Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
What information can be obtained from the tears of a patient with primary open angle glaucoma? / Tamkovich, Svetlana; Grigor'eva, Alina; Eremina, Alena и др.
в: Clinica Chimica Acta, Том 495, 01.08.2019, стр. 529-537.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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TY - JOUR
T1 - What information can be obtained from the tears of a patient with primary open angle glaucoma?
AU - Tamkovich, Svetlana
AU - Grigor'eva, Alina
AU - Eremina, Alena
AU - Tupikin, Alexey
AU - Kabilov, Marcel
AU - Chernykh, Valerii
AU - Vlassov, Valentin
AU - Ryabchikova, Elena
N1 - Publisher Copyright: © 2019 Elsevier B.V.
PY - 2019/8/1
Y1 - 2019/8/1
N2 - Since tears are a biological fluid, they have a potential diagnostic value for ophthalmic diseases. The aim of this study was to compare tear supernatants and pellets obtained from patients suffering from primary open angle glaucoma (POAG) and healthy persons (HPs) using transmission electron microscopy (TEM) and molecular biological methods. Tear supernatants and pellets were prepared using ultrafiltration and ultracentrifugation and were examined by negative staining and immunogold labelling TEM. DNA of the pellets was isolated, quantified and sequenced using a MiSeq (Illumina, USA) genomic sequencer with the Reagent Kit v3 (600 cycles, Illumina, USA). MicroRNA was isolated and quantified from the pellets; miR-146b, miR-16 and miR-126 were detected using TaqMan MicroRNA Assays (Applied Biosystems, USA). TEM of tear supernatants from both POAG patients and HPs revealed identical constituents: spherical or cup-shaped vesicles, “non-vesicles”, cell debris and macromolecular aggregates. Pellets of POAG patients and HPs contained small extracellular vesicles (sEVs) non-labelled vesicles and “non-vesicles”; pellets of sick persons also contained sEVs with “a capsule”. POAG-patient tear pellets showed elevated concentrations of genomic ds-DNA and SINE-repeats, and different expressions of miR-146b, miR-16 and miR-126 and a different set of bacterial DNA in comparison with pellets obtained from the tears of HPs. The data obtained indicate that the tears of HPs and POAG patients could serve as an object for TEM studies and as a source of sEV-containing preparations (pellets), which, in turn, could be used for the isolation and study of genomic ds-DNA and RNA. Our data provide the basis for using tears for diagnostic applications.
AB - Since tears are a biological fluid, they have a potential diagnostic value for ophthalmic diseases. The aim of this study was to compare tear supernatants and pellets obtained from patients suffering from primary open angle glaucoma (POAG) and healthy persons (HPs) using transmission electron microscopy (TEM) and molecular biological methods. Tear supernatants and pellets were prepared using ultrafiltration and ultracentrifugation and were examined by negative staining and immunogold labelling TEM. DNA of the pellets was isolated, quantified and sequenced using a MiSeq (Illumina, USA) genomic sequencer with the Reagent Kit v3 (600 cycles, Illumina, USA). MicroRNA was isolated and quantified from the pellets; miR-146b, miR-16 and miR-126 were detected using TaqMan MicroRNA Assays (Applied Biosystems, USA). TEM of tear supernatants from both POAG patients and HPs revealed identical constituents: spherical or cup-shaped vesicles, “non-vesicles”, cell debris and macromolecular aggregates. Pellets of POAG patients and HPs contained small extracellular vesicles (sEVs) non-labelled vesicles and “non-vesicles”; pellets of sick persons also contained sEVs with “a capsule”. POAG-patient tear pellets showed elevated concentrations of genomic ds-DNA and SINE-repeats, and different expressions of miR-146b, miR-16 and miR-126 and a different set of bacterial DNA in comparison with pellets obtained from the tears of HPs. The data obtained indicate that the tears of HPs and POAG patients could serve as an object for TEM studies and as a source of sEV-containing preparations (pellets), which, in turn, could be used for the isolation and study of genomic ds-DNA and RNA. Our data provide the basis for using tears for diagnostic applications.
KW - Bacterial DNA
KW - miRNA
KW - Small extracellular vesicles
KW - Tears
UR - http://www.scopus.com/inward/record.url?scp=85066785984&partnerID=8YFLogxK
U2 - 10.1016/j.cca.2019.05.028
DO - 10.1016/j.cca.2019.05.028
M3 - Article
C2 - 31153869
AN - SCOPUS:85066785984
VL - 495
SP - 529
EP - 537
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
SN - 0009-8981
ER -
ID: 20533154