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osu1273854679.pdf (11.43 MB)
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Comparison of Symptoms, Signs, Composition, and Tear Film Dynamics in Sjögrens vs. Non-Sjögrens Subjects
Author Info
Paulett, Christopher Lewis
Permalink:
http://rave.ohiolink.edu/etdc/view?acc_num=osu1273854679
Abstract Details
Year and Degree
2010, Master of Science, Ohio State University, Vision Science.
Abstract
Purpose: To compare dry eye symptoms, signs, and proteins, which are identified in tear samples of Sjögrens syndrome (SS) patients to an age and sex matched non-Sjögrens syndrome (non-SS) control. Methods: Twelve SS and twelve non-SS underwent a dry eye exam. The procedures that followed were tear film interferometry (Tear film thickness and tear film thinning rates), osmolarity, protein tear collection, lipid tear collection, The Ocular Surface Disease Index (OSDI)™ and patient demographic forms, Schirmer test (type 1), fluorescein tear break up time (TBUT), fluorescein corneal staining, and lissamine green conjunctival staining. All clinical data were quantified and analyzed using Microsoft Excel™ and SPSS™. Proteins extracted from the tear samples and Schirmer strips were identified and quantified by using iTRAQ™ and Orbitrap with electrospray liquid chromatography tandem-mass spectrometry (LC-MS/MS). The data was then compared to a large protein database using the Protein Information Resource (PIR) website. RayBiotech™ microarrays were also performed to identify inflammatory mediators present in the tear samples. Results: Fluorescein corneal staining, lissamine green conjunctival staining, OSDI™, and tear film thinning rates were significant in differentiating between the SS and non-SS groups. Tear film thickness, Schirmer strip wetting length, and TBUT were not effective methods to differentiate between the two groups. Alpha-enolase, S100 A9, S100 A4, and S100 A11 are all proteins which are differentially expressed in an up-regulated manner with increased dry eye symptoms in this study. Lipocalin, PIP, Lysozyme C, and Lactoferin are proteins which are differentially expressed in a down-regulated maner and decrease as the severity of dry eye increases. IL-2, IL-6, INF- γ, and IL-10 are cytokines which seem to be increased as the severity of dry eye increases. Conclusion: An efficient technique to identify proteins in tear samples collected via Schirmer strips and microcapillary tubes, is to use combined iTRAQ™ and LC-MS/MS. RayBiotech™ microarrays have proven to be an efficient way to quantify the amount of a specific inflammatory protein in a tear sample. The microassay is used to identify a broad number of proteins in the tear samples by testing for forty different cytokines. Using custom micoroarrays which target a smaller number of inflammatory mediators of interest would be recommended for future studies. In the future using a larger sample size, masking clinicians, and keeping strict control of enviromental conditions when performing osmolarity measurment may improve the studied outcome. The proteomic analyses performed here are very informative and warrant future research in dry eye and Sjögren’s syndrome patients.
Committee
Kelly Nichols, O.D. M.P.H. Ph.D. (Advisor)
Peter Ewen King-Smith, Ph.D. (Committee Member)
Kari B. Green-Church, Ph.D. (Committee Member)
Pages
130 p.
Subject Headings
Biomedical Research
;
Ophthalmology
Keywords
Sj&246
;
grens syndrome
;
Dry Eye
;
iTRAQ
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Citations
Paulett, C. L. (2010).
Comparison of Symptoms, Signs, Composition, and Tear Film Dynamics in Sjögrens vs. Non-Sjögrens Subjects
[Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1273854679
APA Style (7th edition)
Paulett, Christopher.
Comparison of Symptoms, Signs, Composition, and Tear Film Dynamics in Sjögrens vs. Non-Sjögrens Subjects.
2010. Ohio State University, Master's thesis.
OhioLINK Electronic Theses and Dissertations Center
, http://rave.ohiolink.edu/etdc/view?acc_num=osu1273854679.
MLA Style (8th edition)
Paulett, Christopher. "Comparison of Symptoms, Signs, Composition, and Tear Film Dynamics in Sjögrens vs. Non-Sjögrens Subjects." Master's thesis, Ohio State University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=osu1273854679
Chicago Manual of Style (17th edition)
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Document number:
osu1273854679
Download Count:
532
Copyright Info
© 2010, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.