Project description:We collected some vitreous humors from rat eyes through a photolabile paper based chip which experienced different natural recovery processes at day 0, 0.25, 1, 3, 7 day after eye stroke by laser induced in rat. These mirs samples were collected from extracelluar vesicles of vitreous humor in rat. We used microarray to find some new miRNAs that may participate in the natural recovery process after experience eye stroke.
Project description:Comprehensive proteomic analyses of vitreous humor using liquid chromatography with tandem mass spectrometry were performed in 10 patients with VRL, 10 control patients with idiopathic epiretinal membrane or macular hole, and 10 patients with ocular sarcoidosis.
Project description:In this study, we investigated the proteome of normal human vitreous humor using high resolution Fourier transform mass spectrometry. We identified 1269 proteins. This largest catalog of vitreous proteins to date should facilitate biomedical research into pathological conditions of the eye including diabetic retinopathy, retinal detachment and cataract.
Project description:Purpose: To study the proteome of the subretinal fluid (SRF) from rhegmatogenous retinal detachment (RRD) in search for novel markers for improved diagnosis and prognosis of RRD. Methods: Human undiluted SRF obtained during vitrectomy for primary RRD using a 41G needle (n = 24) was analyzed and compared to vitreous humor from 2-days post-mortem eyes (n = 20). Sample preparation underwent nanoflow liquid chromatography-tandem mass spectrometry (LC-MS/MS). Label-free quantification (LFQ) using MaxQuant was used to determine differential expressed proteins between SRF and vitreous humor. The intensity-based absolute quantification (iBAQ) was used to rank proteins according to their molar fractions within groups. Identification of proteins beyond the quantitative level was performed using the Mascot search engine. Results: The protein concentration of the control vitreous humor was lower and more consistent (1.2 ± 0.4 mg) than that of the SRF (17.9 ± 22 mg). The iBAQ analysis showed high resemblance between SRF and vitreous humor, except for crystallins solely identified in vitreous humor. The LFQ analysis found 37 protein misregulations between SRF and vitreous humor of which the blood coagulation pathway was found to be enriched using the PANTHER Classification System. Combined, the iBAQ, LFQ and Mascot analysis found an overlap only in chitinase-3-like protein 1 and galectin-3-binding protein unique to the SRF. Conclusions: The proteome of the SRF was highly represented by proteins involved in proteolysis. Such proteins can possibly serve as targets in modulating the effects of SRF in RD.
Project description:Lens epithelial explants consist of lens epithelial cells (P8 FVB/N mice) grown in vitro on their native basement membrane, the lens capsule. For decades, biologists have used lens epithelial explants to study lens fiber cell differentiation. However, the global change in the accessibility of the chromatin and transcriptome during the process of explanting and culture is unknown. Therefore, P8 FVB/N lens epithelial explants cultured in either unsupplemented media or media containing 50% bovine vitreous humor for one or five days were collected. Chromatin and RNA was collected for ATAC-sequencing and RNA-sequencing respectively. Differentially accessible regions and differentially expressed genes were identified for each condition to provide a genome wide view of chromatin architecture and gene expression during fiber cell differentiation in vitro. Vitreous humor generally increased chromatin accessibility in promoter regions of genes associated with fiber differentiation and immune response, and this was associated with increased transcript levels for these genes. In contrast, vitreous had relatively little effect on the accessibility of most of the genes highly expressed in the lens epithelium despite dramatic reductions in the transcript levels of these genes.
Project description:Lens epithelial explants consist of lens epithelial cells (P8 FVB/N mice) grown in vitro on their native basement membrane, the lens capsule. For decades, biologists have used lens epithelial explants to study lens fiber cell differentiation. However, the global change in the accessibility of the chromatin and transcriptome during the process of explanting and culture is unknown. Therefore, P8 FVB/N lens epithelial explants cultured in either unsupplemented media or media containing 50% bovine vitreous humor for one or five days were collected. Chromatin and RNA was collected for ATAC-sequencing and RNA-sequencing respectively. Differentially accessible regions and differentially expressed genes were identified for each condition to provide a genome wide view of chromatin architecture and gene expression during fiber cell differentiation in vitro. Vitreous humor generally increased chromatin accessibility in promoter regions of genes associated with fiber differentiation and immune response, and this was associated with increased transcript levels for these genes. In contrast, vitreous had relatively little effect on the accessibility of most of the genes highly expressed in the lens epithelium despite dramatic reductions in the transcript levels of these genes.
Project description:S. aureus SA564 and SA564-codY-mutant were grown in bovine aqueous humor, bovine vitreous humor and a chemically defined medium. Samples were extracted in midlog phase and affymetrix microarray processing was performed.
Project description:<p><strong>INTRODUCTION:</strong> Application of metabolomic methods to forensic studies may expand the limits of the post-mortem interval (PMI) estimation, and improve the accuracy of the estimation. To this end, it is important to determine which tissue is the most suitable for analysis, and which compounds are the most promising candidates for PMI estimation.</p><p><strong>OBJECTIVES:</strong> This work is aimed at the comparison of human serum, aqueous humor (AH), and vitreous humor (VH) as per- spective tissues for metabolomic-based PMI estimation, at the determination of most promising PMI biomarkers, and at the development of method of PMI estimation based on the measurement of concentrations of PMI biomarkers.</p><p><strong>METHODS:</strong> Quantitative metabolomic pro ling of samples of the human serum, AH, and VH taken at di erent PMIs has been performed with the use of NMR spectroscopy.</p><p><strong>RESULTS:</strong> It is found that the metabolomic changes in anatomically isolated ocular uids are slower and smoother than that in blood. A good positive time correlation (Pearson coe cient r > 0.5) was observed for several metabolites, including hypox- anthine, choline, creatine, betaine, glutamate, and glycine. A model for PMI estimation based on concentrations of several metabolites in AH and VH is proposed.</p><p><strong>CONCLUSIONS:</strong> The obtained results demonstrate that the metabolomic analysis of AH and VH is more suitable for the PMI estimation than that of serum. The compounds with good positive time correlation can be considered as potential PMI biomarkers.</p><p><br></p>
Project description:Retinal detachment is a severe eye condition characterized by the detachment of the neurosensory retina from the retinal pigment epithelium and caused by retinal tears. Pars plana vitrectomy is the elective surgical procedure during which vitreous humor, a fluid which shapes the eye globe and supports mechanically the adherence of the retina to the posterior chamber of the eye, is collected. Therefore, profiling of the vitreous humor proteome of subjects diagnosed with retinal detachment is supposed to provide molecular evidence on the pathobiology of the disease and on its secondary complications, such as proliferative vitreo-retinopathy, which predispose to recurrent RD (observed in 20% of cases), a sight threatening condition. Herein, we investigated the perturbations of vitreous proteome between subjects affected by primary retinal detachment and controls by shot-gun proteomics approaches. Proteomic datasets were first analyzed and searched for global proteome changes. Thereafter, starting from the assumption that the disease could be sustained by altered proteolytic processing of structural and non-structural elements of the VH, mining of N- and C-termini was performed to uncover endogenous proteolytic events underscoring the disease condition. The search retrieved evidence of a wide repertoire of either previously characterized or uncharacterized proteolytic events. Most notably, comparison of the N- and C-termini landscape between experimental groups highlighted robust alterations in the repertoire of cleaved proteins. Further strengthened by immunoblotting studies on a select panel of proteins, the data envisage that retinal detachment is promoted or characterized, as a secondary process, by proteolytic-based alterations of structural and non-structural components involved in the regulation of immune process, proteases inhibition and most notably, angiogenesis.