-
Molecular Biology Reports Dec 2023MicroRNA (miRNA) expression analysis has been shown to provide them as biomarkers in several eye diseases and has a regulatory role in pathogenesis. However, miRNA...
BACKGROUND
MicroRNA (miRNA) expression analysis has been shown to provide them as biomarkers in several eye diseases and has a regulatory role in pathogenesis. However, miRNA expression analysis in the vitreous humor (VH) of intraocular tuberculosis (IOTB) is not studied. Thus, we aim to find miRNA expression signatures in the VH of IOTB patients to identify their regulatory role in disease pathogenesis and to find them as potential biomarkers for IOTB.
METHODS AND RESULTS
First, we profiled miRNAs in VH of three IOTB and three Macular hole (MH) samples as controls through small-RNA deep sequencing using Illumina Platform. In-house bioinformatics analysis identified 81 dysregulated miRNAs in IOTB. Further validation in VH of IOTB (n = 15) compared to MH (n = 15) using Real-Time quantitative PCR (RT-qPCR) identified three significantly upregulated miRNAs, hsa-miR-150-5p, hsa-miR-26b-5p, and hsa-miR-21-5p. Based on the miRNA target prediction, functional network analysis, and RT-qPCR analysis of target genes, the three miRNAs downregulating WNT5A, PRKCA, MAP3K7, IL7, TGFB2, IL1A, PRKCB, TNFA, and TP53 genes involving MAPK signaling pathway, PI3K-AKT signaling pathway, WNT signaling pathway, Cell cycle, TGF-beta signaling pathway, Long-term potentiation, and Sphingolipid signaling pathways, have a potential role in disease pathogenesis. The ROC analysis of RT-qPCR data showed that hsa-miR-150-5p with AUC = 0.715, hsa-miR-21-5p with AUC = 0.789, and hsa-miR-26b-5p with AUC = 0.738; however, the combination of hsa-miR-21-5p and hsa-miR-26b-5p with AUC = 0.796 could serve as a potential biomarker for IOTB.
CONCLUSIONS
This study provides the first report on miRNA expression signatures detected in VH for IOTB pathogenesis and also provides a potential biomarker for IOTB.
Topics: Humans; Vitreous Body; Phosphatidylinositol 3-Kinases; Gene Expression Profiling; MicroRNAs; Biomarkers
PubMed: 37906423
DOI: 10.1007/s11033-023-08819-1 -
Die Ophthalmologie Jun 2024An epiretinal membrane (ERM) is a frequently occurring disease affecting the macula, which can be associated with visual impairment and metamorphopsia, depending on the... (Review)
Review
An epiretinal membrane (ERM) is a frequently occurring disease affecting the macula, which can be associated with visual impairment and metamorphopsia, depending on the severity and location. A distinction is made between an idiopathic form caused by age-related changes of the vitreous body and a secondary form associated with diseases of the posterior segment. The development of fibrocellular epiretinal membranes formed by dedifferentiation of intraretinal and extraretinal cells at the level of the vitreomacular interface plays a major role in the pathogenesis. The diagnostics and indications for surgical treatment of ERM are based on the visual acuity, evidence of metamorphopsia, ophthalmoscopic findings and optical coherence tomography (OCT) of the macula. In addition to the possibility of observation of the course where benign spontaneous courses are not uncommon, pars plana vitrectomy (PPV) with peeling of the ERM and internal limiting membrane (ILM) to prevent recurrences is the treatment of choice in symptomatic patients. The prognosis after surgical treatment is very good. In approximately two thirds of the cases, an improvement in visual acuity and/or a reduction of metamorphopsia can be achieved, with a number of predictive, primarily OCT-based factors enabling a prediction of the functional prognosis. Comprehensive patient education regarding the generally long duration of postoperative rehabilitation and the possibility of persistent symptoms or visual deterioration despite successful membrane removal is essential.
Topics: Humans; Epiretinal Membrane; Vitrectomy; Tomography, Optical Coherence; Vision Disorders; Visual Acuity
PubMed: 38831204
DOI: 10.1007/s00347-024-02055-z -
International Ophthalmology May 2024To evaluate the levels of anxiety and depression in patients with symptomatic vitreous floaters and to determine the possible correlations of psychological implications...
PURPOSE
To evaluate the levels of anxiety and depression in patients with symptomatic vitreous floaters and to determine the possible correlations of psychological implications with the symptoms duration and possible improvement, the degree of posterior vitreous detachment, and the discomfort severity.
METHODS
Ninety patients complaining for floaters and fifty-seven age- and gender-matched healthy-control subjects were recruited. Every participant underwent a complete ophthalmological examination, including funduscopy and optical coherence tomography scans, while clinical and demographic data were also gathered. The Patient Health Questionnaire-9 (PHQ-9), the Zung Depression Inventory-Self-Rating Depression Scale (Zung SDS), and the Hospital Anxiety and Depression Scale (HADS) were completed by everyone.
RESULTS
Between the studied groups, no significant differences were detected regarding the clinical and demographic data (p > 0.05). The patients with floaters had significantly higher scores of PHQ-9, Zung SDS, HADS Anxiety, and HADS Depression (p < 0.001). After adjustment for several confounders, PHQ-9 (p = 0.041), Zung SDS (p = 0.003), and HADS Anxiety (p = 0.036) values remained significantly impaired. Among the patients, PHQ-9 and Zung SDS scores were significantly elevated in the patients with floaters duration less than 4 weeks (p < 0.05). Finally, anxiety and depression were significantly correlated with the symptoms duration and intensity, with the floater-associated discomfort, and with the stage of posterior vitreous detachment.
CONCLUSION
Vitreous floaters have a negative impact on patients' psychological status, by the terms of enhanced depressive and anxiety levels. To the best of our knowledge, our study is the first in the literature to elaborate the aforementioned association, by assessing three different questionnaires simultaneously.
Topics: Humans; Male; Female; Middle Aged; Vitreous Body; Depression; Adult; Anxiety; Eye Diseases; Tomography, Optical Coherence; Surveys and Questionnaires; Aged; Case-Control Studies; Vitreous Detachment; Vision Disorders
PubMed: 38713290
DOI: 10.1007/s10792-024-03006-y -
Legal Medicine (Tokyo, Japan) Jun 2024Ocular tension decreases with increasing postmortem interval (PMI) and eyes collapse with extreme progression of postmortem change; however, time-related changes in...
BACKGROUND
Ocular tension decreases with increasing postmortem interval (PMI) and eyes collapse with extreme progression of postmortem change; however, time-related changes in postmortem computed tomography (PMCT) findings have not been clarified. This study aimed to quantitatively evaluate the vitreous volume and CT values of the vitreous body to clarify time-related changes in PMCT.
METHODS
We retrospectively reviewed PMCT images of the eyes of subjects who underwent autopsy at our institution between July 2023 and February 2024. They were classified into four PMI groups: PMI-1: < 1 day, PMI-2: 1-2 days, PMI-3: 3-6 days, PMI-4: ≥ 7 days. The vitreous volumes and mean CT values of the vitreous chambers were measured. Additionally, the presence of intraocular gas and crystalline lens dislocation was observed.
RESULTS
The PMCT images of 131 eyes of 66 patients (41 males, 25 females) were analyzed. The mean age was 63.7 (22-99) years. The volume of the vitreous cavity was significantly decreased at PMI-3 and PMI-4, which indicated scleral depression. Intraocular gas appeared in PMI-4. Lens dislocation began at PMI-3 and was more frequently observed at PMI-4 and in the higher-temperature environment group.
CONCLUSION
Ocular findings on PMCT are useful for estimating the time of death.
PubMed: 38936142
DOI: 10.1016/j.legalmed.2024.102477 -
Veterinary Ophthalmology Mar 2024To describe a repeatable method of measuring ocular structures and to establish ocular biometry reference ranges in adult domestic rabbits (Oryctolagus cuniculus)...
OBJECTIVE
To describe a repeatable method of measuring ocular structures and to establish ocular biometry reference ranges in adult domestic rabbits (Oryctolagus cuniculus) without medical history or imaging findings consistent with ophthalmic disease using a 64-slice multidetector computed tomography scanner.
PROCEDURE
In this retrospective and observational anatomic study, 100 eyes from 50 rabbits without medical history or imaging findings consistent with ophthalmic disease who received a head computed tomography scan were selected for measurement of globe length, width, and height using 3D multiplanar reconstruction. Lens width and length, the anteroposterior length of the anterior and vitreous chambers, and attenuation of the lens and vitreous chamber were collected. These parameters were compared against age, sex, weight, body condition, and ear conformation.
RESULTS
A reference guide was established, with globe width being the largest dimension (18.03 ± 0.81 mm), followed by height (17.18 ± 0.69 mm) and then length (16.64 ± 0.66 mm). Increased weight was associated with an increase in globe height (p = 2.43 × 10 ), length (p = 1.63 × 10 ), and width (p = 7.0 × 10 ). Increased age was associated with increased lens attenuation (p = 1.28 × 10 ) and increased transverse lens width (p = 1.64 × 10 ). Inter- and intra-observer agreement was excellent.
CONCLUSIONS
CT is a reliable modality for measurement of ocular biometry dimensions in rabbits. These reference values can be applied to aid in identifying diseases that affect the dimensions of the ocular structures in rabbits over 18 months of age.
PubMed: 38468158
DOI: 10.1111/vop.13209 -
Scientific Reports Aug 2023The exact link between systemic and ocular endogenous corticoids (steroidome) is unclear and whether the ocular steroidome is altered in CSCR eyes is unknown. The aims...
The exact link between systemic and ocular endogenous corticoids (steroidome) is unclear and whether the ocular steroidome is altered in CSCR eyes is unknown. The aims of this study were to analyze the human steroidome in the aqueous humor as a function of age, sex and time of the day, to correlate systemic and ocular steroidome and to analyze the ocular steroidome in long lasting complex inactive CSCR. Based on our results, we present two CSCR cases treated by the combination of oral mineralocorticoid antagonist and glucocorticoids drops. In a cross-sectional study, aqueous humor (AH) was collected between 8am and 6 pm from 50 unaffected individuals (25 men and 25 women) and from 14 patients with chronic CSCR, during cataract surgery. In addition, simultaneous serum and AH were collected from 27 individuals undergoing cataract surgery and, simultaneous AH and vitreous were collected from 9 patients undergoing cataract and vitrectomy to estimate corticoids levels in the different compartments. The steroidome was determined using a LC-MS/MS method that quantifies 13 endogenous corticoids from the gluco, mineralocorticoid and androgen pathways. In AH and vitreous, the highest corticoid level is reached by cortisol (F), that represents less than 10% of F serum level. The cortisol levels in the serum did not correlate with ocular cortisol levels. Serum and ocular cortisone (E) levels correlate, although less than 5% of circulating E reaches the eye. The only mineralocorticoids measured in the AH were corticosterone (B) and its inactive form, the 11-desoxycorticosterone (A). There was no influence of circadian rhythm on cortisol ocular levels and there was no correlation between the age or the sex and the level of F, E, A, and B. In eyes with chronic inactive CSCR, the levels of the active glucocorticoid form F was lower than in control eyes and the F/E ratio was reduced by 50% but the B/A ratio was higher indicating imbalance towards active mineralocorticoids. Base on this observation, we propose to combine an antagonist of the mineralocorticoid receptor together with topical glucocorticoids in two CSCR patients, resistant to all other treatments, with favorable outcome. Our results indicate that the ocular psteroidome is highly regulated suggesting a local metabolism of ocular corticoids. In eyes with long-lasting complex inactive CSCR, the steroidome analysis shows lower active glucocorticoids and higher active mineralocorticoids.
Topics: Male; Humans; Female; Central Serous Chorioretinopathy; Glucocorticoids; Mineralocorticoids; Hydrocortisone; Chromatography, Liquid; Cross-Sectional Studies; Tandem Mass Spectrometry; Cataract
PubMed: 37644063
DOI: 10.1038/s41598-023-41126-0 -
Neuropharmacology Jan 2024To investigate whether Tafluprost could promote optic nerve regeneration in mice after optic nerve crush (ONC) and determine the underlying molecular mechanism.
PURPOSE
To investigate whether Tafluprost could promote optic nerve regeneration in mice after optic nerve crush (ONC) and determine the underlying molecular mechanism.
METHODS
Tafluprost was injected into the vitreous body immediately after ONC. The level of Zn in the inner plexiform layer (IPL) of the retina was stained using autometallography (AMG). The number of survival retinal ganglion cells (RGCs) was determined via dual staining with RGC markers Tuj1 and RBPMS. Individual axons that regenerated to 0.25, 0.5, 0.75 and 1 mm were manually counted in the whole-mount optic nerve labeled by cholera toxin B fragment (CTB). Immunofluorescence and Western blot were performed to detect protein expression levels. Pattern electroretinogram was used to evaluate RGCs function.
RESULTS
Tafluprost promoted RGC survival in a dose-dependent manner with an optimal concentration of 1 μM. Tafluprost significantly decreased ZnT-3 expression and Zn accumulation in the IPL of retina. Tafluprost stimulated intense axonal regeneration and maintained RGCs function compared to control. Mechanistically, Tafluprost and Zn elimination treatment (TPEN or ZnT-3 deletion) can activate the mTOR pathway with an improved percentage of pS6 RGCs in the retina. However, rapamycin, a specific inhibitor of the mTOR1, inhibited the activation of the mTOR pathway and abolished the regenerative effect mediated by Tafluprost. Tafluprost also inhibited the upregulation of p62, LC3 and Beclin-1, attenuated the overactivation of microglia/macrophages and downregulated the expression of TNFα and IL-1β.
CONCLUSIONS
Our results suggest that Tafluprost promoted axon regeneration via regulation of the Zn-mTOR pathway, and provide novel research directions for glaucomatous optic nerve injury mechanisms.
Topics: Mice; Animals; Axons; Nerve Regeneration; Optic Nerve; Optic Nerve Injuries; TOR Serine-Threonine Kinases; Zinc; Nerve Crush
PubMed: 37832634
DOI: 10.1016/j.neuropharm.2023.109746 -
Scientific Reports Apr 2024The mechanisms underlying myopia remain not fully understood. We proposed to examine the function and underlying mechanisms of miR-204-5p in myopia development. The...
The mechanisms underlying myopia remain not fully understood. We proposed to examine the function and underlying mechanisms of miR-204-5p in myopia development. The miR-204-5p expression level was assessed in the vitreous humor (VH) of a cohort consisting of 11 patients with high myopia (HM) and 16 control patients undergoing vitrectomy. Then the functional implications of miR-204-5p in ARPE-19 cells were assessed. Thioredoxin-interacting protein (TXNIP) was found as a possible target of miR-204-5p through mRNA sequencing, and its interaction with miR-204-5p was confirmed employing luciferase assay and western blotting. Furthermore, the miR-204-5p function in regulating oxidative stress was examined by measuring reactive oxygen species (ROS) accumulation. The results indicated a significant reduction of miR-204-5p in the VH of HM patients. Overexpression of miR-204-5p suppressed cell proliferation, migration, invasion, and apoptosis in ARPE-19 cells. The direct targeting of miR-204-5p on TXNIP has been confirmed, and its downregulation mediated the miR-204-5p impacts on ARPE-19 cells. Moreover, miR-204-5p overexpression significantly reduced ROS accumulation by targeting TXNIP. Our findings revealed the possible contribution of the miR-204-5p/TXNIP axis in myopia development by regulating oxidative stress, which may provide new targets to combat this prevalent and debilitating condition.
Topics: Humans; MicroRNAs; Oxidative Stress; Carrier Proteins; Myopia; Reactive Oxygen Species; Female; Cell Line; Cell Proliferation; Apoptosis; Male; Cell Movement; Adult
PubMed: 38684840
DOI: 10.1038/s41598-024-60688-1 -
Acta Biomaterialia Nov 2023Retinal vascular diseases such as neovascular age-related macular degeneration (nAMD) are the leading cause of blindness worldwide. They can be treated with intravitreal...
Retinal vascular diseases such as neovascular age-related macular degeneration (nAMD) are the leading cause of blindness worldwide. They can be treated with intravitreal injections of anti-vascular endothelial growth factor (anti-VEGF) agents by inhibiting VEGF which is a major agent of abnormal blood vessel growth. However, because of drug's short half-life, clinical treatment often requires monthly repeated intravitreal injections, causing treatment burden and undertreatment. Among various kinds of drug carriers, in situ forming hydrogels have been studied as potential intravitreal drug carriers for the high drug loading, easy injection, controlled drug release, and protection of encapsulated drugs from the environment. However, gelation time, crosslinking degree, and drug release patterns following injection of a liquid that will be subsequently gelled in situ are susceptible to be hindered by dilution of the hydrogel precursor solution with body fluids (e.g., blood or vitreous). Here, we report an injectable pre-crosslinked hydrogel rod to overcome the limitations of in situ forming hydrogels and to extend intravitreal half-life of anti-VEGF for reducing intraocular injection frequency. Hydrogel rods can be simply prepared using in situ forming hydrogels, and injectable using a designed rod injector. The adjustable crosslinking degree of hydrogel rods easily controlled bevacizumab release profiles in a sustained manner. Compared with in situ forming hydrogels, hydrogel rods effectively reduced initial burst release, and showed sustained release with long-term drug efficacy in vitro. From the 4-month in vivo pharmacokinetic analysis, following the intravitreal injection of hydrogel rods, the half-life of bevacizumab in the vitreous and retina was significantly extended, and drug elimination to aqueous humor was effectively reduced. Finally, intraocular stability, degradation, and inflammatory response of hydrogel rods were evaluated. We expect that the hydrogel rod can be a potential drug delivery system for the treatment of nAMD and other conditions that need long-term and local sustained drug administration. STATEMENT OF SIGNIFICANCE: Herein, we report an injectable pre-crosslinked hydrogel rod based on an in situ forming hydrogel to achieve intravitreal long-acting anti-VEGF delivery to reduce injection frequency and improve the long-term visual outcomes of patients with retinal vascular diseases. Hydrogel rods were readily prepared using removable molds and injected using customized injectors. Compared to the in situ forming hydrogel, hydrogel rods showed significantly reduced initial burst release, controllable release profiles for several months, physical stability, and a long-acting anti-angiogenic effect. Animal studies demonstrated that the hydrogel rods dramatically prolonged the intraocular drug half-life while significantly reducing drug elimination for up to four months. Moreover, the biodegradability and safety of the hydrogel rods suggest their suitability as an advanced intravitreal DDS for treating retinal vascular diseases.
Topics: Animals; Humans; Bevacizumab; Hydrogels; Angiogenesis Inhibitors; Retina; Intravitreal Injections; Drug Carriers; Vascular Diseases
PubMed: 37739248
DOI: 10.1016/j.actbio.2023.09.025 -
Journal of Visualized Experiments : JoVE Sep 2023A critical challenge in translational research is establishing a viable and efficient interface between patient care in the operating room (OR) and the research...
A critical challenge in translational research is establishing a viable and efficient interface between patient care in the operating room (OR) and the research laboratory. Here, we developed a protocol for acquiring high-quality liquid biopsies for molecular analyses from the aqueous humor and the vitreous from patients undergoing eye surgery. In this workflow, a Mobile Operating Room Lab Interface (MORLI) cart equipped with a computer, a barcode scanner, and lab instruments, including onboard cold storage, is used to obtain and archive human biological samples. A web-based data privacy-compliant database enables annotating each sample over its lifetime, and a cartesian coordinate system allows tracking each barcoded specimen in storage, enabling quick and accurate retrieval of samples for downstream analyses. Molecular characterization of human tissue samples not only serves as a diagnostic tool (e.g., to distinguish between infectious endophthalmitis and other non-infectious intraocular inflammation) but also represents an important component of translational research, allowing the identification of new drug targets, development of new diagnostic tools, and personalized therapeutics.
Topics: Humans; Biological Specimen Banks; Vitreous Body; Endophthalmitis; Aqueous Humor; Liquid Biopsy
PubMed: 37747194
DOI: 10.3791/65804