Anterior Chamber Of The Eye Research Articles

Small-molecule inhibitors of the CD40–CD40L costimulatory interaction are effective in pancreatic islet transplantation and prevention of type 1 diabetes models

As part of our work to develop small-molecule inhibitors (SMIs) of the CD40-CD40L(CD154) costimulatory protein-protein interaction, here, we describe the ability of two of our most promising SMIs, DRI-C21041 and DRI-C21095, to prolong the survival and function of islet allografts in two murine models of islet transplantation (under the kidney capsule and in the anterior chamber of the eye) and to prevent autoimmune type 1 diabetes (T1D) onset in NOD mice. In both transplant models, a significant portion of islet allografts (50%-80%) remained intact and functional long after terminating treatment, suggesting the possibility of inducing operational immune tolerance via inhibition of the CD40-CD40L axis. SMI-treated mice maintained the structural integrity and function of their islet allografts with concomitant reduction in immune cell infiltration as evidenced by direct longitudinal imaging in situ. Furthermore, in female NODs, three-month SMI treatment reduced the incidence of diabetes from 80% to 60% (DRI-C21041) and 25% (DRI-C21095). These results (i) demonstrate the susceptibility of this TNF superfamily protein-protein interaction to small-molecule inhibition, (ii) confirm the in vivo therapeutic potential of these SMIs of a critical immune checkpoint, and (iii) reaffirm the therapeutic promise of CD40-CD40L blockade in islet transplantation and T1D prevention. Thus, CD40L-targeting SMIs could ultimately lead to alternative immunomodulatory therapeutics for transplant recipients and prevention of autoimmune diseases that are safer, less immunogenic, more controllable (shorter half-lives), and more patient-friendly (i.e., suitable for oral administration, which makes them easier to administer) than corresponding antibody-based interventions.

Physiological activation of liver X receptor provides protection against ocular inflammation in uveitic glaucoma.

Virus-induced trabeculitis is considered a significant cause of uveitic glaucoma, being marked by a sudden increase in intraocular pressure and relatively mild inflammation in the anterior chamber of the eye. In previous proteome analyses of aqueous humor (AH) derived from Cytomegalovirus (CMV) uveitic glaucoma patients, we observed the liver X receptor (LXR) pathway to be among the most prominently activated canonical pathways. In the present study, we explored the role of the LXR pathway in the etiology of glaucoma in association with ocular inflammation. LXRα/β and ABCA1, the downstream targets of LXR, were distributed throughout the conventional AH outflow pathway of the human eye, and their increased levels in human trabecular meshwork cells in response to CMV infection and Lipopolysaccharide treatment. Treatment with an LXR agonist (T091317) suppressed LPS-induced inflammation and this response was reversed under the deficiency of LXRα/LXRβ. Furthermore, in the rat endotoxin uveitis model, the LXR agonist significantly reduced infiltrating cells and expression of proinflammatory cytokines in the iris and retina. These results reveal upregulation of LXR-ABCA1 under inflammatory insult in the conventional AH outflow pathway, and activation of LXR exhibiting an anti-inflammatory effect, implying its essential physiological protective role in glaucoma associated with ocular inflammation.

Glaucoma-Protective Human Single-Nucleotide Polymorphism in the Angpt2 Locus Increased ANGPT2 Expression and Schlemm Canal Area in Mice-Brief Report.

The ANGPT (angiopoietin)-TEK (tyrosine kinase, endothelial) vascular signaling pathway plays a key role in the formation of Schlemm canal, and loss-of-function mutations in the TEK or ANGPT1 gene are associated with primary congenital glaucoma in children. In genome-wide association studies, an association was identified between protection from primary open-angle glaucoma and the single-nucleotide polymorphism rs76020419 (G>T), located within a predicted miR-145-binding site in the 3' untranslated region of ANGPT2. To date, the functional impact of this variant in the anterior chamber of the eye remains largely unexplored. MT (mutant) mice harboring an orthologous rs76020419 minor allele (T) were generated using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat-associated 9). Plasma and tissue samples, including eyes, were collected, and ANGPT2 expression was quantified using ELISA. Anterior segments from eyes were collected from WT (wild-type) and MT mice, and Schlemm canal area was quantified. In the MT group, higher ANGPT2 concentrations were observed in the plasma, lungs, kidneys, and eyes (P=0.0212, P<0.001, P=0.0815, and P=0.0215, respectively). Additionally, the Schlemm canal was larger in MT mice compared with WT mice (P=0.0430). The rs76020419 minor allele (T) is associated with increased levels of ANGPT2 and a larger Schlemm canal in mice. These findings suggest a potential protective mechanism in glaucoma.

Histamine H3 receptor antagonist/nitric oxide donors as novel promising therapeutic hybrid-tools for glaucoma and retinal neuroprotection

Glaucoma is a degenerative optic neuropathy in which the degeneration of optic nerve and blindness occur. The main cause is a malfunction of ciliary processes (protrusions of the ciliary bodies) resulting in increased intraocular pressure (IOP). Ocular hypertension (OHT) causes ischemic events leading to retinal ganglion cell (RGC) depletion and blindness.Histaminergic and nitrergic systems are involved in the regulation of IOP. Therefore, we developed novel hybrid compounds that target histamine H3 receptor (H3R) with nitric oxide (NO) releasing features (ST-1989 and ST-2130). After H3R binding was proven in vitro, we investigated their effects in two OHT models in New Zealand White rabbits. Compound ST-1989 showed the highest NO elevation, together with antioxidative and anti-inflammatory features partly superior to the co-administered H3R antagonist (ciproxifan) and NO donor (molsidomine). This hybrid compound demonstrated IOP reduction in both OHT models induced by intravitreal injection of hypertonic saline and carbomer into the anterior chamber of the eye, respectively. Ocular perfusion and photoreceptor neuroprotection were evaluated in a model of ischemia/reperfusion (I/R) of the ophthalmic artery induced by repeated sub-tenon injections of endothelin-1 (ET-1), twice a week for six weeks. Compound ST-1989 counteracts retinal degeneration reducing ophthalmic artery resistance index and increasing photoreceptor responses, thus rescuing RGCs.Our results indicate that compound ST-1989 is a promising molecule with long-lasting hypotensive effects and good effectiveness in reducing inflammation, oxidative stress, and RGCs apoptosis. In conclusion, these hybrid compounds could be a novel strategy to combat glaucomatous blindness and RGC depletion for ocular diseases involving retinal damage.

Pigment Dispersion Contributes to Ocular Immune Privilege in a DBA/2J Mouse Model of Pigmentary Glaucoma.

To investigate the effects of anterior chamber pigment dispersion on ocular immune privilege and the possible mechanisms involved in a DBA/2J mouse model of pigmentary glaucoma. DBA/2J mice were utilized as a pigment dispersion model, and age-matched C57BL/6J mice were used as the control group in this study. Proteins in the aqueous humor (AH) and serum were quantified using the bicinchoninic acid assay. Immune cells in the AH were detected using hematoxylin and eosin staining and immunocytochemistry. The expression of TGF-β2 in the AH and cytokine levels (IL-10, IFN-γ) in serum were measured using ELISA. Anterior chamber-associated immune deviation (ACAID) was induced in DBA/2J mice by injecting antigens into the anterior chamber. Delayed-type hypersensitivity (DTH) assays were used to assess the induction of ACAID. In DBA/2J mice, before and after pigment dispersion, following anterior chamber injection of pigment particles, and after ACAID modeling, the expression of regulatory T cells (Tregs) was detected using flow cytometry. Compared to C57BL/6J mice, the protein concentration, immune cell count, and TGF-β2 levels in the AH were elevated in DBA/2J mice. Protein concentration and IL-10 levels in serum were increased, while IFN-γ levels were decreased in DBA/2J. Additionally, the expression of Treg cells in the spleen of DBA/2J mice was significantly increased after pigment dispersion and anterior chamber injection of pigment particles. At 3 and 6 months, DTH responses in DBA/2J mice were not inhibited, thus preventing ACAID induction. However, the opposite was observed at 9 months in DBA/2J mice. Furthermore, the ACAID group exhibited an augmented expression of Treg cells. Dispersion of pigment particles in the anterior chamber of the eye enhances the state of ocular immune privilege by influencing the immunosuppressive microenvironment and inducing more Treg cells to reestablish ACAID.

Sex-dependent intra-islet structural rearrangements affecting alpha-to-beta cell interactions lead to adaptive enhancements of Ca2+ dynamics in prediabetic beta cells

Aims/hypothesisPrediabetic pancreatic beta cells can adapt their function to maintain normoglycaemia for a limited period of time, after which diabetes mellitus will manifest upon beta cell exhaustion. Understanding sex-specific beta cell compensatory mechanisms and their failure in prediabetes (impaired glucose tolerance) is crucial for early disease diagnosis and individualised treatment. Our aims were as follows: (1) to determine the key time points of the progression from beta cells’ functional adaptations to their failure in vivo; and (2) to mechanistically explain in vivo sex-specific beta cell compensatory mechanisms and their failure in prediabetes.MethodsIslets from male and female transgenic Ins1CreERT2-GCaMP3 mice were transplanted into the anterior chamber of the eye of 10- to 12-week-old sex-matched C57BL/6J mice. Recipient mice were fed either a control diet (CD) or western diet (WD) for a maximum of 4 months. Metabolic variables were evaluated monthly. Beta cell cytoplasmic free calcium concentration ([Ca2+]i) dynamics were monitored in vivo longitudinally by image fluorescence of the GCaMP3 reporter islets. Global islet beta cell [Ca2+]i dynamics in line with single beta cell [Ca2+]i analysis were used for beta cell coordination studies. The glucagon receptor antagonist L-168,049 (4 mmol/l) was applied topically to the transplanted eyes to evaluate in vivo the effect of glucagon on beta cell [Ca2+]idynamics. Human islets from non-diabetic women and men were cultured for 24 h in either a control medium or high-fat/high-glucose medium in the presence or absence of the glucagon receptor antagonist L-168,049. [Ca2+]i dynamics of human islets were evaluated in vitro after 1 h exposure to Fura-10.ResultsMice fed a WD for 1 month displayed increased beta cell [Ca2+]i dynamics linked to enhanced insulin secretion as a functional compensatory mechanism in prediabetes. Recruitment of inactive beta cells in WD-fed mice explained the improved beta cell function adaptation observed in vivo; this occurred in a sex-specific manner. Mechanistically, this was attributable to an intra-islet structural rearrangement involving alpha cells. These sex-dependent cytoarchitecture reorganisations, observed in both mice and humans, induced enhanced paracrine input from adjacent alpha cells, adjusting the glucose setpoint and amplifying the insulin secretion pathway. When WD feeding was prolonged, female mice maintained the adaptive mechanism due to their intrinsically high proportion of alpha cells. In males, [Ca2+]i dynamics progressively declined subsequent to glucose stimulation while insulin secretion continue to increase, suggesting uncoordinated beta cell function as an early sign of diabetes.Conclusions/interpretationWe identified increased coordination of [Ca2+]i dynamics as a beta cell functional adaptation mechanisms in prediabetes. Importantly, we uncovered the mechanisms by which sex-dependent beta cell [Ca2+]i dynamics coordination is orchestrated by an intra-islet structure reorganisation increasing the paracrine input from alpha cells on beta cell function. Moreover, we identified reduced [Ca2+]i dynamics coordination in response to glucose as an early sign of diabetes preceding beta cell secretory dysfunction, with males being more vulnerable. Alterations in coordination capacity of [Ca2+]i dynamics may thus serve as an early marker for beta cell failure in prediabetes.Graphical

Lack of evidence for GWAS signals of exfoliation glaucoma working via monogenic loss-of-function mutation in the nearest gene.

Exfoliation syndrome (XFS) is a systemic disease of elastin-rich tissues involving a deposition of fibrillar exfoliative material (XFM) in the anterior chamber of the eye, which can promote glaucoma. The purpose of this study was to create mice with CRISPR/Cas9-induced variations in candidate genes identified from human genome-wide association studies (GWAS) and screen them for indices of XFS. Variants predicted to be deleterious were sought in the Agpat1, Cacna1a, Loxl1, Pomp, Rbms3, Sema6a, and Tlcd5 genes of C57BL/6J mice using CRISPR/Cas9-based gene editing. Strains were phenotyped by slit-lamp, SD-OCT imaging, and fundus exams at 1-5 mos of age. Smaller cohorts of 12-mos-old mice were also studied. Deleterious variants were identified in six targets; Pomp was recalcitrant to targeting. Multiple alleles of some targets were isolated, yielding 12 strains. Across all genotypes and ages, 277 mice were assessed by 902 slit-lamp exams, 928 SD-OCT exams, and 358 fundus exams. Homozygosity for Agpat1 or Cacna1a mutations led to early lethality; homozygosity for Loxl1 mutations led to pelvic organ prolapse, preventing aging. Loxl1 homozygotes exhibited a conjunctival phenotype of potential relevance to XFS. Multiple other genotype-specific phenotypes were variously identified. XFM was not observed in any mice. This study did not detect XFM in any of the strains. This may have been due to species-specific differences, background dependence, or insufficient aging. Alternatively, it is possible that the current candidates, selected based on proximity to GWAS signals, are not effectors acting via monogenic loss-of-function mechanisms.

Biochemical Changes in Anterior Chamber of the Eye in Diabetic Patients-A Review.

This article aims to provide a comprehensive review of the biochemical changes observed in the anterior chamber of the eye in diabetic patients. The increased levels of inflammatory markers, alterations in antioxidant defense mechanisms, and elevated levels of advanced glycation end products (AGEs) in the aqueous humor (AH) are explored. Additionally, the impact of these biochemical changes on diabetic retinopathy progression, increased intraocular pressure, and cataract formation is discussed. Furthermore, the diagnostic and therapeutic implications of these findings are presented. This study explores potential biomarkers for detecting diabetic eye disease at an early stage and monitoring its progression. An investigation of the targeting of inflammatory and angiogenic pathways as a potential treatment approach and the role of antioxidant agents in managing these biochemical changes is performed.

Morphometric analysis of the anterior chamber in patients with refractory glaucoma after micropulse cyclophotocoagulation

PURPOSE. To evaluate the anatomical-topographic parameters of the anterior chamber of the eye before and after micropulse cyclophotocoagulation (mCPC) in patients with refractory glaucoma in pseudophakic and phakic eyes.METHODS. The study included 60 patients (62 eyes) with moderate (7 eyes) and advanced (55 eyes) refractory glaucoma. Study groups were formed as follows: the 1st group consisted of 31 pseudophakic eyes (32 patients), the 2nd group included 31 phakic eyes (30 patients). All patients underwent mCPC according to a modified technique. To assess anatomical parameters patients underwent optical coherence tomography of the anterior segment (AS-OCT). Anterior chamber angle (ACA) and anterior chamber depth (ACD) were measured at specific follow-up time points after mCPC.RESULTS. Hypotensive effect by the end of 12-month follow-up was achieved in the patients of the first group in 83.1% of cases, in the second group — in 80.6% of cases (p&lt;0.05). According to OCT findings, anterior chamber depth in patients with phakic eyes during the 12-month follow-up remained the same as before surgery — 2.43±0.21 (mm). In the other group with pseudophakic eyes, anterior chamber depth also did not change during the observation period, with an average value of 3.27±0.12 (mm). No significant changes (p&gt;0.05) were found in the irido-corneal angle measurements in the 4 meridians over the entire follow-up period in both groups.CONCLUSION. Micropulse cyclophotocoagulation performed at fluence rate F=121 J/cm2 does not result in changes of structures of the anterior segment of the eye. The stability of anterior chamber anatomy as well as the absence of postoperative complications in early and longterm follow-up proved the minimal traumatic effect of this surgery in patients with refractory glaucoma.

Pathological characteristic of the eye changes within dogs ophthalmopathy associated with paraneoplastic syndrome

Ophthalmopathy associated with parneoplastic syndrome is a poorly studied pathology of the dogs eye. They occur as a result of the malignant neoplasm’s systematic exposure on the animal's body. They appear in the form of non-specific clinical signs. Main objective of the study – to study the pathomorphological changes in the dog’s eyes with malignant neo-plasms. The material for the study were 30 eyeballs enucleated in dogs in the terminal stage of cancer. Autopsy material obtained from animals was subjected to standard histological processing. As a result of histological studies, pathomorphological changes in eyeball structures associated with Patnaik G2 and G3 mastocytomas (28.6% of cases), liver and kidney carcinomas (28.6%), lymphoma (28.6%) and spleen hemangiosarcoma (14.2%) were revealed in 46.7% of cases. The main histopathological changes were thickening of the iris due to inflammatory cellular infiltration (57.1%) and vasodilation of blood vessels (42.9%), the presence of fibrinous hemorrhagic exudate in the anterior chamber of the eye (28.6%), edema and inflammatory infiltration of the vascular membrane proper (42.9%), thinning, loss of layering and weak retinal atrophy (42.9%). The revealed histological changes indicate the predominant involvement in the pathological process of the vascular membrane of the eye with a violation of the hematophthalmic barrier, as the main pathogenetic mechanism for the development of endogenous uveitis against the background of malignant neoplasm in the animal body.

Beyond the Dusty Fog: Local Eye Drop Therapy and Potentially New Treatment Alternatives in Pseudoexfoliative Glaucoma.

Pseudoexfoliative glaucoma (PEG) is a type of secondary open-angle glaucoma characterized by the accumulation of whitish-gray material on the trabecular meshwork and lens, leading to an increase in intraocular pressure (IOP) and optic nerve damage. Local eye drop therapy is one of the first-line treatments for PEG, which include prostaglandin analogues, beta-blockers, and alpha-adrenergic agonists to lower IOP. New treatments beyond conventional techniques, however, are constantly being developed. One potential treatment proposed for PEG is based on magnetic phage display, which involves using magnetic nanoparticles conjugated to specific peptides or proteins selected using phage display techniques to remove aggregates in the anterior chamber of the eye or inflammatory cells and cytokines that contribute to PEG pathogenesis. Other potential treatments include microRNAs (miRNAs) that are involved in the regulation of gene expression at the post-transcription stages. Gene therapies, nanotechnology, immunotherapy and methods based on stem cells can also be potentially used to target and treat specific tissues and cells responsible for regulating IOP. In addition, photobiomodulation therapy (PBMT), a non-invasive procedure that utilizes low-level laser therapy to improve cellular function and promote tissue repair, can prove an interesting alternative in treating PEG. The aim of our mini-review is to provide a brief overview of these innovative methods that appear to offer potentially promising treatment options for PEG.

In Vivo Imaging of Liver Spheroids Engrafted in the Anterior Chamber of the Mouse Eye.

Biomedical studies of the liver in mammals are hindered by the lack of methods for in vivo noninvasive longitudinal imaging at cellular resolution. Until now, optical imaging of the liver in situ is possible by intravital imaging, which offers high-resolution imaging at the cellular level but cannot be performed multiple times and, therefore, longitudinally in the same animal. Noninvasive imaging methods, such as bioluminescence, allow repeated imaging sessions on the same animal but do not achieve cell resolution. To address this methodology gap, we have developed a platform for noninvasive in vivo imaging of liver spheroids engrafted in the anterior chamber of the mouse eye. In the workflow described in this study, primary mouse liver spheroids are generated in vitro and transplanted into the anterior chamber of the eye of recipient mice, where they engraft on the iris. The cornea acts as a natural body window through which we can image the engrafted spheroids by conventional confocal microscopy. The spheroids survive for months in the eye, during which the cells can be studied in contexts of health and disease, as well as being monitored in response to different stimuli over repeated imaging sessions using appropriate fluorescent probes. In this protocol, we provide a breakdown of the necessary steps to implement this imaging system and explain how to best harness its potential.

Abstract 3633: Multi-parametric characterization of extracellular vesicles in the aqueous humor of uveal melanoma eyes undergoing brachytherapy

Abstract Introduction: Aqueous humor (AH), fluid in the anterior chamber of the eye, is a liquid biopsy source of tumor information in uveal melanoma (UM). Extracellular vesicles (EVs) have recently been characterized in the AH of retinoblastoma eyes, however, have not been investigated in UM. EVs are small membrane-enclosed particles secreted by virtually every cell type with a role in intercellular communications. We hypothesize that tumor-derived EVs are detectable in the AH of UM eyes and that understanding the type of vesicles present may help us better understand their function, particularly for cancer. Small extracellular vesicles (sEVs) have been shown to promote tumorigenesis, treatment resistance, and metastasis, leading to the discovery of clinically-relevant biomarkers for multiple cancer types. As the diversity and role of sEVs in the AH of UM patients has never previously been reported, this project is a first step towards investigating new biomarkers of clinical utility. Our aim is to use single particle analysis to characterize sEV subpopulations in the AH of UM patients by quantifying their size, concentration, and phenotypes based on cell surface markers, specifically tetraspanin co-expression patterns. Methods: sEVs were analyzed from paired pre- and post-brachytherapy AH samples collected from 19 UM patients. AH samples from 5 glaucoma patients served as the control cohort. Unprocessed AH underwent Nanoparticle Tracking Analysis for size quantification and Single Particle-Interferometric Reflectance Imaging Sensor analysis for fluorescent particle count and tetraspanin immunophenotyping; counts were subsequently converted to percentages for analysis. Statistics were performed using Mann-Whitney U, ANOVA, and Tukey’s tests. Results: AH derived from UM eyes (pre-brachytherapy) displayed a broader sEV size distribution than glaucoma eyes, with more diverse sEV subpopulations. Both tumor and nontumor AH samples demonstrated a dominant CD63+ sEV subpopulation. A significantly higher percentage of CD63/81+ sEVs (P= 0.0217) and CD9/63/81+ sEVs (P= 0.0146) were found in post-brachytherapy samples compared to pre-brachytherapy. Conclusions: sEV subpopulations in UM eyes demonstrate increased heterogeneity in comparison to nontumor eyes. Our results are consistent with prior studies reporting that the CD63+ sEV subtype is an AH-specific biomarker enriched across multiple ocular diseases. The increase in CD63/81+ sEVs and CD9/63/81+ sEVs after brachytherapy is presumably a result of radiation-induced release of these vesicles, suggesting a tumor-derived origin of these subpopulations. CD63/81+ sEV subpopulation was previously reported to also be a tumor-derived subpopulation in AH of retinoblastoma eyes. Further study of the cargo carried by these sEV subpopulations may uncover molecular mechanisms pivotal to UM and other ocular cancers. Citation Format: Shreya Sirivolu, Chen-Ching Peng, Paolo Neviani, Jesse L. Berry, Liya Xu. Multi-parametric characterization of extracellular vesicles in the aqueous humor of uveal melanoma eyes undergoing brachytherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3633.

Comparative analysis of the anterior chamber changes after intravitreal injections of aflibercept and brolucizumab

BACKGROUND: Currently, the information is insufficient about the effect of multiple intravitreal injections (IVI) of brolucizumab in comparison with aflibercept on the parameters of iridolenticular diaphragm. AIM: The aim of this study is using optical coherence tomography of the anterior segment (AS-OCT) to evaluate changes in the parameters of the anterior chamber in patients with multiple intravitreal injections of aflibercept and brolucizumab in the treatment of the neovascular form of age-related macular degeneration (nAMD). MATERIALS AND METHODS: In total, 60 patients (60 eyes) were included into the study. The follow-up period duration was 12 months. Patients were divided into 2 groups: group 1 — 30 patients receiving intravitreal injections of aflibercept, group 2 — 30 patients receiving intravitreal injections of brolucizumab. Using a Revo NX tomograph, AS-OCT was performed, the depth of the anterior chamber and the dimensions of the anterior chamber angle were examined. The measurement was performed before intravitreal injections, one month after the first, one month after the third intravitreal injections, and one year after treatment. RESULTS: In both groups, a shift of the iridolenticular diaphragm was noted according to AS-OCT data — a significant decrease in depth of the anterior chamber, anterior chamber angle parameters before treatment and after 4 months of treatment, as well as after a year of treatment. In group 1, in contrast to group 2, a greater number of intravitreal injections led to a greater decrease in the depth of the anterior chamber (rxy = 0.415, p = 0.02) and to a greater narrowing of the anterior chamber angle on the nasal and temporal sides (rxy = 0.37, p = 0.049 and rxy = 0.424, p = 0.02, respectively). CONCLUSIONS: For the first time, changes of the anterior chamber of the eye were determined against the background of multiple intravitreal injections of aflibercept and brolucizumab in the treatment of the neovascular form of age-related macular degeneration using AS-OCT.

Pharmacologic activity of mGLUR4 agonist in a model of primary open-angle glaucoma: A preclinical experimental study

Background. Worldwide, more than 67 million people suffer from glaucoma. In Russia, this number exceeds 1.08 million people. Annually, the number of primary cases increases by 3–4%. The increasing prevalence of glaucoma intensifies the search for neuroprotectants that can reduce the loss of retinal ganglion cells, thereby impeding the progression of the disease. Objective. To study of the pharmacological activity of mGLUR4 agonist, ZC64-0001 substance, on a model of primary open-angle glaucoma. Methods. The conducted preclinical study involved 60 sexually mature male Wistar rats, weighing 180–220 g. The observation period was 73 days. The animals were divided into 6 groups with 10 animals in each. Group 1 included intact animals (without any manipulations); Group 2 included negative control animals with the water injected in the anterior chamber of the eye; in Group 3–6, primary open-angle glaucoma was modelled by injecting 1% hyaluronic acid solution into the anterior chamber of the eye once every 7 days from day 1 to day 62 of the study. Animals in Group 4 were treated intragastrically with H-[(4-chlorophenyl)methyl]-1,6-dihydro-4-methoxy-1-(2-methylphenyl)-6-oxo-3-pyridazinecarboxamide under the laboratory code of ZC64-0001 at a dose of 10 mg/kg. Animals in Group 5 received Mexidol as a comparison drug intramuscularly at a dose of 25.7 mg/kg. Animals in Group 6 were treated with a Timolol instillation at a dose of 0.009 ml/kg. The studied compounds were administered from day 63 of the study once a day for 10 days. The evaluated indicators included the level of microcirculation in the retina, the amplitude of a-wave and b-wave of the electroretinogram, and the number of retinal ganglionic layer nuclei in the setting of the conducted treatment. Statistical processing of the data was performed using the Statistica 10.0 software (StatSoft, USA). Differences were determined at the significance level of p ˂ 0.05. Results. ZC64-0001 increased the level of microcirculation relative to the group with modelled glaucoma by 11.5%, with this indicator being statistically significantly different from that both in the group with modelled glaucoma and the intact group ( р &lt; 0.05). In the group of animals receiving ZC64-0001, the amplitude of a-wave increased relative to the primary glaucoma group by 17.7%, with this index being statistically different from the groups of intact animals and negative control (р &lt; 0.05). The b-wave amplitude increased by 34.4% relative to the group with modelled glaucoma, being statistically different from the intact group, negative control group, pathology modelled group, and comparison drug groups ( р &lt; 0.05). Administration of ZC64-0001 increased the number of retinal ganglion cell nuclei relative to the group with modelled glaucoma by 41.0%, which had a statistically significant difference from all the studied groups ( р &lt; 0.05). Conclusion. The ZC64-0001 compound demonstrated high neuroprotective properties in a model of primary open-angle glaucoma, leading to an improvement in retinal microcirculation, an increase in the wave amplitude according to the conducted electrophysiological study, and an increase in the number of ganglion cell nuclei.

N-acetylserotonin alleviates retinal ischemia-reperfusion injury via HMGB1/RAGE/NF-κB pathway in rats.

To observe the effects of N-acetylserotonin (NAS) administration on retinal ischemia-reperfusion (RIR) injury in rats and explore the underlying mechanisms involving the high mobility group box 1 (HMGB1)/receptor for advanced glycation end-products (RAGE)/nuclear factor-kappa B (NF-κB) signaling pathway. A rat model of RIR was developed by increasing the pressure of the anterior chamber of the eye. Eighty male Sprague Dawley were randomly divided into five groups: sham group (n=8), RIR group (n=28), RIR+NAS group (n=28), RIR+FPS-ZM1 group (n=8) and RIR+NAS+ FPS-ZM1 group (n=8). The therapeutic effects of NAS were examined by hematoxylin-eosin (H&E) staining, and retinal ganglion cells (RGCs) counting. The expression of interleukin 1 beta (IL-1β), HMGB1, RAGE, and nod-like receptor 3 (NLRP3) proteins and the phosphorylation of nuclear factor-kappa B (p-NF-κB) were analyzed by immunohistochemistry staining and Western blot analysis. The expression of HMGB1 protein was also detected by enzyme-linked immunosorbent assay (ELISA). H&E staining results showed that NAS significantly reduced retinal edema and increased the number of RGCs in RIR rats. With NAS therapy, the HMGB1 and RAGE expression decreased significantly, and the activation of the NF-κB/NLRP3 pathway was antagonized along with the inhibition of p-NF-κB and NLRP3 protein expression. Additionally, NAS exhibited an anti-inflammatory effect by reducing IL-1β expression. The inhibitory of RAGE binding to HMGB1 by RAGE inhibitor FPS-ZM1 led to a significant decrease of p-NF-κB and NLRP3 expression, so as to the IL-1β expression and retinal edema, accompanied by an increase of RGCs in RIR rats. NAS may exhibit a neuroprotective effect against RIR via the HMGB1/RAGE/NF-κB signaling pathway, which may be a useful therapeutic target for retinal disease.

Ocular Toxocariasis in The Anterior Chamber of The Eye

Introduction : Ocular toxocariasis (OT) is a zoonotic infection caused by Toxocara canis and Toxocara cati. Dogs and cats, the definitive hosts of the roundworm, pass unembryonated eggs via their feces into the environment (often into the soil). Transmission to humans occurs through ingestion of soil or contaminated food, or the fecal– oral route.&#x0D; Case Illustration : A 14 years old male presented with pain and redness in the right eye 2 months prior to the visit. Complaints are redness, itching, and pain especially in the night on the right eye. The patient had habits of eating raw meat and walking around in dirt without any footwear. Ophthalmologic examination showed a toxocara sp. larvae in the anterior chamber of the right eye. This patient treated with surgical removal of the worm, given antihelminth and topical corticosteroid.&#x0D; Discussion : Toxocariasis results from infection with Toxocara cati (an intestinal parasite of cats) or Toxocara canis (an intestinal parasite of dogs). Children acquire the disease by close association with pets and by eating dirt (pica) contaminated with Toxocara ova. The ingested ova form larvae that penetrate the intestinal mucosa and gain access to the systemic circulation and finally to the eye. Systemic or local corticosteroids should be given when there is evidence of significant intraocular inflammation.&#x0D; Conclusion : This case described a patient who was diagnosed with a living worm as foreign bodies in the right eye.

Topical Immunosuppressant Agent Cyclosporine (CsA) as Therapy for Ocular Alkali Chemical Injury: Is It Really Beneficial?

Introduction : Alkali chemicals can cause more severe ocular injury than acidic chemicals, because the alkali can rapidly penetrate the cornea into the anterior chamber of the eye and damage the iris, ciliary body, lens and trabecular meshwork, which can cause significant visual impairment. A topical immunosuppressant Cyclosporine (which also known as Cyclosporin A or CsA) was used in the treatments.&#x0D; Case Illustration : A 3-year-old male patient presented painful eyes 3 days after an accidental ocular injury with Calcium Hydroxide. The ophthalmic examination revealed based on Roper-Hall grading, the injuries indicated grade III-IV ODS. The patient's BCVA OD 2/60, and OS 20/200. The patient was hospitalized and underwent intensive conservative pharmacological therapy, including topical CsA. Corneal healing was examined by serial corneal Optical coherence tomography (OCT) and showed improvement of the corneal haze throughout the follow-ups. From the last follow-up, the BCVA OD 20/70, OS 20/40.&#x0D; Discussion : Topical CsA reduces the number of CD3-positive cells (T lymphocytes) and the production of pro- inflammatory cytokines, which was associated with the decreased expression of matrix metalloproteinase-9, inducible nitric oxide synthase, VEGF and active caspase-3. Corneal neovascularization is a clinical condition caused by alkali burn that, when left untreated, leads to significant visual impairment. VEGF-A is one of the most important mediators of angiogenesis, leading to neovascularization. Hence, topical CsA 0.1% eye drops may play a role in the treatment of alkali corneal injury.&#x0D; Conclusion : This case report showed that topical CsA gave a significant corneal wound healing, combined with the mainstay of alkali chemical injury.

AHP DB: a reference database of proteins in the human aqueous humor.

The aqueous humor (AH) is a low-viscosity biofluid that continuously circulates from the posterior chamber to the anterior chamber of the eye. Recent advances in high-resolution mass-spectrometry workflows have facilitated the study of proteomic content in small-volume biofluids like AH, highlighting the potential clinical implications of the AH proteome. Nevertheless, in-depth investigations into the role of AH proteins in ocular diseases have encountered challenges due to limited accessibility to these workflows, difficulties in large-scale AH sample collection and the absence of a reference AH proteomic database. In response to these obstacles, and to promote further research on the involvement of AH proteins in ocular physiology and pathology, we have developed the web-based Aqueous Humor Proteomics Database (AHP DB). The current version of AHP DB contains proteomic data from 307 human AH samples, which were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The database offers comprehensive information on 1683 proteins identified in the AH samples. Furthermore, relevant clinical data are provided for each analyzed sample. Researchers also have the option to download these datasets individually for offline use, rendering it a valuable resource for the scientific community. Database URL: https://ahp.augusta.edu/.

A modern approach to diagnostic methods of the cornea and aqueous humor of anterior chamber of the eye in patients with anterior uveitis, associated with spondyloarthritis

Anterior uveitis is the dominant clinical extra-articular manifestation of spondyloarthritis associated with the carriage of the human leukocyte antigen allele (HLA-B27), which may precede joint and spinal involvement. In 20–30% of cases, uveitis occurs with frequent relapses – up to 5–6 exacerbations per year, often has a protracted character and is poorly amenable to local anti-inflammatory therapy, which leads to a decrease in vision and a deterioration in the quality of life. Timely and accurate assessment of the severity of intraocular inflammation plays an important role in making a diagnosis and in choosing the right treatment tactics for treating anterior uveitis in patients with HLA-B27-associated spondyloarthritis. The current standard clinical approach to assessing inflammation is the presence and number of inflammatory cells in the anterior chamber aqua humor according to the SUN classification using slit lamp imaging. However, the assessment of SUN is very subjective. Moreover, the individual variability in anterior chamber cell count, which is the main diagnostic criterion for uveitis, can be exacerbated by certain testing conditions, such as the brightness of the slit lamp light bar, light beam size calibration, light source intensity. Advances in optical imaging technology, the application of the latest technologies and the improvement of already known methods offer new opportunities for an objective, accurate and quantitative assessment of inflammation. The article describes the possibilities of studying the cornea and the anterior chamber of the eye using anterior segment optical coherence tomography, confocal microscopy of the cornea and keratotopometry.