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Validation of the Flush Method as an Alternative to Basal or Reflex Tear Collection

Purpose: To validate the more easily applicable "flush" tear collection technique as a viable alternative to basal and reflex tear collection. Materials and Methods: Total protein content (TPC) and immunoglobulin A (IgA) concentrations were determined in the basal, reflex, and flush tears...

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Published in:	Current eye research 2011-03, Vol.36 (3), p.198-207
Main Authors:	Markoulli, Maria, Papas, Eric, Petznick, Andrea, Holden, Brien
Format:	Article
Language:	English
Subjects:	Adolescent Adult Basal tears Blinking - physiology Electrophoresis, Polyacrylamide Gel Eye Proteins - analysis Female Flush tears Human tear film Humans Immunoglobulin A, Secretory - analysis Male Mass Spectrometry Middle Aged Ophthalmology - methods Reflex tears Specimen Handling - methods Tear collection Tear proteins Tears - chemistry Young Adult
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creator	Markoulli, Maria Papas, Eric Petznick, Andrea Holden, Brien
description	Purpose: To validate the more easily applicable "flush" tear collection technique as a viable alternative to basal and reflex tear collection. Materials and Methods: Total protein content (TPC) and immunoglobulin A (IgA) concentrations were determined in the basal, reflex, and flush tears of 16 healthy non-contact lens wearers. The overall protein profile was established using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Results: Collection-rates were 4.6 ± 6.7 μl/min, 13.9 ± 11.1 μl/min, and 25.7 ± 12.4 μl/min for the basal, reflex, and flush tears, respectively. TPC was 7.14 ± 2.22 mg/mL, 6.01 ± 2.11 mg/mL, and 3.79 ± 1.51mg/ mL for basal, reflex, and flush tears, respectively, with flush tears being significantly less concentrated than basal (p = 0.001) and reflex (p = 0.008). IgA concentration was 1.04 ± 0.29 mg/ mL, 0.64 ± 0.26 mg/mL, and 0.65 ± 0.23 mg/mL for basal, reflex, and flush tears, respectively, with basal tears being significantly more concentrated (p
doi_str_mv	10.3109/02713683.2010.542867
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Materials and Methods: Total protein content (TPC) and immunoglobulin A (IgA) concentrations were determined in the basal, reflex, and flush tears of 16 healthy non-contact lens wearers. The overall protein profile was established using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Results: Collection-rates were 4.6 ± 6.7 μl/min, 13.9 ± 11.1 μl/min, and 25.7 ± 12.4 μl/min for the basal, reflex, and flush tears, respectively. TPC was 7.14 ± 2.22 mg/mL, 6.01 ± 2.11 mg/mL, and 3.79 ± 1.51mg/ mL for basal, reflex, and flush tears, respectively, with flush tears being significantly less concentrated than basal (p = 0.001) and reflex (p = 0.008). IgA concentration was 1.04 ± 0.29 mg/ mL, 0.64 ± 0.26 mg/mL, and 0.65 ± 0.23 mg/mL for basal, reflex, and flush tears, respectively, with basal tears being significantly more concentrated (p < 0.001). As a percentage of TPC, IgA represented 19.8 ± 14.9%, 11.4 ± 3.9%, and 19.8 ± 8.7% for basal, reflex, and flush, respectively. The flush was not significantly different to basal (p = 1.00) but significantly greater than reflex (p = 0.02). SDS-PAGE showed the same tear profiles for basal and flush tears. MS identified the most abundant proteins in all tear types. Conclusions: The flush method allows much faster collection than basal secretion sampling but returns essentially the same spectrum of proteins in similar proportions. This behavior is confirmation that the flush technique has utility as a more convenient alternative to basal tear sampling in studies involving composition analysis.</description><identifier>ISSN: 0271-3683</identifier><identifier>EISSN: 1460-2202</identifier><identifier>DOI: 10.3109/02713683.2010.542867</identifier><identifier>PMID: 21275520</identifier><language>eng</language><publisher>England: Informa Healthcare</publisher><subject>Adolescent ; Adult ; Basal tears ; Blinking - physiology ; Electrophoresis, Polyacrylamide Gel ; Eye Proteins - analysis ; Female ; Flush tears ; Human tear film ; Humans ; Immunoglobulin A, Secretory - analysis ; Male ; Mass Spectrometry ; Middle Aged ; Ophthalmology - methods ; Reflex tears ; Specimen Handling - methods ; Tear collection ; Tear proteins ; Tears - chemistry ; Young Adult</subject><ispartof>Current eye research, 2011-03, Vol.36 (3), p.198-207</ispartof><rights>2011 Informa Healthcare USA, Inc. 2011</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-a2d050432baa10008b072b8b4dfde8ed9c2e57341bea1412be1a60c8911e93283</citedby><cites>FETCH-LOGICAL-c417t-a2d050432baa10008b072b8b4dfde8ed9c2e57341bea1412be1a60c8911e93283</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21275520$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Markoulli, Maria</creatorcontrib><creatorcontrib>Papas, Eric</creatorcontrib><creatorcontrib>Petznick, Andrea</creatorcontrib><creatorcontrib>Holden, Brien</creatorcontrib><title>Validation of the Flush Method as an Alternative to Basal or Reflex Tear Collection</title><title>Current eye research</title><addtitle>Curr Eye Res</addtitle><description>Purpose: To validate the more easily applicable "flush" tear collection technique as a viable alternative to basal and reflex tear collection. Materials and Methods: Total protein content (TPC) and immunoglobulin A (IgA) concentrations were determined in the basal, reflex, and flush tears of 16 healthy non-contact lens wearers. The overall protein profile was established using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Results: Collection-rates were 4.6 ± 6.7 μl/min, 13.9 ± 11.1 μl/min, and 25.7 ± 12.4 μl/min for the basal, reflex, and flush tears, respectively. TPC was 7.14 ± 2.22 mg/mL, 6.01 ± 2.11 mg/mL, and 3.79 ± 1.51mg/ mL for basal, reflex, and flush tears, respectively, with flush tears being significantly less concentrated than basal (p = 0.001) and reflex (p = 0.008). IgA concentration was 1.04 ± 0.29 mg/ mL, 0.64 ± 0.26 mg/mL, and 0.65 ± 0.23 mg/mL for basal, reflex, and flush tears, respectively, with basal tears being significantly more concentrated (p < 0.001). As a percentage of TPC, IgA represented 19.8 ± 14.9%, 11.4 ± 3.9%, and 19.8 ± 8.7% for basal, reflex, and flush, respectively. The flush was not significantly different to basal (p = 1.00) but significantly greater than reflex (p = 0.02). SDS-PAGE showed the same tear profiles for basal and flush tears. MS identified the most abundant proteins in all tear types. Conclusions: The flush method allows much faster collection than basal secretion sampling but returns essentially the same spectrum of proteins in similar proportions. This behavior is confirmation that the flush technique has utility as a more convenient alternative to basal tear sampling in studies involving composition analysis.</description><subject>Adolescent</subject><subject>Adult</subject><subject>Basal tears</subject><subject>Blinking - physiology</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Eye Proteins - analysis</subject><subject>Female</subject><subject>Flush tears</subject><subject>Human tear film</subject><subject>Humans</subject><subject>Immunoglobulin A, Secretory - analysis</subject><subject>Male</subject><subject>Mass Spectrometry</subject><subject>Middle Aged</subject><subject>Ophthalmology - methods</subject><subject>Reflex tears</subject><subject>Specimen Handling - methods</subject><subject>Tear collection</subject><subject>Tear proteins</subject><subject>Tears - chemistry</subject><subject>Young Adult</subject><issn>0271-3683</issn><issn>1460-2202</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNp9kE1LxDAQhoMoun78A5HcPHVN0vTroujiFyiCrl7DtJ3SStpokqr7722pK3jxNDA87zvDQ8ghZ_OQs-yEiYSHcRrOBRtWkRRpnGyQGZcxC4RgYpPMRiQYmR2y69wrY-NCbpMdwUUSRYLNyNML6KYE35iOmor6GumV7l1N79HXpqTgKHT0XHu03UB9IPWGXoADTY2lj1hp_KJLBEsXRmssxqJ9slWBdnjwM_fI89XlcnET3D1c3y7O74JC8sQHIEoWMRmKHIAPv6U5S0Se5rKsSkyxzAqBURJKniNwyUWOHGJWpBnnmIUiDffI8dT7Zs17j86rtnEFag0dmt6pNOJZGDE-knIiC2ucs1ipN9u0YFeKMzXaVGubarSpJptD7OjnQJ-3WP6G1voG4GwCmq4ytoVPY3WpPKy0sZWFrmjcWP_vidM_DTWC9nUBFtWr6Qfn2v3_4zetuJWz</recordid><startdate>201103</startdate><enddate>201103</enddate><creator>Markoulli, Maria</creator><creator>Papas, Eric</creator><creator>Petznick, Andrea</creator><creator>Holden, Brien</creator><general>Informa Healthcare</general><general>Taylor & Francis</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201103</creationdate><title>Validation of the Flush Method as an Alternative to Basal or Reflex Tear Collection</title><author>Markoulli, Maria ; Papas, Eric ; Petznick, Andrea ; Holden, Brien</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-a2d050432baa10008b072b8b4dfde8ed9c2e57341bea1412be1a60c8911e93283</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Adolescent</topic><topic>Adult</topic><topic>Basal tears</topic><topic>Blinking - physiology</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Eye Proteins - analysis</topic><topic>Female</topic><topic>Flush tears</topic><topic>Human tear film</topic><topic>Humans</topic><topic>Immunoglobulin A, Secretory - analysis</topic><topic>Male</topic><topic>Mass Spectrometry</topic><topic>Middle Aged</topic><topic>Ophthalmology - methods</topic><topic>Reflex tears</topic><topic>Specimen Handling - methods</topic><topic>Tear collection</topic><topic>Tear proteins</topic><topic>Tears - chemistry</topic><topic>Young Adult</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Markoulli, Maria</creatorcontrib><creatorcontrib>Papas, Eric</creatorcontrib><creatorcontrib>Petznick, Andrea</creatorcontrib><creatorcontrib>Holden, Brien</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Current eye research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Markoulli, Maria</au><au>Papas, Eric</au><au>Petznick, Andrea</au><au>Holden, Brien</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Validation of the Flush Method as an Alternative to Basal or Reflex Tear Collection</atitle><jtitle>Current eye research</jtitle><addtitle>Curr Eye Res</addtitle><date>2011-03</date><risdate>2011</risdate><volume>36</volume><issue>3</issue><spage>198</spage><epage>207</epage><pages>198-207</pages><issn>0271-3683</issn><eissn>1460-2202</eissn><abstract>Purpose: To validate the more easily applicable "flush" tear collection technique as a viable alternative to basal and reflex tear collection. Materials and Methods: Total protein content (TPC) and immunoglobulin A (IgA) concentrations were determined in the basal, reflex, and flush tears of 16 healthy non-contact lens wearers. The overall protein profile was established using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Results: Collection-rates were 4.6 ± 6.7 μl/min, 13.9 ± 11.1 μl/min, and 25.7 ± 12.4 μl/min for the basal, reflex, and flush tears, respectively. TPC was 7.14 ± 2.22 mg/mL, 6.01 ± 2.11 mg/mL, and 3.79 ± 1.51mg/ mL for basal, reflex, and flush tears, respectively, with flush tears being significantly less concentrated than basal (p = 0.001) and reflex (p = 0.008). IgA concentration was 1.04 ± 0.29 mg/ mL, 0.64 ± 0.26 mg/mL, and 0.65 ± 0.23 mg/mL for basal, reflex, and flush tears, respectively, with basal tears being significantly more concentrated (p < 0.001). As a percentage of TPC, IgA represented 19.8 ± 14.9%, 11.4 ± 3.9%, and 19.8 ± 8.7% for basal, reflex, and flush, respectively. The flush was not significantly different to basal (p = 1.00) but significantly greater than reflex (p = 0.02). SDS-PAGE showed the same tear profiles for basal and flush tears. MS identified the most abundant proteins in all tear types. Conclusions: The flush method allows much faster collection than basal secretion sampling but returns essentially the same spectrum of proteins in similar proportions. This behavior is confirmation that the flush technique has utility as a more convenient alternative to basal tear sampling in studies involving composition analysis.</abstract><cop>England</cop><pub>Informa Healthcare</pub><pmid>21275520</pmid><doi>10.3109/02713683.2010.542867</doi><tpages>10</tpages></addata></record>
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subjects	Adolescent Adult Basal tears Blinking - physiology Electrophoresis, Polyacrylamide Gel Eye Proteins - analysis Female Flush tears Human tear film Humans Immunoglobulin A, Secretory - analysis Male Mass Spectrometry Middle Aged Ophthalmology - methods Reflex tears Specimen Handling - methods Tear collection Tear proteins Tears - chemistry Young Adult
title	Validation of the Flush Method as an Alternative to Basal or Reflex Tear Collection
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