Lens Tissue Research Articles

An isotopically-labelled temporal mass spectrometry imaging data analysis workflow to reveal glucose spatial metabolism patterns in bovine lens tissue

Application of stable isotopically labelled (SIL) molecules in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI) over a series of time points allows the temporal and spatial dynamics of biochemical reactions to be tracked in a biological system. However, these large kinetic MSI datasets and the inherent variability of biological replicates presents significant challenges to the rapid analysis of the data. In addition, manual annotation of downstream SIL metabolites involves human input to carefully analyse the data based on prior knowledge and personal expertise. To overcome these challenges to the analysis of spatiotemporal MALDI-MSI data and improve the efficiency of SIL metabolite identification, a bioinformatics pipeline has been developed and demonstrated by analysing normal bovine lens glucose metabolism as a model system. The pipeline consists of spatial alignment to mitigate the impact of sample variability and ensure spatial comparability of the temporal data, dimensionality reduction to rapidly map regional metabolic distinctions within the tissue, and metabolite annotation coupled with pathway enrichment modules to summarise and display the metabolic pathways induced by the treatment. This pipeline will be valuable for the spatial metabolomics community to analyse kinetic MALDI-MSI datasets, enabling rapid characterisation of spatio-temporal metabolic patterns from tissues of interest.

TRB3 Promotes Cataract Progression through Endoplasmic Reticulum Stress-mediated Mitochondrial Dysfunction and Cell Apoptosis.

Cataracts are characterized as a disease affecting lens opacity. Endoplasmic reticulum (ER) stress can cause lens epithelial cell (LEC) dysfunction, affecting normal lens transparency and function, but the role of Tribbles 3 (TRB3), an inducible gene of ER stress, in cataracts is poorly understood. This study explored how TRB3 promotes cataract progression through ER stress. We administered a subcutaneous injection of sodium selenite at a dosage of 3.46 mg/kg to rats to create an animal model of cataracts. Additionally, we exposed rat LEC cells to 0.01 μM tunicamycin (TM) for 24 h to establish a cell model of ER stress. The detection of related genes and proteins was performed via RT‒qPCR and Western blot techniques. Flow cytometry, along with JC-1, TUNEL, and HE staining, was employed to assess damage to cells and lens tissues. This study revealed that TRB3 was abnormally highly expressed in both a cataract rat model and an ER stress cell model. Knocking down TRB3 has a similar effect as treatment with an ER stress inhibitor, effectively reversing the ER stress and apoptosis induced by TM. This effect includes increasing the mitochondrial membrane potential in LEC cells, lowering reactive oxygen species (ROS) levels, increasing ATP production, suppressing the expression of the apoptosis-related proteins Bax and C-caspase-3, increasing Bcl-2 expression, and decreasing apoptosis. Furthermore, TRB3 knockdown improved the pathological conditions of rat lenses and inhibited mitochondrial dysfunction and cell apoptosis to relieve the development of cataracts in rats. Mechanistically, CHOP promotes the expression of TRB3 by binding to the TRB3 promoter, thereby activating ER stress, leading to mitochondrial dysfunction and cell apoptosis in LEC cells and accelerating the development of cataracts. According to our findings, targeting TRB3 expression inhibition could emerge as a novel approach for cataract therapy.

MicroRNA-204-5p Attenuates Oxidative Stress, Apoptosis and Inflammation by Targeting TXNIP in Diabetic Cataract.

Diabetic cataract (DC) is a major cause of blindness in diabetic patients and it is characterized by early onset and rapid progression. MiR-204-5p was previously identified as one of the top five down-regulated miRNAs in human DC lens tissues. We aimed to determine the expression of miR-204-5p in human lens epithelial cells (HLECs) and explore its effects and mechanisms in regulating the progression of DC. The expression of miR-204-5p in the anterior capsules of DC patients and HLECs was examined by RT-qPCR. Bioinformatics tools were then used to identify the potential target of miR-204-5p. The relationship between miR-204-5p and the target gene was confirmed through a dual luciferase reporter assay. Additionally, the regulatory mechanism of oxidative stress, apoptosis, and inflammation in DC was investigated by overexpressing miR-204-5p using miR-204-5p agomir. The expression of miR-204-5p was downregulated in the anterior capsules of DC patients and HLECs. Overexpression of miR-204-5p reduced ROS levels, pro-apoptosis genes (Bid, Bax, caspase-3), and IL-1β production in HG-treated HLECs. TXNIP was the direct target of miR-204-5p by dual luciferase reporter assay. Therefore, this study demonstrated that miR-204-5p effectively reduced oxidative damage, apoptosis, and inflammation in HLECs under HG conditions by targeting TXNIP. Targeting miR-204-5p could be a promising therapeutic strategy for the potential treatment of DC.

Multi-tissue transcriptome-wide association study identifies novel candidate susceptibility genes for cataract.

Cataract is the leading cause of blindness among the elderly worldwide. Twin and family studies support an important role for genetic factors in cataract susceptibility with heritability estimates up to 58%. To date, 55 loci for cataract have been identified by genome-wide association studies (GWAS), however, much work remains to identify the causal genes. Here, we conducted a transcriptome-wide association study (TWAS) of cataract to prioritize causal genes and identify novel ones, and examine the impact of their expression. We performed tissue-specific and multi-tissue TWAS analyses to assess associations between imputed gene expression from 54 tissues (including 49 from the Genotype Tissue Expression (GTEx) Project v8) with cataract using FUSION software. Meta-analyzed GWAS summary statistics from 59,944 cataract cases and 478,571 controls, all of European ancestry and from two cohorts (GERA and UK Biobank) were used. We then examined the expression of the novel genes in the lens tissue using the iSyTE database. Across tissue-specific and multi-tissue analyses, we identified 99 genes for which genetically predicted gene expression was associated with cataract after correcting for multiple testing. Of these 99 genes, 20 (AC007773.1, ANKH, ASIP, ATP13A2, CAPZB, CEP95, COQ6, CREB1, CROCC, DDX5, EFEMP1, EIF2S2, ESRRB, GOSR2, HERC4, INSRR, NIPSNAP2, PICALM, SENP3, and SH3YL1) did not overlap with previously reported cataract-associated loci. Tissue-specific analysis identified 202 significant gene-tissue associations for cataract, of which 166 (82.2%), representing 9 unique genes, were attributed to the previously reported 11q13.3 locus. Tissue-enrichment analysis revealed that gastrointestinal tissues represented one of the highest proportions of the Bonferroni-significant gene-tissue associations (21.3%). Moreover, this gastrointestinal tissue type was the only anatomical category significantly enriched in our results, after correcting for the number of tissue donors and imputable genes for each reference panel. Finally, most of the novel cataract genes (e.g., Capzb) were robustly expressed in iSyTE lens data. Our results provide evidence of the utility of imputation-based TWAS approaches to characterize known GWAS risk loci and identify novel candidate genes that may increase our understanding of cataract etiology. Our findings also highlight the fact that expression of genes associated with cataract susceptibility is not necessarily restricted to lens tissue.

Biochemical component analysis of human myopic corneal stroma using the Raman spectrum.

This study aimed to measure the Raman spectrum of the human corneal stroma lens obtained from small incision lenticule extraction surgery (SMILE) in Asian myopic eyes using a confocal Raman micro-spectrometer built in the laboratory. Forty-three myopic patients who underwent SMILE with equivalent diopters between - 4.00 and - 6.00 D were selected, and the right eye data were collected. Corneal stroma lenses were obtained during surgery, and the Raman spectra were measured after air drying. The complete Raman spectrum of human myopic corneal stroma lens tissue was obtained within the range of 700-4000cm-1. Thirteen characteristic peaks were found, with the stronger peaks appearing at 937cm-1, corresponding to proline, valine, and the protein skeleton of the human myopic corneal stroma lens; 1243cm-1, corresponding to collagen protein; 1448cm-1, corresponding to the collagen protein and phospholipids; and 2940cm-1, corresponding to the amino acid and lipids, which was the strongest Raman peak. These results demonstrated that Raman spectroscopy has much potential as a fast, cost-effective, and reliable diagnostic tool in the diagnosis and treatment of eye diseases, including myopia, keratoconus, and corneal infection.

Uric acid–driven NLRP3 inflammasome activation triggers lens epithelial cell senescence and cataract formation

Excessive uric acid (UA) is associated with age-related cataract. A previous study showed that a high UA level in the aqueous humor stimulated the senescence of lens epithelial cells (LECs), leading to cataract progression. To better understand the underlying mechanisms, we investigated UA-driven senescence in human lens tissue samples obtained during surgery, rat lens organ cultures, and in vivo experiments, using senescence-associated β-galactosidase (SA-β-gal) staining, electronic microscopy, Western blotting, and histological analyses. Initially, we identified markedly higher expressions of NLRP3 and caspase-1 in the lens capsules of hyper-uricemic patients compared to normo-uricemic patients. This increase was accompanied by a significant rise in the SA-β-gal positive rate. We next built a cataract model in which rat lenses in an organ culture system were treated with an increasing dosage of UA. Notably, opacification was apparent in the lenses treated with 800 μM of UA starting on the fifth day. Mechanistically, UA treatment not only significantly induced the expression of NLRP3, caspase-1, and IL-1β, but also upregulated the levels of SA-β-gal and the senescence regulators p53 and p21. These effects were fully reversed, and lens opacification was ameliorated by the addition of MCC950, a selective NLRP3 antagonist. Moreover, an in vivo model showed that intravitreal UA injection rapidly induced cataract phenotypes within 21 days, an effect significantly mitigated by co-injection with MCC950. Together, our findings suggest that targeting the UA-induced NLRP3 inflammasome with MCC950 could be a promising strategy for preventing cataract formation associated with inflammageing.

On the gamma radiation response of commercially available 3D printing materials for medical dosimetry

3D printing technology has rapidly spread for decades, allowing the fabrication of medical implants and human phantoms and revolutionizing healthcare. The objective of this study is to evaluate some radiological properties of commercially available 3D printing materials as potential tissue mimicking materials. Among fifteen materials, we compared their properties with nine human tissues. In all materials and tissues, exposure and energy absorption buildup factors were calculated for photon energies between 0.015 and 15 MeV and penetration depths up to 40 mean free path. Furthermore, the Geant4 Monte Carlo toolkit (version 10.5) was used to simulate their percentage depth dose distributions. In addition, equivalent atomic numbers, effective atomic numbers, attenuation coefficients, and CT numbers have been examined. All parameters were considered in calculating the average relative error (σ), which was used as a statistical comparison tool. With σ between 6 and 7, we found that Polylactic Acid (PLA) was capable of simulating eye lenses, blood, soft tissue, lung, muscle, and brain tissues. Moreover, Polymethacrylic Acid (PMAA) material has a σ value of 4 when modeling adipose and breast tissues, respectively. Aside from that, variations in 3D printing materials’ infilling percentage can affect their CT numbers. We therefore suggest the PLA for mimicking soft tissue, muscle, brain, eye lens, lung and blood tissues, with an infill of between 92.7 and 94.3 percent. We also suggest an 89 percent infill when simulating breast tissue. Furthermore, with a 96.7 percent infill, the PMAA faithfully replicates adipose tissue. Additionally, we found that a 59 percent infill of Fe-PLA material is comparable to cortical bone. Due to the benefits of creating individualized medical phantoms and equipment, the results might be seen as an added value for both patients and clinicians.

The α-crystallin Chaperones Undergo a Quasi-ordered Co-aggregation Process in Response to Saturating Client Interaction

Small heat shock proteins (sHSPs) are ATP-independent chaperones vital to cellular proteostasis, preventing protein aggregation events linked to various human diseases including cataract. The α-crystallins, αA-crystallin (αAc) and αB-crystallin (αBc), represent archetypal sHSPs that exhibit complex polydispersed oligomeric assemblies and rapid subunit exchange dynamics. Yet, our understanding of how this plasticity contributes to chaperone function remains poorly understood. Using biochemical and biophysical analyses combined with single-particle electron microscopy (EM), we examined structural changes in αAc, αBc and native heteromeric lens α-crystallins (αLc) in their apo-states and at varying degree of chaperone saturation leading to co-aggregation, using lysozyme and insulin as model clients. Quantitative single-particle analysis unveiled a continuous spectrum of oligomeric states formed during the co-aggregation process, marked by significant client-triggered expansion and quasi-ordered elongation of the sHSP oligomeric scaffold, whereby the native cage-like sHSP assembly displays a directional growth to accommodate saturating conditions of client sequestration. These structural modifications culminated in an apparent amorphous collapse of chaperone-client complexes, resulting in the creation of co-aggregates capable of scattering visible light. Intriguingly, these co-aggregates maintain internal morphological features of highly elongated sHSP oligomers with striking resemblance to polymeric α-crystallin species isolated from aged lens tissue. This mechanism appears consistent across αAc, αBc and αLc, albeit with varying degrees of susceptibility to client-induced co-aggregation. Importantly, our findings suggest that client-induced co-aggregation follows a distinctive mechanistic and quasi-ordered trajectory, distinct from a purely amorphous process. These insights reshape our understanding of the physiological and pathophysiological co-aggregation processes of α-crystallins, carrying potential implications for a pathway toward cataract formation.

RNA‐binding proteins and their downstream regulatory networks in lens development and cataract

Development of the ocular lens involves dynamic control over gene expression, perturbations of which can cause congenital cataract. Thus far, several key proteins in the lens development program that are involved in signalling, transcription, and more recently, post‐transcriptional control – all distinct regulatory mechanisms that together determine the proteome – have been identified. To understand the pathology of congenital cataract, it is critical to continue to identify key regulatory genes that play an important role in the lens, and define their relationships with other regulators in the form of networks. We have characterized eye/lens defects in animal knockout models for several conserved RNA‐binding proteins (RBPs) such as Celf1, Caprin2, Rbm24 and Tdrd7 and demonstrated their important role in mediating post‐transcriptional networks in the lens. However, there are still many mechanistic events that remain to be defined in the lens. For example, while it is known that abundant levels of specific mRNAs (e.g. crystallins) need to be made in the lens, it is currently unknown whether regulators that control the ubiquitously important “transcription elongation” process, are specifically required for achieving optimal levels of such highly expressed transcripts in fibre cells. Furthermore, how these transcriptional and post‐transcriptional regulatory pathways coordinately function to achieve optimal transcriptome and proteome in lens epithelial and fibre cells is unknown. Here, we applied the bioinformatics tool iSyTE to identify the transcription elongation factor Ell2 and the RNA‐binding protein (RBP) Elavl1 in the lens. Lens‐specific conditional knockout mouse models for Elavl1 and Ell2 were generated using Pax6GFPCre. Lens tissue from Elavl1cKO and Ell2cKO mice was characterized by immunostaining, RNA‐sequencing (RNA‐seq) and RT‐qPCR. Elavl1cKO and Ell2cKO lenses exhibit morphological defects that are detected in embryonic and early postnatal stages, respectively. RNA‐seq identifies mis‐expression of many genes in both cKO lenses. Ell2cKO lenses show significantly reduced transcript levels of a cohort of fibre‐expressed genes such as crystallins, while Elavl1cKO lenses show abnormal mRNA levels of several novel factors in the lens, including Ell2. These data demonstrate that the new iSyTE‐predicted genes, the RBP, Elavl1, and transcription elongation factor, Ell2, mediate regulation of key genes in lens development, and provide new insights into the molecular pathology of cataract and lens defects in animals deficient in these genes. Importantly, in addition to Elavl1cKO lenses, Ell2 is found to be mis‐expressed in mouse cKO lenses for another RBP, Celf1, suggesting a crosstalk between transcriptional and post‐transcriptional regulatory networks that governs optimal transcriptome and proteome in lens development.

Sestrin 2 protects human lens epithelial cells from oxidative stress and apoptosis induced by hydrogen peroxide by regulating the mTOR/Nrf2 pathway.

We aimed to explore the effect and potential mechanism of Sestrin 2 (SESN2) in human lens epithelial cells (HLECs). To mimic the oxidative stress environment, SAR01/04 cells were treated with 200μM hydrogen peroxide (H2O2) for 24h. Cell viability and apoptosis were checked by cell counting kit-8 and flow cytometry. Western blot was taken to check the protein changes of SESN2, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), mechanistic target of rapamycin (mTOR), phosphorylated (p)-mTOR, ribosomal protein S6 kinase B1 (p70S6K), p-p70S6K, and nuclear factor erythroid 2-related factor 2 (Nrf2). Superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and reactive oxygen species (ROS) were detected via the corresponding reagent kit. The levels of interleukin (IL)-1β, IL-18, and tumor necrosis factor (TNF)-α were measured using enzyme-linked immunosorbent assay. SESN2 was down-regulated in cataract lens tissue and up-regulated in SAR01/04 cells treated with H2O2. Under treatment of H2O2, up-regulation of SESN2 improved cell viability, enhanced the activity of SOD and CAT, inhibited cell apoptosis, and reduced the levels of MDA, ROS, IL-1β, IL-18, and TNF-α, while down-regulation of SESN2 caused the contrary effects. Further bioinformatics analysis suggested that SESN2 regulated the mTOR signaling pathway. Treatment of H2O2 inhibited p-mTOR and p-p70S6K protein expression, while overexpression of SESN2 increased p-mTOR and p-p70S6K protein expression in the H2O2 group and down-regulation of SESN2 further decreased p-mTOR and p-p70S6K protein expression in the H2O2 group. Additionally, H2O2 increased Nrf2 protein expression, and overexpression of SESN2 further increased Nrf2 protein expression in the H2O2 group. Importantly, rapamycin (an inhibitor of mTOR signaling pathway) and knockdown of Nrf2 reversed the promotive effects of SESN2 on cell viability and the inhibitive effects of SESN2 on cell apoptosis, oxidative stress, and inflammatory reaction. SESN2 protected HLECs damage induced by H2O2, which was related to the activation of mTOR/Nrf2 pathway.

Identification of new antioxidant drugs in a human in vitro model of ageing‐associated cataract

Aims/Purpose: Cataracts are a significant cause of global blindness, impacting millions (10.8 million patients), and their prevalence is expected to increase over the coming decades as our populations age. Oxidative stress disrupts ROS (reactive oxygen species) balance in lens fibre cells, leading to lens protein degradation and accumulation, developing age‐related cataracts. The lens has protective mechanisms, including high GSH levels and antioxidant enzymes like catalase, safeguarding lens health. While surgery is currently the only effective treatment, non‐invasive drug‐based approaches show potential by reducing risks and complications. With its potent antioxidant activity, resveratrol may protect lens cells and slow cataract development. However, the limited ability of resveratrol to penetrate lens tissue restricts its direct effects. Therefore, designing new molecules to improve ADME (absorption, distribution, metabolism and excretion) properties is required.Methods: In this study, we used an in vitro model of cataracts by inducing oxidative stress and cell death by adding hydrogen peroxide (H2O2) and luperox to lens epithelial cells to evaluate the protective effects of new molecules against oxidative stress and compared them with resveratrol. Our study assessed the cell viability, intracellular ROS level, mitochondrial function, catalase activity and GSH/GSSH level.Results: These compounds preserved cell viability, reduced ROS levels, maintained mitochondrial function and enhanced catalase activity and GSH levels in lens epithelial cells exposed to H2O2. The novel drugs, CAB‐C1 and CAB‐C2, demonstrated beneficial effects on cellular health.Conclusions: Developing drugs for cataract treatment enables early intervention, particularly for individuals with mild or moderate cataracts who may not require surgery yet. Such advancements might improve patients' quality of life and have implications for ocular science and age‐related eye diseases.

Genetic analysis of medaka fish illuminates conserved and divergent roles of Pax6 in vertebrate eye development.

Landmark discovery of eye defects caused by Pax6 gene mutations in humans, rodents, and even fruit flies combined with Pax6 gene expression studies in various phyla, led to the master control gene hypothesis postulating that the gene is required almost universally for animal visual system development. However, this assumption has not been broadly tested in genetically trackable organisms such as vertebrates. Here, to determine the functional role of the fish orthologue of mammalian Pax6 in eye development we analyzed mutants in medaka Pax6.1 gene generated by genome editing. We found that transcription factors implicated in vertebrate lens development (Prox1a, MafB, c-Maf, FoxE3) failed to initiate expression in the presumptive lens tissue of Pax6.1 mutant fish resulting in aphakia, a phenotype observed previously in Pax6 mutant mice. Surprisingly, the overall differentiation potential of Pax6.1-deficient retinal progenitor cells (RPCs) is not severely compromised, and the only cell types affected by the absence of Pax6.1 transcription factor are retinal ganglion cells. This is in stark contrast to the situation in mice where the Pax6 gene is required cell-autonomously for the expansion of RPCs, and the differentiation of all retina cell types. Our results provide novel insight into the conserved and divergent roles of Pax6 gene orthologues in vertebrate eye development indicating that the lens-specific role is more evolutionarily conserved than the role in retina differentiation.

Mass Stopping Power and Range of Alpha Particles in Biological Human Body (Water and Eye Lens Tissue)

Mass Stopping Power and Range of Alpha Particles in Biological Human Body (Water and Eye Lens Tissue)

Lens epithelium cell ferroptosis mediated by m6A-lncRNA and GPX4 expression in lens tissue of age-related cataract

BackgroundIn the present study, we explored the role of N6-methyladenosine (m6A) modification of long non-coding RNAs (lncRNAs) and its association with ferroptosis in lens epithelium cells (LECs) of age-related cataract (ARC).MethodsThrough m6A RNA immunoprecipitation sequencing (m6A-RIP-seq) and RNA sequencing (RNA-seq), we identified m6A mediated and differentially expressed lncRNAs (dme-lncRNAs) in ARC patients. Based on bioinformatics analysis, we selected critical dme-lncRNAs and pathways associated with ARC formation to reveal their potential molecular mechanisms. The downregulation of glutathione peroxidase 4 (GPX4), a key component of ferroptosis, was confirmed by real-time RT-PCR (RT-qPCR) and Western blotting in age-related cortical cataract (ARCC) samples. Transmission electron microscopy was used to assess the change in mitochondrial in LECs.ResultsThe analysis revealed a total of 11,193 m6A peaks within lncRNAs, among which 7043 were enriched and 4150 were depleted. Among those, lncRNA ENST00000586817(upstream of the GPX4 gene) was not only significantly upregulated in the LECs of ARCC but also potentially augmented the expression of GPX4 through a cis mechanism. The expression of m6A-modified lncRNA (ENST00000586817) was correlated with that of GPX4 and was downregulated in ARC patients. The TEM results indicated significant mitochondrial changes in ARCC samples. GPX4 downregulation enhanced LEC ferroptosis and decreased viability via RSL3 in SRA01/04 cells.ConclusionsOur results provide insight into the potential function of m6A-modified lncRNAs. M6A-modified lncRNA ENST00000586817 might regulate the expression of GPX4 by a cis mechanism and be implicated in ferroptosis in ARCs.

Methods for quantifying rain-splash dispersal of Neonectria ditissima conidia in apple canopies

Many microorganisms can be dispersed by rain-splash, whereby spores become suspended in water and are spread via droplets. The resulting dispersal gradient is dependent on several factors including rainfall intensity, the nature of the plant canopy and its effects on splash, deposition, redistribution (secondary splash) and filtering. Gradients of spore dispersal with distance are important for understanding epidemics, and the primary dispersal gradient can shape an epidemic for several pathogen generations. However, microorganisms are difficult to trap, identify and enumerate efficiently. This makes it difficult to study the spread and dispersal of pathogens to aid in biosecurity responses and management of epidemics. We used macroconidia of Neonectria ditissima, the causal organism of apple canker, to explore patterns of rain-splash dispersal in tree canopies. We investigated the use of a fluorescent tracer dye, PTSA (1,3,6,8-pyrenetetrasulfonic acid), as a surrogate to conidia capture during natural and artificial rain events, and lens tissue as ‘surrogate leaves’ to recapture tracer dye. Conidia and dye were released from central point sources 2.5 m above the ground and recaptured in passive rainwater traps or artificial ‘leaves’. Quantile regression and exponential models were used to explore variation and dispersal gradients derived for both conidia and dye, with and without tree canopy and with natural or artificial precipitation. Estimated dispersal gradients were steeper with a flatter tail when no tree canopy was present, whereas presence of tree canopy resulted in more variation and shallower predicted dispersal gradients, with fatter tails, predicting potential dispersal to further distances from the source. The majority of conidia and dye were recaptured at less than 1 m from the source, but small concentrations of spores were detected up to 3 m and dye more than 6 m. High variation in natural conditions requires further investigation to fully quantify natural dispersal gradients. Nevertheless, these results show the merit of tracer dye, artificial leaves, and quantile regression as tools to estimate potential dispersal patterns of N. ditissima and other rain-splash dispersed microorganisms, considering rain-splash factors in real canopies and natural situations for predicting inoculum dispersal.

Uncovering the Hidden World of Aqueous Humor Proteins for Discovery of Biomarkers for Marfan Syndrome.

Ectopia lentis is a hallmark of Marfan syndrome (MFS), a genetic connective tissue disorder affecting 1/5000 to 1/10 000 individuals worldwide. Early detection in ophthalmology clinics and timely intervention of cardiovascular complications can be lifesaving. In this study, a modified proteomics workflow with liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based data-independent acquisition (DIA) and field asymmetric ion mobility spectrometry (FAIMS) to profile the proteomes of aqueous humor (AH) and lens tissue from MFS children with ectopia lentis is utilized. Over 2300 and 2938 comparable proteins are identified in AH and the lens capsule, respectively. Functional enrichment analyses uncovered dysregulation of complement and coagulation-related pathways, collagen binding, and cell adhesion in MFS. Through weighted correlation network analysis (WGCNA) and machine learning, distinct modules associated with clinical traits are constructed and a unique biomarker panel (Q14376, Q99972, P02760, Q07507; gene names: GALE, MYOC, AMBP, DPT) is defined. These biomarkers are further validated using advanced parallel reaction monitoring (PRM) in an independent patient cohort. The results provide novel insights into the proteome characterization of ectopia lentis and offer a promising approach for developing a valuable biomarker panel to aid in the early diagnosis of Marfan syndrome via AH proteome.

The role of dietary vitamin A in mechanisms of cataract development in the teleost lumpfish (Cyclopterus lumpus L).

Lumpfish (Cyclopterus lumpus L) are highly prone to cataract development in the wild and in culture. There is evidence that cataract in farmed fish is related to nutrition. However, both the nutrients and the mechanisms involved in cataract development in lumpfish are not clear. Here we investigated the mechanisms involved and the role of dietary vitamin A in cataract development in a cultured lumpfish population. Cultured lumpfish were fed three diets differing only in vitamin A supplementation level (5000, 15,000 and 120,000 IU/kg) over an 18-month period, and fish weight, cataract frequencies and severities were determined. Western blotting and immunohistochemistry were performed on lens tissue to measure the levels of oxidative stress, and apoptosis. The lowest levels of vitamin A significantly reduced cataract frequencies in adult lumpfish and resulted in less severe cataract and increased weight in males. Oxidative stress levels in the lens were positively correlated with vitamin A intake. Apoptosis was observed at high levels in lenses with severe cataract. Oxidative stress and apoptosis levels were the highest in regions of the lens with severe, advanced cataract pathology when compared to regions with no visible pathology. These results suggest that higher vitamin A intake contributes to cataract development through an oxidative stress pathway, and that both oxidative stress and apoptosis are involved in advanced stages of cataract in lumpfish.

Petroselinum crispum Extract Prevents Scopolamine-Induced Lens Damage in Rats.

Alzheimer's disease (AD) is a neurodegenerative disease that occurs especially in advanced ages. It reduces the quality of life of both the patient and their relatives. In addition to its primary effects, AD causes metabolic defects and tissues are damaged due to these effects. Oxidative stress damages cells by disrupting antioxidant/oxidant balance in many tissues, especially due to AD. In individuals with AD and the elderly, lens tissue is damaged due to oxidative stress and may cause vision loss. Therefore, it is very important to investigate herbal products that both prevent/cure AD and reduce AD-related oxidative stress, as they may have fewer side effects. In this study, the protective effects of parsley (Petroselinum crispum) extract on lens tissues of an experimental AD model induced by scopolamine were examined and evaluated through biochemical parameters. The result of biochemical experiments and principal component analysis, was observed that parsley extract had a therapeutic effect by reducing oxidative stress in lens tissues of experimentally induced AD rats. It can be suggested that the phenolic and flavonoid-rich content of parsley extract may have caused the reduction of oxidative damage in lens tissues and can be used to protect lens tissue against oxidative stress due to AD disease.

Некоторые биохимические изменения хрусталика в процессе старения.

The goal is to study some biochemical changes in the lens of rats and humans during aging. Materials and methods. Experiments were carried out on 25 rats. We also examined 84 patients with initial nuclear cataract. Results. diverse dynamics of the content of total lipids in the lenses of rats was revealed. There is a noticeable decrease in the content of products of oxidative modification of lipids in the lens tissue between 12 and 24 months and a slight increase in the period from 24 to 36 months. A 2-fold decrease in the content of the total protein level was revealed when comparing groups of 5 months and 36 months and an increase in total protein content with age. When comparing the parameters of oxidative degradation, cl in the lens tissue of rats and humans with age, similar indicators of dynamics were obtained. Conclusion. The data obtained indicate changes in the parameters of lipids and proteins in the lens tissue of rats during postnatal ontogenesis. The fact that there is a balance between damage and protection of proteins in the transparent lens was revealed. When comparing data on protein oxidation in the lens tissue of rats and humans with age, similar indicators were obtained. Keywords: lens, aging, biochemical changes

Eye lens β-crystallins are predicted by native ion mobility-mass spectrometry and computations to form compact higher-ordered heterooligomers

Eye lens α- and β-/γ-crystallin proteins are not replaced after fiber cell denucleation and maintain lens transparency and refractive properties. The exceptionally high (∼400-500mg/mL) concentration of crystallins in mature lens tissue and multiple other factors impede precise characterization of β-crystallin interactions, oligomer composition, size, and topology. Native ion mobility-mass spectrometry is used here to probe β-crystallin association and provide insight into homo- and heterooligomerization kinetics for these proteins. These experiments include separation and characterization of higher-order β-crystallin oligomers and illustrate the unique advantages of native IM-MS. Recombinantly expressed βB1, βB2, and βA3 isoforms are found to have different homodimerization propensities, and only βA3 forms larger homooligomers. Heterodimerization of βB2 with βA3 occurs ∼3 times as fast as that of βB1 with βA3, and βB1 and βB2 heterodimerize less readily. Ion mobility experiments, molecular dynamics simulations, and PISA analysis together reveal that observed oligomers are consistent with predominantly compact, ring-like topologies.