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Molecular Vision 2023Increased inflammatory factor levels have been reported in the vitreous humor (VH) of diabetic retinopathy and neovascular age-related macular degeneration, ocular...
PURPOSE
Increased inflammatory factor levels have been reported in the vitreous humor (VH) of diabetic retinopathy and neovascular age-related macular degeneration, ocular diseases generally associated with the formation of new retinal blood vessels and leakage. However, the levels of inflammatory mediators are less known in retinal degeneration without neovascularization. Human retinitis pigmentosa (RP) and animal models of light-induced retinal degeneration (LIRD) share several features, such as photoreceptor death and retinal inflammation. Here, we aimed to determine the levels of inflammatory factors in the VH of the LIRD mouse model.
METHODS
LIRD was induced by exposing BALB/c mice to white light (15,000 lx, 2 h), and the mice were recovered for 2 days before analysis (n = 50 mice). We assessed retinal morphology using optical coherence tomography and hematoxylin and eosin staining; retinal cell viability was determined using terminal deoxynucleotidyl transferase dUTP nick-end labeling, and retinal responses were measured based on electroretinogram signals. Total retinal RNAs were extracted and subjected to RNA sequencing analysis. VH samples from control (n = 4) and LIRD mice (n = 9) were assayed in triplicate for a panel of four inflammatory mediators using the Simple Plex Cartridge on an Ella System.
RESULTS
Retinal degeneration, photoreceptor death, infiltration of microglia/macrophages into the photoreceptor layer, and loss of a- and b-waves were obviously detected after LIRD. RNA sequencing revealed that light damage (LD) led to the significant upregulation of inflammatory factors in mouse retinas. In the VH, LD increased the total protein concentration. Dramatic induction of CCL2 (~3000 fold) and IL6 (~10 fold) was detected in VH in response to LD. Increased but not significant levels of TNFα and IL1β were also detected in light-exposed VH.
CONCLUSIONS
Given that the LIRD model mimics RP pathogenesis in some aspects, these results suggest a causative link between retinal degeneration and VH inflammation in RP progression, and the increased CCL2 level in VH may reflect similar elevated CCL2 expression in the degenerative retina.
Topics: Mice; Humans; Animals; Retinal Degeneration; Vitreous Body; Retina; Retinitis Pigmentosa; Inflammation; Disease Models, Animal; Inflammation Mediators
PubMed: 38222454
DOI: No ID Found -
Forensic Science International Dec 2019In the US, the use of synthetic opioids (e.g. fentanyl and derivatives) has become an increasing health issue with thousands of overdose deaths being observed since...
In the US, the use of synthetic opioids (e.g. fentanyl and derivatives) has become an increasing health issue with thousands of overdose deaths being observed since 2013. With the high mortality rate associated with these substances, postmortem analyses and interpretation of synthetic opioids has become extremely important. However, due to the novelty of these compounds, the available data are limited and provides challenges to toxicologists. The objectives of this study were (1) to develop and validate analytical methods for the determination of synthetic opioids in vitreous humor and brain, and (2) to investigate the postmortem distribution of new synthetic opioids in blood, vitreous humor, and brain tissue. Vitreous humor (0.5mL) and brain tissue (5g) homogenized in water (diluted 1:3, w/w) were extracted by mixed mode cation exchange-reversed phase solid phase extraction. Extracts were analyzed by liquid chromatography tandem mass spectrometry (LC-MSMS). The chromatographic separation was performed by reversed-phase with 0.1% formic acid in water and in acetonitrile as mobile phases in gradient mode, with a total run time of 21min. Data were acquired with ESI+ in dynamic multiple reaction mode (dMRM), monitoring 2 transitions per compound. The methods were succesfully validated following SWGTOX guidelines, with limits of quantification of 0.1ng/mL in vitreous humor and 0.1ng/g in brain. Fifty-eight authentic case samples from the New York City Office of the Chief Medical Examiner (NYC-OCME) were analyzed to assess the distribution and detectability of synthetic opioids in these postmortem samples. Of the fifteen synthetic opioids included in the method, six synthetic opioids and metabolites (4-ANPP, acetylfentanyl, fentanyl, furanylfentanyl, norfentanyl, U-47700) were detected in the authentic cases. Concentrations for most analytes were within the 0.1 to 100ng/mL or ng/g calibration range across all three matrices, with only concentrations from acetylfentanyl and U-47700 exceeding 100ng/mL or ng/g. The highest concentrations were observed in brain (except norfentanyl), followed by blood and vitreous humor. Most analytes were detected in all three matrices in a given case. This was followed by detection of an analyte in combinations of brain and another matrix or brain only. Through the case analyses, vitreous humor and brain demonstrated to be viable alternatives to blood when performing postmortem analyses of synthetic opioids. Brain exhibited a higher detectability for most analytes when compared to blood and vitreous humor.
Topics: Analgesics, Opioid; Brain Chemistry; Chromatography, Liquid; Forensic Toxicology; Humans; Psychotropic Drugs; Solid Phase Extraction; Synthetic Drugs; Tandem Mass Spectrometry; Vitreous Body
PubMed: 31671355
DOI: 10.1016/j.forsciint.2019.109999 -
Progress in Retinal and Eye Research Sep 2019Diabetic retinopathy (DR) is one of the leading causes of visual impairment in the working-age population. DR is a progressive eye disease caused by long-term... (Review)
Review
Diabetic retinopathy (DR) is one of the leading causes of visual impairment in the working-age population. DR is a progressive eye disease caused by long-term accumulation of hyperglycaemia-mediated pathological alterations in the retina of diabetic patients. DR begins with asymptomatic retinal abnormalities and may progress to advanced-stage proliferative diabetic retinopathy (PDR), characterized by neovascularization or preretinal/vitreous haemorrhages. The vitreous, a transparent gel that fills the posterior cavity of the eye, plays a vital role in maintaining ocular function. Structural and molecular alterations of the vitreous, observed during DR progression, are consequences of metabolic and functional modifications of the retinal tissue. Thus, vitreal alterations reflect the pathological events occurring at the vitreoretinal interface. These events are caused by hypoxic, oxidative, inflammatory, neurodegenerative, and leukostatic conditions that occur during diabetes. Conversely, PDR vitreous can exert pathological effects on the diabetic retina, resulting in activation of a vicious cycle that contributes to disease progression. In this review, we recapitulate the major pathological features of DR/PDR, and focus on the structural and molecular changes that characterize the vitreal structure and composition during DR and progression to PDR. In PDR, vitreous represents a reservoir of pathological signalling molecules. Therefore, in this review we discuss how studying the biological activity of the vitreous in different in vitro, ex vivo, and in vivo experimental models can provide insights into the pathogenesis of PDR. In addition, the vitreous from PDR patients can represent a novel tool to obtain preclinical experimental evidences for the development and characterization of new therapeutic drug candidates for PDR therapy.
Topics: Aging; Diabetic Retinopathy; Humans; Vitreous Body
PubMed: 30951889
DOI: 10.1016/j.preteyeres.2019.03.002 -
Experimental Eye Research Nov 2021Vitreous humor (VH) is not considered as a critical structure in the radiotherapy planning process. In the present study, an experimental animal model was performed to...
Vitreous humor (VH) is not considered as a critical structure in the radiotherapy planning process. In the present study, an experimental animal model was performed to examine the effects of radiotherapy on VH. The right eyes of twelve New Zealand rabbits were irradiated to 60 Gy in 3 fractions in accordance with the scheme used in the treatment of uveal melanoma in our clinic, and contralateral (left) eyes were considered as control. Weekly ophthalmologic examination was performed after irradiation, for three months. At the end of the third month, enucleation and vitreous collection were conducted. The vitreous samples were subjected to metabolomic analyses, ELISA analyses, viscosity measurements, and electron microscopic examination. In control and experimental vitreous samples, 275 different metabolites were identified, and 34 were found to differ significantly between groups. In multivariate analyzes, a clear distinction was observed between control and irradiated vitreous samples. Pathway analysis revealed that nine pathways were affected, and these pathways were mainly related to amino acid metabolism. A significant decrease was observed in the expressions of type II, V, and XI collagens in protein level in the ELISA. There was a non-significant decrease in type IX collagen and viscosity. Electron microscopic examination revealed disrupted collagen fibrillar ultra-structure and dispersed collagen fragments in the experimental vitreous. An intact vitreous is essential for a healthy eye. In this study, we observed that radiation causes changes in the vitreous that may have long-term consequences.
Topics: Animals; Body Fluids; Collagen; Male; Melanoma; Neoplasms, Experimental; Rabbits; Uveal Neoplasms; Vitreous Body
PubMed: 34688623
DOI: 10.1016/j.exer.2021.108802 -
Translational Vision Science &... Feb 2020Proliferative vitreoretinopathy (PVR) occurs in 5%-10% of rhegmatogenous retinal detachment cases and is the principle cause for failure of retinal reattachment surgery.... (Review)
Review
PURPOSE
Proliferative vitreoretinopathy (PVR) occurs in 5%-10% of rhegmatogenous retinal detachment cases and is the principle cause for failure of retinal reattachment surgery. Although there are a number of surgical adjunctive agents available for preventing the development of PVR, all have limited efficacy. Discovering predictive molecular biomarkers to determine the probability of PVR development after retinal reattachment surgery will allow better patient stratification for more targeted drug evaluations.
METHODS
Narrative literature review.
RESULTS
We provide a summary of the inflammatory and fibrogenic factors found in ocular fluid samples during the development of retinal detachment and PVR and discuss their possible use as molecular PVR predictive biomarkers.
CONCLUSIONS
Studies monitoring the levels of the above factors have found that few if any have predictive biomarker value, suggesting that widening the phenotype of potential factors and a combinatorial approach are required to determine predictive biomarkers for PVR.
TRANSLATIONAL RELEVANCE
The identification of relevant biomarkers relies on an understanding of disease signaling pathways derived from basic science research. We discuss the extent to which those molecules identified as biomarkers and predictors of PVR relate to disease pathogenesis and could function as useful disease predictors. (http://www.umin.ac.jp/ctr/ number, UMIN000005604).
Topics: Biomarkers; Humans; Retinal Detachment; Risk Factors; Vitreoretinopathy, Proliferative; Vitreous Body
PubMed: 32742753
DOI: 10.1167/tvst.9.3.23 -
Forensic Science International Jan 2021Vitreous humor (VH) is a specimen of great value in forensic investigations and is being used for evaluating possible post-mortem formation of ethanol. Ethyl glucuronide...
BACKGROUND
Vitreous humor (VH) is a specimen of great value in forensic investigations and is being used for evaluating possible post-mortem formation of ethanol. Ethyl glucuronide (EtG) is an ethanol metabolite that has found interest for the same purpose. Both compounds can be measured in VH and because of differences in rate of distribution and elimination they may offer complementary information.
METHODS
VH, femoral blood (FB) and urine were collected from 117 autopsy cases for forensic investigation. Ethanol was measured with headspace gas chromatography, while EtG was measured with liquid chromatography - tandem mass spectrometry.
RESULTS AND CONCLUSION
Ethanol was detected in all matrices in 39 cases, while EtG was present in 62 cases. The VH-FB and the VH-urine ethanol concentrations in the 39 cases were statistically correlated (p < 0.00001). In one case with an ethanol concentration of 0.11 g/L in FB, no ethanol was detected in VH and urine, and no EtG in any specimen, indicating a possible post-mortem formation. EtG was present in VH in more cases than in FB and urine. The correlation between the EtG concentrations in VH and FB was statistically significant (p < 0.0003) as was the case also for VH and urine (p < 0.001). The combined information on ethanol and EtG concentrations in the three matrices can be used to interpret alcohol drinking habit before death. This study confirms the value of using VH as a specimen in forensic investigations regarding recent exposure to ethanol. EtG can be used not only for investigating post-mortem ethanol formation but also for estimating recent alcohol drinking.
Topics: Adult; Aged; Aged, 80 and over; Alcohol Drinking; Biomarkers; Central Nervous System Depressants; Chromatography, Liquid; Ethanol; Female; Forensic Toxicology; Glucuronates; Humans; Male; Middle Aged; Tandem Mass Spectrometry; Vitreous Body; Young Adult
PubMed: 33234349
DOI: 10.1016/j.forsciint.2020.110567 -
Investigative Ophthalmology & Visual... Jan 2023Pathologic myopia (PM) is one of the primary causes of blindness. This study aims to explore the possible relations between the composition of microRNA in vitreous...
PURPOSE
Pathologic myopia (PM) is one of the primary causes of blindness. This study aims to explore the possible relations between the composition of microRNA in vitreous exosomes of patients with PM and the progression of myopic maculopathy.
METHODS
Vitreous humor (VH) samples were collected from patients undergoing retinal surgery. A total of 15 and 12 VH samples were obtained from patients with PM and control, respectively. The PM group was divided into PM-L (G2) and PM-H groups (G3 and G4) in order to explore differentially expressed microRNAs (DEMs) that account for the relatively poor prognosis in G3 and G4 myopic maculopathy. A Weighted Gene Co-Expression Network Analysis (WGCNA) was conducted to find the persistently altered key microRNAs in myopic maculopathy progression. The Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analysis were used.
RESULTS
High purity exosomes were extracted from the vitreous fluid of patients with PM and control. The top five downregulated DEMs of PM-H versus PM-L can reflect the tendency of deterioration of PM-H myopic maculopathy. MiR-143-3p and miR-145-5p, which were found in WGCNA, may participate in the development of myopic maculopathy. These microRNAs all relate to the insulin resistance pathway.
CONCLUSIONS
This is the first study to explore the relations between the progression of myopic maculopathy and vitreous exosomal microRNAs. Vitreous exosomal miR-143-3p and miR-145-5p can be considered biomarkers for patients with PM, and the vitreous exosomal DEM associated with PM-H may represent alarming signals of myopic maculopathy deterioration.
Topics: Humans; MicroRNAs; Myopia, Degenerative; Vitreous Body; Body Fluids; Retinal Diseases; Macular Degeneration; Exosomes
PubMed: 36648415
DOI: 10.1167/iovs.64.1.9 -
International Ophthalmology Jul 2023To research whether serum vascular endothelial growth factor (VEGF) levels could be used to evaluate diabetic retinopathy (DR) progression and to compare vitreous VEGF...
PURPOSE
To research whether serum vascular endothelial growth factor (VEGF) levels could be used to evaluate diabetic retinopathy (DR) progression and to compare vitreous VEGF levels after injections of intravitreal bevacizumab (IVB), ranibizumab (IVR), and triamcinolone acetonide (IVTA) in proliferative diabetic retinopathy (PDR).
METHODS
We enrolled a total of 91 eyes of 89 subjects (70 eyes of 68 diabetics and 21 eyes of 21 non-diabetic controls). The diabetic subjects were divided into three groups as PDR (n = 28), non-proliferative diabetic retinopathy (n = 20), and no-DR (n = 20). Eyes with PDR (n = 31) were injected with IVB (n = 7), IVR (n = 10), or IVTA (n = 6) 3 days before vitrectomy, and eight eyes did not receive an injection. Serum and vitreous samples were collected before vitrectomy and analyzed using ELISA.
RESULTS
We found the severity of retinopathy was not correlated with serum VEGF levels (P = .919, ρ = -0.011). Compared with the controls, vitreous VEGF was higher in the PDR (P < .001), whereas serum VEGF did not differ (P = .99). The controls had lower vitreous VEGF than the IVB, IVR, and no-injection subgroups (P = .01, P < .001, and P = .04, respectively). Vitreous VEGF was similar among the injected and no-injection subgroups (P = .17).
CONCLUSIONS
Serum VEGF levels may not directly reflect retinopathy progression. Neither IVB, IVR nor IVTA could eliminate vitreous VEGF levels within 3 days before vitrectomy.
Topics: Humans; Vascular Endothelial Growth Factor A; Diabetic Retinopathy; Vitreous Body; Bevacizumab; Ranibizumab; Vitrectomy; Enzyme-Linked Immunosorbent Assay; Diabetes Mellitus
PubMed: 36580154
DOI: 10.1007/s10792-022-02620-y -
Vestnik Oftalmologii 2023There are two main age-related changes that can occur in the vitreous body of healthy individuals throughout life: liquefaction (synchesis) and aggregation of collagen...
There are two main age-related changes that can occur in the vitreous body of healthy individuals throughout life: liquefaction (synchesis) and aggregation of collagen fibrils into dense bundles (syneresis). Progressive age-related degradation leads to posterior vitreous detachment (PVD). At present many classifications of PVD exist, in which authors relied either on the morphological features, or on the differences in pathogenesis before and after widespread use of OCT. The course of PVD can be either normal or anomalous. Physiological PVD induced by age-related vitreous changes progresses in specific stages. The review emphasizes that PVD can occur initially not only in the central zone of the retina, but also on the periphery with further spread to the posterior pole. Anomalous PVD can lead to various negative effects on the retina, as well as on the vitreous as a result of traction in the area of vitreoretinal interface.
Topics: Humans; Vitreous Body; Vitreous Detachment; Retina; Tomography, Optical Coherence
PubMed: 37379116
DOI: 10.17116/oftalma2023139031106 -
International Journal of Medical... May 2021We investigated whether nanopore amplicon sequencing of aqueous humor was capable of rapid pathogen identification in infectious endophthalmitis.
OBJECTIVES
We investigated whether nanopore amplicon sequencing of aqueous humor was capable of rapid pathogen identification in infectious endophthalmitis.
METHODS
5 cases of culture-positive bacterial endophthalmitis and 3 cases of fungal endophthalmitis (1 culture-positive and 2 presumed) were included. DNA was extracted from the aqueous humor and vitreous specimen, and PCR of bacterial rDNA (16S) and fungal rDNA (ITS1 and D1/2/3) was performed. Then, nanopore amplicon sequencing was performed for 2 h. The results of amplicon sequencing were compared to those of conventional culture studies.
RESULTS
In all cases, pathogens were identified by amplicon sequencing of aqueous humor specimens. In 3 cases of bacterial endophthalmitis, the identified microbes were confirmed by culture studies of both aqueous humor and vitreous specimens. In 2 cases of bacterial and 1 case of fungal endophthalmitis, the identified pathogens were confirmed only by culture studies of vitreous specimens. In all cases, amplicon sequencing identified pathogen in a shorter turnaround time than culture studies. In 2 cases with negative culture results, amplicon sequencing of aqueous humor identified fungal pathogens.
CONCLUSIONS
Our data demonstrates the potential of amplicon nanopore sequencing using aqueous humor to enable rapid, sensitive and less invasive microbial diagnosis of endophthalmitis.
Topics: DNA, Bacterial; Endophthalmitis; Humans; Nanopore Sequencing; Nanopores; Vitreous Body
PubMed: 33930723
DOI: 10.1016/j.ijmm.2021.151505