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Diagnosis of AMI 1.Typical symptoms (may not be present) 2.Distinctive ECG (may not be present) 3.Abnormal blood tests –Troponin-I –“CK,” “CPK,” or “CK-Mb”

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Presentation on theme: "Diagnosis of AMI 1.Typical symptoms (may not be present) 2.Distinctive ECG (may not be present) 3.Abnormal blood tests –Troponin-I –“CK,” “CPK,” or “CK-Mb”"— Presentation transcript:

1 Diagnosis of AMI 1.Typical symptoms (may not be present) 2.Distinctive ECG (may not be present) 3.Abnormal blood tests –Troponin-I –“CK,” “CPK,” or “CK-Mb”

2 Definition: unstable angina Unstable angina—an acute process of myocardial ischemia that is not of sufficient severity and duration to result in myocardial necrosis. – Do not release biomarkers indicative of myocardial necrosis into the blood.

3 REQUISITI PER UN MARCATORE DI LESIONE DEL MIOCARDIO “IDEALE”  Presente in elevate concentrazioni nel muscolo cardiaco  Contenuto unicamente nella cellula miocardica (SPECIFICITA’ ASSOLUTA)  Assenza dal circolo in condizioni di normalità (ESTREMAMENTE SENSIBILE E PRECISO)  Rilasciato dal muscolo cardiaco danneggiato rapidamente in maniera completa ed in quantita’ proporzionale al danno miocardico  Rilevabile nel plasma in una finestra diagnostica utile a porre diagnosi sia precocemente che tardivamente  Validato dai trials clinici  Saggio ampiamente disponibile e poco costoso

4

5 A causa della loro scarsa sensibilita’ e specificita’, le determinazioni di aspartato amminotransferasi (AST), lattato deidrogenasi (LDH) totale e suoi isoenzimi, CK totale ed attivita’ catalitica del suo isoenzima MB dovrebbero essere considerate come obsolete. Panteghini M et al. G Ital Cardiol 1999;29:810

6 Tools: Cardiac Markers for AMI Diagnosis Creatine Kinase MB isoenzyme (CK-MB) Myocardial specific. 4-10 hrs to achieve diagnostic performance. 2-3 days to return to normal. Myoglobin 2-3 hrs to achieve diagnostic performance. 24 hrs to return to normal. Not specific, abundant in skeletal muscle. Troponin T or I (TnT, TnI) Myocardiac specific. Up to 12 hrs to achieve diagnostic performance. Several days to return to normal.

7 Myoglobin Rapid rise Non-specific. Cannot be used alone to confirm MI

8 CPK-MB  15% of cardiac CPK, small amount in skeletal muscle  MB-Mass Validated as marker for MI. However:  Can increase after muscle injury, muscular diseases.  Can be found in tongue, intestine, diaphragm, uterus, prostate.

9 Le Troponine cardiache Sono costituite da un eterotrimero suddiviso in 3 sub-unità : Troponina C : 18 kD deputata a legare ioni Ca++ Troponina I : 22 kD con funzione inibitoria Troponina T : 37 kD con funzione di legame alla tropomiosina

10 Figure 1. Plot of the appearance of cardiac markers in blood vs time after onset of symptoms. Peak A, early release of myoglobin or CK-MB isoforms after AMI; peak B, cardiac troponin after AMI; peak C, CK-MB after AMI; peak D, cardiac troponin after unstable angina. Data are plotted on a relative scale, where 1.0 is set at the AMI cutoff concentration.

11 New and old diagnostic criteria for MI 2 of 3 Elevated cardiac markers Trop or CK and CKMB Evidence of myocardial damage as assessed by pathologic exam, ECG or imaging modality One of the following WHO Criteria for Diagnosis of MI ESC / ACC Recommendations for diagnosis of MI Chest pain ECG changes Serum markers Alpert et al - ACC Consensus. JACC 2000; 36: 959 - 969

12 Cardiac Analyzer 2nd and 3rd Specimens 6hrs and 12hrs Second or third specimen Measure CKMB Previous Troponin > 99th percentile Measure troponin Troponin > 3.0 ng/ml Cancel remaining tests Continue test algorithm No Yes Delta CKMB > 20% No Measure troponin Yes

13 The Future Early indicators of plaque instability Early indicators of thrombotic activity Markers for IIb/IIIa sensitivity Indicators of cytokine activation

14 Technology in the Future : Family of Near-Patient Diagnostic Sensors

15 Human protein: P05413 - Fatty acid-binding protein, heart (H-FABP) (Heart-type fatty acid- binding protein) (Muscle fatty acid-binding protein) (M-FABP) (Mammary-derived growth inhibitor) (MDGI). EMBL EMBL FORUM Length: 132 aa, molecular weight: 14727 Da, CRC64 checksum: 5FD3C988879AECCCLength: 132 aa, molecular weight: 14727 Da, CRC64 checksum: 5FD3C988879AECCC VDAFLGTWKL VDSKNFDDYM KSLGVGFATR QVASMTKPTT IIEKNGDILT LKTHSTFKNT 60 EISFKLGVEF DETTADDRKV KSIVTLDGGK LVHLQKWDGQ ETTLVRELID GKLILTLTHG 120 TAVCTRTYEK EA 132 // VDAFLGTWKL VDSKNFDDYM KSLGVGFATR QVASMTKPTT IIEKNGDILT LKTHSTFKNT 60 EISFKLGVEF DETTADDRKV KSIVTLDGGK LVHLQKWDGQ ETTLVRELID GKLILTLTHG 120 TAVCTRTYEK EA 132 // SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page GoTo: SOURCE @ Stanford University GeneReport for: X56549 FABP3 fatty acid binding protein 3, muscle and heart (mammary-derived growth inhibitor) UniGene, LocusLink, OMIM, GenAtlas, GeneCard, Ensembl, MapView, Genome Browser UniGeneLocusLinkOMIMGenAtlasGeneCard EnsemblMapViewGenome Browser Aliases MDGI; O-FABP; FABP11; H-FABP FATTY ACID-BINDING PROTEIN, SKELETAL MUSCLE Fatty acid-binding protein 3, muscle fatty acid binding protein 11 FATTY ACID-BINDING PROTEIN, MUSCLE AND HEART Chromosomal Location Chromosome/CytobandChromosome/Cytoband 1p33-p32 Microarray Gene Expression Data Data availableData available Show Gene Expression Data LocusLink Information Locus Link SummaryLocus Link Summary The intracellular fatty acid-binding proteins (FABPs) belongs to a multigene family. FABPs are divided into at least three distinct types, namely the hepatic-, intestinal- and cardiac-type. They form 14-15 kDa proteins and are thought to participate in the uptake, intracellular metabolism and/or transport of long-chain fatty acids. They may also be responsible in the modulation of cell growth and proliferation. Fatty acid- binding protein 3 gene contains four exons and its function is to arrest growth of mammary epithelial cells. This gene is a candidate tumor suppressor gene for human breast cancer. SwissProt Information SwissProt Accession No.SwissProt Accession No. P05413P05413 Fatty acid-binding protein, heart (Homo sapiens); 100% similarity over 133 a.a. FunctionFunction fabp are thought to play a role in the intracellular transport of long-chain fatty acids and their acyl-coa esters. Subcellular LocationSubcellular Location cytoplasmic. SimilaritySimilarity belongs to the fatty-acid binding protein (fabp) family. SwissProt CopyrightSwissProt Copyright This SWISS-PROT entry is copyright. It is produced through a collaboration between the Swiss Institute of Bioinformatics and the EMBL outstation - the European Bioinformatics Institute. There are no restrictions on its use by non-profit institutions as long as its content is in no way modified and this statement is not removed. Usage by and for commercial entities requires a license agreement (See http://www.isb-sib.ch/announce/ or send an email to license@isb-sib.ch).http://www.isb-sib.ch/announce/license@isb-sib.ch Annotations Gene OntologiesGene Ontologies UniGene & EST Expression Information UniGene ClusterUniGene Cluster Hs.112669Hs.112669 from Build No. 173, Released on 2004-09-06Build No. 173 Normalized expression distribution for tissue type Top ten [of 16][Help]Normalized expression distribution for tissue type Top ten [of 16][Help] SAGE (NCBI)SAGE (NCBI) Go to Gene-to-tag Mapping at NCBIGene-to-tag Mapping Upstream Genomic Sequence TRASERTRASER Upstream genomic sequenceUpstream genomic sequence for fatty acid binding protein 3, muscle and heart (mammary-derived growth inhibitor) Representative mRNA Sequences UniGeneUniGene BG336702 LocusLink RefSeqLocusLink RefSeq Alias PubMed Search PubMedPubMed Search PubMed using aliases AND | Search | No Clone Report | Help | Search Help SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page ACTIVATE: SMART analysis Confidently predicted domains, repeats, motifs and features: namebeginendE-value Pfam:Lipocalin31311.50e-36 SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page UniGene Cluster Hs.112669 Homo sapiens Fatty acid binding protein 3, muscle and heart (mammary-derived growth inhibitor) (FABP3) LinksLinks SELECTED PROTEIN SIMILARITIES SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page 9/23 82.6 %: cytoplasmic 13.0 %: nuclear 4.3 %: peroxisomal SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page EMBL/Genbank->Unigene mapping provided by RZPD http://www.rzpd.de EBI-Hinxton-"Uniprot-Swissprot-TrEMBL" database SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page GoTo: SOSUI analysis GoTo: SOSUI analysis This amino acid sequence is of a SOLUBLE PROTEIN. SOURCESOURCE - BLAST -SMART - HomoloGene - ensEMBL - GenomeBrowser - PSORT2 - Entrez - RZPD - CDART - SOSUI - STRINGBLASTSMARTHomoloGeneensEMBLGenomeBrowserPSORT2EntrezRZPDCDARTSOSUISTRINGtop of page OntologyAnnotation Evidenc e Sourc e Molecular Function Lipid transporter activityNRGOA Fatty acid bindingNRGOA Biological Process Negative regulation of cell proliferation TASGOA TransportIEAGOA Cellular Component CytoplasmNRGOA Soluble fractionNRGOA Tissue Normalized Expression (%) Cluster Clones : Tissue clones Heart:39.9513:35504 Muscle:21.6218:90826 Lung:8.1417:227901 Uterus:5.947:128533 Peripheral_Nervous_System : 5.191:21019 Testis:5.075:107669 Kidney:2.873:113979 Brain:2.397:319574 Bone:2.131:51252 Prostate:1.131:96523 Accession Descriptio n NM_00410 2 NA General information www.uniprot.orgFABH_HUMANFABH_HUMAN | XMLXML Entry nameFABH_HUMAN Accession number P05413, Q99957 CreatedRel. 09, 1-NOV-1988 Sequence updateRel. 21, 1-MAR-1992 Annotation updateRel. 45, 25-OCT-2004 Description and origin of the Protein DescriptionFatty acid-binding protein, heart (H-FABP) (Heart-type fatty acid- binding protein) (Muscle fatty acid-binding protein) (M-FABP) (Mammary-derived growth inhibitor) (MDGI). Gene name(s)FABP3 Organism sourceHomo sapiens (Human). TaxonomyEukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Primates; Catarrhini; Hominidae; Homo. NCBI TaxID9606 References [1]Offner,G.D., Brecher,P., Sawlivich,W.B., Costello,C.E., Troxler,R.F., Characterization and amino acid sequence of a fatty acid-binding protein from human heart. (1988) Biochem. J. 252:191-198 PositionSEQUENCE. Medline88339792 PubMed3421901 [2]Boerchers,T., Hoejrup,P., Nielsen,S.U., Roepstorff,P., Spener,F., Knudsen,J., Revision of the amino acid sequence of human heart fatty acid-binding protein. (1990) Mol. Cell. Biochem. 98:127-133 PositionREVISIONS, AND SEQUENCE. Medline91094793 PubMed2266954 [3]Peeter,R.A., Veerkamp,J.H., Kanda,T., Ono,T., Geurts van Kessel,A., Cloning of the cDNA encoding human skeletal-muscle fatty-acid-binding protein, its peptide sequence and chromosomal localization. (1991) Biochem. J. 276:203-207 PositionSEQUENCE FROM N.A. CommentsTISSUE=Skeletal muscle; Medline91248148 PubMed1710107 [4] Hu,Y.F., Ao,X., Russo,I.H., Russo,J., Submitted MAR-1997 to the EMBL GenBank DDBJ databases PositionSEQUENCE FROM N.A. CommentsTISSUE=Breast; [5]Wu,X., Arlt,M., Goodfellow,P.J., Rottman,J.N., Genomic organization and complete nucleotide sequence of the human cardiac fatty acid binding protein gene (FABP3), and identification of a closely related genomic sequence.Submitted NOV-1994 to the EMBL GenBank DDBJ databases PositionSEQUENCE FROM N.A. [6]Strausberg,R.L., Feingold,E.A., Grouse,L.H., Derge,J.G., Klausner,R.D., Collins,F.S., Wagner,L., Shenmen,C.M., Schuler,G.D., Altschul,S.F., Zeeberg,B., Buetow,K.H., Schaefer,C.F., Bhat,N.K., Hopkins,R.F., Jordan,H., Moore,T., Max,S.I., Wang,J., Hsieh,F., Diatchenko,L., Marusina,K., Farmer,A.A., Rubin,G.M., Hong,L., Stapleton,M., Soares,M.B., Bonaldo,M.F., Casavant,T.L., Scheetz,T.E., Brownstein,M.J., Usdin,T.B., Toshiyuki,S., Carninci,P., Prange,C., Raha,S.S., Loquellano,N.A., Peters,G.J., Abramson,R.D., Mullahy,S.J., Bosak,S.A., McEwan,P.J., McKernan,K.J., Malek,J.A., Gunaratne,P.H., Richards,S., Worley,K.C., Hale,S., Garcia,A.M., Gay,L.J., Hulyk,S.W., Villalon,D.K., Muzny,D.M., Sodergren,E.J., Lu,X., Gibbs,R.A., Fahey,J., Helton,E., Ketteman,M., Madan,A., Rodrigues,S., Sanchez,A., Whiting,M., Young,A.C., Shevchenko,Y., Bouffard,G.G., Blakesley,R.W., Touchman,J.W., Green,E.D., Dickson,M.C., Rodriguez,A.C., Grimwood,J., Schmutz,J., Myers,R.M., Butterfield,Y.S.N., Krzywinski,M.I., Skalska,U., Smailus,D.E., Schnerch,A., Schein,J.E., Jones,S.J.M., Marra,M.A., Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. (2002) Proc. Natl. Acad. Sci. U.S.A. 99:16899-16903 PositionSEQUENCE FROM N.A. CommentsTISSUE=Liver; Medline22388257 DOI10.1073/pnas.242603899 PubMed12477932 [7]Troxler,R.F., Offner,G.D., Jiang,J.W., Wu,B.L., Skare,J.C., Milunsky,A., Wyandt,H.E., Localization of the gene for human heart fatty acid binding protein to chromosome 1p32-1p33. (1993) Hum. Genet. 92:563-566 PositionSEQUENCE OF 14-132 FROM N.A. CommentsTISSUE=Heart; Medline94085953 PubMed8262516 [8]Kovalyov,L.I., Shishkin,S.S., Efimochkin,A.S., Kovalyova,M.A., Ershova,E.S., Egorov,T.A., Musalyamov,A.K., The major protein expression profile and two-dimensional protein database of human heart. (1995) Electrophoresis 16:1160-1169 PositionSEQUENCE OF 31-38. CommentsTISSUE=Heart; Medline96007936 PubMed7498159 [9]Zanotti,G., Scapin,G., Spadon,P., Veerkamp,J.H., Sacchettini,J.C., Three-dimensional structure of recombinant human muscle fatty acid- binding protein. (1992) J. Biol. Chem. 267:18541-18550 PositionX-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS). Medline92406763 PubMed1526991 [10]Young,A.C.M., Scapin,G., Kromminga,A., Patel,S.B., Veerkamp,J.H., Sacchettini,J.C., Structural studies on human muscle fatty acid binding protein at 1.4- A resolution: binding interactions with three C18 fatty acids. (1994) Structure 2:523-534 PositionX-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS). Medline95006319 PubMed7922029 Comments FUNCTION FABP are thought to play a role in the intracellular transport of long-chain fatty acids and their acyl-CoA esters. SUBCELLULAR LOCATION Cytoplasmic. SIMILARITY Belongs to the fatty-acid binding protein (FABP) family. Copyright This SWISS-PROT entry is copyright. It is produced through a collaboration between the Swiss Institute of Bioinformatics and the EMBL outstation - the European Bioinformatics Institute. There are no restrictions on its use by non-profit institutions as long as its content is in no way modified and this statement is not removed. Usage by and for commercial entities requires a license agreement (See http://www.isb-sib.ch/announce/ or send an email to license@isb-sib.ch). Database cross-references EMBL X56549X56549; CAA39889.1; -.CAA39889 Y10255Y10255; CAA71305.1; -.CAA71305 U57623U57623; AAB02555.1; -.AAB02555 U17081U17081; AAC99800.1; -.AAC99800 S67314S67314; AAB29294.1; -.AAB29294 BC007021BC007021; AAH07021.1; -.AAH07021 PIR S15432S15432; FZHUC. PDB 1G5W1G5W; NMR; A=1-132. 1HMR1HMR; X-ray; @=1-132. 1HMS1HMS; X-ray; @=-. 1HMT1HMT; X-ray; @=1-132. 2HMB2HMB; X-ray; @=1-132. HSC-2DPAGE P05413P05413; HUMAN. Genew HGNC:3557; FABP3.3557 H-InvDB HIX0000353; -. MIM 134651134651; -. GO GO:0008289GO:0008289; F:lipid binding; TAS. GO:0008285GO:0008285; P:negative regulation of cell proliferation; TAS. InterPro IPR011038IPR011038; Calycin. IPR000463IPR000463; Fatty_acid_BP. IPR000566IPR000566; Lipocln_cytFABP. Pfam PF00061PF00061; Lipocalin; 1.Lipocalin PRINTS PR00178PR00178; FATTYACIDBP. PROSITE PS00214PS00214; FABP; 1. Keywords 3D-structure3D-structure; Acetylation; Direct protein sequencing; Lipid-binding; Phosphorylation; Transport;AcetylationDirect protein sequencingLipid-bindingPhosphorylationTransport Features KeyBeginEndLengthDescriptionDescription INIT_MET001 MOD_RES111N-acetylvaline.N-acetylvaline. 19 1Phosphotyrosine (by Tyr-kinases) (By similarity).Phosphotyrosine (by Tyr-kinases) (By similarity). CONFLICT111V -> A (in Ref. 3).V -> A (in Ref. 3). 104 1L -> K (in Ref. 1).L -> K (in Ref. 1). CONFLICT124 1C -> S (in Ref. 1).C -> S (in Ref. 1). CONFLICT129 1E -> Q (in Ref. 2).E -> Q (in Ref. 2). HELIX243 STRAND6149 HELIX16238 TURN24 1 HELIX27348 TURN35 1 STRAND39457 TURN46472 STRAND48547 STRAND60645 TURN66672 STRAND70734 TURN75762 STRAND79879 TURN88892 STRAND90978 TURN98992 STRAND10010910 TURN1101112 STRAND1121198 TURN1201212 STRAND1221309

16 Tools: FABP in AMI Diagnosis FABP Characteristics Small cytoplasmic protein, abundant in heart and muscle. Cardiac specific isoforms Normal levels below 1ng/ml. Most sensitive early cardiac marker. <3 hrs to achieve diagnostic performance. 12-24 hrs to return to normal.

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