Dataset Information

The Shape of Posterior Sclera as a Biometric Signature in Open-angle Glaucoma: An Intereye Comparison Study.

ABSTRACT: PURPOSE:To characterize intereye differences in posterior segment parameters and determine their significance in open-angle glaucoma patients with unilateral damage. METHODS:Both eyes from 65 subjects without any nerve damage and 43 patients undergoing treatment for unilateral open-angle glaucoma were included in this study. A 12.0×9.0×2.6?mm volume of the posterior segment in each eye was scanned with swept-source optical coherence tomography. Coronally reconstructed optical coherence tomography images were analyzed to determine the deepest point of the eye (DPE), which we then calculated the distance (Disc-DPE distance), depth (Disc-DPE depth), angle (Disc-DPE angle) from the optic disc center to the DPE. Posterior pole shape was analyzed measuring the posterior pole-cross-sectional area, posterior pole-horizontal width (PP-HW), and posterior pole-vertical width) of the posterior pole. These measurements and their intereye absolute difference (IAD; absolute difference in measurements between the right and left eyes) values were compared between the healthy and unilateral glaucomatous patients. RESULTS:The posterior sclera measurements, including the Disc-DPE distance, Disc-DPE depth, and posterior pole-cross-sectional area, were significantly different between the unilateral glaucoma eyes and contralateral healthy eyes (P=0.043, P=0.035, and P=0.049, respectively). By contrast, none of the intereye differences in optic nerve head parameters were significant in the unilateral glaucoma patients. In comparison with the IAD values, the baseline intraocular pressure and PP-HW of the posterior segment showed significant differences between the healthy and the unilateral glaucoma patients (P=0.019 and P=0.036, respectively). A multivariate analysis showed that a larger baseline intraocular pressure IAD [odds ratio (OR), 1.381; P=0.009)] and larger PP-HW IAD (OR, 1.324; P=0.032) were significantly associated with the presence of glaucoma. CONCLUSIONS:Compared with the fellow healthy eyes, glaucomatous eyes had larger and more steeply curved posterior poles, which represent a structural variation of the posterior sclera that might be associated with glaucomatous optic neuropathy.

SUBMITTER: Kim YC

PROVIDER: S-EPMC7647446 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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The Shape of Posterior Sclera as a Biometric Signature in Open-angle Glaucoma: An Intereye Comparison Study.

Kim Yong Chan YC Koo Yong Ho YH Bin Hwang Hyung H Kang Kui Dong KD

Journal of glaucoma 20201001 10

<h4>Purpose</h4>To characterize intereye differences in posterior segment parameters and determine their significance in open-angle glaucoma patients with unilateral damage.<h4>Methods</h4>Both eyes from 65 subjects without any nerve damage and 43 patients undergoing treatment for unilateral open-angle glaucoma were included in this study. A 12.0×9.0×2.6 mm volume of the posterior segment in each eye was scanned with swept-source optical coherence tomography. Coronally reconstructed optical cohe ...[more]

PMID: 32555059

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Project description:The total RNA from control and POAG optic nerve lamina cribrosa region was isolated and subjected to analysis on U133A 2.0 affymetrix microarray chip. The results from these two analysis was compared to determine whether PAD II RNA level is altered. Purpose of the study. This study was done to determine which RNA transcripts undergo significant changes in the optic nerve between normal and cadaver primary open-angle glaucoma donors. Using donor tissues from NDRI we have performed proteomic analyses (which will be published shortly), we intend a comparison between the proteomic and RNA changes for select proteins in our study. Sample Collection and Preparation of Labeled Copy RNA; The total RNA from optic nerve was obtained using TRIZOL (Invitrogen Inc., Carlsbad, CA) with modification of recommended protocols. The optic nerve from donor eyes were carefully excised and minced into small pieces first using a scissor and then a scalpel. Prior to use tissue was washed with DEPC water and all solutions were prepared in DEPC water. The minced tissue was placed in a glass homogenizer with 1 ml TRIZOL per 100 mg of tissue and homogenized in a glass homogenizer DUALL 20 (Kimble Kontes Glass Co, Vineland, NJ) with 10 strokes cycles each at room temperature and after freezing with liquid nitrogen for 1 min for 40 cycles. This RNA was extracted with chloroform, isoamylalcohol and precipitated with sodium citrate/sodium chloride and isopropanol. The final air-dried RNA precipitate was suspended in DEPC water and stored in â??80oC prior to use. RNA was prepared for hybridization according to the GeneChip Expression Analysis Technical Manual (Affymetrix, Santa Clara, CA). The hybridization was performed at Gene Expression Array Core Facility at Case Western Reserve University (www.geacf.net). The RNA sample was cleaned using a column from a Qiagen RNeasy kit, (part # 74104 ) in accordance with the manufacturerâ??s instructions. The volume of DEPC water used to elute the RNA from the Qiagen column was dictated by the nominal mass of the loaded RNA sample. Samples were eluted in 100ml of water, precipitated with 0.5 volume of 7.5M Ammonium Acetate and 2.5 volumes of 100% ethanol at -20oC for 1 hr and centrifuged for 10 min at room temperature. The pellet was drained and washed twice in 500Âµl of 70% ethanol. Supernatant was aspirated. The pellet was dried briefly (less than 1 min) in a speeedvac (Savant) and re-suspended in a small volume (~ 12â??15ml) of DEPC water. A small aliquot (2ml) of the sample was used for quantitation. Cleaned RNA samples were annotated as â??clnRNAâ?? and stored at -20oC. cDNA Synthesis; The protocol recommended by Affymetrix Inc. for cDNA synthesis was used at all times. The reaction was primed by annealing an oligo-dT primer coupled to a T7 RNA polymerase promoter to the RNA sample. Whenever possible, 8 mg of clnRNA were reverse transcribed using Superscript II reverse transcriptase in a 20ml reaction at 42oC for 1 hr. Second strand synthesis was carried out immediately in a total reaction volume of 150ml in the presence of E. coli DNA polymerase I, RNAse H and DNA ligase. The reaction mixture incubated for 2 hrs at 16oC and for a further 5 min in the presence of T4 DNA pol.. The reaction was terminated by the addition of EDTA. The sample was cleaned with a Qiagen cDNA clean-up column according to the manufacturers directions. The elution buffer volume was 14ml. The eluted samples were stored overnight at -20oC. In-Vitro Transcription (IVT) reaction; In a 0.5ml microfuge tube, complementary RNA (cRNA) was generated in an IVT reaction using a BioArray High Yield ENZO kit (Affymetrix). The 40ml reaction contained 10ml of cDNA sample and was incubated at 37oC in the heating block for 4-5hrs. Samples were mixed by flicking, pulse-centrifuged and returned to the incubator once every 40 minutes. Upon completion of the reaction, IVT samples were adjusted to a volume of 100ml with DEPC-treated water and cleaned using Qiagen RNA clean-up columns. Samples were eluted in 45ml of DEPC-treated water. A 2ml-aliquot was used for quantitation. Corrected concentrations of cRNA (minus the input clnRNA) were determined. A standard fragmentation reaction volume was 40ml in a 0.5 ml microfuge tube and used 20mg of overall IVT RNA in 1X fragmentation buffer (40mM Tris acetate, pH 8.1, 100mM KOAc, 30mM MgOAc). The reaction was incubated at 94oC for 35min in a heat block. After the 35 min incubation the fragmented samples were placed on ice. The standard hybridization cocktail volume was 300ml. 150ml of 2X hybridization buffer was added (final concentrations : 100mM MES, 1M [Na+], 20mM EDTA, 0.01% Tween 20) Herring sperm and acetylated BSA were added to a final concentration of 0.1 and 0.5mg/ml respectively. The amount of fragmentation reaction containing 15mg of cRNA was added to the cocktail and the remaining volume was made up with molecular biology grade water. A 15ml aliquot of a 20X mixture of in-vitro transcripts of bacterial genes BioB BioC BioD and cre as external controls (spikes) were added to the cocktail to give final concentrations of 1.5, 5, 25 and 100pM respectively. Control oligonucleotide (5ml) was added to a final concentration of 50pM. Array Hybridization; Sample Hyridization cocktails (300ml) were removed from storage at â??20oC and thawed in a 45oC temperature block. The samples were incubated at 45oC for 5 min, incubated at 45oC for 5 min and then centrifuged at 15,000g for 5 min. 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Dataset Information

The Shape of Posterior Sclera as a Biometric Signature in Open-angle Glaucoma: An Intereye Comparison Study.

Publications

The Shape of Posterior Sclera as a Biometric Signature in Open-angle Glaucoma: An Intereye Comparison Study.

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