the macropores within the HPHs. The resulting hierarchically macro-micronanoporous structures demonstrate a substantial improvement in the viability, adhesion, and proliferation of human umbilical vein endothelial cells (HUVECs) when incubated with the hydrogels. These findings present a significant advancement in the fabrication of hierarchically macro-micronanoporous hydrogels, with potential applications in the fields of tissue engineering and organoid development.

Profiling of RNA-binding protein binding sites by in situ reverse transcription-based sequencing.

RNA-binding proteins (RBPs) regulate diverse cellular processes by dynamically interacting with RNA targets. However, effective methods to capture both stable and transient interactions between RBPs and their RNA targets are still lacking, especially when the interaction is dynamic or samples are limited. Here we present an assay of reverse transcription-based RBP binding site sequencing (ARTR-seq), which relies on in situ reverse transcription of RBP-bound RNAs guided by antibodies to identify RBP binding sites. ARTR-seq avoids ultraviolet crosslinking and immunoprecipitation, allowing for efficient and specific identification of RBP binding sites from as few as 20 cells or a tissue section. Taking advantage of rapid formaldehyde fixation, ARTR-seq enables capturing the dynamic RNA binding by RBPs over a short period of time, as demonstrated by the profiling of dynamic RNA binding of G3BP1 during stress granule assembly on a timescale as short as 10 minutes.

LhCLIP reveals the in vivo RNA-RNA interactions recognized by hnRNPK.

RNA-RNA interactions play a crucial role in regulating gene expression and various biological processes, but identifying these interactions on a transcriptomic scale remains a challenge. To address this, we have developed a new biochemical technique called pCp-biotin labelled RNA hybrid and ultraviolet crosslinking and immunoprecipitation (lhCLIP) that enables the transcriptome-wide identification of intra- and intermolecular RNA-RNA interactions mediated by a specific RNA-binding protein (RBP). Using lhCLIP, we have uncovered a diverse landscape of intermolecular RNA interactions recognized by hnRNPK in human cells, involving all major classes of noncoding RNAs (ncRNAs) and mRNA. Notably, hnRNPK selectively binds with snRNA U4, U11, and U12, and shapes the secondary structure of these snRNAs, which may impact RNA splicing. Our study demonstrates the potential of lhCLIP as a user-friendly and widely applicable method for discovering RNA-RNA interactions mediated by a particular protein of interest and provides a valuable tool for further investigating the role of RBPs in gene expression and biological processes.

The role of Transcription Factor IIH complex in nucleotide excision repair.

DNA damage occurs throughout life from a variety of sources, and it is imperative to repair damage in a timely manner to maintain genome stability. Thus, DNA repair mechanisms are a fundamental part of life. Nucleotide excision repair (NER) plays an important role in the removal of bulky DNA adducts, such as cyclobutane pyrimidine dimers from ultraviolet light or DNA crosslinking damage from platinum-based chemotherapeutics, such as cisplatin. A main component for the NER pathway is transcription factor IIH (TFIIH), a multifunctional, 10-subunit protein complex with crucial roles in both transcription and NER. In transcription, TFIIH is a component of the pre-initiation complex and is important for promoter opening and the phosphorylation of RNA Polymerase II (RNA Pol II). During repair, TFIIH is important for DNA unwinding, recruitment of downstream repair factors, and verification of the bulky lesion. Several different disease states can arise from mutations within subunits of the TFIIH complex. Most strikingly are xeroderma pigmentosum (XP), XP combined with Cockayne syndrome (CS), and trichothiodystrophy (TTD). Here, we summarize the recruitment and functions of TFIIH in the two NER subpathways, global genomic (GG-NER) and transcription-coupled NER (TC-NER). We will also discuss how TFIIH's roles in the two subpathways lead to different genetic disorders.

Enhancing water flux and rejection performance through UV crosslinking: Optimization of surface modification of polyacrylonitrile (PAN)/acrylonitrile butadiene rubber (NBR) blend membrane using benzophenone as a crosslinking agent

Polyacrylonitrile (PAN)/acrylonitrile butadiene rubber (NBR) blend membranes were prepared by the phase inversion method using dimethylformamide as solvent. The blend membrane with a ratio of 85:15 was modified by ultraviolet (UV) radiation crosslinking of NBR using benzophenone as the crosslinking agent. The blend membrane was characterized through Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), atomic force microscopy (AFM), porosity, contact angel, tensile strength, pure water flux, and rejection measurements. The blend membrane showed an increased pure water flux and decreased BSA rejection. However, the addition of benzophenone and UV radiation reduced the porosity and flux of the membrane, while improving the tensile strength and BSA rejection. The response surface method (RSM) with central composite design (CCD) using Design Expert software was employed to optimize the modification conditions of the PAN/NBR blend membrane, and the suggested membrane contained 2.5 wt% benzophenone and was exposed to UV radiation for 2 h. This membrane showed a pure water flux of 95.3 L/m2h, BSA rejection of 81.1%, and a tensile strength of 4.01 MPa, which is a 45% increase in water flux compared to the neat PAN membrane while showing better rejection and preserving the desired mechanical properties.

Tendon-derived matrix crosslinking techniques for electrospun multi-layered scaffolds.

Tendon tears are common and healing often occurs incompletely and by fibrosis. Tissue engineering seeks to improve repair, and one approach under investigation uses cell-seeded scaffolds containing biomimetic factors. Retention of biomimetic factors on the scaffolds is likely critical to maximize their benefit, while minimizing the risk of adverse effects, and without losing the beneficial effects of the biomimetic factors. The aim of the current study was to evaluate cross-linking methods to enhance the retention of tendon-derived matrix (TDM) on electrospun poly(ε-caprolactone) (PCL) scaffolds. We tested the effects of ultraviolet (UV) or carbodiimide (EDC:NHS:COOH) crosslinking methods to better retain TDM to the scaffolds and stimulate tendon-like matrix synthesis. Initially, we tested various crosslinking configurations of carbodiimide (2.5:1:1, 5:2:1, and 10:4:1 EDC:NHS:COOH ratios) and UV (30 s 1 J/cm2 , 60 s 1 J/cm2 , and 60 s 4 J/cm2 ) on PCL films compared to un-crosslinked TDM. We found that no crosslinking tested retained more TDM than coating alone (Kruskal-Wallis: p > .05), but that human adipose stem cells (hASCs) spread most on the 60 s 1 J/cm2 UV- and 2.5:1:1 EDC-crosslinked films (Kruskal-Wallis: p < .05). Next, we compared the effects of 60 s 1 J/cm2 UV- and 2.5:1:1 EDC-crosslinked to TDM-coated and untreated PCL scaffolds on hASC-induced tendon-like differentiation. UV-crosslinked scaffolds had greater modulus and stiffness than PCL or TDM scaffolds, and hASCs spread more on UV-crosslinked scaffolds (ANOVA: p < .05). Fourier transform infrared spectra revealed that UV- or EDC-crosslinking TDM did not affect the peaks at wavenumbers characteristic of tendon. Crosslinking TDM to electrospun scaffolds improves tendon-like matrix synthesis, providing a viable strategy for improving retention of TDM on electrospun PCL scaffolds.

First Experience of Combined Treatment of Corneal and Graft Ulcers in Preschool Children Using Local Ultraviolet Crosslinking (Clinical Cases)

Purpose: to evaluate the results of a new combined method for the treatment of corneal and graft ulcers in preschool children, including local ultraviolet crosslinking (UVA CL) of corneal collagen and amniotic membrane transplantation (AMT).Methods. 1 patient 4 years old with a corneal ulcer and transplant disease after penetrating subtotal keratoplasty and 1 child 6 years old with a corneal ulcer of herpetic etiology was under observation for 1 year. The treatment included 3–5 consecutive procedures of local UVA CL of the cornea lasting 5 min. and amniotic membrane transplantation (AMT) after the first CL procedure. Biomicroscopy with fluorescein test and optical coherence tomography (OCT Spectralis, Heidelberg Engineering, Germany) were used to dynamically monitor the clinical course of corneal and graft ulcers. Local CL was performed using a new Keratolink device (OA Elatomsky Instrument Plant, Russia). Immediately after the 1st CL procedure, a TAM operation was performed under operating conditions under anesthesia, after which 2–4 more local CL procedures were performed sequentially every 3 days.Results. During treatment with each subsequent CL procedure, the clinical condition of the eyes improved: inflammation was relieved, photophobia, lacrimation, blepharospasm disappeared, a gradual increase in the epithelium from the periphery of the cornea with the formation of a gentle translucent opacification was noted, vascular activity decreased, and visual acuity significantly increased. After 2–10 months, in both cases there are no active complaints, the eyes are calm, infiltrates were not determined, in their place a gentle cloud-like translucent opacity formed, including in a child after keratoplasty. Vessels are empty.Conclusion. Combined treatment of corneal and graft ulcers in children (local UVA CL together with TAM) demonstrated high therapeutic efficacy, ensured stable corneal epithelialization in a short time and improved visual functions at the end of the inflammatory process. This method can significantly expand the possibilities of treating ulcerative lesions of the cornea and graft in preschool children.

RBP-Tar – a searchable database for experimental RBP binding sites

Background: RNA-binding proteins (RBPs) play a critical role in regulating gene expression by binding to specific sites on RNA molecules. Identifying these binding sites is crucial for understanding the many functions of RBPs in cellular function, development and disease. Current experimental methods for identifying RBP binding sites, such as ultra-violet (UV) crosslinking and immunoprecipitation (CLIP), and especially the enhanced CLIP (eCLIP) protocol, were developed to identify authentic RBP binding sites experimentally. Methods: To make this data more accessible to the scientific community, we have developed RBP-Tar (https://ncbr.muni.cz/RBP-Tar), a web server and database that utilises eCLIP data for 167 RBPs mapped on the human genome. The web server allows researchers to easily search and retrieve binding site information by genomic location and RBP name. Use case: Researchers can produce lists of all known RBP binding sites on a gene of interest, or produce lists of binding sites for one RBP on different genomic loci. Conclusions: Our future goal is to continue to populate the web server with additional experimental datasets from CLIP experiments as they become available and processed, making it an increasingly valuable resource for the scientific community.