Comparison of non-collagen protein and total creatine as reference for determination of energy stores in endomyocardial biopsies
Objective: The aim was to establish a reliable reference system for biochemical measurements in endomyocardial biopsies. Methods: Myocardial tissue samples were obtained from pigs before and after cardioplegic arrest and reperfusion. Non-collagen protein content was evaluated as a non-specific refer...
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| Veröffentlicht in: | Cardiovascular research 1993-12, Vol.27 (12), p.2113-2117 |
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| container_title | Cardiovascular research |
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| creator | Bøtker, Hans Erik Kimose, Hans Henrik Helligsø, Per Thomassen, Anne Rahbek Nielsen, Torsten Toftegaard |
| description | Objective: The aim was to establish a reliable reference system for biochemical measurements in endomyocardial biopsies. Methods: Myocardial tissue samples were obtained from pigs before and after cardioplegic arrest and reperfusion. Non-collagen protein content was evaluated as a non-specific reference system and compared with total creatine content representing a specific myocardial reference system. The influence of base strength, extraction temperature, and extraction time on protein yields was determined in tissue precipitates redissolved in NaOH. Interference from protein of collagenous origin was excluded by hydroxyproline determinations. Variability of myocardial ATP content in relation to non-collagen protein and total creatine was compared in endomyocardial biopsies taken before and after cardioplegic arrest and reperfusion. Results: The two methods showed comparable analytical precision. Apart from an interference in 1.0 mol·litre−1 NaOH for extended extraction periods at high temperatures, myocardial protein yields increased with increasing base strength, extraction temperature, and extraction time. During cardioplegic arrest and reperfusion heart weight increased due to oedema. Simultaneously, myocardial non-collagen protein content decreased. No change in total creatine was found during cardioplegic arrest but there was a significant loss of creatine after reperfusion. Comparison of variability in myocardial ATP content with non-collagen protein or total creatine as reference systems revealed no difference. Conclusions: Determination of non-collagen protein can be optimised with standardised conditions for protein extraction in tissue precipitates. Employment of total creatine as a reference system does not reduce variability of myocardial metabolite determinations in endomyocardial biopsies compared with non-collagen protein. Loss of myocardial creatine may in itself provide additional information about myocardial injury but this makes it unsuitable as a reference system for measuring metabolic changes during reperfusion. Multiple biopsies seem necessary for estimation of myocardial energy stores. Cardiovascular Research 1993; 27: 2113-2117 |
| doi_str_mv | 10.1093/cvr/27.12.2113 |
| format | Article |
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Methods: Myocardial tissue samples were obtained from pigs before and after cardioplegic arrest and reperfusion. Non-collagen protein content was evaluated as a non-specific reference system and compared with total creatine content representing a specific myocardial reference system. The influence of base strength, extraction temperature, and extraction time on protein yields was determined in tissue precipitates redissolved in NaOH. Interference from protein of collagenous origin was excluded by hydroxyproline determinations. Variability of myocardial ATP content in relation to non-collagen protein and total creatine was compared in endomyocardial biopsies taken before and after cardioplegic arrest and reperfusion. Results: The two methods showed comparable analytical precision. Apart from an interference in 1.0 mol·litre−1 NaOH for extended extraction periods at high temperatures, myocardial protein yields increased with increasing base strength, extraction temperature, and extraction time. During cardioplegic arrest and reperfusion heart weight increased due to oedema. Simultaneously, myocardial non-collagen protein content decreased. No change in total creatine was found during cardioplegic arrest but there was a significant loss of creatine after reperfusion. Comparison of variability in myocardial ATP content with non-collagen protein or total creatine as reference systems revealed no difference. Conclusions: Determination of non-collagen protein can be optimised with standardised conditions for protein extraction in tissue precipitates. Employment of total creatine as a reference system does not reduce variability of myocardial metabolite determinations in endomyocardial biopsies compared with non-collagen protein. Loss of myocardial creatine may in itself provide additional information about myocardial injury but this makes it unsuitable as a reference system for measuring metabolic changes during reperfusion. Multiple biopsies seem necessary for estimation of myocardial energy stores. Cardiovascular Research 1993; 27: 2113-2117</description><identifier>ISSN: 0008-6363</identifier><identifier>EISSN: 1755-3245</identifier><identifier>DOI: 10.1093/cvr/27.12.2113</identifier><identifier>PMID: 8313416</identifier><identifier>CODEN: CVREAU</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>Animals ; assay evaluation ; ATP ; Biological and medical sciences ; Biopsy ; cardioplegia ; creatine ; Creatine - analysis ; endomyocardial biopsy ; Energy Metabolism - physiology ; Investigative techniques, diagnostic techniques (general aspects) ; Medical sciences ; Myocardial Reperfusion Injury - metabolism ; Myocardium - chemistry ; myoglobin ; non-collagen protein ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; pig ; Proteins - analysis ; reference system ; reperfusion ; Respiratory system ; Swine</subject><ispartof>Cardiovascular research, 1993-12, Vol.27 (12), p.2113-2117</ispartof><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c427t-e8b2ef22726a420517579898c2e7b8fc0ccfb01b1bb8ccf13d201d5f9ffa15d3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,782,786,27933,27934</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3864483$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8313416$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bøtker, Hans Erik</creatorcontrib><creatorcontrib>Kimose, Hans Henrik</creatorcontrib><creatorcontrib>Helligsø, Per</creatorcontrib><creatorcontrib>Thomassen, Anne Rahbek</creatorcontrib><creatorcontrib>Nielsen, Torsten Toftegaard</creatorcontrib><title>Comparison of non-collagen protein and total creatine as reference for determination of energy stores in endomyocardial biopsies</title><title>Cardiovascular research</title><addtitle>Cardiovasc Res</addtitle><description>Objective: The aim was to establish a reliable reference system for biochemical measurements in endomyocardial biopsies. Methods: Myocardial tissue samples were obtained from pigs before and after cardioplegic arrest and reperfusion. Non-collagen protein content was evaluated as a non-specific reference system and compared with total creatine content representing a specific myocardial reference system. The influence of base strength, extraction temperature, and extraction time on protein yields was determined in tissue precipitates redissolved in NaOH. Interference from protein of collagenous origin was excluded by hydroxyproline determinations. Variability of myocardial ATP content in relation to non-collagen protein and total creatine was compared in endomyocardial biopsies taken before and after cardioplegic arrest and reperfusion. Results: The two methods showed comparable analytical precision. Apart from an interference in 1.0 mol·litre−1 NaOH for extended extraction periods at high temperatures, myocardial protein yields increased with increasing base strength, extraction temperature, and extraction time. During cardioplegic arrest and reperfusion heart weight increased due to oedema. Simultaneously, myocardial non-collagen protein content decreased. No change in total creatine was found during cardioplegic arrest but there was a significant loss of creatine after reperfusion. Comparison of variability in myocardial ATP content with non-collagen protein or total creatine as reference systems revealed no difference. Conclusions: Determination of non-collagen protein can be optimised with standardised conditions for protein extraction in tissue precipitates. Employment of total creatine as a reference system does not reduce variability of myocardial metabolite determinations in endomyocardial biopsies compared with non-collagen protein. Loss of myocardial creatine may in itself provide additional information about myocardial injury but this makes it unsuitable as a reference system for measuring metabolic changes during reperfusion. Multiple biopsies seem necessary for estimation of myocardial energy stores. Cardiovascular Research 1993; 27: 2113-2117</description><subject>Animals</subject><subject>assay evaluation</subject><subject>ATP</subject><subject>Biological and medical sciences</subject><subject>Biopsy</subject><subject>cardioplegia</subject><subject>creatine</subject><subject>Creatine - analysis</subject><subject>endomyocardial biopsy</subject><subject>Energy Metabolism - physiology</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Medical sciences</subject><subject>Myocardial Reperfusion Injury - metabolism</subject><subject>Myocardium - chemistry</subject><subject>myoglobin</subject><subject>non-collagen protein</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>pig</subject><subject>Proteins - analysis</subject><subject>reference system</subject><subject>reperfusion</subject><subject>Respiratory system</subject><subject>Swine</subject><issn>0008-6363</issn><issn>1755-3245</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kM9rHCEYhqWkpNu0194KHkJvs_HHODrHsCRNYEsp5BB6Ecf5DCYzulE3ZG_90-uyy55U3scH3hehb5QsKen5lX1LV0wuKVsySvkHtKBSiIazVpyhBSFENR3v-Cf0Oefn-hRCtufoXHHKW9ot0L9VnDcm-RwDjg6HGBobp8k8QcCbFAv4gE0YcYnFTNgmMMUHwCbjBA4SBAvYxYRHKJBmH2p8MEGA9LTDucQEGVcLhDHOu2hNGn1VDT5usof8BX10Zsrw9XheoIfbm4fVXbP-_fN-db1ubMtkaUANDBxjknWmZUTUlrJXvbIM5KCcJda6gdCBDoOqV8pHRugoXO-coWLkF-jHQVtLvW4hFz37bKE2DRC3WcuuujuhKrg8gDbFnGtJvUl-NmmnKdH7xXVdXDOpKdP7xeuH70fzdphhPOHHiWt-ecxNtmZyyQTr8wnjqmtbtdc0B8znAu-n2KQX3Ukuhb57_Kt_rW__9OtHqjn_D92Qm8M</recordid><startdate>19931201</startdate><enddate>19931201</enddate><creator>Bøtker, Hans Erik</creator><creator>Kimose, Hans Henrik</creator><creator>Helligsø, Per</creator><creator>Thomassen, Anne Rahbek</creator><creator>Nielsen, Torsten Toftegaard</creator><general>Oxford University Press</general><scope>BSCLL</scope><scope>IQODW</scope><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>19931201</creationdate><title>Comparison of non-collagen protein and total creatine as reference for determination of energy stores in endomyocardial biopsies</title><author>Bøtker, Hans Erik ; Kimose, Hans Henrik ; Helligsø, Per ; Thomassen, Anne Rahbek ; Nielsen, Torsten Toftegaard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c427t-e8b2ef22726a420517579898c2e7b8fc0ccfb01b1bb8ccf13d201d5f9ffa15d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Animals</topic><topic>assay evaluation</topic><topic>ATP</topic><topic>Biological and medical sciences</topic><topic>Biopsy</topic><topic>cardioplegia</topic><topic>creatine</topic><topic>Creatine - analysis</topic><topic>endomyocardial biopsy</topic><topic>Energy Metabolism - physiology</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Medical sciences</topic><topic>Myocardial Reperfusion Injury - metabolism</topic><topic>Myocardium - chemistry</topic><topic>myoglobin</topic><topic>non-collagen protein</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>pig</topic><topic>Proteins - analysis</topic><topic>reference system</topic><topic>reperfusion</topic><topic>Respiratory system</topic><topic>Swine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bøtker, Hans Erik</creatorcontrib><creatorcontrib>Kimose, Hans Henrik</creatorcontrib><creatorcontrib>Helligsø, Per</creatorcontrib><creatorcontrib>Thomassen, Anne Rahbek</creatorcontrib><creatorcontrib>Nielsen, Torsten Toftegaard</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><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>Cardiovascular research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bøtker, Hans Erik</au><au>Kimose, Hans Henrik</au><au>Helligsø, Per</au><au>Thomassen, Anne Rahbek</au><au>Nielsen, Torsten Toftegaard</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison of non-collagen protein and total creatine as reference for determination of energy stores in endomyocardial biopsies</atitle><jtitle>Cardiovascular research</jtitle><addtitle>Cardiovasc Res</addtitle><date>1993-12-01</date><risdate>1993</risdate><volume>27</volume><issue>12</issue><spage>2113</spage><epage>2117</epage><pages>2113-2117</pages><issn>0008-6363</issn><eissn>1755-3245</eissn><coden>CVREAU</coden><abstract>Objective: The aim was to establish a reliable reference system for biochemical measurements in endomyocardial biopsies. Methods: Myocardial tissue samples were obtained from pigs before and after cardioplegic arrest and reperfusion. Non-collagen protein content was evaluated as a non-specific reference system and compared with total creatine content representing a specific myocardial reference system. The influence of base strength, extraction temperature, and extraction time on protein yields was determined in tissue precipitates redissolved in NaOH. Interference from protein of collagenous origin was excluded by hydroxyproline determinations. Variability of myocardial ATP content in relation to non-collagen protein and total creatine was compared in endomyocardial biopsies taken before and after cardioplegic arrest and reperfusion. Results: The two methods showed comparable analytical precision. Apart from an interference in 1.0 mol·litre−1 NaOH for extended extraction periods at high temperatures, myocardial protein yields increased with increasing base strength, extraction temperature, and extraction time. During cardioplegic arrest and reperfusion heart weight increased due to oedema. Simultaneously, myocardial non-collagen protein content decreased. No change in total creatine was found during cardioplegic arrest but there was a significant loss of creatine after reperfusion. Comparison of variability in myocardial ATP content with non-collagen protein or total creatine as reference systems revealed no difference. Conclusions: Determination of non-collagen protein can be optimised with standardised conditions for protein extraction in tissue precipitates. Employment of total creatine as a reference system does not reduce variability of myocardial metabolite determinations in endomyocardial biopsies compared with non-collagen protein. Loss of myocardial creatine may in itself provide additional information about myocardial injury but this makes it unsuitable as a reference system for measuring metabolic changes during reperfusion. Multiple biopsies seem necessary for estimation of myocardial energy stores. Cardiovascular Research 1993; 27: 2113-2117</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><pmid>8313416</pmid><doi>10.1093/cvr/27.12.2113</doi><tpages>5</tpages></addata></record> |
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| subjects | Animals assay evaluation ATP Biological and medical sciences Biopsy cardioplegia creatine Creatine - analysis endomyocardial biopsy Energy Metabolism - physiology Investigative techniques, diagnostic techniques (general aspects) Medical sciences Myocardial Reperfusion Injury - metabolism Myocardium - chemistry myoglobin non-collagen protein Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques pig Proteins - analysis reference system reperfusion Respiratory system Swine |
| title | Comparison of non-collagen protein and total creatine as reference for determination of energy stores in endomyocardial biopsies |
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